Characterization and Astrocyte-Specific Knockout of Acyl-CoA-Binding Protein Using a Cre-LoxP Model

open_access
Open Access
Author:
Mykut, Carley
Area of Honors:
Biology
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
Grayson Oren Sipe, Thesis Supervisor
Benoit A Dayrat, Thesis Honors Advisor
Keywords:
neuroscience
astrocyte
anxiety
knock-out
Abstract:
ABSTRACT Post-traumatic stress disorder (PTSD) is a complex neuropsychiatric condition characterized by anxiety and the disarray of many other neuronal mechanisms. While most research is focused on neurons, astrocytes are an emerging area for many researchers. One astrocyte-specific protein, Acyl-CoA-binding protein (ACBP), which is also known as diazepam binding inhibitor (DBI), has been linked to stress and anxiety regulation. Within this study, we evaluated the effectiveness of an inducible Cre-recombinase system for astrocyte-specific knockout and characterized its distribution across brain regions and cell types. Adult transgenic mice expressing Aldh1l1-CreERT2 were crossed with floxed DBI mice and administered tamoxifen to induce recombination. Brain tissue from the primary motor cortex (M1), caudate & putamen (CPu), and hippocampus (CA1) was analyzed using immunohistochemistry and confocal analysis to quantify DBI expression in astrocytes, neurons, and microglia. Results demonstrated a significant difference in DBI expression in astrocytes for Cre+ animals compared to control animals (Cre-) (p < 0.05). This confirmed the specificity of the Lox/P model for astrocytes. Furthermore, no significant changes in DBI expression for neurons and microglia were present. Likewise, further regional analysis revealed the ubiquitous nature of the DBI protein across brain regions studied: M1, CPu, and CA1. These findings validate the inducible Cre/LoxP system utilized to be astrocyte specific. This model now provides a foundation for further behavioral studies investigating the role of astrocyte-specific DBI expression in stress and anxiety regulation.