Showing 1 - 10 of 23 Items

Date: 2007-07-01

Creator: Patsy S. Dickinson, Jake S. Stevens, Szymon Rus, Henry R. Brennan, Christopher C., Goiney, Christine M. Smith, Lingjun Li, David W. Towle, Andrew E. Christie

Access: Open access

In arthropods, a group of peptides possessing a -Y(SO3H)GHM/ LRFamide carboxy-terminal motif have been collectively termed the sulfakinins. Sulfakinin isoforms have been identified from numerous insect species. In contrast, members of this peptide family have thus far been isolated from just two crustaceans, the penaeid shrimp Penaeus monodon and Litopenaeus vannamei. Here, we report the identification of a cDNA encoding prepro-sulfakinin from the American lobster Homarus americanus. Two sulfakinin-like sequences were identified within the open-reading frame of the cDNA. Based on modifications predicted by peptide modeling programs, and on homology to the known isoforms of sulfakinin, particularly those from shrimp, the mature H. americanus sulfakinins were hypothesized to be pEFDEY(SO3H)GHMRFamide (Hoa-SK I) and GGGEY(SO3H)DDY(SO3H)GHLRFamide (Hoa-SK II). Hoa-SK I is identical to one of the previously identified shrimp sulfakinins, while Hoa-SK II is a novel isoform. Exogenous application of either synthetic Hoa-SK I or Hoa-SK II to the isolated lobster heart increased both the frequency and amplitude of spontaneous heart contractions. In preparations in which spontaneous contractions were irregular, both peptides increased the regularity of the heartbeat. Our study provides the first molecular characterization of a sulfakinin-encoding cDNA from a crustacean, as well as the first demonstration of bioactivity for native sulfakinins in this group of arthropods.

• Restriction End Date: 2027-06-01

Date: 2022-01-01

Creator: Emily Yuan-ann Pan

Access: Access restricted to the Bowdoin Community

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.

Date: 2009-12-15

Creator: J. S. Stevens, C. R. Cashman, C. M. Smith, K. M. Beale, D. W., Towle, A. E. Christie, P. S. Dickinson

Access: Open access

pQDLDHVFLRFamide is a highly conserved crustacean neuropeptide with a structure that places it within the myosuppressin subfamily of the FMRFamide-like peptides. Despite its apparent ubiquitous conservation in decapod crustaceans, the paracrine and/or endocrine roles played by pQDLDHVFLRFamide remain largely unknown. We have examined the actions of this peptide on the cardiac neuromuscular system of the American lobster Homarus americanus using four preparations: the intact animal, the heart in vitro, the isolated cardiac ganglion (CG), and a stimulated heart muscle preparation. In the intact animal, injection of myosuppressin caused a decrease in heartbeat frequency. Perfusion of the in vitro heart with pQDLDHVFLRFamide elicited a decrease in the frequency and an increase in the amplitude of heart contractions. In the isolated CG, myosuppressin induced a hyperpolarization of the resting membrane potential of cardiac motor neurons and a decrease in the cycle frequency of their bursting. In the stimulated heart muscle preparation, pQDLDHVFLRFamide increased the amplitude of the induced contractions, suggesting that myosuppressin modulates not only the CG, but also peripheral sites. For at least the in vitro heart and the isolated CG, the effects of myosuppressin were dose-dependent (10 -9 to 10-6mol l-1 tested), with threshold concentrations (10-8-10-7 mol l-1) consistent with the peptide serving as a circulating hormone. Although cycle frequency, a parameter directly determined by the CG, consistently decreased when pQDLDHVFLRFamide was applied to all preparation types, the magnitudes of this decrease differed, suggesting the possibility that, because myosuppressin modulates the CG and the periphery, it also alters peripheral feedback to the CG.

• Embargo End Date: 2027-05-19

Date: 2022-01-01

Creator: Usira Ahmed Ali

Access: Embargoed

Date: 2020-01-01

Creator: Emily R Oleisky

Access: Access restricted to the Bowdoin Community

Date: 2020-01-01

Creator: Audrey J. Muscato

Access: Open access

Central pattern generators (CPGs) produce patterned outputs independent of sensory input. The cardiac neuromuscular system of the American lobster (Homarus americanus) is driven by a CPG called the cardiac ganglion (CG), which is composed of nine neurons, making it a model system of study. Modulation of CPGs allows for functional flexibility. One neuropeptide family that modulates the CG is C-type allatostatin (AST-C I-III). Previous research has shown variation in the responses of the CG across the three isoforms and among individuals. First, we investigated why AST-C I and III elicit responses that are more similar to each other than they are to the responses elicited by AST-C II. We hypothesized that an amino acid difference in the conserved sequence was responsible for the observed variation in responses. We synthesized isoforms of AST-C that replaced the endogenous amino acid and recorded responses to these isoforms. The identity of one particular amino acid in the conserved sequence seems to be responsible for variations in responses in frequency. Next, we focused on variation among individuals in their responses to AST-C I and III. We hypothesized that the mechanism behind this individual variation is differential expression of AST-C receptors and/or their downstream targets. We recorded physiological responses of the cardiac system to AST-C and then sequenced CG RNA from the same lobsters. Differential expression of one of the AST-C receptors and a number of downstream factors is correlated with physiological response. These findings inspire further experimentation investigating molt cycle as the underlying cause.

