Assessment of recombinant porcine follicle-stimulating hormone receptor using a novel polyclonal ectodomain antibody

Follicle‐stimulating hormone (FSH) receptors (FSHR) are critically involved in mediating the responses of granulosa cells and Sertoli cells to FSH. The dynamic changes in cell surface FSH receptors (FSHR) in response to FSH remain unclear in part because of the heavy reliance on ligand‐binding methodologies. This study was designed to determine the molecular and cellular properties of recombinant porcine FSHR using a novel, high‐affinity purified polyclonal antibody to the ectodomain of the pFSHR. A full‐length porcine FSHR cDNA was cloned and sequenced and recombinant pFSHR protein was stably expressed in a clonal cell line of Chinese hamster ovary cells (pFSHR‐CHO). Recombinant receptor was stably expressed in an ovarian cell line with a density similar to that of porcine ovarian cells. A specific polyclonal antibody was generated in chickens to a 100‐amino acid fragment of the pFSHR ectodomain. Immunoblotting, immunoprecipitation, indirect immunofluorescence cytochemistry and immunoelectron microscopy ...