# Neuroimmune mechanisms mediating effects of maternal ethanol consumption on offspring’s hypocretin neuron transcriptome and behavior: translation from zebrafish to rodent

> **NIH NIH F32** · ROCKEFELLER UNIVERSITY · 2020 · $65,310

## Abstract

Project Summary
Compelling evidence suggests that ethanol exposure early in life, even at low levels, disturbs the development
of neurons in the brain and increases the risk for developing alcohol use disorder (AUD). Low-dose maternal
ethanol (ME) consumption in both zebrafish (ZF) and rats has revealed conserved neurobehavioral effects
across species, with ME consumption significantly increasing in the offspring the expression of the hypothalamic
orexigenic peptide hypocretin/orexin (Hcrt), a potent stimulator of consummatory behavior, and the consumption
of ethanol and anxiety-related behaviors. While there is evidence linking neuroimmune signaling to AUD and
showing ethanol in rats to stimulate various inflammatory chemokines and cytokines in glial cells, there are few
studies examining neuroimmune factors within neurons and investigating how they coexpress with
neuropeptides and affect the development of these neurons and the behaviors they control. Focusing on
hypothalamic Hcrt neurons and neuroimmune systems that exist within these neurons, this proposal will use a
dual-species approach to test the following hypothesis: Maternal ethanol consumption disturbs specific
neuroimmune transcripts in Hcrt neurons of the offspring which, in turn, stimulate the development of these
neurons and contribute to an increased propensity for greater ethanol consumption and preference in the
offspring. In Aim 1, transgenic Hcrt:EGFP ZF will be used, an advantageous vertebrate due to its external
development, genetic tractability, optical transparency and small size, to first evaluate ME’s effects on the well-
studied chemokine CXCL12a and its receptor CXCR4b within Hcrt neurons of the offspring. Then, the Hcrt
transcriptome will be sequenced using RNA-seq, and bioinformatic analyses will be performed to identify the top
differentially expressed neuroimmune gene induced by ethanol exposure. This analysis will be followed by
CRISPR/Cas9 gene editing, which will be used to determine in ZF offspring the functional role of the most
strongly affected candidate gene as well as CXCL12a and CXCR4b in altering the development of Hcrt neurons,
to be examined using quantitative live imaging, and also in stimulating ethanol consumption and related
behaviors. In Aim 2, the ZF findings will be translated directly to the rat model, first by confirming results obtained
in the ZF and then by knocking down through injection into rat embryo brain of AAV delivered shRNA for the
target neuroimmune gene identified in Aim 1. To determine the functional role of this target gene in mediating
the effects of ME on neuronal development and behavior, Hcrt neurons will be examined in adolescent rat
offspring using iDISCO brain clearing, and ethanol consumption will be measured using the intermittent access
two-bottle choice paradigm. With this research being in line with multiple objectives of the NIAAA strategic plan,
the proposed studies which will provide extensive training with differ...

## Key facts

- **NIH application ID:** 9927482
- **Project number:** 5F32AA027702-02
- **Recipient organization:** ROCKEFELLER UNIVERSITY
- **Principal Investigator:** Adam Collier
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $65,310
- **Award type:** 5
- **Project period:** 2019-05-01 → 2022-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9927482

## Citation

> US National Institutes of Health, RePORTER application 9927482, Neuroimmune mechanisms mediating effects of maternal ethanol consumption on offspring’s hypocretin neuron transcriptome and behavior: translation from zebrafish to rodent (5F32AA027702-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9927482. Licensed CC0.

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