INTRAFOLLICULAR REGULATION OF OVARIAN TGF-B1 EXPRESSION

卵巢 TGF-B1 表达的卵泡内调节

• 批准号: 2885805
• 负责人: JEFFREY V. MAY
• 金额: $ 10万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2885805
• 关键词: animal tissue; cell cell interaction; female; follicle stimulating hormone; gel mobility shift assay; gene expression; graafian follicles; granulosa cell; growth factor receptors; hormone receptor; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; luteinizing hormone; menstrual cycle; messenger RNA; northern blottings; ovulation; paracrine; receptor expression; tissue /cell culture; transforming growth factors; translation factor; western blottings

Transforming growth factor beta1 (TGF-beta1) has been identified as a potentially important intraovarian growth factor due to its pronounced effects upon theca and granulosa cell function and the localization of the growth of work regarding TGF-beta1 and folliculogenesis, there remains a notable deficit in our knowledge regarding its regulation at the cellular level and how this relates to ovarian function. Solving the riddle regarding TGF- beta1 function and regulation represents the long term goals of this project. Using both cultured cells and intact hemi-follicles from the porcine ovary, a model for the regulation of TGF-beta1 expression has been formulated. Data suggest that although both cell types express TGF-beta1mRNA, only theca cells express and secrete the growth factor. Neither FSH, LH, nor cAMP stimulates TGF-beta section in either isolated cell type in culture. In small hemi-follicles, however, FSH but not LH stimulates TGF-beta1 secretion as does cAMP. We hypothesize that theca cells express and secrete TGF-beta1 during follicle development but that it is under the direction of FSH acting upon granulosa cells. We further hypothesize that a FSH-stimulated granulosa cell product regulates theca cell TGF-beta expression by activating the PLA/PKC signal transduction system. To further delineate the inter-follicular regulation of TGF-beta1 it will be established whether TGF- beta1 auto expression occurs in granulosa but not theca cells, whether this is mediated by selective expression of TGF-beta type I and II receptors, and whether auto-expression or receptor expression change with follicle development. The extent to which FSH and TGF-beta1 reciprocally regulate the expression of the other receptor in granulosa cells will be determined. The extent to which granulosa cells can directly regulate theca cell TGF- beta1 expression via secretory factors will be tested by co-culture experiments. Whether follicular TGF-beta1 expression switches from FSH to LH governance with follicle development will be determined. Lastly, the selective granulosa cell expression of a cytosolic protein capable of binding to a regulatory stem-loop structure of the TGF-beta1 mRNA 5'-untranslated region which inhibits translation will be investigated. This may be the mechanism by which granulosa cells differentially express TGF-beta1 mRNA but not the growth factor. It is believed that completion of the studies described herein will provide novel and important information regarding inter-follicular, paracrine regulation of TGF-beta1, which will provide insight into its role during follicle development.

转化生长因子Beta1（TGF-BETA1）由于其对theca和颗粒细胞功能的明显影响以及有关TGF-BETA1和TGF-BETA1和folliculicoening的工作的定位，已被确定为一个潜在的重要载体内生长因子，因此在我们在细胞层面和该功能方面的知识方面仍然存在明显的缺陷。解决有关TGF-BETA1功能和法规的谜语代表了该项目的长期目标。使用养猪卵巢中的培养细胞和完整的半流体，已经制定了调节TGF-BETA1表达的模型。数据表明，尽管两种细胞类型都表达了TGF-BETA1MRNA，但只有theca细胞表达并分泌生长因子。 FSH，LH和CAMP均未刺激培养物中任何一种孤立的细胞类型中的TGF-β截面。然而，在小的半流体中，FSH但没有LH刺激TGF-BETA1分泌。我们假设Theca细胞在卵泡发育过程中表达并分泌TGF-BETA1，但它在FSH作用于颗粒细胞上的指导下。我们进一步假设，FSH刺激的颗粒细胞产物通过激活PLA/PKC信号转导系统来调节THECA细胞TGF-β表达。为了进一步描述TGF-BETA1的散热性调节，它将确定TGF-BETA1自动表达是否发生在颗粒中，而不是在TheCA细胞中发生，无论是通过选择性表达I型I型和II受体的选择性介导的，以及自动表达还是随着卵巢发育的变化而介导的。 FSH和TGF-BETA1相互调节颗粒细胞中其他受体的表达的程度。颗粒细胞可以通过分泌因子直接调节theca细胞TGF-BETA1表达的程度将通过共培养实验测试。是否会确定卵泡TGF-BETA1表达从FSH转向LH治理，并确定卵泡发育。最后，将研究抑制抑制翻译的TGF-BETA1 mRNA 5'-非翻译区的胞质蛋白的选择性颗粒细胞表达。这可能是颗粒细胞差异表达TGF-BETA1 mRNA而不是生长因子的机制。据信，本文所述的研究完成将提供有关TGF-BETA1的嗜热间，旁分泌调节的新颖和重要信息，这将洞悉其在卵泡发育中的作用。