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Neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease are linked to mitochondrial dysfunction and the underexpression of TOM40, a protein with chaperone-like qualities that is responsible for transporting precursor proteins into the mitochondria. Overexpression of TOM40 is reported to partially restore mitochondrial dysfunction and decrease the accumulation of neurotoxic aggregates of α-synuclein. Our goal is to develop an effective method for delivery of TOM40 protein to the brain. Previous studies have used lentiviruses to carry TOM40 into the hippocampus of α-synuclein transgenic mice. The disadvantage of lentiviral transfection is the random insertions of the target gene into the host genome, which could cause toxic effects. Synthetic phospholipid vesicles containing TOM40 were considered as an alternative delivery method, but these "liposomes" elicit not only toxicity, but also an immune response. Thus, development of a safer delivery method of TOM40 protein is needed. We investigated exosomes, which are extracellular vesicles originating from multivesicular endosomes filled with protein, lipid, or RNA cargoes for cell-cell communication. Since exosomes are created from host cells, they are non-immunogenic and may be a more desirable method. Expression constructs have been made for the production of TOM40 protein within or on the surface of exosomes. In order to target the delivery of TOM40 to the brain, we attached peptides to the surface of the exosomes, which specifically interact with receptors on neural cells. We attempted to confirm the functionality of the expression constructs through immunocytochemistry followed by flow cytometry and Western blotting.

Flashbulb memories are formed through widely shared events that have affected the culture and community. The “flash” in flashbulb memories refers to the specific details that individuals have developed in their memories. The presented research focuses on the specific event of college students at the University of Central Florida (UCF) hearing about university closure due to the COVID-19 pandemic. This research study sought to identify the variables that have affected the accuracy of flashbulb memories formation regarding the event of college students hearing about university closure because of the pandemic. The variables of the students’ relevance to the university, location, source of hearing about the university’s closure, and political group they belong to were investigated to determine if they affected the accuracy of the memories that were developed. An online survey was sent to students inquiring about the specifics and details that they remembered when finding out that the university was closed and was going to move to remote instruction. Participants included 226 college students who filled out the survey between February 3, 2021, and July 21, 2021. The survey included questions regarding the experiences of students when they learned about COVID-19 and university closure. Data revealed that a greater relevance to the event can cause a greater amount of rehearsal and recall of memories. The rehearsal and recall of memories are crucial variables to developing accurate flashbulb memories. This study contributes to the lack of research in flashbulb memories associated with pandemics. The study will be an addition to determining variables that have affected the accuracy of flashbulb memories.

Cannabis is widely consumed by humans for pharmacological effects that are mediated by THC, though there is little evidence that THC is a primary reinforcer in non-human animal models. We hypothesized that THC may have potent reinforcement enhancing effects, comparable to other drugs (e.g., nicotine and caffeine) which are also widely consumed by humans, but difficult to establish as primary reinforcers in non-humans. In three experiments with male and female rats saccharin (SACC) or a visual stimulus (VS) served as reinforcers for operant behavior. We explored several pharmacological parameters of THC on responding for SACC or VS, including THC dose, intervals between THC injections and testing, and intervals between test sessions. THC acts as a reinforcement enhancer for both SACC and a VS across a range of doses and intervals. Daily THC injections systematically reduced behavior, possibly due to accumulation of THC bioavailability.

