Showing 201 - 250 of 274 Items

Human protein Sam68 relocalization and interaction with poliovirus RNA polymerase in infected cells

Date: 1996-03-19

Creator: A. E. Mcbride, A. Schlegel, K. Kirkegaard

Access: Open access

A HeLa cDNA expression library was screened for human polypeptides that interacted with the poliovirus RNA-dependent RNA polymerase, 3D, using the two-hybrid system in the yeast Saccharomyces cerevisiae. Sam68 (Src- associated in mitosis, 68 kDa) emerged as the human cDNA that, when fused to a transcriptional activation domain, gave the strongest 3D interaction signal with a LexA-3D hybrid protein. 3D polymerase and Sam68 coimmunoprecipitated from infected human cell lysates with antibodies that recognized either protein. Upon poliovirus infection. Sam68 relocalized from the nucleus to the cytoplasm, where poliovirus replication occurs. Sam68 was isolated from infected cell lysates with an antibody that recognizes poliovirus protein 2C, suggesting that it is found on poliovirus-induced membranes upon which viral RNA synthesis occurs. These data, in combination with the known RNA- and protein-binding properties of Sam68, make Sam68 a strong candidate for a host protein with a functional role in poliovirus replication.


Xanthophyll cycle activity in two prominent arctic shrub species

Date: 2017-01-01

Creator: T.S. Magney, B.A. Logan, J.S. Reblin, N.T. Boelman, J.U.H., Eitel, H.E. Greaves, K.L. Griffin, C.M. Prager, L.A. Vierling

Access: Open access



Live imaging and biophysical modeling support a button-based mechanism of somatic homolog pairing in Drosophila

Date: 2021-06-01

Creator: Myron Child, Jack R. Bateman, Amir Jahangiri, Armando Reimer, Nicholas C., Lammers, Nica Sabouni, Diego Villamarin, Grace C. McKenzie-Smith, Justine E. Johnson, Daniel Jost, Hernan G. Garcia

Access: Open access

3D eukaryotic genome organization provides the structural basis for gene regulation. In Drosophila melanogaster, genome folding is characterized by somatic homolog pairing, where homologous chromosomes are intimately paired from end to end; however, how homologs identify one another and pair has remained mysterious. Recently, this process has been proposed to be driven by specifically interacting “buttons” encoded along chromosomes. Here, we turned this hypothesis into a quantitative biophysical model to demonstrate that a button-based mechanism can lead to chromosome-wide pairing. We tested our model using live-imaging measurements of chromosomal loci tagged with the MS2 and PP7 nascent RNA labeling systems. We show solid agreement between model predictions and experiments in the pairing dynamics of individual homologous loci. Our results strongly support a button-based mechanism of somatic homolog pairing in Drosophila and provide a theoretical framework for revealing the molecular identity and regulation of buttons.


Positive Effects of Nonnative Invasive Phragmites australis on Larval Bullfrogs

Date: 2012-08-30

Creator: Mary Rogalski

Access: Open access



islet reveals segmentation in the amphioxus hindbrain homolog

Date: 2000-04-01

Creator: William R. Jackman, James A. Langeland, Charles B. Kimmel

Access: Open access

The vertebrate embryonic hindbrain is segmented into rhombomeres. Gene expression studies suggest that amphioxus, the closest invertebrate relative of vertebrates, has a hindbrain homolog. However, this region is not overtly segmented in amphioxus, raising the question of how hindbrain segmentation arose in chordate evolution. Vertebrate hindbrain segmentation includes the patterning of cranial motor neurons, which can be identified by their expression of the LIM-homeodomain transcription factor islet1. To learn if the amphioxus hindbrain homolog is cryptically segmented, we cloned an amphioxus gene closely related to islet1, which we named simply islet. We report that amphioxus islet expression includes a domain of segmentally arranged cells in the ventral hindbrain homolog. We hypothesize that these cells are developing motor neurons and reveal a form of hindbrain segmentation in amphioxus. Hence, vertebrate rhombomeres may derive from a cryptically segmented brain present in the amphioxus/vertebrate ancestor. Other islet expression domains provide evidence for amphioxus homologs of the pineal gland, adenohypophysis, and endocrine pancreas. Surprisingly, homologs of vertebrate islet1-expressing spinal motor neurons and Rohon-Beard sensory neurons appear to be absent. (C) 2000 Academic Press.


Rapid oligo-galacturonide induced changes in protein phosphorylation in arabidopsis

Date: 2016-04-01

Creator: Bruce D. Kohorn, Divya Hoon, Benjamin B. Minkoff, Michael R. Sussman, Susan L., Kohorn

Access: Open access

The wall-associated kinases (WAKs)1 are receptor protein kinases that bind to long polymers of cross-linked pectin in the cell wall. These plasma-membrane-associated protein kinases also bind soluble pectin fragments called oligo-galacturonides (OGs) released from the wall after pathogen attack and damage. WAKs are required for cell expansion during development but bind water soluble OGs generated from walls with a higher affinity than the wall-associated polysaccharides. OGs activate a WAKdependent, distinct stress-like response pathway to help plants resist pathogen attack. In this report, a quantitative mass-spectrometric-based phosphoproteomic analysis was used to identify Arabidopsis cellular events rapidly induced by OGs in planta. Using N14/ N15 isotopic in vivo metabolic labeling, we screened 1,000 phosphoproteins for rapid OG-induced changes and found 50 proteins with increased phosphorylation, while there were none that decreased significantly. Seven of the phosphosites within these proteins overlap with those altered by another signaling molecule plants use to indicate the presence of pathogens (the bacterial "elicitor" peptide Flg22), indicating distinct but overlapping pathways activated by these two types of chemicals. Genetic analysis of genes encoding 10 OG-specific and two Flg22/OG-induced phosphoproteins reveals that null mutations in eight proteins compromise the OG response. These phosphorylated proteins with genetic evidence supporting their role in the OG response include two cytoplasmic kinases, two membrane-associated scaffold proteins, a phospholipase C, a CDPK, an unknown cadmium response protein, and a motor protein. Null mutants in two proteins, the putative scaffold protein REM1.3, and a cytoplasmic receptor like kinase ROG2, enhance and suppress, respectively, a dominant WAK allele. Altogether, the results of these chemical and genetic experiments reveal the identity of several phosphorylated proteins involved in the kinase/ phosphatase-mediated signaling pathway initiated by cell wall changes.


Cell wall-associated kinases and pectin perception

Date: 2016-01-01

Creator: Bruce D. Kohorn

Access: Open access

The pectin matrix of the angiosperm cell wall is regulated in both synthesis and modification and greatly influences the direction and extent of cell growth. Pathogens, herbivory and mechanical stresses all influence this pectin matrix and consequently plant form and function. The cell wall-associated kinases (WAKs) bind to pectin and regulate cell expansion or stress responses depending upon the state of the pectin. This review explores the WAKs in the context of cell wall biology and signal transduction pathways.


Transvection-based gene regulation in Drosophila is a complex and plastic trait

Date: 2014-01-01

Creator: Xinyang Bing, Teresa Z. Rzezniczak, Jack R. Bateman, Thomas J.S. Merritt

Access: Open access

Transvection, a chromosome pairing-dependent form of trans-based gene regulation, is potentially widespread in the Drosophila melanogaster genome and varies across cell types and within tissues in D. melanogaster, characteristics of a complex trait. Here, we demonstrate that the trans-interactions at the Malic enzyme (Men) locus are, in fact, transvection as classically defined and are plastic with respect to both genetic background and environment. Using chromosomal inversions, we show that trans-interactions at the Men locus are eliminated by changes in chromosomal architecture that presumably disrupt somatic pairing. We further show that the magnitude of transvection at the Men locus is modified by both genetic background and environment (temperature), demonstrating that transvection is a plastic phenotype. Our results suggest that transvection effects in D. melanogaster are shaped by a dynamic interplay between environment and genetic background. Interestingly, we find that cis-based regulation of the Men gene is more robust to genetic background and environment than trans-based. Finally, we begin to uncover the nonlocal factors that may contribute to variation in transvection overall, implicating Abd-B in the regulation of Men in cis and in trans in an allele-specific and tissue-specific manner, driven by differences in expression of the two genes across genetic backgrounds and environmental conditions.


