Zhang C, Shimada K, Saito N, Kansaku N
Laboratory of Animal Physiology, School of Agricultural Sciences, Nagoya University, Japan.
Gen Comp Endocrinol. 1997 Mar;105(3):402-9. doi: 10.1006/gcen.1996.6843.
Expression of luteinizing hormone receptor (LHR) and follicle-stimulating hormone receptor (FSHR) mRNAs was demonstrated in the granulosa and theca layers of the large preovulatory follicles of the chicken ovary by Northern hybridization and reverse transcription-polymerase chain reaction (RT-PCR). Northern hybridization results showed multiple LHR and FSHR mRNA transcripts and the predominant species were 2.3 and 2.5 kb, respectively. The highest abundance of LHR mRNA was found in the granulosa layer of the largest follicle (F1 follicle), while the abundance remained low in the granulosa layers of the third (F3) and fifth largest (F5) follicles. FSH mRNA abundance was the highest in the granulosa layer of F5 follicle, but decreased in the granulosa layers of F3 and F1 follicles. In the theca layers of all the three follicles examined LHR and FSHR mRNAs were extremely low. These results were confirmed by RT-PCR experiment which involved coamplification of LHR or FSHR mRNA and beta-actin mRNA as the internal control. The LHR PCR product was sequenced and indicated 92.2% homology with the corresponding region of the quail LHR cDNA. This study indicated that the marked increase in expression of LHR mRNA in granulosa layer of the F1 follicle might be important for LHR protein synthesis and succeeding bonus progesterone production in F1 follicle destined to ovulation. However, higher expression of FSHR mRNA in the granulosa layer of the less mature follicles may be involved in the differentiation and maturation of granulosa cells in these follicles.
通过Northern杂交和逆转录-聚合酶链反应（RT-PCR），在鸡卵巢排卵前大卵泡的颗粒层和卵泡膜层中证实了促黄体生成素受体（LHR）和促卵泡激素受体（FSHR）mRNA的表达。Northern杂交结果显示有多个LHR和FSHR mRNA转录本，主要的转录本分别为2.3 kb和2.5 kb。在最大卵泡（F1卵泡）的颗粒层中发现LHR mRNA丰度最高，而在第三大（F3）和第五大（F5）卵泡的颗粒层中丰度仍然较低。FSHR mRNA丰度在F5卵泡的颗粒层中最高，但在F3和F1卵泡的颗粒层中降低。在所检测的所有三个卵泡的卵泡膜层中，LHR和FSHR mRNA极低。RT-PCR实验证实了这些结果，该实验涉及将LHR或FSHR mRNA与作为内对照的β-肌动蛋白mRNA共同扩增。对LHR PCR产物进行测序，结果表明其与鹌鹑LHR cDNA的相应区域有92.2%的同源性。本研究表明，F1卵泡颗粒层中LHR mRNA表达的显著增加可能对LHR蛋白合成以及随后注定要排卵的F1卵泡中孕酮的产生很重要。然而，在较不成熟卵泡的颗粒层中FSHR mRNA的较高表达可能参与了这些卵泡中颗粒细胞的分化和成熟。
