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日本鰻魚紅體與卵巢組織在發育過程中其PTEN(phosphatase a...

國立高雄大學生物科技研究所

日本鰻魚紅體與卵巢組織在發育過程中其PTEN(phosphatase and tensin homologue deleted on chromosome 10)Homolog和EDF-1(Endothelial differentiation-related factor-1)表現與影響其表現因子之研究 = Investigation of PTEN Homolog and EDF-1expression inJapanese eel(Anguilla japonica)rete mirabile and ovary tissues during development and the possible regulation studies

紀錄類型:	書目-語言資料,印刷品 : 單行本
並列題名:	Investigation of PTEN Homolog and EDF-1expression inJapanese eel(Anguilla japonica)rete mirabile and ovary tissues during development and the possible regulation studies
作者:	陳雅玫,
其他團體作者:	國立高雄大學
出版地:	[高雄市]
出版者:	撰者;
出版年:	2009[民98]
面頁冊數:	145面圖、表 : 30公分;
標題:	卵巢發育指數
標題:	EDF-1
附註:	指導教授：黃永森
附註:	參考書目：面
摘要註:	PTEN (phosphatase and Tensin homolog deleted on chromosome 10，亦稱MMAC1 or TEP1)為一種磷酸酶，擷抗磷脂酰肌醇激酶（PI3K；phosphoinositide 3-kinase）所引起的生理活性。內皮分化因子(Endothlial differentiating-related Factor-1；EDF-1)，其與血管內皮細胞分化相關。我們選殖日本鰻的EDF-1基因序列，並探討鰻魚在發育過程中紅體和卵巢PTEN long、PTEN short和EDF-1基因的表現。在不同體型的養殖鰻之紅體發育(RMI值)會隨著其體重增加而增加。而PTEN和EDF-1mRNA表現量會隨著其紅體發育(RMI值)增加而增加表現量的趨勢。催熟過程中，對於調控PTEN的作用不同，當紅體(RMI值)組織發育，其PTEN mRNA表現量和蛋白質都會增加，而當卵巢(GSI值)發育，PTEN long基因表現量與卵巢組織(GSI值)發育成負相關，但是PTEN short基因表現量與卵巢組織(GSI值)發育的相關性較不一致，而PTEN蛋白質會受到抑制。而當組織不發育時，會減少EDF-1 mRA表現量，組織發育時，則會增加EDF-1 mRA表現量。而添加睪固酮會抑制PTEN和EDF-1表現量，但會增加PTEN蛋白質免疫反應。我們推測添加睪固酮之混合液進行催熟，會抑制PTEN和EDF-1基因表現量，使得卵巢發育不會受到抑制。離體(in vitro)實驗中，長期培養紅體細胞，對於PTEN基因而言似乎約2~3週就會有一個週期性的波動，而EDF-1表現量維持恆定。我們以PPAR agonists、生長因子、類固醇和金屬離子處理紅體細胞，並進一步觀察PTEN long與PTEN short的基因表現和蛋白質調控以及EDF-1基因表現量是否具有相關性。在PPAR agonists(α、β/δ、γ)刺激作用下，會增加PTEN基因和蛋白質與EDF-1基因表現量。ECGs會抑制PTEN基因表現，但會增加EDF-1表現量。IGF-1劑量為10-7M時，會明顯增加PTEN long免疫染色反應強度。Insulin會增加PTEN long基因表現量。類固醇(T、E2、P4)會增加PTEN和EDF-1基因表現量，並且T、P4會增加PTEN long免疫染色反應的強度。Zn長時間作用下，會抑制PTEN long免疫反應強度。CoCl2長時間作用下，會增加PTEN和EDF-1表現量。Cu2+長時間作用下，會增加PTEN表現量，但會抑制PTEN long免疫反應的強度。另外，免疫染色反應結果顯示，PTEN long蛋白質會在紅體細胞的細胞核周圍表現，我們推測可能是在高爾基氏體表現，而PTEN short在紅體細胞的免疫反應比較沒有專一性，似乎會在全細胞中都有表現。總合以上結果，在鰻魚的紅體細胞中，PTEN long和PTEN short的作用不同，而且我們推測PTEN long與PI3K訊號傳遞路徑相關。而PTEN short的表現都沒有太大差異，我們推測PTEN short的功能似乎與染色體穩定性、DNA修復、細胞週期停止和細胞內穩定性相關。而EDF-1基因會受到生長因子、類固醇和二價離子所調控。 ABSTRACTPTEN (phosphatase and Tensin homolog deleted on chromosome 10, also called MMAC1 or TEP1) is a phosphatase，PTEN against the bioactivity provoked by PI3K (phosphoinositide 3-kinase). EDF-1 (Endothelial differentiating-related Factor-1) is a transcriptional factor involved into angiogenesis and the differentiation of vascular endothelial cells. Used Japanese as the model, EDF-1 and two forms of PTEN genes have been cloned, we investigated the gene expression profiles in the gas glands as well as the ovary during the development of eel. The increase of RMI (rete mirabile index) was positive with the increase of body weight, the expression levels of both PTEN and EDF-1 were also increased with the increase of RMI. In the eels stimulated by catfish pituitary extracts, the tissue masses were also promoted, the expression levels as well as cellular proteins content was increased, the expression levels of PTEN-long form was decreased with increase of GSI (gonadal somatic index), while there was no correlation between GSI and the expression levels of PTEN-short form, the samples with higher PTEN-short form expression levels had the lower cellular proteins content. The expression levels of EDF-1 were increased in the developing ovary while the expression levels of EDF-1 were decreased in the non-hyperplasia tissue. The supplement of testosterone (T) decreased the expression levels of both PTEN as well as EDF-1, but the cellular proteins content of those were increased, it seemed that CPE supplement with T inhibited the expression levels of both PTEN and EDF-1 leaded the ovarian development. In vitro experiments showed that there was a fluctuation on the expression levels of PTEN for every 2-3 weeks to the long-term primary culture of gas glands cells as long as 8 weeks, this result was not observed in EDF-1. The primary culture of gas glands cells was treated with PPAR agonists, growth factors, steroids, and metal ions to see their effects on the different gene expression levels as well as correlation on gene expression and cellular proteins content. The PTEN production and EDF-1 expression levels were stimulated by PPAR agonists (α、β/δ、γ); 。ECGs inhibited the expression levels of PTEN but increased those of EDF-1. 10-7 M of IGF-1 increased cellular proteins content of PTEN-long; Insulin stimulated the expression of PTEN-long form. Steroids (T、E2、P4) stimulated the expression levels of PTEN and EDF-1, and the cellular proteins content of PTEN-long form was increased by T and P4. the production of PTEN-long form was effected by Zn, Co, and Cu. Our cellular immunofluorescence staining showed that The PTEN-long form had a strong staining intensity around the nuclear, this area is presumed the Golgi apparatus, while the immuno-staining on PTEN-short form was more ubiquitous and widely distributed. In summary, from our data, we speculated that the physiological functions of PTEN-long and PTEN-short form are different; PTEN-long form seems to involve in the PI3K signal transduction pathway; the expression levels of PTEN-short were not effected by various treatments. Growth factors, steroids, and metal ions regulates the expression of EDF-1.