Taming the giant within

Date: 2019-01-01

Creator: Jack R. Bateman, David J. Anderson

Access: Open access




The cell wall-associated kinases, WAKs, as pectin receptors

Date: 2012-05-08

Creator: Bruce D. Kohorn, Susan L. Kohorn

Access: Open access

The wall-associated kinases, WAKs, are encoded by five highly similar genes clustered in a 30-kb locus in Arabidopsis. These receptor-like proteins contain a cytoplasmic serine threonine kinase, a transmembrane domain, and a less conserved region that is bound to the cell wall and contains a series of epidermal growth factor repeats. Evidence is emerging that WAKs serve as pectin receptors, for both short oligogalacturonic acid fragments generated during pathogen exposure or wounding, and for longer pectins resident in native cell walls. This ability to bind and respond to several types of pectins correlates with a demonstrated role for WAKs in both the pathogen response and cell expansion during plant development. © 2012 Kohorn and Kohorn.


Mutations in a signal sequence for the thylakoid membrane identify multiple protein transport pathways and nuclear suppressors

Date: 1994-07-01

Creator: Tracy A. Smith, Bruce D. Kohorn

Access: Open access

The apparatus that permits protein translocation across the internal thylakoid membranes of chloroplasts is completely unknown, even though these membranes have been the subject of extensive biochemical analysis. We have used a genetic approach to characterize the translocation of Chlamydomonas cytochrome f, a chloroplast-encoded protein that spans the thylakoid once. Mutations in the hydrophobic core of the cytochrome f signal sequence inhibit the accumulation of cytochrome f, lead to an accumulation of precursor, and impair the ability of Chlamydomonas cells to grow photosynthetically. One hydrophobic core mutant also reduces the accumulation of other thylakoid membrane proteins, but not those that translocate completely across the membrane. These results suggest that the signal sequence of cytochrome f is required and is involved in one of multiple insertion pathways. Suppressors of two signal peptide mutations describe at least two nuclear genes whose products likely describe the translocation apparatus, and selected second- site chloroplast suppressors further define regions of the cytochrome f signal peptide.


Direct selection for sequences encoding proteases of known specificity

Date: 1991-06-15

Creator: Tracy A. Smith, Bruce D. Kohorn

Access: Open access

We have developed a simple genetic selection that could be used to isolate eukaryotic cDNAs encoding proteases that cleave within a defined amino acid sequence. The selection was developed by using the transcription factor GAL4 from Saccharomyces cerevisiae as a selectable marker, a cloned protease from tobacco etch virus (TEV), and an 18-amino acid TEV protease target sequence. In yeast, TEV protease cleaves its target even when the target is fused to internal regions of the GAL4 protein. This cleavage separates the DNA binding domain from the transcription activation domain of GAL4, rendering it transcriptionally inactive. The proteolytic cleavage can be detected phenotypically by the inability of cells to metabolize galactose. Cells expressing the TEV protease can also be selected on the suicide substrate 2-deoxygalactose. DNA binding studies show that the TEV protease decreases the activity of the GAL4/target fusion protein. Because another protease target sequence of 55 amino acids can be inserted into GAL4 without any loss of transcriptional activity, this assay offers the opportunity to use high-efficiency cDNA cloning and expression vectors to select coding sequences of other proteases from various species. The assay could also be used to help define both target specificities and functional domains of proteases.


Comparing enhancer action in cis and in trans

Date: 2012-08-01

Creator: Jack R. Bateman, Justine E. Johnson, Melissa N. Locke

Access: Open access

Studies from diverse systems have shown that distinct interchromosomal interactions are a central component of nuclear organization. In some cases, these interactions allow an enhancer to act in trans, modulating the expression of a gene encoded on a separate chromosome held in close proximity. Despite recent advances in uncovering such phenomena, our understanding of how a regulatory element acts on another chromosome remains incomplete. Here, we describe a transgenic approach to better understand enhancer action in trans in Drosophila melanogaster. Using phiC31-based recombinase-mediated cassette exchange (RMCE), we placed transgenes carrying combinations of the simple enhancer GMR, a minimal promoter, and different fluorescent reporters at equivalent positions on homologous chromosomes so that they would pair via the endogenous somatic pairing machinery of Drosophila. Our data demonstrate that the enhancer GMR is capable of activating a promoter in trans and does so in a variegated pattern, suggesting stochastic interactions between the enhancer and the promoter when they are carried on separate chromosomes. Furthermore, we quantitatively assessed the impact of two concurrent promoter targets in cis and in trans to GMR, demonstrating that each promoter is capable of competing for the enhancer's activity, with the presence of one negatively affecting expression from the other. Finally, the single-cell resolution afforded by our approach allowed us to show that promoters in cis and in trans to GMR can both be activated in the same nucleus, implying that a single enhancer can share its activity between multiple promoter targets carried on separate chromosomes. © 2012 by the Genetics Society of America.


Wall-associated kinase 1 (WAK1) is crosslinked in endomembranes, and transport to the cell surface requires correct cell-wall synthesis

Date: 2006-06-01

Creator: Bruce D. Kohorn, Masaru Kobayashi, Sue Johansen, Henry Perry Friedman, Andy, Fischer, Nicole Byers

Access: Open access

The Arabidopsis thaliana wall-associated kinases (WAKs) bind to pectin with an extracellular domain and also contain a cytoplasmic protein kinase domain. WAKs are required for cell elongation and modulate sugar metabolism. This work shows that in leaf protoplasts a WAK1-GFP fusion protein accumulates in a cytoplasmic compartment that contains pectin. The WAK compartment contains markers for the Golgi, the site of pectin synthesis. The migration of WAK1-GFP to the cell surface is far slower than that of a cell surface receptor not associated with the cell wall, is influenced by the presence of fucose side chains on one or more unidentified molecules that might include pectin, and is dependent upon cellulose synthesis on the plasma membrane. WAK is crosslinked into a detergent-insoluble complex within the cytoplasmic compartment before it appears on the cell surface, and this is independent of fucose modification or cellulose synthesis. Thus, the assembly and crosslinking of WAKs may begin at an early stage within a cytoplasmic compartment rather than in the cell wall itself, and is coordinated with synthesis of surface cellulose.


