Showing 31 - 40 of 257 Items
Context-dependent protein stabilization by methionine-to-leucine substitution shown in T4 lysozyme
Date: 1998-01-01
Creator: Leigh Ann Lipscomb, Nadine C. Gassner, Sheila D. Snow, Aimee M. Eldridge, Walter A., Baase, Devin L. Drew, Brian W. Matthews
Access: Open access
- The substitution of methionines with leucines within the interior of a protein is expected to increase stability both because of a more favorable solvent transfer team as well as the reduced entropic cost of holding a leucine side chain in a defined position. Together, these two terms are expected to contribute about 1.4 kcal/mol to protein stability for each Met → Leu substitution when fully buried. At the same time, this expected beneficial effect may be offset by steric factors due to differences in the shape of leucine and methionine. To investigate the interplay between these factors, all methionines in T4 lysozyme except at the amino-terminus were individually replaced with leucine. Of these mutants, M106L and M120L have stabilities 0.5 kcal/mol higher than wild-type T4 lysozyme, while M6L is significantly destabilized (-2.8 kcal/mol). M102L, described previously, is also destabilized (-0.9 kcal/mol). Based on this limited sample it appears that methionine-to-leucine substitutions can increase protein stability but only in a situation where the methionine side chain is fully or partially buried, yet allows the introduction of the leucine without concomitant steric interference. The variants, together with methionine-to-lysine substitutions at the same sites, follow the general pattern that substitutions at rigid, internal sites tend to be most destabilizing, whereas replacements at more solvent-exposed sites are better tolerated.
Environmental issues in russia
Date: 2008-12-01
Creator: Laura A. Henry, Vladimir Douhovnikoff
Access: Open access
- This review examines the literature available on the state of the environment and environmental protection in the Russian Federation. As the largest country on Earth, rich in natural resources and biodiversity, Russia's problems and policies have global consequences. Environmental quality and management are influenced by the legacy of Soviet economic planning and authoritarian governance, as well as by Russia's post-Soviet economic recession and current strategies of economic development. Russia achieved a reduction in some pollutants owing to the collapse of industrial production in the 1990s, but many environmental indicators suggest growing degradation. Russia has signed on to a number of international environmental agreements, but its record on implementation is mixed, and it discourages environmental activism. Scholarship on the Russian environment is a limited, but growing, field, constrained by challenges of data availability, yet it offers great potential for testing scientific and social scientific hypotheses. ©2008 by Annual Reviews.
Destabilization of cortical dendrites and spines by BDNF
Date: 1999-01-01
Creator: Hadley Wilson Horch, Alex Krüttgen, Stuart D. Portbury, Lawrence C. Katz
Access: Open access
- Particle-mediated gene transfer and two-photon microscopy were used to monitor the behavior of dendrites of individual cortical pyramidal neurons coexpressing green fluorescent protein (GFP) and brain-derived neurotrophic factor (BDNF). While the dendrites and spines of neurons expressing GFP alone grew modestly over 24-48 hr, coexpressing BDNF elicited dramatic sprouting of basal dendrites, accompanied by a regression of dendritic spines. Compared to GFP-transfected controls, the newly formed dendrites and spines were highly unstable. Experiments utilizing Trk receptor bodies, K252a, and overexpression of nerve growth factor (NGF) demonstrated that these effects were mediated by secreted BDNF interacting with extracellular TrkB receptors. Thus, BDNF induces structural instability in dendrites and spines, which, when restricted to particular portions of a dendritic arbor, may help translate activity patterns into specific morphological changes.
