|Title:||菜鴨卵巢濾泡細胞生長及類固醇生成調控機制之研究(2/2)||Authors:||吳兩新||Keywords:||菜鴨;卵巢濾泡;類固醇內泌素;Tsaiya duck;ovarian follicle;steroid hormone||Issue Date:||2004||Publisher:||臺北市：國立臺灣大學動物科學技術學系暨研究所||Abstract:||
本研究首先製備類固醇合成酵素P450scc、3β-HSD 與P450aromatase 三種抗體，並利用菜鴨（Tsaiya duck;Anas platyrhynchos var. domestica）卵巢濾泡細胞，建立卵巢濾泡三細胞分離技術，採用體外培養卵巢濾泡三細胞，以瞭解其孕酮、睪固酮與雌二醇等類固醇，以及3β-HSD、17β-HSD 與P450aromatase 酵素活性，以瞭解菜鴨卵巢濾泡細胞類固醇內泌素之分泌模式。並進一步瞭解生長因子對類固醇分泌的影響。
一、P450scc、3β-HSD 與P450aromatase 三種抗體可應用於西方墨染法，以供辨識牛、豬、羊、鴨等之類固醇內泌素生成組織。
二、菜鴨卵巢濾泡中之粒性細胞以合成孕酮為主，睪固酮次之，其基礎分泌量及3β-HSD 與17β-HSD 酵素活性，以即將排卵的F1 最高，漸次下降至成熟度最低的F4/5。雌二醇分泌量極低，且濾泡大小間無顯著差異。內膜細胞主要合成睪固酮及雌二醇，其基礎分泌量及17β-HSD 與P450aromatase 酵素活性，皆以最小濾泡F4/5 為最高，並隨著濾泡的逐漸成熟而下降。內膜細胞並無孕酮分泌能力。濾泡中之外膜細胞以合成雌二醇為主，其基礎分泌量及P450aromatase 酵素活性以F2 為最高;而外膜細胞幾乎無孕酮及睪固酮分泌能力。
三、EGF 及IGF-1 會刺激粒性細胞分泌孕酮;TGF-β會抑制之;而FGF 則沒有影響。
The purpose of this study was to prepare the steroidogenic enzymes-- P450scc ,
3β-HSD, and P450arom, and also to establish the isolation techniques of ovarian
follicular granulosa, theca interna and theca externa cells in Tsaiya duck (Anas
platyrhynchos var. domestica). Furthermore, in order to establish steroid hormone
secretion model of ovarian follicular cells in Tsaiya duck, progesterone, testosterone
and estradiol secretion as well as expression of 3β-HSD, 17β-HSD and P450arom in
follicular cells were studied by in vitro cell culture system. Finally, the effect of
growth factors on steroids production also will be examined.
Results showed as follow:
1. The polyclonal antibodies of P450scc、3β-HSD and P450aromatase obtained
can recognize the steroid-producing tissues of cattle, pig, goat and duck by using
2. The progesterone was the main steroid produced by granulosa cell, and
testosterone was the second one. The progesterone level and enzyme activities of
3β-HSD and 17β-HSD of granulosa cell increased gradually with the follicle
maturity. The estradiol secretion from granulosa cell was low, and was not
significantly different between follicles. The main steroids produce from theca
interna cell was testosterone and estradiol. The F4/5 follicles exhibited the highest
basal testosterone and estradiol secretion as well as enzyme activities of 17β-HSD
and P450arom from theca interna cell. Both of them were declined with the follicle
maturity. Besides, theca interna did not secrete progesterone. Estradiol was the main
steroid secreted by theca externa. Theca externa of F2 presented the highest estradiol
secretion and P450arom activity. Moreover, theca externa did not secrete
progesterone and testosterone.
3. The EGF and IGF-1 could stimulate the progesterone production from
granulose cells in vitro, and TGF-β did not. The FGF has no any effect in this part.
This study demonstrated the different abilities of steroid secretion among
ovarian follicular granulosa, theca interna and theca externa cells in Tsaiya duck by
individual in vitro culture system. However, the interactions between different
layers are required further study.
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