Hypoxia (i.e., low oxygen (O2) levels) is a common environmental challenge for several aquatic species, including fish and invertebrates. To survive or escape these conditions, these animals have developed novel biological mechanisms, some regulated by neuropeptides. By utilizing mass spectrometry, this study aims to provide a global perspective of neuropeptides in the blue crab, Callinectes sapidus, and their changes over time (0, 1, 4, and 8 hours) due to severe hypoxia (~10% O2 water saturation) stress using a 4-plex dimethyl labeling strategy to increase throughput. Using both electrospray ionization and matrix-assisted laser desorption/ionization provided complementary coverage– 88 neuropeptides were identified with only five overlapping between the two ionization techniques. Interesting trends include (1) an overall decrease in expression due to hypoxia exposure, (2) a return to basal levels after 4 or 8 hours of exposure following an initial response, (3) changes only after 4+ hours exposure, and (4) an oscillating pattern. Overall, this study boosts the power of multiplexed quantitation to understand the large-scale changes due to severe hypoxia stress over time. Furthermore, improved sample preparation strategies that enable comparative matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) of the crustacean neuropeptides under hypoxia and hypercapnia stress conditions will be presented.

Temporal and spatial study of neuropeptidomic changes in response to hypoxia via a multi-faceted mass spectrometry platform

2019 Fall National Meeting

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