# Elucidating the Neuropeptidome Implicated in Crustacean Feeding Processes through Multiplexed Data-Independent Acquisition Mass Spectrometry

> **NIH NIH F31** · UNIVERSITY OF WISCONSIN-MADISON · 2024 · $37,165

## Abstract

Project Summary/Abstract
Feeding processes are intertwined in a number of human health investigations, though the precise mechanisms
through which these operate remains elusive. Neuropeptides are signaling molecules that provide a dynamic
fingerprint of neuronal function, serving as a rich resource for pathway elucidation and biomarker discovery.
Despite this potential, neuropeptide analyses are met with many hurdles, stemming from their low in vivo
abundance and a lack of computational resources for confident identification and accurate quantification. It is
also imperative that the investigation of feeding processes through neuropeptides is conducted in a global and
unbiased manner, as neuropeptide activity is heavily co-modulated by other neuropeptides. My proposed aims
to bridge these gaps to elucidate feeding mechanisms through holistic investigation of the neuropeptidome. This
task will be accomplished through use of a crustacean model organism, Cancer borealis, revered as the premier
model for neuroendocrine investigations of this nature for its simpler, yet well-characterized, nervous system
with established neuropeptide homology to humans. A multipronged approach through strategic employment of
data-independent acquisition (DIA) mass spectrometry, key for overcoming the inherent bias of the mass
spectrometer toward highly abundant analytes, will be used to profile and subsequently quantify neuropeptides
from C. borealis neuroendocrine tissue extract. As neuropeptide identifications are not immediately amenable
to existing software packages designed for protein analysis, partially due to the endogenous nature in which they
are analyzed, a software suite, named EndoGenius, will be designed and optimized specifically for identification
and quantification of neuropeptides obtained from DIA experiments (Aim 1). In parallel, a spectral library of
crustacean neuropeptides will be built for identification of neuropeptides from DIA experiments, a strategy to
maximize the extent of neuropeptide elucidation and overall coverage (Aim 2). To improve resolution of
neuropeptide abundance alterations and enable multiplexed analysis, isobaric tagging of neuropeptides will be
utilized together with DIA mass spectrometry, providing a complete picture of neuropeptide profile changes in
response to feeding (Aim 3). Each aim is designed for specific application to feeding samples, collected at four
time points, contributing to the overall neuropeptidome associated with these critical behavioral functions. I
hypothesize that these methods will comprehensively resolve the temporal profile of neuropeptidome
changes induced following feeding activity with high levels of quantitative accuracy. Following these
advancements, the findings can be used to elucidate feeding regulatory mechanisms as well as neuropeptide
biomarkers that are associated with feeding disorders.

## Key facts

- **NIH application ID:** 10996413
- **Project number:** 1F31GM156104-01
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** Lauren Fields
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $37,165
- **Award type:** 1
- **Project period:** 2025-01-01 → 2026-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10996413

## Citation

> US National Institutes of Health, RePORTER application 10996413, Elucidating the Neuropeptidome Implicated in Crustacean Feeding Processes through Multiplexed Data-Independent Acquisition Mass Spectrometry (1F31GM156104-01). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/10996413. Licensed CC0.

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