Browsing by Subject "Neurosciences"
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Item Bioinformatic Analysis and Design of a GLUT3 Construct to Examine Cell Surface Expression(2019-10-25) Ogunwole, Sandra; Rameau, Gerald; Dehzangi, Abdollah; Wachira, James; Li, Yuejin; Biology; Master of ScienceGlucose transporter type 3 (GLUT3) expression at cell surface plays a vital homeostatic function in neurons. However, the mechanism involved has yet to be fully examined. We devise a strategy to tag a rat GLUT3 cDNA clone. This will be used to assess GLUT3 localization at cell surface. To install the tag, we attempt to generate a point mutation at the first exofacial loop, which would result in a unique Hpa1 restriction site in GLUT3. In another strategy, to compare total versus surface GLUT3, we propose an HA-GLUT3-eGFP fusion protein. Bioinformatic analyses suggest that this protein would be functional. GLUT3 movement to plasma membrane is carried out through vesicles. These vesicles interact with many trafficking proteins that regulate GLUT3 surface expression. Data mining using the STRING database has identified potential GLUT3-interacting proteins. Future work will focus on expressing tagged-GLUT3 and examining interactions with these proteins.Item Connections To Histamine Neurons That Underlie Wakefulness(2010) Turnbull, Erica Louise; Hoffman, Gloria E.; Biology; Master of ScienceHistamine neurons are located in the posterior hypothalamus of the brain, specifically the tuberomammillary nuclei. These neurons play a fundamental role in arousal. This project addresses the question of what regulates histamine neurons in order for them to effect arousal. There are two main signals that prompt wakefulness: the daily rhythm of the biological clock (suprachiasmatic nucleus, SCN, that contain vasopressin and vasoactive intestinal polypetide, VIP) and arousal after external stimuli (brainstem reticular activating system: which express the noradrenergic markers, dopamine beta hydroxylase (DBH) and neuropeptide Y (NPY)). Understanding if either or both systems innervate histamine neurons could provide an explanation of how histamine neurons promote wakefulness. Using immunocytochemical techniques to co-localize histamine neurons and axons from neurons within the SCN (vasopressin or VIP) and brainstem catecholamine neurons (DBH or NPY), this project aims to identify in rat brain whether either or both of the two possible regulatory systems innervate the histamine neurons. The results of the study showed that vasopressin neurons of the SCN send axons to the dorsal and ventral tuberomammillary histamine neurons; however, neurons containing VIP had no axon projections in this area. The catecholamine neurons containing both dopamine beta hydroxylase and neuropeptide Y sent axons to the ventral and dorsal tuberomammillary nuclei. Collectively, these data indicate that both dorsal and ventral histamine neurons are connected to regions of the brain that are involved in timing and arousal.Item Effects Of Neonatal Stress Exposure On Bdnf Protein In Adolescent (Pnd 30) Mouse Brain(2017) Mpamugo, Jessica; Hohmann, Christine F. F; Biology; Master of ScienceTraumatic childhood experiences are a major risk factor for anxiety disorders and depression. Brain-derived neurotrophic factor (BDNF) is critical to the developing brain; reduced BDNF levels have been observed in these disorders. Studies show that early life stress programs the development of the hypothalamic-pituitary-adrenal (HPA) axis, resulting in sustained activation of the HPA axis and behavioral changes. A link between elevated levels of corticosteroids (CORT), and decreased BDNF protein also has been observed. Thus, early life trauma and stress may contribute to functional changes in brain structure and neuroplasticity via alteration in BDNF protein levels. Our laboratory developed a model for short duration, early life, maternal separation (MS) stress using Balb/CByJ mice, employing a split-litter design, where half the pups were removed from their dam for 1 hr./day from PND 2 to PND 7 and exposed for 30 minutes to cold (4oC) or hot (37o C) stress on alternating days. Control group included littermates (LMC) who stayed with the dam and age-matched (AMC) litters raised without any intervention. This MS paradigm resulted in altered CORT release, anxiety and social behavior, as well as changes in brain structure and plasticity. Altered levels of mature BDNF (mBDNF) in the cortex and hippocampus of stressed (STR) mice were seen as well. This thesis, examines if BDNF level changes can be seen in adolescence, a typical age of onset for depression; altered social behavior in adolescence is seen in our model. We hypothesized that mBDNF levels would be lower and precursor BDNF (pro-BDNF) would be higher in cortex and hippocampus of stressed adolescent mice, at postnatal day 30, compared to the control group. I have examined the cortex and hippocampus of STR (N=20), LMC (N=10), and AMC (N=18) male and female mice who were sacrificed at PND 30. Brains were extracted and dissected on ice and stored at -70o C until they were processed for Western blot analysis of mBDNF and pro-BDNF. Multiple technical difficulties were encountered throughout this study hampering data generation and results returned inconclusive. I will discuss this process of trouble shooting my experiments and lessons learned from this study.Item Effects Of Neonatal Stress On Inflammatory Responses In The Brain Of Adult Balb/Cbyj Mice.