The pyloric neural circuit of the herbivorous crab Pugettia producta shows limited sensitivity to several neuromodulators that elicit robust effects in more opportunistically feeding decapods
Date: 2008-05-01
Creator: Patsy S. Dickinson, Elizabeth A. Stemmler, Andrew E. Christie
Access: Open access
- Modulation of neural circuits in the crustacean stomatogastric nervous system (STNS) allows flexibility in the movements of the foregut musculature. The extensive repertoire of such resulting motor patterns in dietary generalists is hypothesized to permit these animals to process varied foods. The foregut and STNS of Pugettia producta are similar to those of other decapods, but its diet is more uniform, consisting primarily of kelp. We investigated the distribution of highly conserved neuromodulators in the stomatogastric ganglion (STG) and neuroendocrine organs of Pugettia, and documented their effects on its pyloric rhythm. Using immunohistochemistry, we found that the distributions of Cancer borealis tachykinin-related peptide I (CabTRP I), crustacean cardioactive peptide (CCAP), proctolin, red pigment concentrating hormone (RPCH) and tyrosine hydroxylase (dopamine) were similar to those of other decapods. For all peptides except proctolin, the isoforms responsible for the immunoreactivity were confirmed by mass spectrometry to be the authentic peptides. Only two modulators had physiological effects on the pyloric circuit similar to those seen in other species. In non-rhythmic preparations, proctolin and the muscarinic acetylcholine agonist oxotremorine consistently initiated a full pyloric rhythm. Dopamine usually activated a pyloric rhythm, but this pattern was highly variable. In only about 25% of preparations, RPCH activated a pyloric rhythm similar to that seen in other species. CCAP and CabTRP I had no effect on the pyloric rhythm. Thus, whereas Pugettia possesses all the neuromodulators investigated, its pyloric rhythm, when compared with other decapods, appears less sensitive to many of them, perhaps because of its limited diet.
Mass spectrometric identification of pEGFYSQRYamide: A crustacean peptide hormone possessing a vertebrate neuropeptide Y (NPY)-like carboxy-terminus
Date: 2007-05-15
Creator: Elizabeth A. Stemmler, Emily A. Bruns, Noah P. Gardner, Patsy S. Dickinson, Andrew E., Christie
Access: Open access
- In invertebrates, peptides possessing the carboxy (C)-terminal motif -RXRFamide have been proposed as the homologs of vertebrate neuropeptide Y (NPY). Using matrix assisted laser desorption/ionization mass spectrometry, in combination with sustained off-resonance irradiation collision-induced dissociation and chemical and enzymatic reactions, we have identified the peptide pEGFYSQRYamide from the neuroendocrine pericardial organ (PO) of the crab Pugettia producta. This peptide is likely the same as that previously reported, but misidentified, as PAFYSQRYamide in several earlier reports (e.g. [Li, L., Kelley, W.P., Billimoria, C.P., Christie, A.E., Pulver, S.R., Sweedler, J.V., Marder, E. 2003. Mass spectrometric investigation of the neuropeptide complement and release in the pericardial organs of the crab, Cancer borealis. J. Neurochem. 87, 642-656; Fu, Q., Kutz, K.K., Schmidt, J.J., Hsu, Y.W., Messinger, D.I., Cain, S.D., de la Iglesia, H.O., Christie, A.E., Li, L. 2005. Hormone complement of the Cancer productus sinus gland and pericardial organ: an anatomical and mass spectrometric investigation. J. Comp. Neurol. 493, 607-626.]). The -QRYamide motif contained in pEGFYSQRYamide is identical to that present in many vertebrate members of the NPY superfamily. Mass spectrometric analysis conducted on the POs of several other decapods showed that pEGFYSQRYamide is present in three other brachyurans (Cancer borealis, Cancer irroratus and Cancer productus) as well as in one species from another decapod infraorder (Lithodes maja, an anomuran). Thus, our findings show that at least some invertebrates possess NPY-like peptides in addition to those exhibiting an -RXRFamide C-terminus, and raise the question as to whether the invertebrate -QRYamides are functionally and/or evolutionarily related to the NPY superfamily. © 2007 Elsevier Inc. All rights reserved.
