Doyle L K, Hogg C O, Watson E D, Donadeu F X
The Roslin Institute and Royal (Dick) School of Veterinary Studies, Roslin BioCentre, University of Edinburgh, Midlothian, UK.
Reproduction. 2008 Nov;136(5):589-98. doi: 10.1530/REP-07-0507. Epub 2008 Aug 7.
The response of follicles to IGF1 was compared between the transition into the ovulatory season (transitional period) and the ovulatory season (ovulatory period) in eight mares using a cross-over experimental design within periods. Granulosa cells were collected from follicles 15-24 or 25-34 mm and expression of IGF1R, IGF2R, FSHR, LHCGR and PAPPA was determined by qPCR. In addition, 10 mg IGF1 or vehicle were injected into the largest follicle (transitional period) or the second largest follicle (ovulatory period) of a follicular wave before the beginning of diameter deviation between the two largest follicles (mean diameters at injection 19.2 and 20.0 mm during transitional and ovulatory periods respectively). Follicular fluid was collected 24 h after injection for determination of free IGF1, IGFBP, inhibin A and oestradiol levels. Granulosa cells from follicles 25-34 mm, but not follicles 15-24 mm, expressed higher levels of IGF1R (P=0.01), FSHR (P<0.007) and LHCGR (P=0.09) during the ovulatory period than during the transitional period, whereas IGF2R expression was higher in transitional than ovulatory follicles (P=0.06). Follicular IGFBP2 levels were not different (P>0.1) between periods and treatments, whereas IGFBP5 levels were higher (P<0.05) during the ovulatory period. Finally, IGF1 injection before the beginning of deviation induced an approximately twofold increase (P=0.01) in follicular inhibin A levels during each period and did not affect oestradiol (P>0.1). These results suggest that, as during ovulatory waves, equine follicles during transitional waves are responsive to IGF1 before the beginning of deviation and that, therefore, inadequate IGF1 responsiveness before deviation may not underlie the deficient development of dominant follicles during transition.
采用交叉实验设计,在八个母马的不同时期,比较了进入排卵季节(过渡期)和排卵季节(排卵期)时卵泡对胰岛素样生长因子1(IGF1)的反应。从直径15 - 24毫米或25 - 34毫米的卵泡中收集颗粒细胞,通过定量PCR测定IGF1R、IGF2R、促卵泡激素受体(FSHR)、促黄体生成素/绒毛膜促性腺激素受体(LHCGR)和妊娠相关血浆蛋白A(PAPPA)的表达。此外,在两个最大卵泡直径出现偏差之前(过渡期注射时平均直径为19.2毫米,排卵期注射时平均直径为20.0毫米),向卵泡波的最大卵泡(过渡期)或第二大卵泡(排卵期)注射10毫克IGF1或赋形剂。注射后24小时收集卵泡液,测定游离IGF1、胰岛素样生长因子结合蛋白(IGFBP)、抑制素A和雌二醇水平。在排卵期,直径25 - 34毫米的卵泡颗粒细胞,而非直径15 - 24毫米的卵泡颗粒细胞,IGF1R(P = 0.01)、FSHR(P < 0.007)和LHCGR(P = 0.09)的表达水平高于过渡期,而IGF2R的表达在过渡期卵泡中高于排卵期卵泡(P = 0.06)。各时期和处理之间卵泡IGFBP2水平无差异(P > 0.1),而IGFBP5水平在排卵期较高(P < 0.05)。最后,在偏差开始前注射IGF1,在每个时期均可使卵泡抑制素A水平升高约两倍(P = 0.01),且不影响雌二醇水平(P > 0.1)。这些结果表明,与排卵波期间一样,过渡期卵泡波中的马卵泡在偏差开始前对IGF1有反应,因此,偏差前IGF1反应不足可能不是过渡期优势卵泡发育缺陷的原因。