本研究主要在探討點帶石斑 (Epinephelus coioides) 促性腺激素釋素受體 (gonadotropin-releasing hormone receptor, GnRH-R) 與 neuropeptide Y ，在腦區與腦下垂體的表現情況，同時透過不同激素注射實驗了解其調控機制。本實驗選殖出片段GnRH-R，包含部分 TMD2 到 TMD6 序列。組織分佈方面，GnRH-R 在腦下垂體的表現量明顯的高於其它組織。不同年齡石斑腦及腦下垂體 GnRH-R 的表現量都在第四年之後會有顯著的增加，顯示此時已開始為繁殖行為做準備，而三年齡石斑腦下垂體GnRH-R不同季節表現差異並不顯著，不同性類固醇 (E2、T) 注射對二年齡與三年齡石斑，E2能有效的增加腦下垂體 GnRH-R 表現，而 T 則只對三年齡石斑有誘導效果，顯示點帶石斑對於性類固醇的感受性會因注射種類與年齡有所不同。在不同腦區中，二年齡石斑前中腦 GnRH-R 表現量注射E2、T組會低於控制組，性類固醇的注射抑制了二年齡石斑前中腦區 GnRH-R 的表現。三年齡石斑注射 T 能有效的誘導前中腦區 GnRH-R 表現，注射 E2 則無顯著效果。後腦區方面，二年齡與三年齡石斑都不會受到性類固醇注射影響，性類固醇調控的區域仍是在前中腦區，這可能與大量類固醇受體的分佈位置有關。以 LHRH 與 pimozide (多巴胺抑制劑) 注射二年齡石斑，LHRH 只能有效的增加石斑腦下垂體 GnRH-R 表現量，對其它腦區則無影響，pimozide 則無促進表現，兩者共同注射亦無法增加 GnRH-R 表現量。顯示，外加 LHRH，亦能增加腦下垂體 GnRH-R 表現，而多巴胺 (dopamine) 對 GnRH-R 表現影響並不明顯。
NPY 的研究中，NPY 主要分佈在不同腦區，前腦表現量最多，在鰓、肝臟、腸道等組織中也有發現，在鰓組織中 NPY 含量僅次於腦區的表現，顯示，NPY 在相關滲透壓功能調節上可能也有影響。不同年齡石斑腦中 NPY 表現，隨著年齡增加而下降，確實影響因素尚不清楚。注射不同性類固醇對二年齡與三年齡石斑，腦中之 NPY 表現並沒有顯著影響，這與許多過去研究結果有差異，推測是因為實驗設計上，腦分區上的不精細所造成，而使得其變化無法顯現。石斑腦中 NPY 的表現也不因注射 LHRH 與 pimozide 而有所變化。

The aim of the present research was to investigate the expression profiles of GnRH-R and neuropeptide Y (NPY) genes in brain and pituitary of Orange-spotted Grouper, Epinephelus coioides and also to understand the regulatory mechanism by administering different sex steroids. GnRH-R (TMD2 to TMD6) was partially involved cloned in this study. Tissue distribution analysis revealed a significant expression of GnRH-R in pituitary compared to others tissues. The expression of GnRH-R in brain and pituitary of groupers at different ages showed a significant increased during the fourth year, probably indicating the time of maturation. However, there was no significant difference in the expression of GnRH-R during different seasons. Treatment of two and three year old groupers with different sex steroids revealed an increase in the expression of GnRH-R in pituitary by E2 in both the age groups tested, while T could induce the expression of GnRH-R only in three year old groupers. The result, thus, indicates that the sensitivity of grouper to sex steroid is dependent on the age and the kind of steroid adminstered. In different sections of brain, the GnRH-R expression was in general lower in the group treated with E2 or T compared to the control group. The expression of the gene was more or less the same in two year and three year old control groups. This result may have been caused by suppression of GnRH-R expression in forebrain and midbrain after sex steroid injection. Administration of T induced a significant increase in the expression of GnRH-R in forebrain and midbrain, while E2 treatment did not have a similar effect. In hindbrain, the expression profile GnRH-R was not affected by sex steroid treatment in both two year and three year old groupers. The results suggest that sex steroids can only regulate the expression of GnRH-R in the forebrain and midbrain, probably due to the wide distribution of steroid’s receptor in these regions. LHRH and pimozide injections to two year old groupers showed an increase in the expression of GnRH-R in pituitary after LHRH treatment while there was no stimulatory effect on other sections of the brain. In contrast, treating the fish with pimozide alone or pimozide together with LHRH did not stimulate GnRH-R expression in brain. Thus, the study suggests that LHRH can significantly increase the expression of GnRH-R in pituitary while dopamine has no stimulatory effect.
Studies on NPY showed that the gene was distributed in different sections of brain especially in the forebrain but it was also present in gills, liver, intestine … etc. The presence of NPY in gills, though less compared to that in brain, suggests that NPY might play an important role in osmosis regulation. The expression of NPY decreased with increase in age which may be due to the effect of other regulatory factors. Treatment of two and three year old groupers with different sex steroids did not effect the expression of NPY significantly in brain, which is different from other published reports. This may be due to the difference in the zoning of brain regions. In the present study, forebrain and midbrain were sampled together for analyses. The expression of NPY in brain did not change by treating the fish with LHRH or pimozide.

中文摘要…………………………………………………………………………iii
英文摘要…………………………………………………………………………v
目錄………………………………………………………………………………vii
圖目錄………………………………………………………………………… viii
緒言………………………………………………………………………………1
材料方法…………………………………………………………………………11
結果………………………………………………………………………………24
討論………………………………………………………………………………29
結論………………………………………………………………………………36
參考文獻…………………………………………………………………………37
個人履歷…………………………………………………………………………92

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