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The effect of estrogen on gonadal cells transdifferentiation in the testis of protandrous black porgy, Acanthopagrus schlegelii
|Authors: ||Kuo, Wei-Lun|
|Contributors: ||NTOU:Department of Aquaculture|
|Keywords: ||錯置卵細胞;kcnq1 isoform;heparan sulfate sulfotransferase|
ectopic oocytes;kcnq1 isoform;heparan sulfate sulfotransferase
|Issue Date: ||2018-08-22T06:06:47Z
|Abstract: ||黑鯛為的精巢與卵巢中不會有錯置的生殖細胞。幼魚經E2處理精巢會不發育，停止處理後新生的精巢中會發現錯置卵細胞，因此錯置卵細胞被認為是E2處理之次要產物。錯置卵細胞周圍的Sertoli cell會隨著錯置卵細胞成長而轉型成follicle cell，其中錯置卵細胞的figla有延長大量表現的現象，顯示錯置卵細胞中figla大量表現可能與適應錯誤環境以及生殖體細胞轉型有關。 本研究將探討錯置卵細胞為高濃度E2直接或間接造成之結果，本實驗利用藥物追蹤停餵E2後精巢內細胞新生之狀況。停餵後血漿中E2濃度皆非常低，停餵三週後開始發現錯置卵細胞存在於精巢中。根據BrdU染色，精巢內是體細胞先進行新生。根據上述，雌性生殖細胞可於低濃度E2下分化。根據先前離體培養實驗，轉殖不同濃度figla卵細胞的qPCR結果，發現2個與figla表現高度相關的基因 (kcnq1 isoform和hs6st1a)。kcnq1於卵巢的表現顯著高於精巢以及在雌性生殖細胞的表現顯著高於體細胞。kcnq1的表現隨卵細胞卵徑增加有上升的趨勢；kcnq1 isoform於卵巢的表現顯著高於精巢以及在雌性生殖細胞的表現顯著高於體細胞。kcnq1 isoform的表現隨卵細胞卵徑增加有下降的趨勢。從黑鯛轉錄體中選殖出9個heparan sulfate sulfotransferase基因 (hs2st1a、hs3st1、hs3st3a、hs3st3b1b、hs3st4、hs6st1a、hs6st2、hs6st3a、hs6st3b)。hs2st1a、hs3st3a、hs3st3b1b、hs3st4、hs6st1a、hs6st3a於卵巢的表現顯著高於精巢以及在雌性生殖細胞的表現顯著高於體細胞。hs2st1a和hs6st3a的表現和卵徑呈現正相關； hs3st3a1和hs3st3b1b的表現和卵徑呈現負相關，我們推測這些基因可能與錯置卵細胞適應錯誤環境及體細胞轉變有關。|
The digonic gonad (testis and ovary) of protandrous black porgy are separated by connective tissue. No intersex (ectopic located germ cells) was observed in either part. However, ectopic oocytes were observed in the regenerated testis after E2 withdrawal in E2-treated fish. These ectopic oocytes were altered the sexual fate of oocytes-surrounded cells from male (Sertoli cells) to female (follicle cells) through the Figla signaling. However, Figla is transcription factor. Thus, Figla signaling needs the oocytes-releasing factors to alter the male characteristics. In this study, we would like to understand the origin of ectopic oocytes in regenerated testis and mechanism of Figla signaling. Base on histology, no ectopic oocytes were observed in regressed testis in the E2-treated fish. These ectopic oocytes were observed after 3 weeks of E2 withdrawal in E2-treated fish. Furthermore, the expression of vtg in liver was significantly low during testis regeneration in E2-terminated fish compared with the E2-treated (6 mg E2/kg fed and 1 mg E2/kg fed) fish. We also analyze the expression profiles of the potential downstream genes in Figla signaling, including the hs6st1a and kcnq1 isoform. In further, we also analyze the expression profiles of the 9 heparan sulfate sulfotransferase family genes in testis and ovary. Our data shows that kcnq1 isoform and six hsst genes highly expressed in ovary compare with testis. These genes were significantly expressed in oocytes compare with somatic cells. The expression of kcnq1 isoform and four hsst genes were difference in differt oocyte diameter. Taking togrther, our data demonstrates that ectopic female germ cells are differentiated under the low E2 levels. Our results shows five genes may include in the downstream of the Figla signaling. These finding might helps us to understand the early transition from gonochorism to hermaphroditism while germ cells facing the incorrect microenvironment.
|Appears in Collections:||[水產養殖學系] 博碩士論文|
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