Aortic pressure and heart rate in the lobster Homarus americanus are modulated by mechanical feedback and neuropeptides
Date: 2021-01-01
Creator: Grace Marie Hambelton
Access: Open access
- Baroreceptors are stretch receptors located in the aorta of mammals; in response to increased afterload, they elicit a decrease in heart rate, creating a negative feedback loop that lowers blood pressure. Although lobsters (Homarus americanus) do not have baroreceptors like mammals, closely related land crabs have been shown to have baroreceptor-like responses. Heart contraction is also regulated by the Frank-Starling response, where increasing stretch or preload increases the contractile force of the heart. In addition to these types of biomechanical modulations, lobsters use a central pattern generator, the cardiac ganglion, to maintain synchronicity of the heartbeat. The heart is also controlled by the central nervous system via neuromodulators, such as myosuppressin, which has been shown to increase active force and decrease frequency in isolated lobster hearts. We performed experiments on a lobster heart with the main arteries still intact, and varied the preload by stretching anterior arteries, and the afterload by elevating the dorsal abdominal artery. We added myosuppressin to modulate the cardiac ganglion output and muscle contraction. We found that the baroreceptor-like response is most directly modulated by active force, whereas frequency could be a secondary control. Increasing preload does increase active force, but that does not correlate to a higher cardiac output, which shows that how hard the heart pumps is not what determines how effectively it is pumping. Additionally, we found that myosuppressin has a much stronger effect on frequency than active force, and so with myosuppressin, frequency becomes the main determinant of cardiac output.
Molecular characterization of putative neuropeptide, amine, diffusible gas and small molecule transmitter biosynthetic enzymes in the eyestalk ganglia of the American lobster, Homarus americanus
Date: 2018-12-01
Creator: Andrew E. Christie, Meredith E. Stanhope, Helen I. Gandler, Tess J. Lameyer, Micah G., Pascual, Devlin N. Shea, Andy Yu, Patsy S. Dickinson, J. Joe Hull
Access: Open access
- The American lobster, Homarus americanus, is a model for investigating the neuromodulatory control of physiology and behavior. Prior studies have shown that multiple classes of chemicals serve as locally released/circulating neuromodulators/neurotransmitters in this species. Interestingly, while many neuroactive compounds are known from Homarus, little work has focused on identifying/characterizing the enzymes responsible for their biosynthesis, despite the fact that these enzymes are key components for regulating neuromodulation/neurotransmission. Here, an eyestalk ganglia-specific transcriptome was mined for transcripts encoding enzymes involved in neuropeptide, amine, diffusible gas and small molecule transmitter biosynthesis. Using known Drosophila melanogaster proteins as templates, transcripts encoding putative Homarus homologs of peptide precursor processing (signal peptide peptidase, prohormone processing protease and carboxypeptidase) and immature peptide modifying (glutaminyl cyclase, tyrosylprotein sulfotransferase, protein disulfide isomerase, peptidylglycine-α-hydroxylating monooxygenase and peptidyl-α-hydroxyglycine-α-amidating lyase) enzymes were identified in the eyestalk assembly. Similarly, transcripts encoding full complements of the enzymes responsible for dopamine [tryptophan-phenylalanine hydroxylase (TPH), tyrosine hydroxylase and DOPA decarboxylase (DDC)], octopamine (TPH, tyrosine decarboxylase and tyramine β-hydroxylase), serotonin (TPH or tryptophan hydroxylase and DDC) and histamine (histidine decarboxylase) biosynthesis were identified from the eyestalk ganglia, as were those responsible for the generation of the gases nitric oxide (nitric oxide synthase) and carbon monoxide (heme oxygenase), and the small molecule transmitters acetylcholine (choline acetyltransferase), glutamate (glutaminase) and GABA (glutamic acid decarboxylase). The presence and identity of the transcriptome-derived transcripts were confirmed using RT-PCR. The data presented here provide a foundation for future gene-based studies of neuromodulatory control at the level of neurotransmitter/modulator biosynthesis in Homarus.
