Author ORCID Identifier

https://orcid.org/0000-0001-9813-0978

Defense Date

2024

Document Type

Thesis

Degree Name

Master of Science

Department

Pharmacology & Toxicology

First Advisor

Sammanda Ramamoorthy

Abstract

Rationale: Serotonin (5-HT) is a crucial neurotransmitter involved in regulating various physiological processes. Dysregulation of the serotonergic system has been implicated in several neuropsychiatric disorders. Though numerous 5-HT receptors exist, the 5-HT transporter (SERT) is responsible for 5-HT clearance in the brain and periphery. 5-HT1Bautoreceptors regulate 5-HT release and interact with glycogen synthase kinase-3β (GSK3β), which can modulate SERT function via specific Ser44/48 phosphorylation sites. While these findings suggest a potential interplay between 5-HT1Band GSK3β-mediated SERT phosphorylation in regulating extracellular 5-HT, the primary causal molecular link remains unknown.

Objectives: To investigate if 5-HT1B activation by the agonist CP-94253 influences SERT-mediated 5-HT clearance through GSK3β-dependent - SERT-Ser44/48 phosphorylation, and to examine regional and sex differences.

Methods: 5-HT uptake assay and kinetic analysis performed with region-specific crude synaptosomes from female wild-type mice and SERT-A44/A48 knock-in mice, following CP-94253 treatment.

Results: In wild-type female mice, CP-94253 treatment showed a non-significant enhancement of SERT-mediated 5-HT uptake in the striatum and hippocampus. Kinetic analyses revealed a significant increase in the Vmax of 5-HT uptake in the hippocampus. However, SERT S44A/S48A knock-in mice exhibited elevated SERT-mediated uptake even without treatment, and CP-94253 did not further increase uptake in these mice, suggesting the importance of these phosphorylation sites.

Conclusions: The loss of CP-94253-induced changes in uptake kinetics in the SERT S44A/S48A knock-in mice, in both the striatum and hippocampus, reinforces the critical role of these SERT phosphorylation sites in mediating the 5-HT1B receptor-GSK3β regulation of SERT function and 5-HT reuptake dynamics.

Rights

© Sanyukta Jalihalkar

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

8-8-2024

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