MEES Theses and Dissertations
Permanent URI for this collectionhttp://hdl.handle.net/1903/19655
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Item The Gonadotropin Releasing Hormone-3 System in Zebrafish: Early Development and Regulation(2008-12-15) Abraham, Eytan; Zohar, Yonathan; Marine-Estuarine-Environmental Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)The objective of this study was to expand our understanding of the early development of forebrain Gonadotropin Releasing Hormone (GnRH) neurons in vertebrates in general and in fish in particular. The correct migration during early development of the hypophysiotropic GnRH neurons from the olfactory region to the hypothalamus is crucial for normal gonadal development and reproduction. We developed a Tg(GnRH3:EGFP) zebrafish line in which EGFP is specifically expressed in GnRH3 neurons. Using this line, we have studied in detail the early spatiotemporal development of the GnRH3 system in vivo. In addition, we have studied various factors, including GnRH3, Netrins and Hedgehog to better understand some of the mechanisms that mediate this complex axophilic neuron migration event. Lastly, we have conducted targeted GnRH3 neuron ablation experiments in view of determining the embryonic origin of POA-hypothalamic GnRH3 neurons and the effect of lack of GnRH3 neurons in the CNS. Our findings show that: 1) GnRH neurons first differentiate and express GnRH3 at 24-26 hours post fertilization (hpf) and immediately thereafter begin to extend fibers. 2) GnRH3 neurons project a complex network of fibers, prior the GnRH3 soma migration, to various CNS regions, and to the pituitary. 3) GnRH3 soma begin migrating towards the hypothalamus at 3 days post fertilization (dpf), passing through the terminal nerve (TN), lateral telencephalon, and reaching the hypothalamus by 12 dpf. 4) expression of GnRH3 itself is necessary for the normal early differentiation and fiber extensions of GnRH3 neurons. 5) Netrin1a is directly involved as a chemoattractant in GnRH3 fiber organization and subsequently, in GnRH3 soma migration to the hypothalamus. 6). Netrin2 is required for normal early ZF embryogenesis. 7). Sonic hedgehog a does not serve as a specific factor in the development of the GnRH3 system. 8). GnRH3 neuron regeneration capacity is temporally limited. 9). Successful ablation of olfactory GnRH3 neurons during development results in lack of GnRH3 neurons in the entire sexually mature brain as well as abnormal gonadal development and inability to reproduce. This study expands our understanding vis-à-vis the early events that occur during GnRH3 system development and that regulate this complex process. In a broader sense these findings augment current knowledge regarding the regulation of long range tangential neuron migration during development.Item HARMFUL ALGAL BLOOM STRESSORS ALTER BEHAVIOR AND BRAIN ACTIVITY IN THE KILLIFISH, FUNDULUS HETEROCLITUS.(2005-04-15) Salierno, James D; Kane, Andrew S; Marine-Estuarine-Environmental Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Harmful algal bloom (HAB) events are increasing in severity and frequency worldwide, and are known to severely impact fish populations. Impacts of HABs on fish, as well as other organisms, occur through toxic and physical stress. Behavioral and central nervous system (CNS) alterations can have direct consequences to the fitness and survival of individuals and populations. This study investigated and characterized alterations in social and swimming behaviors and brain activity in mummichog (Fundulus heteroclitus) exposed to HAB stressors. The mummichog is an ecologically important estuarine fish species exposed to a variety of HAB events in the wild. A behavioral analysis system was developed to study swimming and social behavior of fish and an immunohistochemistry technique was used to investigate alterations in neuronal activity as evidenced by c-Fos protein expression. HAB stressors included excitatory (domoic acid, brevetoxin) and inhibitory (saxitoxin) neurotoxic agents as well as direct exposures to the dinoflagellate Pfiesteria shumwayae and the diatom Chaetoceros concavicornis. P. shumwayae and C. concavicornis are HAB species that are known to induce mortality through physical trauma to fish. Brevetoxin exposure increased swimming and social behaviors whereas saxitoxin decreased these behaviors. The effects of saxitoxin on swimming and social behaviors were consistent with exposure to a fish anesthetic, MS-222. Similarly, it was found, through c-Fos expression, that the excitatory HAB neurotoxins brevetoxin and domoic acid, increased neuronal activity while saxitoxin decreased activity. Exposure to P. shumwayae and C. concavicornis, resulted in significant dose related increases in neuronal activity. Stressor-specific neuronal activity was greatest in the optic lobe, but was also found in the telencephalon with physical stressors increasing activity greater than chemical stressors. Results demonstrate that sublethal exposures to HAB neurotoxins can alter swimming and social behavior in mummichog and exposures to both neurotoxins and algae can alter neuronal activity. Alterations in brain activity, and knowledge of specific regions within the brain activated during stress, can provide insights into the control of fish behavior. Ultimately, HAB exposure related changes in neuronal signaling may alter behaviors, resulting in individual and population level alterations during HAB events.