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系統識別號 U0026-1108201421083900
論文名稱(中文) 老化與跑步運動透過不同的機制影響海馬迴神經新生
論文名稱(英文) Aging and running exercise affect adult hippocampal neurogenesis via different mechanisms
校院名稱 成功大學
系所名稱(中) 基礎醫學研究所
系所名稱(英) Institute of Basic Medical Sciences
學年度 102
學期 2
出版年 103
研究生(中文) 羅振鵬
研究生(英文) Chen-Peng Lo
學號 S58931578
學位類別 博士
語文別 英文
論文頁數 50頁
口試委員 指導教授-郭余民
召集委員-莊季瑛
口試委員-楊尚訓
口試委員-蔡坤哲
口試委員-林靜茹
口試委員-林惠菁
中文關鍵字 老化  運動  成年期海馬迴新生  神經幹細胞  增殖  成熟 
英文關鍵字 aging  exercise  adult hippocampal neurogenesis  neural stem cells  proliferation  maturation 
學科別分類
中文摘要 海馬迴神經新生終生都持續地在進行。海馬迴新生成的神經細胞被認為會參與該腦區神經迴路的修補,以及記憶的形成與清除。新生神經細胞的總數可由以下幾個因子決定:1)神經幹細胞的數目,2)神經幹細胞的增殖速率,3)新生成的細胞走向神經分化的比例,和4)新生成神經細胞的存活率。目前已知海馬迴新生神經細胞的數目,會隨著年齡增加而減少;反之,跑步運動則會增加其數目。但是,老化與運動是透過何種機制而影響新生神經細胞的數目,目前尚未有定論。本研究的目的是要探討老化與運動分別是透過哪一個決定因子來影響海馬迴新生神經細胞的數目。我們使用不同年齡(2、5、8、11和20個月大)的小鼠,給予為期一個月的跑步機運動訓練,並於隔天犧牲。犧牲前兩小時,給予一針溴脫氧尿核苷(BrdU)腹腔注射,用以標定新生細胞。我們以BrdU和雙皮脂蛋白(DCX,未成熟神經細胞的特定蛋白)來標定新生神經細胞。結果顯示,當小鼠年齡達9個月時,其海馬迴新生神經細胞的數目比3個月時下降超過95%以上。以絲巢蛋白(nestin)標定神經幹細胞,發現隨著年齡增長,神經幹細胞的數目只有稍微減少,而運動訓練則延緩此減少趨勢。神經幹細胞之增殖速率,在9個月大前顯著下降,且運動無法改變此一下降趨勢。新生細胞分化為神經細胞的比例隨著老化而降低,但運動訓練後會增加。新生細胞的存活率並不受到年齡與運動的影響。老化顯著地延緩新生神經細胞的成熟,而運動訓練可以有效的加速其成熟。降低腦神經滋養因子接受器TrkB的表現量,會顯著地減少新生細胞與未成熟神經細胞的數量。總結以上結果,本研究發現造成海馬迴新生神經細胞數目隨著老化而下降的主要原因,是神經幹細胞的增殖速率大幅降低。雖然運動可以增加神經幹細胞的數目,與增加新生細胞分化為神經細胞的比例,但是並無法回復老化後神經幹細胞增殖速率的巨幅衰減。所以,運動對成年期海馬迴神經新生的總數,影響有限。相較之下,運動對新生神經細胞的成熟,有很強的促進效果。TrkB能影響成年期海馬迴神經新生,但其調控的機制仍有待進一步的研究。
英文摘要 Adult hippocampal neurogenesis continues throughout life and has been associated with the formation, exhibition and clearance of certain types of memory. The number of newborn neuron is determined by 1) the number of neural stem/progenitor cell (NSC), 2) proliferation of NSCs, 3) neuron lineage specification, and 4) survival of the newborn neurons. The number of adult hippocampal neurogenesis declines during aging, but can be increased by running exercise (Ex). However, it remains unclear which of the determinants are affected by aging and Ex. Here, we characterized the four determinants in different ages (2, 5, 8, 11, 20 months) of mice that received one month of Ex training or remained sedentary (Sed). An intraperitoneal injection of bromodeoxyuridine (BrdU) was given two hours before sacrifice to label the newly proliferated cells. The results showed that the numbers of newborn neuron, double labeled by BrdU and DCX (a specific marker for immature neuron), were massively decreased (>95%) by the time the mice were 9-month-old, while Ex increased the numbers of newborn neuron. The numbers of nestin+ NSC were mildly reduced during aging and Ex delayed the decline of NSC numbers during aging. The proliferation rates (numbers of BrdU+ cell/ numbers of nestin+ cell) were greatly decreased by the time the mice were 9-month-old and were not changed by Ex. The rates of neuron lineage specification (numbers of BrdU+DCX+ cell/ numbers of BrdU+ cell) were decreased during aging, but increased by Ex. The survival rate (numbers of BrdU+ cell at four weeks after BrdU injection/ numbers of BrdU+ cell at two hours after BrdU injection) was not affected by age or Ex. Aging greatly reduced newborn neuron maturation (numbers of DCX+ dendritic branch and length of DCX+ dendrite), while Ex potently enhanced newborn neuron maturation. Knocking down the receptor of brain-derived neurotrophic factor (TrkB) reduced the numbers of both newborn cell and immature neuron. In conclusion, age-associated decline of hippocampal neurogenesis is mainly caused by reduction of NSC proliferation rate. Ex increases adult hippocampal neurogenesis by sustaining the numbers of NSC and increasing the rates of neuron lineage specification. Because Ex does not restore the massive decline of NSC proliferation rate, the effect of Ex on adult hippocampal neurogenesis during aging is limited. Rather, Ex greatly enhances the maturation of newborn neurons. TrkB is involved in adult hippocampal neurogenesis, but its underlying mechanism requires further investigation.
論文目次 中文摘要....................................................I
Abstract..................................................II
誌謝......................................................IV
Contents...................................................V
Figure contents .........................................VII
Abbreviations.............................................IX
I.Introduction.............................................1
1.Adult Neurogenesis...................................1
2.Adult hippocampal neurogenesis and aging.............2
3.Adult hippocampal neurogenesis and exercise..........2
4.The role of BDNF and TrkB in adult hippocampal neurogenesis...............................................3
5.Markers of neurogenesis..............................4
II.Objective...............................................6
III.Specific aims..........................................7
IV.Experimental designs....................................8
V.Materials and methods...................................10
1.Animals.............................................10
2.Treadmill exercise (Ex).............................10
3.BrdU administration and brain tissue processing.....11
4.Immunohistochemistry................................11
5.Cell counting.......................................12
6.Quantification of dendritic branch and length of DCX+ newborn neurons...........................................12
7.Intracranial injection of short hairpin (sh) RNA expressing lentivirus.....................................13
8.Statistical analysis................................13
VI.Results................................................14
1.Effects of aging and Ex on the rate of adult hippocampal neurogenesis..................................14
2.Effects of aging and Ex on the numbers of NSC in the adult hippocampus.........................................15
3.Effects of aging and Ex on proliferation, neuron lineage specification and survival of adult hippocampal neurogenesis..............................................16
4.Effects of aging and Ex on the maturation of newborn neurons...................................................17
5.Effects of TrkB pathway on adult hippocampal neurogenesis..............................................17
VII.Discussion............................................18
VIII.Conclusion...........................................23
IX.References.............................................24
X.Figures.................................................31
XII.Appendix..............................................49
XIII.Publications.........................................50
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