Date: 2020-10-01

Creator: Emily R. Oleisky, Meredith E. Stanhope, J. Joe Hull, Andrew E. Christie, Patsy S., Dickinson

Access: Open access

The American lobster, Homarus americanus, cardiac neuromuscular system is controlled by the cardiac ganglion (CG), a central pattern generator consisting of four premotor and five motor neurons. Here, we show that the premotor and motor neurons can establish independent bursting patterns when decoupled by a physical ligature. We also show that mRNA encoding myosuppressin, a cardioactive neuropeptide, is produced within the CG. We thus asked whether myosuppressin modulates the decoupled premotor and motor neurons, and if so, how this modulation might underlie the role(s) that these neurons play in myosuppressin's effects on ganglionic output. Although myosuppressin exerted dose-dependent effects on burst frequency and duration in both premotor and motor neurons in the intact CG, its effects on the ligatured ganglion were more complex, with different effects and thresholds on the two types of neurons. These data suggest that the motor neurons are more important in determining the changes in frequency of the CG elicited by low concentrations of myosuppressin, whereas the premotor neurons have a greater impact on changes elicited in burst duration. A single putative myosuppressin receptor (MSR-I) was previously described from the Homarus nervous system. We identified four additional putative MSRs (MSR-II-V) and investigated their individual distributions in the CG premotor and motor neurons using RT-PCR. Transcripts for only three receptors (MSR-II-IV) were amplified from the CG. Potential differential distributions of the receptors were observed between the premotor and motor neurons; these differences may contribute to the distinct physiological responses of the two neuron types to myosuppressin. NEW & NOTEWORTHY Premotor and motor neurons of the Homarus americanus cardiac ganglion (CG) are normally electrically and chemically coupled, and generate rhythmic bursting that drives cardiac contractions; we show that they can establish independent bursting patterns when physically decoupled by a ligature. The neuropeptide myosuppressin modulates different aspects of the bursting pattern in these neuron types to determine the overall modulation of the intact CG. Differential distribution of myosuppressin receptors may underlie the observed responses to myosuppressin.

Date: 2009-02-01

Creator: Patsy S. Dickinson, Elizabeth A. Stemmler, Elizabeth E. Barton, Christopher R. Cashman, Noah P., Gardner, Szymon Rus, Henry R. Brennan, Timothy S. McClintock, Andrew E. Christie

Access: Open access

Recently, cDNAs encoding prepro-orcokinins were cloned from the crayfish Procambarus clarkii; these cDNAs encode multiple copies of four orcokinin isoforms as well as several other peptides. Using the translated open reading frames of the P. clarkii transcripts as queries, five ESTs encoding American lobster Homarus americanus orthologs were identified via BLAST analysis. From these clones, three cDNAs, each encoding one of two distinct prepro-hormones, were characterized. Predicted processing of the deduced prepro-hormones would generate 13 peptides, 12 of which are conserved between the 2 precursors: the orcokinins NFDEIDRSGFGFN (3 copies), NFDEIDRSGFGFH (2 copies) and NFDEIDRSGFGFV (2 copies), FDAFTTGFGHN (an orcomyotropin-related peptide), SSEDMDRLGFGFN, GDY(SO3)DVYPE, VYGPRDIANLY and SAE. Additionally, one of two longer peptides (GPIKVRFLSAIFIPIAAPARSSPQQDAAAGYTDGAPV or APARSSPQQDAAAGYTDGAPV) is predicted from each prepro-hormone. MALDI-FTMS analyses confirmed the presence of all predicted orcokinins, the orcomyotropin-related peptide, and three precursor-related peptides, SSEDMDRLGFGFN, GDYDVYPE (unsulfated) and VYGPRDIANLY, in H. americanus neural tissues. SAE and the longer, unshared peptides were not detected. Similar complements of peptides are predicted from P. clarkii transcripts; the majority of these were detected in its neural tissues with mass spectrometry. Truncated orcokinins not predicted from any precursor were also detected in both species. Consistent with previous studies in the crayfish Orconectes limosus, NFDEIDRSGFGFN increased mid-/hindgut motility in P. clarkii. Surprisingly, the same peptide, although native to H. americanus, did not affect gut motility in this species. Together, our results provide the framework for future investigations of the regulation and physiological function of orcokinins/orcokinin precursor-related peptides in astacideans. © 2008 Elsevier Inc. All rights reserved.

Date: 2021-01-01

Creator: Audrey Elizabeth Jordan

Access: Open access

Networks of neurons known as central pattern generators (CPGs) generate rhythmic patterns of output to drive behaviors like locomotion. CPGs are relatively fixed networks that produce consistent patterns in the absence of other inputs. The heart contractions of the Homarus americanus are neurogenic and controlled by the CPG known as the cardiac ganglion. Neuromodulators can enable flexibility in CPG motor output, and also on muscle contractions by acting on the neuromuscular junction and the muscle itself. A tissue-specific transcriptome gleaned from the cardiac ganglion and cardiac muscle of the American lobster was used to predict the sites and sources of a variety of crustacean neuromodulators. If corresponding receptors were predicted to be expressed in the cardiac muscle, then it was hypothesized that the neuropeptide had peripheral effects. One peptide for which a cardiac muscle receptor was identified is myosuppressin. Myosuppressin has been shown to have modulatory effects at the cardiac neuromuscular system of the American lobster. In previous research, myosuppressin had modulatory effects on the periphery of cardiac neuromuscular system alone. It remains an open question of whether myosuppressin acts on the cardiac muscle directly, if it is exerting its effects at the neuromuscular junction (NMJ), or both. To test this, I performed physiological experiments on the isolated NMJ. Myosuppressin did not modulate the amplitude of the excitatory junction potentials. Since no modulatory effects were seen at the NMJ, the cardiac muscle was isolated from the cardiac ganglion and then glutamate-evoked contractions were stimulated. I showed that myosuppressin increased glutamate-evoked contraction amplitude. These data suggest myosuppressin exerts its peripheral effects at the cardiac muscle and not the NMJ.