Endocannabinoids (eCBs) are small molecules that are capable of modulating synaptic plasticity of both excitatory and inhibitory synapses in the brain. While eCBs bind to transient receptor potential vanilloid 1 (TRPV1) and cannabinoid receptor 1 (CB1) in the central nervous system, we recently identified a form of non-CB1, non-TRPV1 mediated long term depression activated by the eCB anandamide at CA1 hippocampal stratum radiatum interneurons. GPR55, an orphan G-protein receptor, has been identified in the hippocampus and is capable of activation by eCBs, making it a good candidate for mediating this non-CB1, non-TRPV1 form of synaptic plasticity. Here we performed whole-cell patch clamp recordings from CA1 stratum radiatum interneurons in rat brain slices to investigate the effect of GPR55 agonist O-1602 on excitatory synapses. We also performed field recordings from CA1 pyramidal cells in rats and GPR55 knockout mice and littermate controls to investigate the effect of GPR55 agonists O-1602 and lysophosphatidylinositol (LPI) on both basal output and electrically induced long-term depression and long-term potentiation in the hippocampus. Application of O-1602 in rats depressed long-term potentiation in CA1 pyramidal cells, and depressed excitatory glutamatergic transmission onto some interneurons. O-1602 had no effect on long-term depression of CA1 pyramidal cells. GPR55 +/+ mice showed an increase in long-term potentiation in the presence of LPI compared to GPR55-/- littermates. GPR55-/-mice had no change in long-term potentiation when exposed to O-1602, though there was an increase in post-tetanic potentiation with O-1602. In order to examine whether GPR55 has a role in formation of spatial memory, GPR55 -/- mice were compared to littermate controls during a Morris water maze behavioral task, with a reversal task after 7 days of training. GPR55-/-mice did not perform in a different manner on either the training task or the reversal, though there may be a trend of difference in training worth investigating further. This study illustrates a novel pathway for synaptic plasticity modulation through GPR55 in the hippocampus, and may therefore provide valuable insight into both the effects of synthetic and endogenous cannabinoids on the brain and the processes underlying learning and memory.

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The specification and development of nervous system diversity is a driving question in the field of Neurobiology. The overarching goals of the projects described in this thesis are to describe tools to aid in the description of nervous system development and to show the use of the described tools to study nervous system development in the nematode Caenorhabditis elegans. The first chapter of this thesis describes a complete map of the male C. elegans nervous system using a tool developed in the lab to uniquely label all neurons in the C. elegans nervous system, NeuroPAL. The second chapter of this thesis largely focuses on a well-studied homeobox gene, unc-86, and its role in fate transformations in dopaminergic and GABAergic neuron types. These two seemingly disparate projects are united in their effort to investigate nervous system development and neuronal fate determination. NeuroPAL is a multicolor transgene that uniquely labels all neurons of the C. elegans hermaphrodite nervous system and here I show it can be used to disambiguate all 93 neurons of the male-specific nervous system. I demonstrate the wide utility of NeuroPAL to visualize and characterize numerous features of the male-specific nervous system, including mapping the expression of gfp-tagged reporter genes and neuron fate analysis. NeuroPAL can be used in combination with any gfp-tagged reporters to unambiguously map the expression of any gene of interest in the male, or hermaphrodite, nervous system. Furthermore, NeuroPAL is used in mutants of several developmental patterning genes to confirm previously described defects in neuronal identity acquisition. Additionally, I show that NeuroPAL can be used to uncover novel neuronal fate losses and identity transformations in these mutants because of the unique labeling of every neuron. Lastly, we show that even though the male-specific neurons are generated throughout all four larval stages, the neurons only terminally differentiate in the fourth and final larval stage, termed ‘just-in-time’ differentiation. In the second part of this thesis, I describe a few examples of mutant analysis of homeobox gene family members and describe their function in the C. elegans nervous system. I focus largely on a couple potential examples of homeotic fate transformations in mutants of the POU homeobox gene, unc-86. In unc-86 mutants, I describe the ectopic expression of multiple GABAergic terminal identity features in one cell in the head of C. elegans. I raise the hypothesis that this cell may be a transformation of a non-GABAergic ring interneuron, RIH, into that of its GABAergic sister cell, AVL, in unc-86 mutants. While ectopic dopaminergic neurons were previously described in unc-86 mutants, I expand the study to show the ectopic expression of all dopaminergic synthesis and packaging genes. I show support that all non-dopaminergic anterior deirid neurons, ADA, AIZ, FLP, and RMG, lose the expression of some of their wild type terminal fate genes and transform to a fate like that of their dopaminergic sister cell, ADE, as assessed by NeuroPAL expression. Taken together, these studies describe tools and methods for studying nervous system development as well as describe many examples of cell fate transformations.