Methylphenidate is a psychostimulant used to treat attention deficit hyperactivity disorder. Neurogenesis occurs throughout adulthood within the dentate gyrus of the hippocampus and can be altered by psychoactive medications; however, the impact of methylphenidate on neurogenesis is not fully understood. We investigated the effects of chronic low (1 mg/kg) and high (10 mg/kg) intraperitoneal doses of methylphenidate on neurogenesis in mouse hippocampus following 28 days and 56 days of treatment. Interestingly, methylphenidate, at both doses, increased neurogenesis. However, if methylphenidate treatment was not continued, the newly generated cells did not survive after 28 days. If treatment was continued, the newly generated neurons survived only in the mice receiving low-dose methylphenidate. To investigate the mechanism for this effect, we examined levels of proteins linked to cell proliferation in the hippocampus, including brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), vascular endothelial growth factor (VEGF), tropomyosin receptor kinase B (TrkB), and beta-catenin. BDNF or GDNF levels were not significantly different between groups. However, hippocampal VEGF, TrkB, and beta-catenin were significantly increased in mice receiving low-dose methylphenidate for 28 days compared to controls. Interestingly, high-dose methylphenidate significantly decreased beta-catenin after 28 days and decreased VEGF, beta-catenin, and TrkB after 56 days compared to controls. Thus, low-dose methylphenidate appears to increase cell proliferation and cell survival in the hippocampus, and these effects may be mediated by increase in VEGF, TrkB, and beta-catenin. While high dose methylphenidate may initially increase neuronal proliferation, newly generated neurons are unable to survive long-term, possibly due to decrease in VEGF, TrkB and beta-catenin.

Alzheimer’s disease (AD) is widely accepted as being linked with abnormal atrophy of the hippocampus. In the nonhuman-focused literature, the hippocampus has been identified as one of the prominent regions of interest with mechanisms of adult neurogenesis from aerobic exercise. Several human studies over the past decade have shown the effect of exercise that improves cardiorespiratory fitness on the size and function of the hippocampus in participants. However, the size of hippocampal subfields, especially the dentate gyrus (DG), has not been examined in humans even though various animal studies have identified the DG subfield as the primary region of adult neurogenesis induced by aerobic exercise. The point of this investigation, therefore, was to investigate the effect of an exercise intervention on the size of the DG subfield and the related subfield of cornu ammonis (CA) 3. The hypothesis was that an endurance training intervention, designed to improve cardiorespiratory fitness, would increase the volume of the DG and CA3 subfields of the hippocampus more than a resistance training intervention, designed to increase strength, flexibility, and balance, and that improvement in cardiorespiratory fitness would positively correlate with the change in volumes of these subfields. For this investigation, 32 participants (young adults from age 20 to 33 with sedentary lifestyles) were selected from a data set collected for an ongoing study by the Brain Plasticity and Neuroimaging (BPN) Laboratory at Boston University School of Medicine (Boston, MA, USA). The fitness data and T1-weighted and T2-weighted structural magnetic resonance imaging (MRI) data were used in the analysis. FreeSurfer v6.0 software was used to extract volumetric data of the hippocampal subfields using a hippocampal subfield segmentation algorithm. Analysis of variance (ANOVA) with repeated measures and linear regression were used to analyze the statistical significance of the results. The change in volumes for the whole hippocampus, DG, and CA3 did not show any statistically significant differences after endurance training compared with after resistance training. The effect of exercise on the volume of the CA3 subfield appeared to be asymmetrical from left to right, with heavier impact on the left CA3 than on the right CA3. There was no statistically significant correlation between the change in cardiorespiratory fitness and the change in volume of any of the regions analyzed. However, the left whole hippocampus showed a slight trend (p = 0.078; R = 0.317) of weak positive correlation between its volume change and the cardiorespiratory fitness change of the participants. This result was consistent with the previous human literature. Although statistically not significant, most data showed that the endurance training group saw more preservation or increase in volume. This result is encouraging and should be explored further to validate the efficacy of cardiorespiratory exercise as a possible prevention mechanism against AD for young adults later in life.