Miniature of Investigating the Role of Heterochromatin Protein 1 (HP1) in the Formation of Chromosome Compartments
Investigating the Role of Heterochromatin Protein 1 (HP1) in the Formation of Chromosome Compartments
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  • Restriction End Date: 2025-06-01

    Date: 2020-01-01

    Creator: Diego Andres Villamarin

    Access: Access restricted to the Bowdoin Community



      Species identification based on a semi-diagnostic marker: Evaluation of a simple conchological test for distinguishing blue mussels Mytilus edulis L. And M. trossulus Gould

      Date: 2021-07-01

      Creator: Vadim Khaitov, Julia Marchenko, Marina Katolikova, Risto Väinölä, Sarah E., Kingston, David B. Carlon, Michael Gantsevich, Petr Strelkov

      Access: Open access

      Cryptic and hybridizing species may lack diagnostic taxonomic characters leaving researchers with semi-diagnostic ones. Identification based on such characters is probabilistic, the probability of correct identification depending on the species composition in a mixed population. Here we test the possibilities of applying a semi-diagnostic conchological character for distinguishing two cryptic species of blue mussels, Mytilus edulis and M. trossulus. These ecologically, stratigraphically and economically important molluscs co-occur and hybridize in many areas of the North Atlantic and the neighboring Arctic. Any cues for distinguishing them in sympatry without genotyping would save much research effort. Recently these species have been shown to statistically differ in the White Sea, where a simple character of the shell was used to distinguish two mussel morphotypes. In this paper, we analyzed the associations between morphotypes and species-specific genotypes based on an abundant material from the waters of the Kola Peninsula (White Sea, Barents Sea) and a more limited material from Norway, the Baltic Sea, Scotland and the Gulf of Maine. The performance of the “morphotype test” for species identification was formally evaluated using approaches from evidence-based medicine. Interspecific differences in the morphotype frequencies were ubiquitous and unidirectional, but their scale varied geographically (from 75% in the White Sea to 15% in the Baltic Sea). In addition, salinity-related variation of this character within M. edulis was revealed in the Arctic Barents Sea. For every studied region, we established relationships between the proportions of the morphotypes in the populations as well as between the proportions of the morphotypes in samples and the probabilities of mussels of different morphotypes being M. trossulus and M. edulis. We provide recommendations for the application of the morphotype test to mussels from unstudied contact zones and note that they may apply equally well to other taxa identified by semi-diagnostic traits.


      Miniature of Egg Size, Breeding Phenology, and Parental Investment in Leach’s Storm Petrels
      Egg Size, Breeding Phenology, and Parental Investment in Leach’s Storm Petrels
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          Date: 2020-01-01

          Creator: James L. O'Shea

          Access: Access restricted to the Bowdoin Community



            The guanine nucleotide exchange factor trio mediates axonal development in the Drosophila embryo

            Date: 2000-01-01

            Creator: Jack Bateman, Huidy Shu, David Van Vactor

            Access: Open access

            Recent analysis of Rho subfamily GTPases in Drosophila revealed roles for Rac and Cdc42 during axonogenesis. Here, we describe the identification and characterization of the Drosophila counterpart of Trio, a guanine nucleotide exchange factor (GEF) that associates with the receptor phosphatase LAR and regulates GTPase activation in vertebrate cells. Mutants deficient in trio activity display defects in both central and peripheral axon pathways reminiscent of pheno-types observed in embryos deficient in small GTPase function. Double mutant analysis shows that trio interacts with Rac in a dose-sensitive manner but not with Rho. Moreover, reduction of trio activity potentiates the phenotype of mutations in the LAR homolog Dlar, suggesting that these proteins collaborate in orchestrating the cytoskeletal events that underlie normal axonogenesis.


            Adjusting to Global change through clonal growth and epigenetic variation

            Date: 2016-07-26

            Creator: Richard S. Dodd, Vladimir Douhovnikoff

            Access: Open access

            The earth is experiencing major changes in global and regional climates and changes are predicted to accelerate in the future. Many species will be under considerable pressure to evolve, to migrate, or be faced with extinction. Clonal plants would appear to be at a particular disadvantage due to their limited mobility and limited capacity for adaptation. However, they have outlived previous environmental shifts and clonal species have persisted for millenia. Clonal spread offers unique ecological advantages, such as resource sharing, risk sharing, and economies of scale among ramets within genotypes. We suggest that ecological attributes of clonal plants, in tandem with variation in gene regulation through epigenetic mechanisms that facilitate and optimize phenotype variation in response to environmental change may permit them to be well suited to projected conditions.


            Similarities and differences in circuit responses to applied Gly 1 -SIFamide and peptidergic (Gly 1 -SIFamide) neuron stimulation

            Date: 2019-03-01

            Creator: Dawn M. Blitz, Andrew E. Christie, Aaron P. Cook, Patsy S. Dickinson, Michael P., Nusbaum

            Access: Open access

            Similarities and differences in circuit responses to applied Gly 1 -SIFamide and peptidergic (Gly 1 -SIFamide) neuron stimulation. J Neurophysiol 121: 950 –972, 2019. First published January 16, 2019; doi:10.1152/jn.00567.2018.—Microcircuit modulation by peptides is well established, but the cellular/synaptic mechanisms whereby identified neurons with identified peptide transmitters modulate microcircuits remain unknown for most systems. Here, we describe the distribution of GYRKPPFNGSIFamide (Gly 1 -SIFamide) immunoreactivity (Gly 1 -SIFamide-IR) in the stomatogastric nervous system (STNS) of the crab Cancer borealis and the Gly 1 -SIFamide actions on the two feeding-related circuits in the stomatogastric ganglion (STG). Gly 1 -SIFamide-IR localized to somata in the paired commissural ganglia (CoGs), two axons in the nerves connecting each CoG with the STG, and the CoG and STG neuropil. We identified one Gly 1 -SIFamide-IR projection neuron innervating the STG as the previously identified modulatory commissural neuron 5 (MCN5). Brief (~10 s) MCN5 stimulation excites some pyloric circuit neurons. We now find that bath applying Gly 1 -SIFamide to the isolated STG also enhanced pyloric rhythm activity and activated an imperfectly coordinated gastric mill rhythm that included unusually prolonged bursts in two circuit neurons [inferior cardiac (IC), lateral posterior gastric (LPG)]. Furthermore, longer duration (±30 s) MCN5 stimulation activated a Gly 1 -SIFamide-like gastric mill rhythm, including prolonged IC and LPG bursting. The prolonged LPG bursting decreased the coincidence of its activity with neurons to which it is electrically coupled. We also identified local circuit feedback onto the MCN5 axon terminals, which may contribute to some distinctions between the responses to MCN5 stimulation and Gly 1 -SIFamide application. Thus, MCN5 adds to the few identified projection neurons that modulate a well-defined circuit at least partly via an identified neuropeptide transmitter and provides an opportunity to study peptide regulation of electrical coupled neurons in a functional context. NEW & NOTEWORTHY Limited insight exists regarding how identified peptidergic neurons modulate microcircuits. We show that the modulatory projection neuron modulatory commissural neuron 5 (MCN5) is peptidergic, containing Gly 1 -SIFamide. MCN5 and Gly 1 -SIFamide elicit similar output from two well-defined motor circuits. Their distinct actions may result partly from circuit feedback onto the MCN5 axon terminals. Their similar actions include eliciting divergent activity patterns in normally coactive, electrically coupled neurons, providing an opportunity to examine peptide modulation of electrically coupled neurons in a functional context.


            The receptor tyrosine phosphatase Dlar and integrins organize actin filaments in the Drosophila follicular epithelium

            Date: 2001-09-04

            Creator: Jack Bateman, R. Srekantha Reddy, Haruo Saito, David Van Vactor

            Access: Open access

            Background: Regulation of actin structures is instrumental in maintaining proper cytoarchitecture in many tissues. In the follicular epithelium of Drosophila ovaries, a system of actin filaments is coordinated across the basal surface of cells encircling the oocyte. These filaments have been postulated to regulate oocyte elongation; however, the molecular components that control this cytoskeletal array are not yet understood. Results: We find that the receptor tyrosine phosphatase (RPTP) Dlar and integrins are involved in organizing basal actin filaments in follicle cells. Mutations in Dlar and the common β-integrin subunit mys cause a failure in oocyte elongation, which is correlated with a loss of proper actin filament organization. Immunolocalization shows that early in oogenesis Dlar is polarized to membranes where filaments terminate but becomes generally distributed late in development, at which time β-integrin and Enabled specifically associate with actin filament terminals. Rescue experiments point to the early period of polar Dlar localization as critical for its function. Furthermore, clonal analysis shows that loss of Dlar or mys influences actin filament polarity in wild-type cells that surround mutant tissues, suggesting that communication between neighboring cells regulates cytoskeletal organization. Finally, we find that two integrin α subunits encoded by mew and if are required for proper oocyte elongation, implying that multiple components of the ECM are instructive in coordinating actin fiber polarity. Conclusions: Dlar cooperates with integrins to coordinate actin filaments at the basal surface of the follicular epithelium. To our knowledge, this is the first direct demonstration of an RPTP's influence on the actin cytoskeleton.