日本鰻魚紅體與卵巢組織在發育過程中其PTEN(phosphatase and tensin homologue deleted on chromosome 10)Homolog和EDF-1(Endothelial differentiation-related factor-1)表現與影響其表現因子之研究 = Investigation of PTEN Homolog and EDF-1expression inJapanese eel(Anguilla japonica)rete mirabile and ovary tissues during development and the possible regulation studies
陳, 雅玫

日本鰻魚紅體與卵巢組織在發育過程中其PTEN(phosphatase and tensin homologue deleted on chromosome 10)Homolog和EDF-1(Endothelial differentiation-related factor-1)表現與影響其表現因子之研究 = Investigation of PTEN Homolog and EDF-1expression inJapanese eel(Anguilla japonica)rete mirabile and ovary tissues during development and the possible regulation studies / 陳雅玫撰 - [高雄市] : 撰者, 2009[民98]. - 145面 ; 圖、表 ; 30公分.
指導教授：黃永森參考書目：面.
卵巢發育指數EDF-1

日本鰻魚紅體與卵巢組織在發育過程中其PTEN(phosphatase and tensin homologue deleted on chromosome 10)Homolog和EDF-1(Endothelial differentiation-related factor-1)表現與影響其表現因子之研究 = Investigation of PTEN Homolog and EDF-1expression inJapanese eel(Anguilla japonica)rete mirabile and ovary tissues during development and the possible regulation studies
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330 $a PTEN (phosphatase and Tensin homolog deleted on chromosome 10，亦稱MMAC1 or TEP1)為一種磷酸酶，擷抗磷脂酰肌醇激酶（PI3K；phosphoinositide 3-kinase）所引起的生理活性。內皮分化因子(Endothlial differentiating-related Factor-1；EDF-1)，其與血管內皮細胞分化相關。我們選殖日本鰻的EDF-1基因序列，並探討鰻魚在發育過程中紅體和卵巢PTEN long、PTEN short和EDF-1基因的表現。在不同體型的養殖鰻之紅體發育(RMI值)會隨著其體重增加而增加。而PTEN和EDF-1mRNA表現量會隨著其紅體發育(RMI值)增加而增加表現量的趨勢。催熟過程中，對於調控PTEN的作用不同，當紅體(RMI值)組織發育，其PTEN mRNA表現量和蛋白質都會增加，而當卵巢(GSI值)發育，PTEN long基因表現量與卵巢組織(GSI值)發育成負相關，但是PTEN short基因表現量與卵巢組織(GSI值)發育的相關性較不一致，而PTEN蛋白質會受到抑制。而當組織不發育時，會減少EDF-1 mRA表現量，組織發育時，則會增加EDF-1 mRA表現量。而添加睪固酮會抑制PTEN和EDF-1表現量，但會增加PTEN蛋白質免疫反應。我們推測添加睪固酮之混合液進行催熟，會抑制PTEN和EDF-1基因表現量，使得卵巢發育不會受到抑制。離體(in vitro)實驗中，長期培養紅體細胞，對於PTEN基因而言似乎約2~3週就會有一個週期性的波動，而EDF-1表現量維持恆定。我們以PPAR agonists、生長因子、類固醇和金屬離子處理紅體細胞，並進一步觀察PTEN long與PTEN short的基因表現和蛋白質調控以及EDF-1基因表現量是否具有相關性。在PPAR agonists(α、β/δ、γ)刺激作用下，會增加PTEN基因和蛋白質與EDF-1基因表現量。ECGs會抑制PTEN基因表現，但會增加EDF-1表現量。IGF-1劑量為10-7M時，會明顯增加PTEN long免疫染色反應強度。Insulin會增加PTEN long基因表現量。類固醇(T、E2、P4)會增加PTEN和EDF-1基因表現量，並且T、P4會增加PTEN long免疫染色反應的強度。Zn長時間作用下，會抑制PTEN long免疫反應強度。CoCl2長時間作用下，會增加PTEN和EDF-1表現量。Cu2+長時間作用下，會增加PTEN表現量，但會抑制PTEN long免疫反應的強度。另外，免疫染色反應結果顯示，PTEN long蛋白質會在紅體細胞的細胞核周圍表現，我們推測可能是在高爾基氏體表現，而PTEN short在紅體細胞的免疫反應比較沒有專一性，似乎會在全細胞中都有表現。總合以上結果，在鰻魚的紅體細胞中，PTEN long和PTEN short的作用不同，而且我們推測PTEN long與PI3K訊號傳遞路徑相關。而PTEN short的表現都沒有太大差異，我們推測PTEN short的功能似乎與染色體穩定性、DNA修復、細胞週期停止和細胞內穩定性相關。而EDF-1基因會受到生長因子、類固醇和二價離子所調控。 