Showing 1 - 10 of 26 Items

Date: 2023-01-01

Creator: Kenneth Garcia

Access: Open access

Neuromodulation allows for the flexibility of neural circuit dynamics and the outputs they produce. Studies of the stomatogastric nervous system (STNS) have expanded our knowledge on the actions of neuromodulators, small molecules that most often activate G-protein coupled receptors and reconfigure circuit activity and composition. In these systems, modulation has been found to occur at every level, from sensory-motor coupling to neuromuscular transmission (Harris-Warrick and Marder 1991). Neuromodulators have complex effects on motor output; they can alter the firing of individual neurons while also modulating muscle properties, neuromuscular transmission, and sensory neuron response to muscle activity (Fort et al. 2004). We investigated this further by recording the motor output produced by the gastric mill rhythm of the lobster STNS under neuromodulator conditions. How is this neuromuscular system as a whole modulated to produce motor flexibility? We hypothesized that these neuromodulators act on individual receptors of component neurons of central pattern generator (CPG)-effector system themselves and at the periphery, coordinately altering muscle contraction by altering all levels of the crustacean neuromuscular system. Application of NRNFLRFamide, RPCH, oxotremorine, and proctolin to the gastric mill 4 (gm4) muscles of the Cancer crab showed that neuromodulators that have been found to have variable, yet significant effects on the activity of the neurons of the STNS directly alter the activity of the gm4 muscles as well, suggesting that coordination of peripheral actions and direct neuronal modulation regulates patterned motor output.

Date: 2014-08-01

Creator: Tricia Hartley

Access: Open access

In many animals, there are groups of neurons, known as central pattern generators (CPGs), which are capable of controlling major everyday life functions. CPGs are responsible for functions that require patterned rhythmic activity, such as the heartbeat, digestion and locomotion. A CPG called the cardiac ganglion, consisting of only nine neurons, controls the rhythmic beating of the heart of the American lobster, Homarus americanus, by stimulating the muscle cells of the heart.My summer consisted of two separate projects in Patsy Dickinson’s neurophysiology lab, both studying the interaction of the cardiac ganglion with neuropeptides. These neuropeptides, GYSDRNYLRFamide (GYS) and SGRNFLRFamide (SGRN) are released hormonally into the cardiac neuromuscular system. The overarching goal of both projects was to determine the role of these neuropeptides in the lobster’s cardiac neuromuscular system.For my first project, I studied the interaction of the neuropeptide GYS with the stretch receptors of the lobster heart. Previous research has found these stretch receptors to be a form of excitatory feedback from the lobster heart to the cardiac ganglion, as heartbeat amplitude and frequency increase as heart is stretched. Further, the dendrites along the cardiac ganglion have been found to be stretch-sensitive, meaning when these dendrites were cut, this excitatory response is no longer observed. By stretching the heart with the dendrites intact and with GYS and next when the dendrites were cut and with GYS, the goal of this project was to determine if GYS would alter the feedback of the stretch receptors back to the cardiac ganglion to change heartbeat frequency and amplitude. Unfortunately, the intricacy involved in being able to cut the dendrites while allowing the heart to continue to beat proved very difficult and I moved on to my next project.The goal of my next project was to examine the interactions of the neuropeptides GYS and SGRN with the decreased and increased presence of nitric oxide, the second form of feedback from the heart muscle to the cardiac ganglion. Previous research shows nitric oxide as having an inhibitory effect, decreasing heartbeat amplitude and frequency. By applying both GYS and SGRN to both the isolated cardiac ganglion and the whole heart in the presence of both a nitric oxide inhibitor and donor, the hope is to be able to determine the interaction of these peptides with and without the presence of the feedback of nitric oxide. Because I started this project later in the summer, with the assistance of Sophie Janes’ data, I have been able to look at the effects of GYS on the whole heart, in addition to the combination of GYS with L-NA, a nitric oxide inhibitor. So far, the data has shown that the combination of GYS with L-NA causes less of a decrease in heartbeat frequency than GYS alone, which shows a significant decrease. We predict this is because GYS enhances the nitric oxide pathway, while L-NA is blocking the nitric oxide pathway, thus giving insight into the role of GYS within the lobster’s cardiac neuromuscular system. For my senior independent study I hope to continue this research and be able to continue to compile data for both SGRN and GYS on the isolated cardiac ganglion as well as on the whole heart, with a nitric oxide inhibitor and donor. Final Report of research funded by a Doherty Coastal Studies Research Fellowship.

