Neuropeptides, consisting of short chains of amino acids, are a large class of signaling molecules that regulate animal behavior by interacting with their respective cell-membrane receptors. The American lobster, Homarus americanus, is a well-understood model system used to study the modulation and control of rhythmic movements in animals. Mass spectrometry has played an essential role in the identification of neuropeptides in H. americanus; however, very few neuropeptide receptors have been identified. One example of neuropeptide receptors of interest are receptors for C-type allatostatins (AST-Cs), which have been found to modulate cardiac function in crustaceans. When AST-C is perfused into the heart of the American lobster H. americanus, heart contraction amplitude increases for some animals while it decreases for others. A working hypothesis to explain the differential response is that there is a change in the expression of AST-C receptors. In this study, we present recent work on the discovery of neuropeptide receptor sequences from H. americanus, determined using data-mining techniques applied to recently generated transcriptomes for neural tissues from the American lobster. To validate the in-silico sequence predictions, we report on efforts to use proteomics techniques, executed using a chip-based nanoESI Q-TOF-MS/MS system, to detect receptors from individual H. americanus brain tissues. The efficiency of different organic solvent and detergent-based extraction techniques, coupled with filter aided sample preparation (FASP) approaches for protein digestion, are reported for the detection of specific H. americanus receptor proteins.

Proteomics and transcriptomics: Techniques for the discovery of neuropeptide receptors in the American lobster, Homarus americanus

253rd National Meeting (2017)

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