Data showing that third‑trimester‑equivalent alcohol exposure does not alter adult cerebellar cytoarchitecture or behavioral flexibility in mice

Description

Developmental alcohol exposure can disrupt cerebellar development and contribute to motor and cognitive deficits associated with Fetal Alcohol Spectrum Disorder (FASD). This study investigated the long-term effects of third-trimester-equivalent alcohol exposure on cerebellar cytoarchitecture and behavioral flexibility in adult male mice. VGAT-Venus transgenic mouse pups received a single subcutaneous ethanol injection (3.5 g/kg) on postnatal day 6. In adulthood, cerebellar cytoarchitecture was assessed in parasagittal sections containing Lobule VI and Crus I, and behavioral flexibility was evaluated using a pairwise visual discrimination and reversal learning task. No significant differences between experimental groups were observed in cerebellar cytoarchitecture and in discrimination and reversal learning performance. The Excel files included here are organized into several tabs (worksheets) as follows: 1. Individual IHC counts: contains the IHC cell-counting data foreach individual mouse. 2. IHC Summary: Provides a summary of the IHC counting data across all mice. 3. Individual mouse data: each worksheet contains both discrimination and reversal learning data for an individual mouse. 4. Behavioral Summary: The final worksheet provides a summary of the discrimination and reversal learning data across all mice.

Steps to reproduce

Ethanol Exposure Paradigm: Vesicular GABA transporter (VGAT)-Venus transgenic mouse pups received a single subcutaneous ethanol (3.5 g/kg) or saline injection on postnatal day 6. Immunohistochemistry Data: Male VGAT mice were deeply anesthetized and transcardially perfused with 4% paraformaldehyde. Following brain extraction and sectioning, parasagittal brain sections containing Lobule VI and Crus I were incubated for 20 minutes with 4’,6-diamidino-2-phenylindole (DAPI, dilution 1:500, Thermo-Fischer Scientific, USA). The sections were rinsed with 1X PBS and mounted on Superfrost Plus microscope (VWR, Radnor, PA) using Fluoromont-G mounting media (Thermo Fisher Scientific, USA), covered with coverslips (VWR, Radnor, PA), and stored at 4 ºC until imaging acquisition. Images were acquired using AxioScan.Z1 slide scanner (Zeiss Germany) at 20× magnification, and selecting appropriate fluorescence filters to detect DAPI and EYFP-positive neurons. Counting cells were performed using Fiji and QuPath’s Cell Detection algorithm (Version 0.5.1.). Behavioral Data: We assessed behavioral flexibility using custom-made acrylic chambers housed within sound- and light-attenuating boxes (Med Associates, St. Albans, VT). Briefly, adult male VGAT mice were trained on a pairwise visual discrimination task. Trial initiation presented two novel equi-luminescent visual stimuli (Fan or Marbles), one per response window; the rewarded stimulus (S+) was counterbalanced across experimental groups. Each session consisted of 30 first-presentation trials. Mice that met the discrimination criterion, completing 30 first-presentation trials with an average of 85% correct responses (excluding correction errors) within 30 minutes on two consecutive days, were moved to the reversal phase. During the reversal, the stimuli were switched, with the previously correct stimulus labeled as incorrect, and vice versa. Mice were trained in daily 30-trial sessions until achieving ≥ 85% correct responses (excluding correction errors) across two consecutive days. Behavioral data were acquired and recorded using K-Limbic Software Package V1.20.2 (Conclusive Solutions, Sawbridgeworth, UK).

Institutions

• University of New Mexico Health Sciences Center

  New Mexico, Albuquerque

Categories

Funders

• National Institutes of Health

  United States Department of Health and Human Services

  Maryland, Bethesda

  Grant ID: R21-AA030639

Licence