Characterization of equine oviductal proteins synthesized and released at estrus and at day 4 after ovulation in bred and nonbred mares.
Proteins synthesized and released in vitro by oviducts collected from horse mares during estrus and at day 4 after ovulation for bred and nonbred mares were examined by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2-D SDS PAGE) and fluorography. Ampullary and isthmic regions both produced a wide array of nondialyzable proteins in culture. Major proteins or groups of proteins identified according to relative molecular weight (kDa) and apparent isoelectric point (pI) were at 100 kDa, pI 8; 100-200 kDa, pI 6; 150 kDa, pI 4.5; 60-100 kDa, pI 4; and an array of polypeptides at 21-22 kDa, pI 5-6. Oviductal secretory activity, measured by incorporation of radiolabeled amino acids into nondialyzable macromolecules released into incubation medium, was greater (P < .01) for the ampullary than the isthmic oviductal region. No consistent differences were observed in fluorograms between estrus vs. day 4 after ovulation, ampulla vs. isthmus, ipsilateral vs. contralateral to the corpus luteum or largest follicle, oviducts from bred vs. nonbred mares, or mare ages. Dialyzed medium from ampullary and isthmic regions of oviducts was subjected to 1-D or 2-D SDS PAGE followed by western blotting utilizing an antiserum directed against human retinol binding protein (RBP). The family of 21-22 kDA polypeptides was identified as immunoreactive RBP.
McDowell
K J
KJ
Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington 40546-0099.
Adams
M H
MH
Williams
N M
NM
D000818:Animals; D015153:Blotting, Western; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D004971:Estrus / Q000378:metabolism*; D005187:Fallopian Tubes / Q000378:metabolism*; D005260:Female; D006736:Horses; D010060:Ovulation / Q000378:metabolism*; D010779:Photofluorography; D011247:Pregnancy; D011506:Proteins / Q000378:metabolism*
D011506:Proteins