            Functions of the ectodomain and cytoplasmic tyrosine phosphatase domains of receptor protein tyrosine phosphatase Dlar in vivo

            Date: 2003-10-01

            Creator: Neil X. Krueger, R. Sreekantha Reddy, Karl Johnson, Jack Bateman, Nancy, Kaufmann, Daniella Scalice, David Van Vactor, Haruo Saito

            Access: Open access

            The receptor protein tyrosine phosphatase (PTPase) Dlar has an ectodomain consisting of three immunoglobulin (Ig)-like domains and nine fibronectin type III (FnIII) repeats and a cytoplasmic domain consisting of two PTPase domains, membrane-proximal PTP-D1 and C-terminal PTP-D2. A series of mutant Dlar transgenes were introduced into the Drosophila genome via P-element transformation and were then assayed for their capacity to rescue phenotypes caused by homozygous loss-of-function genotypes. The Ig-like domains, but not the FnIII domains, are essential for survival. Conversely, the FnIII domains, but not the Ig-like domains, are required during oogenesis, suggesting that different domains of the Dlar ectodomain are involved in distinct functions during Drosophila development. All detectable PTPase activity maps to PTP-D1 in vitro. The catalytically inactive mutants of Dlar were able to rescue Dlar -/- lethality nearly as efficiently as wild-type Dlar transgenes, while this ability was impaired in the PTP-D2 deletion mutants DlarΔPTP-D2 and Dlarbypass. Dlar-C1929S, in which PTP-D2 has been inactivated, increases the frequency of bypass phenotype observed in Dlar-/- genotypes, but only if PTP-D1 is catalytically active in the transgene. These results indicate multiple roles for PTP-D2, perhaps by acting as a docking domain for downstream elements and as a regulator of PTP-D1.


            Targeted identification of glycosylated proteins in the gastric pathogen helicobacter pylori (Hp)

            Date: 2013-09-01

            Creator: Kanokwan Champasa, Scott A. Longwell, Aimee M. Eldridge, Elizabeth A. Stemmler, Danielle H., Dube

            Access: Open access

            Virulence of the gastric pathogen Helicobacter pylori (Hp) is directly linked to the pathogen's ability to glycosylate proteins; for example, Hp flagellin proteins are heavily glycosylated with the unusual nine-carbon sugar pseudaminic acid, and this modification is absolutely essential for Hp to synthesize functional flagella and colonize the host's stomach. Although Hp's glycans are linked to pathogenesis, Hp's glycome remains poorly understood; only the two flagellin glycoproteins have been firmly characterized in Hp. Evidence from our laboratory suggests that Hp synthesizes a large number of as-yet unidentified glycoproteins. Here we set out to discover Hp's glycoproteins by coupling glycan metabolic labeling with mass spectrometry analysis. An assessment of the subcellular distribution of azide-labeled proteins by Western blot analysis indicated that glycoproteins are present throughout Hp and may therefore serve diverse functions. To identify these species, Hp's azide-labeled glycoproteins were tagged via Staudinger ligation, enriched by tandem affinity chromatography, and analyzed by multidimensional protein identification technology. Direct comparison of enriched azide-labeled glycoproteins with a mock-enriched control by both SDS-PAGE and mass spectrometry-based analyses confirmed the selective enrichment of azide-labeled glycoproteins. We identified 125 candidate glycoproteins with diverse biological functions, including those linked with pathogenesis. Mass spectrometry analyses of enriched azide-labeled glycoproteins before and after cleavage of O-linked glycans revealed the presence of Staudinger ligation-glycan adducts in samples only after beta-elimination, confirming the synthesis of O-linked glycoproteins in Hp. Finally, the secreted colonization factors urease alpha and urease beta were biochemically validated as glycosylated proteins via Western blot analysis as well as by mass spectrometry analysis of cleaved glycan products. These data set the stage for the development of glycosylation-based therapeutic strategies, such as new vaccines based on natively glycosylated Hp proteins, to eradicate Hp infection. Broadly, this report validates metabolic labeling as an effective and efficient approach for the identification of bacterial glycoproteins. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.


            Pheromones enhance somatosensory processing in newt brains through a vasotocin-dependent mechanism

            Date: 2008-07-22

            Creator: R. R. Thompson, P. S. Dickinson, J. D. Rose, K. A. Dakin, G. M., Civiello, A. Segerdahl, R. Bartlett

            Access: Open access

            We tested whether the sex pheromones that stimulate courtship clasping in male roughskin newts do so, at least in part, by amplifying the somatosensory signals that directly trigger the motor pattern associated with clasping and, if so, whether that amplification is dependent on endogenous vasotocin (VT). Female olfactory stimuli increased the number of action potentials recorded in the medulla of males in response to tactile stimulation of the cloaca, which triggers the clasp motor reflex, as well as to tactile stimulation of the snout and hindlimb. That enhancement was blocked by exposing the medulla to a V1a receptor antagonist before pheromone exposure. However, the antagonist did not affect medullary responses to tactile stimuli in the absence of pheromone exposure, suggesting that pheromones amplify somatosensory signals by inducing endogenous VT release. The ability of VT to couple sensory systems together in response to social stimulation could allow this peptide to induce variable behavioural outcomes, depending on the immediate context of the social interaction and thus on the nature of the associated stimuli that are amplified. If widespread in vertebrates, this mechanism could account for some of the behavioural variability associated with this and related peptides both within and across species. © 2008 The Royal Society.


            Long-term maintenance of channel distribution in a central pattern generator neuron by neuromodulatory inputs revealed by decentralization in organ culture

            Date: 2001-09-15

            Creator: Adi Mizrahi, Patsy S. Dickinson, Peter Kloppenburg, Valerie Fénelon, Deborah J., Baro, Ronald M. Harris-Warrick, Pierre Meyrand, John Simmers

            Access: Open access

            Organotypic cultures of the lobster (Homarus gammarus) stomatogastric nervous system (STNS) were used to assess changes in membrane properties of neurons of the pyloric motor pattern-generating network in the long-term absence of neuromodulatory inputs to the stomatogastric ganglion (STG). Specifically, we investigated decentralization-induced changes in the distribution and density of the transient outward current, IA, which is encoded within the STG by the shal gene and plays an important role in shaping rhythmic bursting of pyloric neurons. Using an antibody against lobster shal K+ channels, we found shal immunoreactivity in the membranes of neuritic processes, but not somata, of STG neurons in 5 d cultured STNS with intact modulatory inputs. However, in 5 d decentralized STG, shal immunoreactivity was still seen in primary neurites but was likewise present in a subset of STG somata. Among the neurons displaying this altered shal localization was the pyloric dilator (PD) neuron, which remained rhythmically active in 5 d decentralized STG. Two-electrode voltage clamp was used to compare IA in synaptically isolated PD neurons in long-term decentralized STG and nondecentralized controls. Although the voltage dependence and kinetics of IA changed little with decentralization, the maximal conductance of IA in PD neurons increased by 43.4%. This increase was consistent with the decentralization-induced increase in shal protein expression, indicating an alteration in the density and distribution of functional A-channels. Our results suggest that, in addition to the short-term regulation of network function, modulatory inputs may also play a role, either directly or indirectly, in controlling channel number and distribution, thereby maintaining the biophysical character of neuronal targets on a long-term basis.


