Showing 1 - 4 of 4 Items

Date: 2021-02-17

Creator: Kellen Delaney, Mengzhou Hu, Tessa Hellenbrand, Patsy S. Dickinson, Michael P., Nusbaum, Lingjun Li

Access: Open access

The crab Cancer borealis nervous system is an important model for understanding neural circuit dynamics and modulation, but the identity of neuromodulatory substances and their influence on circuit dynamics in this system remains incomplete, particularly with respect to behavioral state-dependent modulation. Therefore, we used a multifaceted mass spectrometry (MS) method to identify neuropeptides that differentiate the unfed and fed states. Duplex stable isotope labeling revealed that the abundance of 80 of 278 identified neuropeptides was distinct in ganglia and/or neurohemal tissue from fed vs unfed animals. MS imaging revealed that an additional 7 and 11 neuropeptides exhibited altered spatial distributions in the brain and the neuroendocrine pericardial organs (POs), respectively, during these two feeding states. Furthermore, de novo sequencing yielded 69 newly identified putative neuropeptides that may influence feeding state-related neuromodulation. Two of these latter neuropeptides were determined to be upregulated in PO tissue from fed crabs, and one of these two peptides influenced heartbeat in ex vivo preparations. Overall, the results presented here identify a cohort of neuropeptides that are poised to influence feeding-related behaviors, providing valuable opportunities for future functional studies.

Date: 2021-09-01

Creator: Solymar Rolón-Martínez, Mohamed R. Habib, Tamer A. Mansour, Manuel Díaz-Ríos, Joshua J.C., Rosenthal, Xiao Nong Zhou, Roger P. Croll, Mark W. Miller

Access: Open access

Freshwater snails of the genus Biomphalaria serve as intermediate hosts for the digenetic trematode Schistosoma mansoni, the etiological agent for the most widespread form of intestinal schistosomiasis. As neuropeptide signaling in host snails can be altered by trematode infection, a neural transcriptomics approach was undertaken to identify peptide precursors in Biomphalaria glabrata, the major intermediate host for S. mansoni in the Western Hemisphere. Three transcripts that encode peptides belonging to the FMRF-NH2-related peptide (FaRP) family were identified in B. glabrata. One transcript encoded a precursor polypeptide (Bgl-FaRP1; 292 amino acids) that included eight copies of the tetrapeptide FMRF-NH2 and single copies of FIRF-NH2, FLRF-NH2, and pQFYRI-NH2. The second transcript encoded a precursor (Bgl-FaRP2; 347 amino acids) that comprised 14 copies of the heptapeptide GDPFLRF-NH2 and 1 copy of SKPYMRF-NH2. The precursor encoded by the third transcript (Bgl-FaRP3; 287 amino acids) recapitulated Bgl-FaRP2 but lacked the full SKPYMRF-NH2 peptide. The three precursors shared a common signal peptide, suggesting a genomic organization described previously in gastropods. Immunohistochemical studies were performed on the nervous systems of B. glabrata and B. alexandrina, a major intermediate host for S. mansoni in Egypt. FMRF-NH2-like immunoreactive (FMRF-NH2-li) neurons were located in regions of the central nervous system associated with reproduction, feeding, and cardiorespiration. Antisera raised against non-FMRF-NH2 peptides present in the tetrapeptide and heptapeptide precursors labeled independent subsets of the FMRF-NH2-li neurons. This study supports the participation of FMRF-NH2-related neuropeptides in the regulation of vital physiological and behavioral systems that are altered by parasitism in Biomphalaria.

Date: 2016-05-01

Creator: Tess Lameyer

Access: Access restricted to the Bowdoin Community

Date: 2019-01-01

Creator: Louis Mendez

Access: Open access

Central pattern generators (CPGs) are neural networks that generate rhythmic motor patterns to allow organisms to perform stereotypical tasks, such as breathing, scratching, flying, and walking. The American lobster, Homarus americanus, is a simple model system whose CPGs are functionally analogous to those in vertebrate models and model complex rhythmic behaviors. CPGs in many Crustacea, including the American lobster, have been studied because of their ability to maintain biological function after isolation in physiologically relevant conditions. The cardiac ganglion (CG) is a CPG consisting of five larger motor neurons and four smaller pacemaker neurons that innervate the cardiac neuromuscular system and generate electrical bursts that drive patterned behaviors. Neuromodulators, such as neuropeptides, are known to modulate neural output in the CPGs of the American lobster. Currently, neuromodulators affecting the cardiac ganglia are thought to be mainly expressed and secreted outside of the cardiac ganglia, acting as extrinsic neuromodulators. However, there is current evidence to support the idea that neuromodulators can be intrinsically expressed within the cardiac ganglion of the American lobster. Preliminary studies using transcriptomic techniques on genomic and transcriptomic information indicate that neuropeptides are likely expressed within the cardiac ganglion. However, little research has been done to determine whether these neuropeptides are expressed in the cardiac ganglion of the American lobster. Therefore, the purpose of this study is to combine bioinformatics and mass spectrometric techniques to determine whether select neuropeptides are present in the cardiac ganglion within the cardiac neuromuscular system of the American lobster, Homarus americanus. Our data mining techniques using protein query sequences obtained from previously annotated brain and eyestalk transcriptomes resulted in the identification of 22 putative neuropeptides preprohormones from 17 neuropeptide families and 20 putative neuropeptide receptors from 17 neuropeptide receptor families in the CG transcriptome. Additionally, 9 putative neuropeptide receptors from 7 neuropeptide receptor families were detected in the cardiac muscle transcriptome. Of the 17 neuropeptide families detected, receptors for 9 of these neuropeptide families were detected in the CG transcriptome. Receptors for 6 of the neuropeptide families were also present in the cardiac muscle transcriptome. Interestingly, receptors for 6 of neuropeptide families detected were not found in either the CG or cardiac muscle transcriptomes, and receptors for 4 neuropeptide families that weren’t detected in the CG transcriptome were found in the cardiac muscle transcriptome. Therefore, our research suggests that neuropeptides are able to modulate CPG activity extrinsically, either though hormonal or local delivery, or intrinsically. Additionally, neuropeptides were extracted from the stomatogastric ganglion and the commissural ganglion using a scaled-down neuropeptide extraction protocol to estimate the number of tissues required to obtain sufficiently strong mass spectrometry signals. Pooled samples with two commissural ganglia and single samples of a commissural ganglion and a stomatogastric ganglion displayed little signal and an increase in larger peptides and impurities relative to single-tissue samples. Therefore, further optimization of the scaled-down neuropeptide extraction protocol must be done prior to analysis of a cardiac ganglion in the American lobster.