ABSTRACTPTEN (phosphatase and Tensin homolog deleted on chromosome 10, also called MMAC1 or TEP1) is a phosphatase，PTEN against the bioactivity provoked by PI3K (phosphoinositide 3-kinase). EDF-1 (Endothelial differentiating-related Factor-1) is a transcriptional factor involved into angiogenesis and the differentiation of vascular endothelial cells. Used Japanese as the model, EDF-1 and two forms of PTEN genes have been cloned, we investigated the gene expression profiles in the gas glands as well as the ovary during the development of eel. The increase of RMI (rete mirabile index) was positive with the increase of body weight, the expression levels of both PTEN and EDF-1 were also increased with the increase of RMI. In the eels stimulated by catfish pituitary extracts, the tissue masses were also promoted, the expression levels as well as cellular proteins content was increased, the expression levels of PTEN-long form was decreased with increase of GSI (gonadal somatic index), while there was no correlation between GSI and the expression levels of PTEN-short form, the samples with higher PTEN-short form expression levels had the lower cellular proteins content. The expression levels of EDF-1 were increased in the developing ovary while the expression levels of EDF-1 were decreased in the non-hyperplasia tissue. The supplement of testosterone (T) decreased the expression levels of both PTEN as well as EDF-1, but the cellular proteins content of those were increased, it seemed that CPE supplement with T inhibited the expression levels of both PTEN and EDF-1 leaded the ovarian development. In vitro experiments showed that there was a fluctuation on the expression levels of PTEN for every 2-3 weeks to the long-term primary culture of gas glands cells as long as 8 weeks, this result was not observed in EDF-1. The primary culture of gas glands cells was treated with PPAR agonists, growth factors, steroids, and metal ions to see their effects on the different gene expression levels as well as correlation on gene expression and cellular proteins content. The PTEN production and EDF-1 expression levels were stimulated by PPAR agonists (α、β/δ、γ); 。ECGs inhibited the expression levels of PTEN but increased those of EDF-1. 10-7 M of IGF-1 increased cellular proteins content of PTEN-long; Insulin stimulated the expression of PTEN-long form. Steroids (T、E2、P4) stimulated the expression levels of PTEN and EDF-1, and the cellular proteins content of PTEN-long form was increased by T and P4. the production of PTEN-long form was effected by Zn, Co, and Cu. Our cellular immunofluorescence staining showed that The PTEN-long form had a strong staining intensity around the nuclear, this area is presumed the Golgi apparatus, while the immuno-staining on PTEN-short form was more ubiquitous and widely distributed. In summary, from our data, we speculated that the physiological functions of PTEN-long and PTEN-short form are different; PTEN-long form seems to involve in the PI3K signal transduction pathway; the expression levels of PTEN-short were not effected by various treatments. Growth factors, steroids, and metal ions regulates the expression of EDF-1.
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