title: Transforming growth factor alpha gene expression and peptide localization in porcine ovarian follicles.
abstract: The present study was undertaken to determine transforming growth factor alpha (TGF alpha) gene expression in various components of medium-sized porcine ovarian follicles by reverse transcription-polymerase chain reaction, and to localize the peptide during folliculogenesis by immunocytochemistry. A strong signal for TGF alpha transcript was detected in cumulus and granulosa cells, whereas the signal in theca cells was very weak but detectable. No TGF alpha mRNA was detected in the oocyte. Immunolocalization studies revealed intense TGF alpha staining in cumulus and granulosa cells of all stages of follicular development. TGF alpha immunostaining was also observed in the oocytes of primordial and primary follicles. In antral follicles, immunostaining was observed only in cumulus and mural granulosa cells, whereas theca cells and oocytes exhibited very little or no detectable staining. TGF alpha immunostaining was absent in follicles undergoing atresia. No TGF alpha immunostaining was observed when the antibody was omitted from the procedure, and the staining was substantially reduced when the primary antibody was preabsorbed with the same mass of TGF alpha peptide, suggesting the specificity of the staining procedure. These results suggest a local follicular production of TGF alpha in the porcine ovary and thus a role for TGF alpha of follicular origin in the regulation of follicular development in an autocrine/paracrine fashion.
authors.affiliation: Department of Obstetrics and Gynaecology, University of Western Ontario, London, Canada.
authors.forename: B
authors.initials: B
authors.lastname: Singh
authors.forename: D T
authors.initials: DT
authors.lastname: Armstrong
mesh_terms: D000818:Animals; D001483:Base Sequence; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006107:Granulosa Cells / Q000737:chemistry; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D009865:Oocytes / Q000737:chemistry; D006080:Ovarian Follicle / Q000737:chemistry / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D012194:RNA-Directed DNA Polymerase; D012756:Sheep; D016211:Transforming Growth Factor alpha / Q000032:analysis / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D016211:Transforming Growth Factor alpha; D012194:RNA-Directed DNA Polymerase; D015252:Deoxyribonucleases, Type II Site-Specific; C040158:GTAC-specific type II deoxyribonucleases