Neural stem cells (NSCs) have been recently identified in the inferior colliculus (IC). These cells are of particular interest, as no casual therapeutic options for impaired neural structures exist. This research project aims to evaluate the neurogenic potential in the rat IC from early postnatal days until adulthood. The IC of rats from postnatal day 6 up to 48 was examined by neurosphere assays and histological sections. In free-floating IC cell cultures, neurospheres formed from animals from early postnatal to adulthood. The amount of generated neurospheres decreased in older ages and increased with the number of cell line passages. Cells in the neurospheres and the histological sections stained positively with NSC markers (Doublecortin, Sox-2, Musashi-1, Nestin, and Atoh1). Dissociated single cells from the neurospheres differentiated and were stained positively for the neural lineage markers β-III-tubulin, glial fibrillary acidic protein, and myelin basic protein. In addition, NSC markers (Doublecortin, Sox-2, CDK5R1, and Ascl-1) were investigated by qRT-PCR. In conclusion, a neurogenic potential in the rat IC was detected and evaluated from early postnatal days until adulthood. The identification of NSCs in the rat IC and their age-specific characteristics contribute to a better understanding of the development and the plasticity of the auditory pathway and might be activated for therapeutic use. / Neuronale Stammzellen wurden kürzlich im unteren Colliculus inferior (CI) identifiziert. Diese Zellen sind von besonderem Interesse, da es keine therapeutischen Optionen für geschädigte neuronale Strukturen gibt. Ziel dieses Forschungsprojekts ist es, das neurogene Potenzial im CI der Ratte von den ersten postnatalen Tagen bis zum Erwachsenenalter zu untersuchen. Der CI von Ratten vom 6. bis zum 48. postnatalen Tag wurde mit Neurosphären-Assays und histologischen Schnitten untersucht. In frei schwimmenden CI-Zellkulturen bildeten sich Neurosphären bei Tieren vom frühen postnatalen Alter bis zum Erwachsenenalter. Die Menge der gebildeten Neurosphären nahm im höheren Alter ab und stieg mit der Anzahl der Zelllinienpassagen. Die Zellen in den Neurosphären und die histologischen Schnitte zeigten eine positive Färbung mit neuronalen Stammzell-Markern (Doublecortin, Sox-2, Musashi-1, Nestin und Atoh1). Dissoziierte Einzelzellen aus den Neurosphären differenzierten und wurden positiv für die neuralen Abstammungsmarker β-III-Tubulin, GFAP und MBP angefärbt. Darüber hinaus wurden neuronalen Stammzell-Marker (Doublecortin, Sox-2, CDK5R1 und Ascl-1) mittels qRT-PCR untersucht. Zusammenfassend lässt sich sagen, dass ein neurogenes Potenzial im CI der Ratte von den frühen postnatalen Tagen bis zum Erwachsenenalter nachgewiesen und bewertet wurde. Die Identifizierung von neuronalen Stammzellen im CI der Ratte und ihre altersspezifischen Merkmale tragen zu einem besseren Verständnis der Entwicklung und der Plastizität der Hörbahn bei und könnten für eine therapeutische Nutzung aktiviert werden.

Depression is associated with reduced declarative memory performance and decreased hippocampal volume. Depression has also been associated with decreased levels of adult neurogenesis in the dentate gyrus. Computational models propose that neurogenesis is critical for the computational process of pattern separation, whereby distinct memory representations are created for very similar stimuli and events. It has been proposed that depression negatively impacts pattern separation abilities; however, a link between depression and performance in pattern separation memory tasks has yet to be investigated. Accordingly, we designed a study to investigate the relationship between pattern separation performance and the severity of depression symptoms. Participants completed a recognition memory test with high pattern separation demands as well as a set of questionnaires to gauge their level of depression. We found a negative relationship between depression scores and pattern separation scores in support of the theory that depression is negatively related to pattern separation performance.

The hippocampus of birds and mammals has been shown to play a crucial role in spatial memory and navigation. The hippocampus exhibits plasticity in adulthood in response to diverse environmental factors associated with spatial demands placed on an animal. The cortical telencephalon of squamate reptiles has been implicated as a functional homologue to the hippocampus. This study sought to experimentally manipulate the navigational demands placed on free-ranging northern Pacific rattlesnakes (Crotalus o. oreganus) to provide direct evidence of the relationship between spatial demands and neuroplasticity in the cortical telencephalon of the squamate brain. Adult male rattlesnakes were radio-tracked for two months, during which one of three treatments was imposed weekly: 225 meter translocation in a random direction, 225 meter walk and release at that day’s capture site (handling control), and undisturbed control. Snakes were then sacrificed and brains were removed and processed for histological analysis of cortical features. The volume of the medial cortex was significantly larger in the translocated group compared to undisturbed controls. No differences in dorsal or lateral cortical volume were detected among the groups. Numbers of 5-Bromo-2’-deoxyuridine (BrdU) -labeled cells in the medial and dorsal cortices three weeks after BrdU injection were not affected by treatment. The activity range was larger in the translocated group compared to handled and undisturbed controls. A causal relationship between increased navigation in a free-ranging reptile and changes in brain morphology was established. The use of translocation as a conservation strategy for reptiles is a controversial topic revisited many times. Previous studies have demonstrated the aberrant movement patterns and mortality caused by translocation and have established that short-distance translocation within an animal’s home range is best for the animal. In conjunction with the neuroplasticity study, we examined the physiological impacts that repeated short-distance translocation and handling have on reptiles. This is essential knowledge if the efficacy of the technique is to be properly evaluated. Baseline and stressed concentrations of corticosterone and testosterone were assayed in blood taken immediately upon capture and following one hour of confinement in a bucket. Neither baseline nor stressed concentrations of either hormone were impacted by translocation or handling. Body condition and change in mass were not affected. Translocated animals had larger MCP activity ranges than handled and undisturbed animals at the 95%, but not 100% levels, while an interaction between time and treatment impacted other movement parameters.Treatment had no effect on a number of behaviors observed during visits to each animal. We suggest that rattlesnakes are quite resistant to potential impacts on their physiology and behavior enacted by frequent short-distance translocation or handling. Additionally, studies that require frequent handling of reptilian subjects are not likely to severely alter stress physiology.