            Quantification of dendritic and axonal growth after injury to the auditory system of the adult cricket gryllus bimaculatus

            Date: 2013-09-27

            Creator: Alexandra Pfister, Amy Johnson, Olaf Ellers, Hadley W. Horch

            Access: Open access

            Dendrite and axon growth and branching during development are regulated by a complex set of intracellular and external signals. However, the cues that maintain or influence adult neuronal morphology are less well understood. Injury and deafferentation tend to have negative effects on adult nervous systems. An interesting example of injury-induced compensatory growth is seen in the cricket, Gryllus bimaculatus. After unilateral loss of an ear in the adult cricket, auditory neurons within the central nervous system (CNS) sprout to compensate for the injury. Specifically, after being deafferented, ascending neurons (AN-1 and AN-2) send dendrites across the midline of the prothoracic ganglion where they receive input from auditory afferents that project through the contralateral auditory nerve (N5). Deafferentation also triggers contralateral N5 axonal growth. In this study, we quantified AN dendritic and N5 axonal growth at 30 h, as well as at 3, 5, 7, 14, and 20 days after deafferentation in adult crickets. Significant differences in the rates of dendritic growth between males and females were noted. In females, dendritic growth rates were non-linear; a rapid burst of dendritic extension in the first few days was followed by a plateau reached at 3 days after deafferentation. In males, however, dendritic growth rates were linear, with dendrites growing steadily over time and reaching lengths, on average, twice as long as in females. On the other hand, rates of N5 axonal growth showed no significant sexual dimorphism and were linear. Within each animal, the growth rates of dendrites and axons were not correlated, indicating that independent factors likely influence dendritic and axonal growth in response to injury in this system. Our findings provide a basis for future study of the cellular features that allow differing dendrite and axon growth patterns as well as sexually dimorphic dendritic growth in response to deafferentation. © 2013 Pfister, Johnson, Ellers and Horch.


            AMGSEFLamide, a member of a broadly conserved peptide family, modulates multiple neural networks in Homarus americanus

            Date: 2019-01-01

            Creator: Patsy S. Dickinson, Evyn S. Dickinson, Emily R. Oleisky, Cindy D. Rivera, Meredith E., Stanhope, Elizabeth A. Stemmler, J. Joe Hull, Andrew E. Christie

            Access: Open access

            Recent genomic/transcriptomic studies have identified a novel peptide family whose members share the carboxyl terminal sequence –GSEFLamide. However, the presence/identity of the predicted isoforms of this peptide group have yet to be confirmed biochemically, and no physiological function has yet been ascribed to any member of this peptide family. To determine the extent to which GSEFLamides are conserved within the Arthropoda, we searched publicly accessible databases for genomic/transcriptomic evidence of their presence. GSEFLamides appear to be highly conserved within the Arthropoda, with the possible exception of the Insecta, in which sequence evidence was limited to the more basal orders. One crustacean in which GSEFLamides have been predicted using transcriptomics is the lobster, Homarus americanus. Expression of the previously published transcriptome-derived sequences was confirmed by reverse transcription (RT)-PCR of brain and eyestalk ganglia cDNAs; mass spectral analyses confirmed the presence of all six of the predicted GSEFLamide isoforms – IGSEFLamide, MGSEFLamide, AMGSEFLamide, VMGSEFLamide, ALGSEFLamide and AVGSEFLamide – in H. americanus brain extracts. AMGSEFLamide, of which there are multiple copies in the cloned transcripts, was the most abundant isoform detected in the brain. Because the GSEFLamides are present in the lobster nervous system, we hypothesized that they might function as neuromodulators, as is common for neuropeptides. We thus asked whether AMGSEFLamide modulates the rhythmic outputs of the cardiac ganglion and the stomatogastric ganglion. Physiological recordings showed that AMGSEFLamide potently modulates the motor patterns produced by both ganglia, suggesting that the GSEFLamides may serve as important and conserved modulators of rhythmic motor activity in arthropods.


            Characterization of the mature form of a β-defensin-like peptide, Hoa-D1, in the lobster Homarus americanus

            Date: 2018-09-01

            Creator: Giap H. Vu, Daniel Do, Cindy D. Rivera, Patsy S. Dickinson, Andrew E., Christie, Elizabeth A. Stemmler

            Access: Open access

            We report on the characterization of the native form of an American lobster, Homarus americanus, β-defensin-like putative antimicrobial peptide, H. americanus defensin 1 (Hoa-D1), sequenced employing top-down and bottom-up peptidomic strategies using a sensitive, chip-based nanoLC-QTOF-MS/MS instrument. The sequence of Hoa-D1 was determined by mass spectrometry; it was found to contain three disulfide bonds and an amidated C-terminus. The sequence was further validated by searching publicly-accessible H. americanus expressed sequence tag (EST) and transcriptome shotgun assembly (TSA) datasets. Hoa-D1, SYVRScSSNGGDcVYRcYGNIINGAcSGSRVccRSGGGYamide (with c representing a cysteine participating in a disulfide bond), was shown to be related to β-defensin-like peptides previously reported from Panulirus japonicas and Panulirus argus. We found Hoa-D1 in H. americanus hemolymph, hemocytes, the supraoesophageal ganglion (brain), eyestalk ganglia, and pericardial organ extracts, as well as in the plasma of some hemolymph samples. Using discontinuous density gradient separations, we fractionatated hemocytes and localized Hoa-D1 to hemocyte sub-populations. While Hoa-D1 was detected in semigranulocytes and granulocytes using conventional proteomic strategies for analysis, the direct analysis of cell lysates exposed evidence of Hoa-D1 processing, including truncation of the C-terminal tyrosine residue, in the granulocytes, but not semigranulocytes. These measurements demonstrate the insights regarding post-translational modifications and peptide processing that can be revealed through the MS analysis of intact peptides. The identification of Hoa-D1 as a widely-distributed peptide in the lobster suggests the possibility that it may be pleiotropic, with functions in addition to its proposed role as an antimicrobial molecule in the innate immune system.


            Characterization of plexinA and two distinct semaphorin1a transcripts in the developing and adult cricket Gryllus bimaculatus

            Date: 2020-03-01

            Creator: Hadley W. Horch, Sara B. Spicer, Isabel I.C. Low, Colby T. Joncas, Eleanor D., Quenzer, Hikmah Okoya, Lisa M. Ledwidge, Harrison P. Fisher

            Access: Open access

            Guidance cues act during development to guide growth cones to their proper targets in both the central and peripheral nervous systems. Experiments in many species indicate that guidance molecules also play important roles after development, though less is understood about their functions in the adult. The Semaphorin family of guidance cues, signaling through Plexin receptors, influences the development of both axons and dendrites in invertebrates. Semaphorin functions have been extensively explored in Drosophila melanogaster and some other Dipteran species, but little is known about their function in hemimetabolous insects. Here, we characterize sema1a and plexA in the cricket Gryllus bimaculatus. In fact, we found two distinct predicted Sema1a proteins in this species, Sema1a.1 and Sema1a.2, which shared only 48% identity at the amino acid level. We include a phylogenetic analysis that predicted that many other insect species, both holometabolous and hemimetabolous, express two Sema1a proteins as well. Finally, we used in situ hybridization to show that sema1a.1 and sema1a.2 expression patterns were spatially distinct in the embryo, and both roughly overlap with plexA. All three transcripts were also expressed in the adult brain, mainly in the mushroom bodies, though sema1a.2 was expressed most robustly. sema1a.2 was also expressed strongly in the adult thoracic ganglia while sema1a.1 was only weakly expressed and plexA was undetectable.


