Copper is a necessary nutrient but
quickly becomes toxic at elevated
levels. To properly handle environmental copper influxes and maintain
metal homeostasis, organisms utilize various methods to chelate, excrete,
and metabolize heavy metals. These mechanisms are believed to involve
complex signaling pathways mediated by neuropeptides. This study incorporates
custom N,N-dimethyl leucine isobaric
tags to characterize the neuropeptidomic changes after different time
points (1, 2, and 4 h) of copper exposure in a model organism, blue
crab, Callinectes sapidus. Using a modified simplex
optimization strategy, the number of identifiable and quantifiable
neuropeptides was increased 3-fold to facilitate a deeper understanding
of the signaling pathways involved in responding to heavy metal exposure.
The time course exposure showed many interesting findings, including
upregulation of inhibitory allatostatin peptides in the pericardial
organs. Additionally, there was evidence of transport of a pigment
dispersing hormone from the sinus glands to the brain. Overall, this
study improves the multiplexing capabilities of neuropeptidomic studies
to understand the temporal changes associated with copper toxicity.