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<p>Over long delays between events, evidence from computational models suggests that neurogenesis may be important for reducing the potential of interference between overlapping memories. These computational models also suggest that at shorter time scales, within a single memory episode, neurogenesis may play a role in binding together elements that share a common spatiotemporal context. Empirical evidence from animal research provides support for both of these hypothesized roles. Interestingly, results from recent research also suggest that depending on the task demands, neurogenesis may either aid or hinder performance.</p> <p>In order to investigate this potential trade-o, we designed the Concentration Memory Task (CMT); a novel spatial memory task which subjected participants to trials where neurogenesis is hypothesized to aid performance and trials where neurogenesis is hypothesized to hinder performance. Furthermore, we tested undergraduates on this novel task and memory tests from the CANTAB battery, and administered neuropsychological mood inventories and a lifestyle questionnaire.</p> <p>Our results suggest that measures on the CMT hypothesized to be dependent upon neurogenesis correlate with and predict performance on putatively neurogenesis-dependent tasks. Furthermore, individuals with potentially suppressed neurogenesis display selective decits on these measures. However, our results failed to provide evidence for a working memory enhancement in these individuals.</p> <p>The results from the present study provide strong encouragement for the continued development of this novel task. We provide evidence that as predicted, individuals with potentially suppressed neurogenesis display increased sensitivity to interference on the CMT. However, we failed to provide evidence that suppressed neurogenesis may enhance working memory performance. This null result may be due to shortcomings in the design of the CMT and a revised protocol that may resolve these shortcomings is discussed. With continued development, the CMT may serve as a tool for detecting early signs of cognitive impairment associated with suppressed neurogenesis.</p> / Master of Science (MSc)

Damage and/or loss of functional neurons can lead to detrimental cognitive and paralyzing effects in humans. Prime examples of such negative situations are well documented in patients with Parkinson's and Alzheimer's disease. In recent years, the utilization of neural stem cells and their derivation into neurons have been the focus of many research endeavors. The main reason for this is because neural stem cells are multi-potent and can differentiate into neurons, astrocytes, and oligodendrocytes. The research that will be detailed in this thesis involves the potential use of inorganic nanostructured materials to efficiently deliver bioactive molecules (i.e., retinoic acid, kinase inhibitors) to cells that can modulate the differentiation potential of certain cells into neurons. Specifically, PC12 (derived from rat pheochromocytoma) cells, as a neural model, was treated with select nanostructured materials with and without neuron inducers (molecules and ions) and the results were analyzed via biochemical assays and live-cell fluorescence microscopy. This thesis will include an in depth look into the cytocompatibility of the tested nanostructured materials that include silica nanoparticles, titanate nanotube microspheres, and carbon microparticles. / Bioengineering / Accompanied by two .avi files.

Hippocampal neurogenesis continues throughout life in mammals. These adult-generated dentate granule cells (DGCs) are generally believed to contribute to hippocampal memory processing and are generated at varying rates in response to neuronal network activity. Deep brain stimulation (DBS) allows clinicians to influence brain activity for therapeutic purposes and raises the possibility of targeted modulation of adult hippocampal neurogenesis. It has recently been shown that DBS may ameliorate cognitive decline associated with Alzheimer’s disease (AD), and while underlying mechanisms are unknown, one possibility is activity-dependent regulation of hippocampal neurogenesis. To this end, whether or not adult-generated DGCs can assume functional roles of developmentally-generated neurons, and stimulation-induced enhanced neurogenesis can benefit memory function in the normal and diseased brain, warrant study. First, we examined separate cohorts of developmentally- and adult-generated DGCs in intact mice and demonstrated similar rates of activation during hippocampus-dependent spatial memory processing, suggesting functional equivalence. Second, we examined the neurogenic and cognitive effects of targeted entorhinal cortex (EC) stimulation in mice using parameters analogous to clinical high frequency DBS. Stimulation increased the generation of DGCs. Moreover, stimulation-induced neurons were functionally recruited by hippocampal spatial memory processing in a cell age-dependent fashion that is consistent with DGC maturation. Importantly, stimulation facilitated spatial memory in the same maturation-dependent manner, and not when stimulation-induced promotion of adult neurogenesis was blocked, suggesting a causal relationship. Finally, we are in the process of testing whether similar stimulation facilitates spatial memory in a transgenic (Tg) disease model of AD that exhibits amyloid neuropathology and cognitive impairment. Preliminary results suggest stimulation promotes neurogenesis and rescues impaired spatial memory in Tg animals. When considered in the context of promising clinical results, this body of work suggests stimulation-induced neurogenesis could provide a novel therapeutic modality in settings where functional hippocampal regenerative therapy is desirable.