Epidermal growth factor and its receptor gene expression and peptide localization in porcine ovarian follicles.
The present study was undertaken to determine the expression of genes for epidermal growth factor (EGF) and its receptor (EGF-R) in various components of medium-sized porcine ovarian follicles by reverse transcription-polymerase chain reaction (RT-PCR), and to localize their peptides during folliculogenesis by immunocytochemistry. A strong band for EGF mRNA transcript was detected in the oocyte, whereas the signal in cumulus, granulosa, and theca cells was very weak but detectable. In contrast, a very strong EGF-R mRNA signal was observed in cumulus, granulosa, and theca cells, whereas the signal in the oocyte was very weak. EGF peptide was localized in the oocyte, cumulus, and granulosa cells of all stages of follicle. In the oocyte, the intensity of immunostaining was more pronounced in primordial and primary follicles, compared to atrial follicles. In large antral follicles, immunostaining was pronounced in granulosa cells, whereas theca cells showed little or no detectable staining for EGF. EGF staining was also observed in the cumulus and granulosa cells of follicles undergoing atresia. EGF-R immunostaining was observed in the oocytes of primordial and primary follicles, and in cumulus, granulosa, and theca cells of all stages of follicle, including atretic follicles. In large antral follicles, the intensity of immunostaining was more pronounced in theca cells than in granulosa cells, and the oocyte showed little or no detectable staining. No immunostaining was observed when the primary antibody was replaced with preimmune serum (EGF), or preabsorbed with the control peptide (EGF-R), confirming the specificity of the staining procedures.(ABSTRACT TRUNCATED AT 250 WORDS)
Singh
B
B
Department of Obstetrics and Gynaecology, University of Western Ontario, London, Canada.
Rutledge
J M
JM
Armstrong
D T
DT
D000818:Animals; D001483:Base Sequence; D004815:Epidermal Growth Factor / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D066246:ErbB Receptors / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D013552:Swine
D012333:RNA, Messenger; D004815:Epidermal Growth Factor; D066246:ErbB Receptors