            How do long-term above-ground biomass dynamics vary between different forest stand types at Harvard Forest?

            Date: 2023-01-01

            Creator: Maya Y. Chandar-Kouba

            Access: Open access

            Monitoring forest carbon storage is necessary in accurately modelling the global carbon cycle. In the Northeast, terrestrial forests represent a major carbon sink with above-ground biomass (AGB) accounting for 40% of stored forest carbon. Therefore, understanding how AGB varies spatiotemporally is essential in predicting future carbon storage. Repeated measurements in permanent, long-term plots provide an opportunity to examine how carbon stored in AGB is changing over time. I used 29 years of data from the Harvard Forest Environmental Monitoring Systems (HF EMS) Site to determine how stand composition, intrinsic factors, and extrinsic environmental factors influenced rates of carbon storage in AGB over time. Using a partition around medoids (PAM) clustering method, I separated the 34 ground plots at the EMS stand into their respective stand types. I found that each stand type at the HF EMS plots accumulates carbon at consistent rates throughout the study period, although rates of carbon accumulation between stands were significantly different. Red Pine stands experience a rapid decline in biomass in 2018 due to the introduction of the Southern Pine Beetle. Across all stand types, sporadic mortality events determine variations in yearly rates of carbon accumulation, although this has little significant influence on total AGB accumulation. Leaf area index (LAI) and foliar N contents have no effect on growth increments. Extrinsic environmental variables had mixed effects on growth and mortality, highlighting the complexities of predicting forest carbon storage under changing climate conditions.


            Miniature of A multifaceted analysis of Semaphorin-induced neuroplasticity in the nervous system of <i>Gryllus bimaculatus</i>
            A multifaceted analysis of Semaphorin-induced neuroplasticity in the nervous system of Gryllus bimaculatus
            Access to this record is restricted to members of the Bowdoin community. Log in here to view.
            • Restriction End Date: 2028-06-01

              Date: 2023-01-01

              Creator: Ean Lev Small

              Access: Access restricted to the Bowdoin Community



                Miniature of Live imaging of somatic homolog pairing in <i>Drosophila</i> supports a button-based mechanism for pairing facilitated by the genetic insulator <i>gypsy</i>
                Live imaging of somatic homolog pairing in Drosophila supports a button-based mechanism for pairing facilitated by the genetic insulator gypsy
                This record is embargoed.
                  • Embargo End Date: 2026-05-18

                  Date: 2023-01-01

                  Creator: Holden D. Hadfield

                  Access: Embargoed



                    Miniature of Characterization of Retinoic Acid Signaling  During Tooth Morphogenesis and Evolution in <i>Danio rerio</i>
                    Characterization of Retinoic Acid Signaling During Tooth Morphogenesis and Evolution in Danio rerio
                    This record is embargoed.
                      • Embargo End Date: 2028-05-17

                      Date: 2023-01-01

                      Creator: Lyn Stephanie Miranda Portillo

                      Access: Embargoed



                        Mitochondrial adaptation in the green crab hybrid zone of the Gulf of Maine

                        Date: 2024-01-01

                        Creator: Jared Lynch

                        Access: Open access

                        The mitochondrial genome has historically been relegated to a neutral genetic marker, but new evidence suggests mitochondrial DNA to be a target for adaptation to environmental stress. The invasive European green crab (Carcinus maenas) exemplifies this in the Gulf of Maine’s hybrid zone, where interbreeding populations exhibit thermal tolerances influenced by mitochondrial genotype. To better understand the mechanism behind this phenomenon, the effect of mitochondrial genotype on mitochondrial activity was tested by measuring mtDNA copy number (mtCN) and the activity of complex I, II, and IV of the electron transport system via high-resolution respirometry. Mitochondria isolated from frozen heart tissue were measured at three temperature points—5°C, 25°C, and 37°C—to represent thermal stresses and a control. It was predicted that cold-adapted haplogroups would exhibit both higher mtCN and increased activity for each complex, either across all temperatures or exclusively at 5°C compared to a warm-adapted haplogroup. Initial comparisons of mitochondria from fresh and frozen tissue at 25°C found lower activity for complex II and IV in frozen extracts, but they continued to be used for convenience. No differences were observed across haplogroups for mtCN or high-resolution respirometry, suggesting that mitochondrial activity does not underlie differences in thermal tolerance. However, temperature greatly influenced activity measurements with complex II and IV exhibiting the highest rates at 37°C while complex I exhibited optimal activity at 25°C. This study represents the first of its kind for C. maenas, providing a foundation for future experiments to continue exploring mitochondria in the context of adaptive evolution.


                        Miniature of Multi-scale effects of environmental stress on Pinus ponderosa
                        Multi-scale effects of environmental stress on Pinus ponderosa
                        Access to this record is restricted to members of the Bowdoin community. Log in here to view.

                            Date: 2025-01-01

                            Creator: Cara Sydney Nova Fields

                            Access: Access restricted to the Bowdoin Community



                              Miniature of Freezing temperatures drive functional trait clustering more than habitat structure in eelgrass communities in the Gulf of Maine
                              Freezing temperatures drive functional trait clustering more than habitat structure in eelgrass communities in the Gulf of Maine
                              This record is embargoed.
                                • Embargo End Date: 2026-05-18

                                Date: 2023-01-01

                                Creator: Bridget Marjorie Patterson

                                Access: Embargoed



                                  Post-translational modification directs nuclear and hyphal tip localization of Candida albicans mRNA-binding protein Slr1

                                  Date: 2017-05-01

                                  Creator: Chaiyaboot Ariyachet, Christian Beißel, Xiang Li, Selena Lorrey, Olivia, Mackenzie, Patrick M. Martin, Katharine O'Brien, Tossapol Pholcharee, Sue Sim, Heike Krebber, Anne E. McBride

                                  Access: Open access

                                  The morphological transition of the opportunistic fungal pathogen Candida albicans from budding to hyphal growth has been implicated in its ability to cause disease in animal models. Absence of SR-like RNA-binding protein Slr1 slows hyphal formation and decreases virulence in a systemic candidiasis model, suggesting a role for post-transcriptional regulation in these processes. SR (serine–arginine)-rich proteins influence multiple steps in mRNA metabolism and their localization and function are frequently controlled by modification. We now demonstrate that Slr1 binds to polyadenylated RNA and that its intracellular localization is modulated by phosphorylation and methylation. Wildtype Slr1-GFP is predominantly nuclear, but also co-fractionates with translating ribosomes. The non-phosphorylatable slr1-6SA-GFP protein, in which six serines in SR/RS clusters are substituted with alanines, primarily localizes to the cytoplasm in budding cells. Intriguingly, hyphal cells display a slr1-6SA-GFP focus at the tip near the Spitzenkörper, a vesicular structure involved in molecular trafficking to the tip. The presence of slr1-6SA-GFP hyphal tip foci is reduced in the absence of the mRNA-transport protein She3, suggesting that unphosphorylated Slr1 associates with mRNA–protein complexes transported to the tip. The impact of SLR1 deletion on hyphal formation and function thus may be partially due to a role in hyphal mRNA transport.


