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Localization of Circadian Clock Neur...
~
Duback, Victoria.
Localization of Circadian Clock Neurons and Investigations of Their Influence on the Expression of Specific Behaviors in the Marine Mollusk, Melibe leonina.
紀錄類型:
書目-語言資料,手稿 : Monograph/item
正題名/作者:
Localization of Circadian Clock Neurons and Investigations of Their Influence on the Expression of Specific Behaviors in the Marine Mollusk, Melibe leonina./
作者:
Duback, Victoria.
面頁冊數:
1 online resource (131 pages)
附註:
Source: Masters Abstracts International, Volume: 57-01.
標題:
Neurosciences. -
電子資源:
click for full text (PQDT)
ISBN:
9780355306552
Localization of Circadian Clock Neurons and Investigations of Their Influence on the Expression of Specific Behaviors in the Marine Mollusk, Melibe leonina.
Duback, Victoria.
Localization of Circadian Clock Neurons and Investigations of Their Influence on the Expression of Specific Behaviors in the Marine Mollusk, Melibe leonina.
- 1 online resource (131 pages)
Source: Masters Abstracts International, Volume: 57-01.
Thesis (M.S.)--University of New Hampshire, 2017.
Includes bibliographical references
Most organisms studied to date express daily behavioral rhythms that are under the influence of some form of circadian clock. While the molecular mechanisms underlying these circadian clocks are well understood, little is known about how these clocks influence the expression of specific behaviors. The overall goal of this thesis research was to identify clock neurons in an organism that is very suitable for investigations of the neural basis of behavior, Melibe leonina. First, we used M. leonina clock protein sequences, obtained in a separate study, to develop antibodies against Melibe-specific clock proteins. These antibodies made it possible to use immunohistochemistry to identify putative clock neurons in the M. leonina brain and buccal ganglia. Specifically, a pair of neurons in the cerebral-pleural ganglia and a single neuron in each buccal ganglion, were immunoreactive to more than one clock protein. M. leonina clock gene sequences were also used to develop primers so that qPCR could be used to measure the expression of clock genes in the brain and buccal ganglia. Clock genes were expressed in both tissues, with the greatest expression in the brain, as well as the most pronounced daily fluctuations. Long-term recordings from isolated buccal ganglia revealed that some buccal nerves are more active in the night than the day, suggesting that there could be clock neurons in the buccal ganglia that are capable of producing a circadian rhythm of activity. This hypothesis is supported by the finding that the hepatic trunks (connected to the stomach), that receive inputs from the buccal ganglia, also exhibit a daily rhythm of contractions. Finally, in order to test the hypothesis that the neurons identified with immunohistochemistry were, in fact, part of the circadian timekeeping network, we determined how deleting them or cutting specific connections, influenced circadian rhythms of locomotion. We found that neither the putative clock cells in the midline of the brain, or the buccal ganglia, are necessary for M. leonina to express a circadian rhythm of locomotion. However, when both the midline cells and the connection between the buccal ganglia and the brain were lesioned at the same time, 80% (4/5) animals were not capable of producing a circadian rhythm of locomotion. Together, these data suggest that M. leonina might have a distributed network of integrated clock neurons in the CNS and while some of these might be capable for driving daily rhythms on their own, together they produce a more robust rhythm. These findings indicate that Melibe leonina is potentially a very useful model system for investigating both the influence of identified clock neurons and the neural networks that drive specific behaviors and the interactions between multiple subpopulations of clock neurons.
Electronic reproduction.
Ann Arbor, Mich. :
ProQuest,
2018
Mode of access: World Wide Web
ISBN: 9780355306552Subjects--Topical Terms:
593561
Neurosciences.
Index Terms--Genre/Form:
554714
Electronic books.
Localization of Circadian Clock Neurons and Investigations of Their Influence on the Expression of Specific Behaviors in the Marine Mollusk, Melibe leonina.
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Most organisms studied to date express daily behavioral rhythms that are under the influence of some form of circadian clock. While the molecular mechanisms underlying these circadian clocks are well understood, little is known about how these clocks influence the expression of specific behaviors. The overall goal of this thesis research was to identify clock neurons in an organism that is very suitable for investigations of the neural basis of behavior, Melibe leonina. First, we used M. leonina clock protein sequences, obtained in a separate study, to develop antibodies against Melibe-specific clock proteins. These antibodies made it possible to use immunohistochemistry to identify putative clock neurons in the M. leonina brain and buccal ganglia. Specifically, a pair of neurons in the cerebral-pleural ganglia and a single neuron in each buccal ganglion, were immunoreactive to more than one clock protein. M. leonina clock gene sequences were also used to develop primers so that qPCR could be used to measure the expression of clock genes in the brain and buccal ganglia. Clock genes were expressed in both tissues, with the greatest expression in the brain, as well as the most pronounced daily fluctuations. Long-term recordings from isolated buccal ganglia revealed that some buccal nerves are more active in the night than the day, suggesting that there could be clock neurons in the buccal ganglia that are capable of producing a circadian rhythm of activity. This hypothesis is supported by the finding that the hepatic trunks (connected to the stomach), that receive inputs from the buccal ganglia, also exhibit a daily rhythm of contractions. Finally, in order to test the hypothesis that the neurons identified with immunohistochemistry were, in fact, part of the circadian timekeeping network, we determined how deleting them or cutting specific connections, influenced circadian rhythms of locomotion. We found that neither the putative clock cells in the midline of the brain, or the buccal ganglia, are necessary for M. leonina to express a circadian rhythm of locomotion. However, when both the midline cells and the connection between the buccal ganglia and the brain were lesioned at the same time, 80% (4/5) animals were not capable of producing a circadian rhythm of locomotion. Together, these data suggest that M. leonina might have a distributed network of integrated clock neurons in the CNS and while some of these might be capable for driving daily rhythms on their own, together they produce a more robust rhythm. These findings indicate that Melibe leonina is potentially a very useful model system for investigating both the influence of identified clock neurons and the neural networks that drive specific behaviors and the interactions between multiple subpopulations of clock neurons.
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