Authors:

Paola A. Colón Rivera^{1, 2}, Arelys Rivas Jiménez^{1, 3}, Daniel F. Ruiz Bolívar^{1, 4}, John A. Florian Alsina^{1, 4}, Cristina Velázquez-Marrero^{1, 3}
Institute of Neurobiology, University of Puerto Rico, Medical Sciences Campus¹
University of Puerto Rico, Cayey Campus²
University of Puerto Rico, Medical Sciences Campus³
University of Puerto Rico, Rio Piedras Campus⁴

Introduction: The expression of β-catenin, as part of the canonical Wnt-signaling pathway, plays a fundamental role in cell processes such as proliferation, differentiation, and homeostasis. The availability of conditional knockouts (K.O.) allow for local and temporal specificity without interrupting normal developmental processes. We will use the B6.129-Ctnnb1^tm2Kem/KnwJ conditional transgenic K.O. strain in a C57 background, to examine the role of β-catenin in alcohol-related behavioral paradigms. Previous work in our lab has shown the importance of Wnt/β-catenin signaling on the development of alcohol molecular tolerance associated with increased voluntary alcohol consumption and extinction learning. Our overarching hypothesis is that activation of the Wnt/β-catenin signaling has a pivotal role in mediating contextual fear extinction deficit induced by alcohol.

Methods: Validation of a local K.O. of β-catenin in key brain regions such as the basal lateral amygdala (BLA) on a transgenic model, will allow us to determine its role in alcohol extinction deficits. The induction of Cre-recombinase by pENN.AAV.CamKII.HI.GFP-Cre.WPRE.SV40 (AAV-Cre) as compared to the control AAV-GFP will be outlined; validating positioning, viral expansion, and quantifying GFP luminescence. Further determination of specific knockdown via immuno-histological quantification of β-catenin will be performed on coronal slices using cryo-preserved tissue samples.

Conclusion: The implementation of this genetic model will conclusively determine the role of Wnt/β-catenin, allowing for novel targets and therapeutic approaches in the development of prevention and treatment alternatives for anxiety disorders associated with binge-alcohol drinking.

IACUC approval number:

A830117

Acknowledgments:

Cristina Velázquez, PhD

The Velázquez Lab Team

1NIH NIAA 1R01AA027808-01A1

HiREC-NIMHDS21MD001830

Keywords: β-catenin, Wnt-signaling, knockouts, extinction, alcohol