                                  State of the Arg: Protein methylation at arginine comes of age

                                  Date: 2001-07-13

                                  Creator: Anne E. McBride, Pamela A. Silver

                                  Access: Open access



                                  Analysis of the yeast arginine methyltransferase Hmt1p/Rmt1p and its in vivo function. Cofactor binding and substrate interactions

                                  Date: 2000-02-04

                                  Creator: Anne E. McBride, Valerie H. Weiss, Heidi K. Kim, James M. Hogle, Pamela A., Silver

                                  Access: Open access

                                  Many eukaryotic RNA-binding proteins are modified by methylation of arginine residues. The yeast Saccharomyces cerevisiae contains one major arginine methyltransferase, Hmt1p/Rmt1p, which is not essential for normal cell growth. However, cells missing HMT1 and also bearing mutations in the mRNA-binding proteins Np13p or Cbp80p can no longer survive, providing genetic backgrounds in which to study Hmt1p function. We now demonstrate that the catalytically active form of Hmt1p is required for its activity in vivo. Amino acid changes in the putative Hmt1p S-adenosyl-L-methionine-binding site were generated and shown to be unable to catalyze methylation of Np13p in vitro and in vivo or to restore growth to strains that require HMT1. In addition these mutations affect nucleocytoplasmic transport of Np13p. A cold- sensitive mutant of Hmt1p was generated and showed reduced methylation of Np13p, but not of other substrates, at 14 °C. These results define new aspects of Hmt1 and reveal the importance of its activity in vivo.


                                  The capacity to act in trans varies among drosophila enhancers

                                  Date: 2016-05-01

                                  Creator: Amanda J. Blick, Ilana Mayer-Hirshfeld, Beatriz R. Malibiran, Matthew A. Cooper, Pieter A., Martino, Justine E. Johnson, Jack R. Bateman

                                  Access: Open access

                                  The interphase nucleus is organized such that genomic segments interact in cis, on the same chromosome, and in trans, between different chromosomes. In Drosophila and other Dipterans, extensive interactions are observed between homologous chromosomes, which can permit enhancers and promoters to communicate in trans. Enhancer action in trans has been observed for a handful of genes in Drosophila, but it is as yet unclear whether this is a general property of all enhancers or specific to a few. Here, we test a collection of well-characterized enhancers for the capacity to act in trans. Specifically, we tested 18 enhancers that are active in either the eye or wing disc of third instar Drosophila larvae and, using two different assays, found evidence that each enhancer can act in trans. However, the degree to which trans-action was supported varied greatly between enhancers. Quantitative analysis of enhancer activity supports a model wherein an enhancer’s strength of transcriptional activation is a major determinant of its ability to act in trans, but that additional factors may also contribute to an enhancer’s trans-activity. In sum, our data suggest that a capacity to activate a promoter on a paired chromosome is common among Drosophila enhancers.


                                  Manipulation of Fgf and Bmp signaling in teleost fishes suggests potential pathways for the evolutionary origin of multicuspid teeth

                                  Date: 2013-03-01

                                  Creator: William R. Jackman, Shelby H. Davies, David B. Lyons, Caitlin K. Stauder, Benjamin R., Denton-Schneider, Andrea Jowdry, Sharon R. Aigler, Scott A. Vogel, David W. Stock

                                  Access: Open access

                                  Teeth with two or more cusps have arisen independently from an ancestral unicuspid condition in a variety of vertebrate lineages, including sharks, teleost fishes, amphibians, lizards, and mammals. One potential explanation for the repeated origins of multicuspid teeth is the existence of multiple adaptive pathways leading to them, as suggested by their different uses in these lineages. Another is that the addition of cusps required only minor changes in genetic pathways regulating tooth development. Here we provide support for the latter hypothesis by demonstrating that manipulation of the levels of Fibroblast growth factor (Fgf) or Bone morphogenetic protein (Bmp) signaling produces bicuspid teeth in the zebrafish (Danio rerio), a species lacking multicuspid teeth in its ancestry. The generality of these results for teleosts is suggested by the conversion of unicuspid pharyngeal teeth into bicuspid teeth by similar manipulations of the Mexican Tetra (Astyanax mexicanus). That these manipulations also produced supernumerary teeth in both species supports previous suggestions of similarities in the molecular control of tooth and cusp number. We conclude that despite their apparent complexity, the evolutionary origin of multicuspid teeth is positively constrained, likely requiring only slight modifications of a pre-existing mechanism for patterning the number and spacing of individual teeth. © 2013 Wiley Periodicals, Inc.


                                  Pleiotropic functions of embryonic sonic hedgehog expression link jaw and taste bud amplification with eye loss during cavefish evolution

                                  Date: 2009-06-01

                                  Creator: Yoshiyuki Yamamoto, Mardi S. Byerly, William R. Jackman, William R. Jeffery

                                  Access: Open access

                                  This study addresses the role of sonic hedgehog (shh) in increasing oral-pharyngeal constructive traits (jaws and taste buds) at the expense of eyes in the blind cavefish Astyanax mexicanus. In cavefish embryos, eye primordia degenerate under the influence of hyperactive Shh signaling. In concert, cavefish show amplified jaw size and taste bud numbers as part of a change in feeding behavior. To determine whether pleiotropic effects of hyperactive Shh signaling link these regressive and constructive traits, shh expression was compared during late development of the surface-dwelling (surface fish) and cave-dwelling (cavefish) forms of Astyanax. After an initial expansion along the midline of early embryos, shh was elevated in the oral-pharyngeal region in cavefish and later was confined to taste buds. The results of shh inhibition and overexpression experiments indicate that Shh signaling has an important role in oral and taste bud development. Conditional overexpression of an injected shh transgene at specific times in development showed that taste bud amplification and eye degeneration are sensitive to shh overexpression during the same early developmental period, although taste buds are not formed until much later. Genetic crosses between cavefish and surface fish revealed an inverse relationship between eye size and jaw size/taste bud number, supporting a link between oral-pharyngeal constructive traits and eye degeneration. The results suggest that hyperactive Shh signaling increases oral and taste bud amplification in cavefish at the expense of eyes. Therefore, selection for constructive oral-pharyngeal traits may be responsible for eye loss during cavefish evolution via pleiotropic function of the Shh signaling pathway. © 2009 Elsevier Inc. All rights reserved.


                                  Drag, drafting, and mechanical interactions in canopies of the red alga Chondrus crispus

                                  Date: 2001-01-01

                                  Creator: A. S. Johnson

                                  Access: Open access

                                  Dense algal canopies, which are common in the lower intertidal and shallow subtidal along rocky coastlines, can alter flow-induced forces in their vicinity. Alteration of flow-induced forces on algal thalli may ameliorate risk of dislodgement and will affect important physiological processes, such as rates of photosynthesis. This study found that the force experienced by a thallus of the red alga Chondrus crispus (Stackhouse) at a given flow speed within a flow tank depended upon (1) the density of the canopy surrounding the thallus, (2) the position of the thallus within the canopy, and (3) the length of the stipe of the thallus relative to the height of the canopy. At all flow speeds, a solitary thallus experienced higher forces than a thallus with neighbors. A greater than 65% reduction in force occurred when the thallus drafted in the region of slower velocities that occurs in the wake region of even a single upstream neighbor, similar to the way racing bicyclists draft one behind the other. Mechanical interactions between thalli were important to forces experienced within canopies. A thallus on the upstream edge of a canopy experienced 6% less force than it did when solitary, because the canopy physically supported it. A thallus in the middle of a canopy experienced up to 83% less force than a solitary thallus, and forces decreased with increasing canopy density. Thus, a bushy morphology that increases drag on a solitary thallus may function to decrease forces experienced by that thallus when it is surrounded by a canopy, because that morphology increases physical support provided by neighbors.