Animal-to-animal variability in the phasing of the crustacean cardiac motor pattern: An experimental and computational analysis
Date: 2013-01-01
Creator: Alex H. Williams, Molly A. Kwiatkowski, Adam L. Mortimer, Eve Marder, Mary Lou, Zeeman, Patsy S. Dickinson
Access: Open access
- The cardiac ganglion (CG) of Homarus americanus is a central pattern generator that consists of two oscillatory groups of neurons: "small cells" (SCs) and "large cells" (LCs). We have shown that SCs and LCs begin their bursts nearly simultaneously but end their bursts at variable phases. This variability contrasts with many other central pattern generator systems in which phase is well maintained. To determine both the consequences of this variability and how CG phasing is controlled, we modeled the CG as a pair of Morris-Lecar oscillators coupled by electrical and excitatory synapses and constructed a database of 15,000 simulated networks using random parameter sets. These simulations, like our experimental results, displayed variable phase relationships, with the bursts beginning together but ending at variable phases. The model suggests that the variable phasing of the pattern has important implications for the functional role of the excitatory synapses. In networks in which the two oscillators had similar duty cycles, the excitatory coupling functioned to increase cycle frequency. In networks with disparate duty cycles, it functioned to decrease network frequency. Overall, we suggest that the phasing of the CG may vary without compromising appropriate motor output and that this variability may critically determine how the network behaves in response to manipulations. © 2013 the American Physiological Society.
Sexual reproduction is more prevalent in continental landscapes in the expanding arctic shrub, Salix glauca
Date: 2019-12-01
Creator: David A. Watts, Vladimir Douhovnikoff, Eric Post
Access: Open access
- The recent expansion of arctic deciduous shrubs has been well documented across a range of habitats, but the phenomenon is not universal. Their spread is often associated with increases in temperature and other abiotic factors, while variation in habitat moisture and herbivory can mediate the location and rate of this rise in abundance. Much less is known about the mode of increase of arctic shrubs. For one such shrub, Salix glauca, we used microsatellite markers to assess the prevalence of clonal growth (i.e. vegetative spread) and sexual reproduction (i.e. recruitment from seed) at sites with maritime and continental climates and differing in the density of large herbivores. We sampled individuals in plots reflecting the spatial scale of expansion in locations where S. glauca recently increased in abundance. The 400 samples collected across the four sites comprised 310 genotypes. Though evidence of sexual recruitment was common across all sites, coastal sites contained both more and larger clonal genotypes. While we expected soil conditions would be influential, the factors that best predicted the likelihood of clonality, genet size and vascular plant cover, suggest the light environment is of primary importance. Furthermore, in spite of the large distances between sites, there was no suggestion of genetic differentiation into distinct populations. These results indicate that differences in climate and herbivory can influence not only where and how extensively deciduous shrubs spread, but how they are likely to do so. We suggest future research integrating how mode of increase is associated with the rate of spread will advance projections of change in arctic ecosystems.
Is variation in susceptibility to Phytophthora ramorum correlated with population genetic structure in coast live oak (Quercus agrifolia)?
Date: 2005-01-01
Creator: Richard S. Dodd, Daniel Hüberli, Vlad Douhovnikoff, Tamar Y. Harnik, Zara, Afzal-Rafii, Matteo Garbelotto
Access: Open access
- California coastal woodlands are suffering severe disease and mortality as a result of infection from Phytophthora ramorum. Quercus agrifolia is one of the major woodland species at risk. This study investigated within- and among-population variation in host susceptibility to inoculation with P. ramorum and compared this with population genetic structure using molecular markers. Susceptibility was assessed using a branch-cutting inoculation test. Trees were selected from seven natural populations in California. Amplified fragment length polymorphism molecular markers were analysed for all trees used in the trials. Lesion sizes varied quantitatively among individuals within populations, with up to an eightfold difference. There was little support for population differences in susceptibility. Molecular structure also showed a strong within-population, and weaker among-population, pattern of variation. Our data suggest that susceptibility of Q. agrifolia to P. ramorum is variable and is under the control of several gene loci. This variation exists within populations, so that less susceptible local genotypes may provide the gene pool for regeneration of woodlands where mortality is high. © New Phytologist (2004).