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: 2019-05-01

Creator: Casey Breslow

Access: Open access

Modulation in neural systems is important for regulating physiology and behavior (Wright et al., 2010). Peptides, hormones, and amines are common neural modulators, acting on many neural systems across species. One group of neural networks that can be regulated are central pattern generators (CPGs), which generate rhythmic neural patterns, which drive behaviors (Marder and Bucher, 2001). Octopamine, and its precursor tyramine, are two amines that have been found to regulate (CPGs) across species (Cooke, 2002; Fussnecker et al., 2006). One role of octopamine in the decapod neurogenic heart is regulating the frequency and the duration of heart beats. However, the precise site of octopamine modulation within the cardiac system is not yet known (Kurumoto and Ebara, 1991). One possible site of action is the cardiac ganglion (CG), the CPG in decapod hearts. The transcripts for the enzymes required to synthesize octopamine from tyramine have been identified and localized in the CG (Christie et al., 2018). This would suggest that octopamine is produced in the CG, where it could have a direct action on those neurons, or it could be released peripherally. We have found individual variation in the response to octopamine and its precursor tyramine, and significant effects of frequency and contraction amplitude in the whole heart.

Date: 2014-08-01

Creator: Xuan Qu

Access: Open access

Central pattern generators are networks of neurons that produce rhythmic and repetitiveoutputs. These outputs control behaviors such as walking, breathing and digestion. In the Americanlobster, central pattern generators control the behavior of muscles in its foregut, which allows thedigestion of a variety of food types. The stomatogastric ganglion (STG) is a bundle of about thirtyneurons in the foregut of American lobsters. It has been studied extensively since each one of theneurons in it is both identifiable and produces simple patterned outputs. The analysis of American lobster’s stomach behaviors and the neural mechanisms controlling them could provide general insights into how rhythmic motor patterns for locomotion are produced. A large number of the neurons in the STG are modulatory neurons that use neuromodulators for at least part of their synaptic receptions. These neuromodulators are released by neurons and cause long-lasting changes in the synaptic efficacies of the targets. At present, many types of neuropeptides have been identified within the crustacean stomatogastric nervous system. The pyrokinins are members of one peptide family, PBAN. PBAN peptides all share the common Cterminalpentapeptide FXPRL-amide, in which X can be S, T, G, N, or V. Previous studies, using immunohistochemistry, have found that there are pyrokinin peptides present in both the STG and the cardiac ganglion (CG) of American lobsters. My research tests five different kinds of pyrokinin peptides, including PevPK1 (DFAFSPRLamide) and PevPK2 (ADFAFNPRLamide) from the shrimp L.vannamei (Torfs et al., 2001; Ma et al., 2010), CabPK1 (TNFAFSPRLamide) and CabPK2(SGGFAFSPRLamide from the crab C.borealis (Saideman et al., 2007;Ma et al., 2009) and Conserved Sequence (FSPRLamide) from the lobster, H.americanus (Ma, et al, 2008). ConservedSequence, the only pyrokinin identified in the American lobster so far, is highly conserved among many other pyrokinin peptides. Therefore, it is believed to be just a fragment with the complete sequence yet to be identified. Thus, we predicted that it might produce a weaker effect on the STG. Previous studies on the pyrokinin peptides have shown that in crabs, CabPK1, CabPK2 and LeucoPK (identified in an insect), all had a virtually identical effect on the CG, suggesting that the differences among these pyrokinin peptides are not important and the receptors for these peptides are the same. However, research done by Bowdoin students in 2011-2012 showed that among PevPK1,PevPK2, CabPK1, CabPK2, and Conserved Sequence, all but Conserved Sequence (not yet tested) had strong effects on the STG. However, only PevPK2 had an effect on the CG. My goal for this summer research was to determine whether or not there are differences between the responses of the STG to the different peptides in order to further determine the cause for the differences between the responses of the CG and those of the STG. The results from the extracellular recordings from the identified neurons in my research have shown that none of the five kinds of pyrokinin peptides affect the pyloric rhythm, which controls the pumping and filtering of food through the pylorus in Americanlobsters. They all, however, excite the gastric mill rhythm, which controls the movements of the teeth that grind up the food before it is transferred into the pylorus. Moreover, there is no significant difference among the effects of these five kinds of pyrokinin peptides. Conserved Sequence, which was predicted to produce a relatively weaker effect, proved to produce virtually identical effect asfour other kinds of pyrokinin peptides. Future research will focus on studying the differences between the STG and CG to determine the cause of the varied responses between them. Final Report of research funded by the Doherty Coastal Studies Research Fellowship.