(2015) Odebode, Gabi A.; Hohmann, Christine F.; Biology; Master of ScienceEarly childhood stress and trauma are risk factors for depression and other mental health disorders. Increased inflammatory cytokine levels are observed in the same mental health disorders. The hypothalamic-pituitary-adrenal (HPA) axis is known to regulate the inflammatory immune response. We hypothesized that early life stress may predispose the brain to neuroinflammation later in life. This thesis investigates the effects of neonatal stress on the production of pro-inflammatory cytokines, including interleukins (IL) 1β, 2, and 6 and tumor necrosis factor-a (TNF-α), in mature Balb/CByJ mice. Half of the litter was removed for the maternal separation stress procedure (STR), while the other half remained with the dam as littermate controls (LMC). A separate group of age-matched, colony-reared pups (AMC) served as an additional control. First, male and female STR, LMC and AMC mice were analyzed for baseline IL-1β, 2, and 6 and TNF-α levels in cerebral cortex and for corticosterone levels in sera, using commercially available ELISA kits. Second, mice received either saline or LPS injections two hours before being sacrificed for collection of brain tissues and blood for cytokine analysis. No significant differences in baseline levels of cytokines and corticosterone were found in the brains of uninjected STR, LMC and AMC mice. By contrast, LPS injections significantly increased production of IL-6 in brain tissue of STR and LMC mice, regardless of sex. In sera, significant increases of IL-6 and decreases of TNF-α were measured, following LPS injections in the STR and LMC mice compared to AMC controls. These data suggest that neonatal experience in STR and LMC mice may have developmentally reprogrammed inflammatory responses in the brain, thus, producing altered cytokine release to an inflammatory agent in adulthood.Item An Examination of the Relationship between a Child's Developmental Age and Early Literacy Learning(Cogent Education, 2016-04-18) Moran, Christine E.; Senseny, Karlen; Student SuccessAmerican students typically attend kindergarten at the chronological age (CA) of five and currently with the implementation of Common Core State Standards, there are expectations that children learn how to read in order to meet these academic standards, despite whether or not they are developmentally ready. This mixed methods study examined age and environmental factors that relate to reading with 83 children from the ages of 4-6½ years. The relationship between developmental age (DA) via the Gesell Developmental Observation-Revised and early literacy learning via Marie's Clay observational tool, Concepts About Print (CAP), were explored. The purpose of the study was to highlight the need for better alignment of educational policies and practices as they relate to child development and to promote more effective synthesis between discoveries in the field of neuroscience about how children learn and what is known about child DAs and stages. The findings revealed a statistically significant relationship between a child's DA and early literacy learning as measured by the CAP. The descriptive statistics revealed that the DA of the children in this study was younger than their CA. Furthermore, a child's DA was found to be the strongest predictor of early literacy learning.Item Neonatal Stress Alters Social Behavior And Bdnf Levels In The Cortex And Hippo-Campus Of Adult Balb/Cbyj Mice(2012) Subedi, Kalpana; Hohmann, Christine F.; Biology; Master of ScienceNeonatal stress and trauma are risk factors for mental health disorders such as autism, schizophrenia, and depression, which are characterized by altered social behavior, and brain-derived neurotrophic factor (BDNF) expression. BDNF has a crucial regulatory role in morphogenesis and neuroplasticity of the developing brain, particularly in the hippocampus and cerebral cortex, which are highly influenced by cortisol/corticosterone (CORT) levels. Previously, our laboratory has shown alteration of development and plasticity of the cerebral cortex as a result of neonatal stress. This thesis examines, whether neonatally stressed Balb/CByJ mice exhibit altered social behavior and BDNF levels in cortex/hippocampus in adulthood. In a split litter design, half of the pups (STR) were removed from dams for 1hr/day from postnatal day (PND) 2 to 7, and exposed for 30 minutes to cold (4 oC) or hot (37 oC) stress on alternating days, whereas littermates (LMC) remained with the dam. A cohort of age-matched, colony-reared mice (AMC) also served as controls. Thirty adult male mice, (10 STR, 10 LMC and 10 AMC) were tested in a three-chambered automated apparatus developed by J. Crawley (Crawley 2004). Detailed analyses of video-taped recordings, using CleverSys software, showed that STR males exhibited significantly less social interaction and exploratory behavior than AMC and LMC males, respectively, with significantly higher repetitive behavior (self-grooming) in STR mice. Following behavioral testing, brains were analyzed for BDNF levels by Western blotting. The results showed significantly lower mature BDNF (mBDNF) levels in the cortex as well as hippocampus of STR mice, compared to LMC. In contrast, a significantly higher precursor BDNF/mBDNF ratio was observed in the hippocampus, and a similar trend was seen in the cortex of STR compared to AMC and LMC mice. Thus, altered processing of BDNF in the adult hippocampus and cortex, following neonatal stress may permanently alter morphology and plasticity in these regions and consequently impair social and exploratory behaviors in adulthood.