Date: 2019-05-01

Creator: Gina Fickera

Access: Access restricted to the Bowdoin Community

• Restriction End Date: 2028-06-01

Date: 2023-01-01

Creator: Emily Grace Herndon

Access: Access restricted to the Bowdoin Community

Date: 2014-05-01

Creator: Mara R Chin-Purcell

Access: Open access

Central pattern generators are neuronal networks that produce reliable rhythmic motor output. A simple pattern generator, known as the cardiac ganglion (CG), controls the heart of the American lobster, Homarus americanus. Previous studies have suggested that stretch feedback relays information to the cardiac ganglion about the degree of filling in the heart, and that this feedback is mediated by stretch-sensitive dendrites extending from CG neurons. I sought to determine the mechanisms behind this stretch feedback pathway. One hundred second extension pyramids were applied to each heart while amplitude and frequency of contractions were recorded; 87% of hearts responded to stretch with a significant increase in frequency of contractions. To ascertain the role of dendrites in this feedback pathway, the accessible branches along the trunk of the CG were severed, de-afferenting the CG. In de-afferented hearts, stretch sensitivity was significantly less than in intact hearts, suggesting that the dendrites extending from the CG are essential for carrying stretch feedback information. To separate the effects of active and passive forces of heart contraction on stretch sensitivity, the CG was de-efferented by severing the motor nerves that induce muscle contraction. Hearts with only anterolateral nerves cut or with all four efferents cut were significantly less stretch sensitive than controls. These results indicate that the CG is sensitive to active stretch of each contraction. Hearts with reduced stretch feedback had more irregular frequency of contractions, indicating that a role of stretch feedback in the cardiac system may be to maintain a regular heart rate.

Date: 2023-01-01

Creator: Isabel Stella Petropoulos

Access: Open access

A substantial factor for behavioral flexibility is modulation — largely via neuropeptides — which occurs at multiple sites including neurons, muscles, and the neuromuscular junction (NMJ). Complex modulation distributed across multiple sites provides an interesting question: does modulation at multiple locations lead to greater dynamics than one receptor site alone? The cardiac neuromuscular system of the American lobster (Homarus americanus), driven by a central pattern generator called the cardiac ganglion (CG), is a model system for peptide modulation. The peptide myosuppressin (pQDLDHVFLRFamide) has been shown in the whole heart to decrease contraction frequency, largely due to its effects on the CG, as well as increase contraction amplitude by acting on periphery of the neuromuscular system, either at the cardiac muscle, the NMJ, or both. This set of experiments addresses the location(s) at which myosuppressin exerts its effects at the periphery. To elucidate myosuppressin’s effects on the cardiac muscle, the CG was removed, and muscle contractions were stimulated with L-glutamate while superfusing myosuppressin. Myosuppressin increased glutamate-evoked contraction amplitude in the isolated muscle, suggesting that myosuppressin exerts its peripheral effects directly on the cardiac muscle. To examine effects on the NMJ, excitatory junction potentials were evoked by stimulating of the motor nerve and intracellularly recording a single muscle fiber both in control saline and in the presence of myosuppressin. Myosuppressin did not modulate the amplitude of EJPs suggesting myosuppressin acts at the muscle and not at the NMJ, to cause an increase in contraction amplitude.

Date: 2020-01-01

Creator: Edward Myron Bull

Access: Open access

Neuropeptides are a class of small peptides that govern various neurological functions, and the American lobster (Homarus americanus) provides a model system for their characterization. Neuropeptides are commonly post-translationally modified (PTM), and one common PTM is glycosylation. Past research in the Stemmler lab has found glycosylated neuropeptides in H. americanus; however, the extent and biological role of this modification has not been well characterized. This study was undertaken to determine the number of glycosylated peptides in the sinus glands of H. americanus and to develop an approach to tag the site of glycosylation using beta-elimination chemistry. LC-MS paired with high pH reverse phase fractionation was used to survey for glycosylated neuropeptides and beta elimination with an amine tag was used as an approach to characterize the site of glycosylation. Our results indicate that high pH fractionation is a useful approach to simplify complex mixtures of neuropeptides and improve glycopeptide detection. Efforts to use beta elimination and tagging to characterize glycosylated neuropeptides have been less successful. Beta elimination of full length peptides resulted in peptide degradation. An approach utilizing chymotrypsin to reduce peptide size coupled with beta elimination and labeling with 2-dimethylaminoethanethiol showed less evidence for degradation, and this approach yielded data isolating two potential serine residues for the site of glycosylation; however, the data was not sufficient to distinguish the two sites. Work to optimize reaction conditions using a glycopeptide standard showed that multiple isomeric products were formed during beta elimination. With the goal of optimizing reaction conditions, future work will further examine reaction kinetics to eventually apply the approach to the entire sinus gland

Date: 2014-05-01

Creator: Elizabeth A Owens

Access: Open access

The American lobster, Homarus americanus, inhabits a large oceanic range spanning from Labrador, Canada to North Carolina, USA. This geographic range varies in temperature by as much as 25ºC, and daily temperature fluctuations of up to 12ºC may occur at a single location depending on season, water depth, and tides. The cardiac system of the lobster is sensitive to these temperature changes, and has been shown to adjust its functioning over a large temperature range. A previous study showed that various functional parameters respond differently to temperature changes, but a stable cardiac output can be maintained over the range of 2-20ºC. The current study showed that the effects of temperature were exerted primarily through changes in the lobster heart central pattern generator, the cardiac ganglion. Similar patterns of change were seen in both semi-intact hearts and isolated cardiac ganglion preparations in response to increasing temperature. Specifically, with increasing temperature, the burst frequency showed a biphasic pattern in which frequency initially increased, then decreased rapidly at high temperatures. The burst duration, duty cycle, and number of spikes per burst generally decreased with increasing temperature, and spike frequency increased over the entire temperature range. Semi-intact hearts and isolated cardiac ganglia showed similar “crash” patterns, characterized by complete loss of function at high temperatures and complete recovery of function when temperature was returned to baseline. Feedback in the semi-intact heart provided some stabilization of bursting activity, but it did not provide the expected protection from high temperatures. The isolated CG had a significantly higher crash temperature than did the semi-intact system. This discrepancy in crash temperatures may be explained by considering factors at the level of the muscle and neuromuscular junction (NMJ), such as stretch and nitric oxide (NO) feedback and the balance of facilitation and depression at the NMJ. Stimulated preparations showed defacilitation of contraction amplitude at high temperatures despite the maintenance of constant burst parameters of stimulation. Therefore, several factors contributing to the relatively low crash temperature of the intact system may be a shift in the balance of facilitation and depression at the NMJ, a depression in ganglion function due to the release of NO by the muscle, or a combination of the two mechanisms.

Date: 2016-05-01

Creator: Michael M Kang

Access: Access restricted to the Bowdoin Community