Expression of cystic fibrosis transmembrane conductance regulator in rat ovary / 华中科技大学学报（医学）（英德文版）

The protein expression of cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated Cl(-) channel, in ovarian stimulated premature female rat ovary during a cycle of follicle development and corpus luteum formation was investigated. Animals were injected with 10 U pregnant Mare's serum gonadotropin (PMSG) and subsequently 10 U hCG 48 h later. Time-dependent immunohistochemistry and Western blotting experiments were performed before and 24, 48, 72 h after hCG treatment. The immunohistochemistry revealed that administration of PMSG stimulated the CFTR expression in thecal cell layer and granulosa cell layer of mature follicles 48 h post injection, coincident with the PMSG-induced peak in follicular estradiol. However, the expression of CFTR in the granulose lutein cell layer and thecal lutein cell layer was time-dependently reduced following hCG injection, in accordance with the gradually increased progestogen level during luteum corpus formation. Western blotting analysis demonstrated that rat ovarian tissue expressed the special CFTR band at 170 kD. It is concluded that cAMP-dependent Cl(-) channels are involved in regulation of follicle development and luteum formation.

Rat luteal cells in tissue culture: a technique for obtaining primary cell culture without enzymatic dissociation.

A technique for obtaining isolated luteal cells without any prior enzymatic dissociation of the rat corpus luteum (CL) has been developed. With a view to obviate any kind of chemical/biophysical trauma to the cells the latter were obtained following simple migration of cells from small pieces of chopped up CL (8-10 day old) put in culture. The cells started migrating in progressively increasing numbers from these tissue pieces within 24 hr leading to monolayer formation by day 10-12 of culture. The cells were found to grow under the described conditions for 35 days without any exogenous hormonal supplementation. The technique is being utilized for characterization of different cell types of the rat CL of pregnancy and the regulatory mechanisms involved in their metabolic function and/or regression.