Date: 2020-01-01

Creator: Grace Bukowski-Thall

Access: Open access

Central pattern generators (CPGs) are neural networks that generate the rhythmic outputs that control behaviors such as locomotion, respiration, and chewing. The stomatogastric nervous system (STNS), which contains the CPGs that control foregut movement, and the cardiac ganglion (CG), which is a CPG that controls heartbeat, are two commonly studied systems in decapod crustaceans. Neuromodulators are locally or hormonally released neuropeptides and amines that change the output patterns of CPGs like the STNS and CG to allow behavioral flexibility. We have hypothesized that neuromodulation provides a substrate for the evolution of behavioral flexibility, and as a result, systems exhibiting more behavioral flexibility are modulated to a greater degree. To examine this hypothesis, we evaluated the extent to which the STNS and the CG are modulated in the majoid crab species Chionoecetes opilio, Libinia emarginata, and Pugettia producta. C. opilio and L. emarginata are opportunistic feeders, whereas P. producta has a highly specialized kelp diet. We predicted that opportunistic feeding crabs that chew and process a wide variety of food types would exhibit greater STNS neuromodulatory capacity than those with a specialized diet. The STNS of L. emarginata and C. opilio responded to the seven endogenous neuromodulators oxotremorine, dopamine, CabTrp Ia, CCAP, myosuppressin, proctolin, and RPCH, whereas the STNS of P. producta only responded to proctolin, oxotremorine, myosuppressin, RPCH (25% of the time), variably to dopamine, and not at all to CabTrp and CCAP. Because P. producta, L. emarginata, and C. opilio all belong to the Majoidea superfamily, their primary distinctions are their feeding habits. For this reason, we further predicted that there would be no relationship between diet and modulatory capacity in the cardiac ganglion (CG) of the neurogenic heart. This would suggest that a lack of STNS modulatory capacity in P. producta relative to L. emarginata and C. opilio is specific to evolved foregut function. Whole-heart recordings from P. producta indicated that, unlike the STNS, the CG responds to CabTrp and CCAP. P. producta hearts also responded to oxotremorine and inconsistently to dopamine and proctolin. The CG of C. opilio was modulated by CabTrp, CCAP, dopamine, proctolin, myosuppressin, and oxotremorine, but not RPCH. The CG of L. emarginata responded to CCAP, and inconsistently to CabTrp, dopamine, and proctolin, but not to myosuppressin, RPCH, and surprisingly oxotremorine. Although cardiac responses were not identical between species, opportunistic and specialist feeders responded more similarly to the modulators tested in the heart than in the STNS. Notably, P. producta responded to each modulator in a similar manner to C. opilio and/or L. emarginata. However, L. emarginata’s surprising lack of cardiac response to oxotremorine suggests that phylogenetic closeness may not control for differences in CG and STNS function between species. Nevertheless, sample sizes of all three species were quite small, and individual differences lead to inconsistencies in the data. As a result, sample size must be enlarged to draw firm conclusions.

Date: 2023-01-01

Creator: Elise Martin

Access: Open access

This project sought to answer the following question: what is the relationship between the extent of neuromodulation in a nervous system, and the behavioral demands on that system? A well-characterized CPG neuronal circuit in decapod crustaceans, the stomatogastric nervous system (STNS), was used as a model circuit to answer this question. The stomatogastric ganglion (STG) in the STNS is responsible for muscular contractions in the stomach that aid in digestion. It has been shown that the neural networks in the STG are subject to neuromodulation. One feature of neuromodulation is that it enables circuit flexibility, which confers upon a system the ability to produce variable outputs in response to specific physiological demands. It was hypothesized that opportunistic feeders require more extensively modulated digestive systems compared to exclusive feeders, because opportunistic feeders require a greater variety of digestive outputs to digest their varied diets. In this study, Chionoecetes opilio and Libinia emarginata, the opportunistic feeders, showed greater neuromodulatory capacity of the STNS than Pugettia producta, the exclusive feeder. The hypothesis that neuromodulatory capacity of the STNS correlates with dietary diversity was supported. The results detailed in this study lend credence to the idea that evolutionary basis for neuromodulatory capacity of a system is related to the behavioral demands on that system.

