Date of Award
Spring 4-23-2026
Document Type
Dissertation
Publication Status
Version of Record
Submission Date
May 2026
Department
Biological Sciences
College Granting Degree
Charles E. Schmidt College of Science
Department Granting Degree
Biological Sciences
Degree Name
Doctor of Philosophy (PhD)
Thesis/Dissertation Advisor [Chair]
Ryohei Yasuda
Thesis/Dissertation Co-Chair
Robert W. Stackman Jr.
Abstract
Protein kinases regulate cellular function through reversible phosphorylation that coordinates signaling across molecular and cellular scales that influence behavior. Although defining isoform-specific kinase functions is essential for both basic biology and medicine, determining when and where a particular isoform is required remains challenging, especially in behaving animals.
To address this limitation, we developed a Light-Operated Kinase Inhibitor (LOKI) strategy that enables molecular specificity with temporal control. Specificity is achieved by genetically tagging endogenous kinase isoforms, while temporal precision is provided by light-dependent and reversible recruitment of an inhibitory module to the tagged kinase. We engineered and characterized LOKI variants targeting two Protein Kinase C (PKC) isoforms implicated in learning and memory and historically difficult to inhibit selectively: PKCα and PKCδ. Quantitative characterization and in vitro application of LOKI-PKCα demonstrated robust, reversible, isoform-specific inhibition and established LOKI as a potentially generalizable kinase inhibition platform. Using LOKI-PKCα in vitro, we found that transient PKCα activity during the induction phase of synaptic plasticity is required for structural long-term potentiation.
Extending this approach in vivo, acute PKCα inhibition in the medial prefrontal cortex (mPFC) impaired formation of auditory sensorimotor association. Complementary experiments in PKCα knockout mice revealed similar behavioral deficits, and region-specific re-expression of PKCα in the mPFC rescued this impairment. Together, these findings demonstrate a temporally and regionally restricted requirement for PKCα in synaptic plasticity and associative learning and establish LOKI as a powerful approach for isoform-selective kinase interrogation both in vitro and in vivo.
Recommended Citation
Menton, Goksu, "A LIGHT-OPERATED KINASE INHIBITOR STRATEGY AND ITS IN VITRO AND IN VIVO APPLICATIONS" (2026). Electronic Theses and Dissertations. 288.
https://digitalcommons.fau.edu/etd_general/288