G-Protein Coupled Receptor (GPCR) signaling is an essential component of neural networking within the brain. Once bound by an agonist, the GDP molecule previously bound to the α subunit during the inactive state is replaced by GTP, activating a signal transduction cascade specific to the receptor. This GDP-GTP replacement mechanism is widely used to identify GPCR activation state in imaging techniques. Currently, the O’Neill lab is developing a new method, named “GTPγScope”, to functionally image GPCRs using fluorescence microscopy techniques. Instead of the radioactively labelled GTPγS analog, GTPγScope uses fluorescent antibodies for detection. We use the α-o antibody which binds to the α subunit of the G-protein. Our research aims to further optimize the GTPγScope protocol to provide reproducible results and uniform (not white-matter dependent) fluorescence under the use of the nociceptin agonist. We hypothesized that reproducibility will increase after the cryosectioning step of the protocol is optimized. This optimization occurred after adjusting drying, cryosectioning, and pipetting methods of the protocol. In our research, we performed immunohistochemistry experiments followed by fluorescence imaging. Studying GPCR location and function is critical to understanding biological mechanisms of various conditions, such as chronic pain. Nociceptin and its receptor (NOP), for example, are critical players within the realm of chronic and/or neuropathic pain and GPCR functioning.

Objective: While prior studies have largely focused on adults, identifying socioeconomic and clinical factors contributing to surgical delay in children is crucial for timely interventions. Methods: Data was extracted from 137 consecutive pediatric patients (ages 0–23) who underwent epilepsy surgery at UCLA between 2021 and 2025. To address missing data, multiple imputation using chained equations was performed. Multivariable Cox proportional hazards regression assessed the impact of sociodemographic predictors on surgical delay. Kaplan-Meier survival analyses with false discovery rate (FDR)-adjusted log-rank tests compared surgical delay across epilepsy diagnosis, etiology, and surgical procedure. Results: Kaplan-Meier survival analysis demonstrated that patients with LGS experienced much longer surgical delay compared to those with infantile spasms and generalized epilepsy (p = 0.0085 and 0.00093, respectively). Patients with structural etiology showed a longer delay compared to other etiologies (p = 0.027). Patients undergoing neuromodulation experienced longer delays relative to other surgical approaches. Multivariable Cox regression revealed no statistically significant independent associations between surgical delay and any sociodemographic factors. Conclusion: This study demonstrated that surgical delay in pediatric epilepsy was influenced most by epilepsy diagnosis and surgery type. Thus, early identification and targeted referral for high-risk groups, such as those with LGS, is needed to reduce surgical delay.

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by synaptic dysfunction and white matter degeneration, most commonly studied in cortical and hippocampal regions. Emerging evidence suggests the cerebellum may also undergo molecular and structural changes that contribute to disease progression. Synaptic loss is a strong correlate of cognitive decline, while myelin disruption impairs neural signaling and network integrity. In this study, we investigated how disease status, age, sex, and hormonal condition influence cerebellar synaptic and myelin integrity in an AD mouse model. Cerebellar tissue was analyzed using immunostaining for synapsin-1 (SYN1) and myelin basic protein (MBP). Mice were grouped by disease (AD vs. wild-type), age (young vs. midlife), sex (male vs. female), and hormonal condition (sham vs. gonadectomized). We observed an age-dependent decrease in SYN1 expression selectively in male AD mice, which was not present in wild-type males. Female mice showed relative preservation of SYN1 across conditions. MBP expression decreased with age and AD, with more pronounced reductions in male mice, suggesting increased myelin vulnerability. Hormonal effects were limited and context-dependent, without consistent trends across groups. Together, these findings support cerebellar involvement in AD pathology and demonstrate that synaptic and myelin alterations are shaped by sex and age, highlighting the importance of biological context in neurodegenerative disease.

Episodic memory is correlated with coordinated theta oscillations in the human cortex. However, transcriptional processes responsible for these oscillations is unknown. Previous in vivo studies are unable to determine these oscillations' fundamental pathways and layer specific activity. Thus, I am utilizing an ex vivo approach to discover layer specific gene expression. My mentor has already utilized electrophysiology to obtain oscillatory recordings of cortex slices. My work will expand on this by identifying differentially expressed genes between and within samples through spatial transcriptomics. Eventually, the data retrieved will be analyzed in R. This will be correlated to the electrophysiology findings using Spearman's rank correlation. Our findings have potential to inform neurodegenerative disease treatments, especially those that degrade memory.

The gut microbiome communicates with the brain via sensory pathways that converge in the nucleus of the solitary tract (NTS), a key site for sensory neuronal input that modulates brain state and behavior. Our preliminary data show that germ-free mice have higher NTS neuronal activation than conventionally colonized controls. Colonization of germ-free mice with conventional microbiome restores NTS activity to baseline, indicating that the NTS actively responds to microbiome status. Acute microbiome depletion with broad-spectrum antibiotics leads to significant increases in NTS activation compared to the vehicle. However, it remains unclear how these neurons respond to microbiome-derived cues. This project asks whether NTS neurons activated following microbiome depletion can be reactivated and are required for microbiome-related behaviors. To address this, we use the TRAP2 genetic mouse line to tag and manipulate NTS neurons activated by microbial depletion. Since these experiments require proper validation, my project focuses on validating viral targeting and neuronal activation in TRAP2 hM3Dq and Casp3 mice. In TRAP2 hM3Dq mice, I administered DCZ to reactivate TRAPed neurons, followed by perfusion, NTS cryosectioning, and immunostaining for DAPI and mCherry and imaging analysis to confirm viral targeting. In TRAP2 Casp3 mice, I validated the viral targeting by quantifying cFos expression in the NTS. Overall, my research provides evidence supporting neural interoception of the gut microbiome and their links to behaviors.