                                  Miniature of Tension production and sarcomere length in lobster (Homarus americanus) cardiac muscles: the mechanisms underlying mechanical anisotropy
                                  Tension production and sarcomere length in lobster (Homarus americanus) cardiac muscles: the mechanisms underlying mechanical anisotropy
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                                      Date: 2019-05-01

                                      Creator: Matthew Maguire

                                      Access: Access restricted to the Bowdoin Community



                                        Complete mitochondrial genomes of the human follicle mites Demodex brevis and D. folliculorum: Novel gene arrangement, truncated tRNA genes, and ancient divergence between species

                                        Date: 2014-12-16

                                        Creator: Michael F. Palopoli, Samuel Minot, Dorothy Pei, Alicia Satterly, Julie, Endrizzi

                                        Access: Open access

                                        Background: Follicle mites of the genus Demodex are found on a wide diversity of mammals, including humans. surprisingly little is known, however, about the evolution of this association. Additional sequence information promises to facilitate studies of Demodex variation within and between host species. Here we report the complete mitochondrial genome sequences of two species of Demodex known to live on humans-Demodex brevis and D. folliculorum-which are the first such genomes available for any member of the genus. We analyzed these sequences to gain insight into the evolution of mitochondrial genomes within the Acariformes. We also used relaxed molecular clock analyses, based on alignments of mitochondrial proteins, to estimate the time of divergence between these two species. Results: Both Demodex genomes shared a novel gene order that differs substantially from the ancestral chelicerate pattern, with transfer RNA (tRNA) genes apparently having moved much more often than other genes. Mitochondrial tRNA genes of both species were unusually short, with most of them unable to encode tRNAs that could fold into the canonical cloverleaf structure. indeed, several examples lacked both D- and T-arms. Finally, the high level of sequence divergence observed between these species suggests that these two lineages last shared a common ancestor no more recently than about 87 mya. Conclusions: Among Acariformes, rearrangements involving tRNA genes tend to occur much more often than those involving other genes. The truncated tRNA genes observed in both Demodex species would seem to require the evolution of extensive tRNA editing capabilities and/or coevolved interacting factors. The molecular machinery necessary for these unusual tRNAs to function might provide an avenue for developing treatments of skin disorders caused by Demodex. The deep divergence time estimated between these two species sets a lower bound on the time that Demodex have been coevolving with their mammalian hosts, and supports the hypothesis that there was an early split within the genus Demodex into species that dwell in different skin microhabitats.


                                        Natural and experimental evolution of sexual conflict within Caenorhabditis nematodes

                                        Date: 2015-05-22

                                        Creator: Michael F. Palopoli, Colin Peden, Caitlin Woo, Ken Akiha, Megan, Ary, Lori Cruze, Jennifer L. Anderson, Patrick C. Phillips

                                        Access: Open access

                                        Background: Although males and females need one another in order to reproduce, they often have different reproductive interests, which can lead to conflict between the sexes. The intensity and frequency of male-male competition for fertilization opportunities is thought to be an important contributor to this conflict. The nematode genus Caenorhabditis provides an opportunity to test this hypothesis because the frequency of males varies widely among species with different mating systems. Results: We find evidence that there is strong inter- and intra-sexual conflict within C. remanei, a dioecious species composed of equal frequencies of males and females. In particular, some C. remanei males greatly reduce female lifespan following mating, and their sperm have a strong competitive advantage over the sperm of other males. In contrast, our results suggest that both types of conflict have been greatly reduced within C. elegans, which is an androdioecious species that is composed of self-fertilizing hermaphrodites and rare males. Using experimental evolution in mutant C. elegans populations in which sperm production is blocked in hermaphrodites (effectively converting them to females), we find that the consequences of sexual conflict observed within C. remanei evolve rapidly within C. elegans populations experiencing high levels of male-male competition. Conclusions: Together, these complementary data sets support the hypothesis that the intensity of intersexual conflict varies with the intensity of competition among males, and that male-induced collateral damage to mates can evolve very rapidly within populations.


                                        Sea star inspired crawling and bouncing

                                        Date: 2020-01-01

                                        Creator: Sina Heydari, Amy Johnson, Olaf Ellers, Matthew J. McHenry, Eva, Kanso

                                        Access: Open access

                                        The oral surface of sea stars is lined with arrays of tube feet that enable them to achieve highly controlled locomotion on various terrains. The activity of the tube feet is orchestrated by a nervous system that is distributed throughout the body without a central brain. How such a distributed nervous system produces a coordinated locomotion is yet to be understood. We develop mathematical models of the biomechanics of the tube feet and the sea star body. In the model, the feet are coupled mechanically through their structural connection to a rigid body. We formulate hierarchical control laws that capture salient features of the sea star nervous system. Namely, at the tube foot level, the power and recovery strokes follow a state-dependent feedback controller. At the system level, a directionality command is communicated through the nervous system to all tube feet. We study the locomotion gaits afforded by this hierarchical control model. We find that these minimally coupled tube feet coordinate to generate robust forward locomotion, reminiscent of the crawling motion of sea stars, on various terrains and for heterogeneous tube feet parameters and initial conditions. Our model also predicts a transition from crawling to bouncing consistently with recent experiments. We conclude by commenting on the implications of these findings for understanding the neuromechanics of sea stars and their potential application to autonomous robotic systems.


                                        Miniature of Photosynthetic phenology of a boreal spruce forest observed at stand and needle scales
                                        Photosynthetic phenology of a boreal spruce forest observed at stand and needle scales
                                        This record is embargoed.
                                          • Embargo End Date: 2025-05-19

                                          Date: 2022-01-01

                                          Creator: Jeremy A. Hoyne Grosvenor

                                          Access: Embargoed



                                            Three members of a peptide family are differentially distributed and elicit differential state-dependent responses in a pattern generator-effector system

                                            Date: 2018-05-01

                                            Creator: Patsy S. Dickinson, Matthew K. Armstrong, Evyn S. Dickinson, Rebecca Fernandez, Alexandra, Miller, Sovannarath Pong, Brian W. Powers, Alixander Pupo-Wiss, Meredith E. Stanhope, Patrick J. Walsh, Teerawat Wiwatpanit, Andrew E. Christie

                                            Access: Open access

                                            C-type allatostatins (AST-Cs) are pleiotropic neuropeptides that are broadly conserved within arthropods; the presence of three AST-C isoforms, encoded by paralog genes, is common. However, these peptides are hypothesized to act through a single receptor, thereby exerting similar bioactivities within each species. We investigated this hypothesis in the American lobster, Homarus americanus, mapping the distributions of AST-C isoforms within relevant regions of the nervous system and digestive tract, and comparing their modulatory influences on the cardiac neuromuscular system. Immunohistochemistry showed that in the pericardial organ, a neuroendocrine release site, AST-C I and/or III and AST-C II are contained within distinct populations of release terminals. Moreover, AST-C I/III-like immunoreactivity was seen in midgut epithelial endocrine cells and the cardiac ganglion (CG), whereas AST-C II-like immunoreactivity was not seen in these tissues. These data suggest that AST-C I and/or III can modulate the CG both locally and hormonally; AST-C II likely acts on the CG solely as a hormonal modulator. Physiological studies demonstrated that all three AST-C isoforms can exert differential effects, including both increases and decreases, on contraction amplitude and frequency when perfused through the heart. However, in contrast to many state-dependent modulatory changes, the changes in contraction amplitude and frequency elicited by the AST-Cs were not functions of the baseline parameters. The responses to AST-C I and III, neither of which is COOH-terminally amidated, are more similar to one another than they are to the responses elicited by AST-C II, which is COOH-terminally amidated. These results suggest that the three AST-C isoforms are differentially distributed in the lobster nervous system/midgut and can elicit distinct behaviors from the cardiac neuromuscular system, with particular structural features, e.g., COOH-terminal amidation, likely important in determining the effects of the peptides. NEW & NOTEWORTHY Multiple isoforms of many peptides exert similar effects on neural circuits. In this study we show that each of the three isoforms of C-type allatostatin (AST-C) can exert differential effects, including both increases and decreases in contraction amplitude and frequency, on the lobster cardiac neuromuscular system. The distribution of effects elicited by the nonamidated isoforms AST-C I and III are more similar to one another than to the effects of the amidated AST-C II.