Date: 2023-01-01

Creator: John T Woolley

Access: Open access

Picrotoxin (PTX) has been employed extensively as a tool within the crustacean stomatogastric nervous system (STNS) for its efficacy in blocking K+ and Cl+ currents gated by both GABA and glutamate. Through blocking some currents in the STNS, PTX allows for examination of other components without their presence. However, effects of PTX are relatively unknown within the lobster’s cardiac ganglion (CG). As an incredibly small nervous system of only nine neurons, the lobster CG presents an excellent model system for studying neural circuits. Given that the chemical synapses in the CG are mediated by glutamate, the present study aimed to investigate the action of PTX in the lobster CG with the intent of better understanding its pharmacological impacts as a potential tool for studying the system. Therefore, this study aimed to establish the effects of PTX on CG responses to the application of exogenous GABA or glutamate. When data from both modulators were pooled, PTX applied at a concentration of 10-5M had significant effects on burst duration but not duty cycle or burst frequency of the CG. PTX did suppress GABA (5x10-5M) mediated inhibition of burst duration and duty cycle. PTX did not have any significant effects on burst duration, duty cycle, or frequency compared to exogenous glutamate application. These results indicate that glutamatergic inhibitory synapses are not present in the CG and PTX partially suppresses only GABAergic responses in this system.

Date: 2022-01-01

Creator: Anthony Yanez

Access: Open access

Central pattern generators are neural circuits that can independently produce rhythmic patterns of electrical activity without central or periphery inputs. They control rhythmic behaviors like breathing in humans and cardiac activity in crustaceans. Rhythmic behaviors must be flexible to respond appropriately to a changing environment; this flexibility is achieved through the action of neuromodulators. The cardiac ganglion of Homarus americanus, the American lobster, is a central pattern generator made up of four premotor neurons and five motor neurons. Membrane currents in each cell type, which can be targeted for modulation by various molecules, generate rhythmic bursts of action potentials. Myosuppressin, a FMRFamide-like peptide, is one such neuromodulator. The currents targeted for neuromodulation by myosuppressin are unknown. I investigated the molecular and physiological underpinnings of the modulatory effect of myosuppressin on motor neurons in the cardiac ganglion. First, using single cell RT-qPCR, I determined that across animals, motor neurons express myosuppressin receptor subtype II at equal levels relative to each other. Using sharp intracellular recordings, I showed that myosuppressin decreased burst frequency and the rate of depolarization during the inter-burst interval. I predicted that this effect resulted from the modulation of either A-type potassium current or calcium-dependent potassium current. Using two-electrode voltage clamp, I found that total outward current did not substantially change after treatment with myosuppressin. This result was surprising and provides grounds for explorations of subtle forms of neuromodulation in simple neural circuits.

Date: 2020-01-01

Creator: William Allen

Access: Open access

Central pattern generators (CPGs) are neural circuits whose component neurons possess intrinsic properties and synaptic connections that allow them to generate rhythmic motor outputs in the absence of descending inputs. The cardiac ganglion (CG) is a nine-cell CPG located in the American lobster, Homarus americanus. Stretch of the myocardium feeds back to the CG through mechano-sensitive dendrites and is thought to play a role in maintaining regularity in the beating pattern of the heart. The novel peptide AMGSEFLamide has been observed to induce irregular beating patterns when applied at high concentrations. This study investigated the interaction between stretch-related feedback and AMGSEFLamide modulation in generating irregular beating patterns in the whole heart of Homarus americanus. It was hypothesized that greater longitudinal stretch of the heart would result in greater regularity in the instantaneous beat frequency, based on previous findings that stretch-sensitive dendrites play a role in the regulation of the heartbeat. Furthermore, it was predicted that the elimination of stretch feedback via deafferentation of the heart would augment the irregularity induced by AMGSEFLamide. Data showed significantly increased irregularity in beating in response to 10-6 M AMGSEFLamide application. Longitudinal stretch did not reliably alter baseline variability in frequency, nor did it influence the modulatory effect of AMGSEFLamide. Deafferentation did not significantly alter baseline irregularity. Deafferented preparations did exhibit a trend of responding to AMGSEFLamide with a greater percent increase in irregularity compared to when afferents were intact, suggesting a potential role of stretch-stabilization in response to modulatory perturbations in the Homarus heart.