Sulfated oviductal glycoproteins in the rabbit: quantitation by competitive enzyme-linked immunosorbent assay.
The rabbit oviductal epithelium synthesizes and secretes a family of antigenically related, sulfated oviductal glycoproteins (SOG). Anti-SOG monoclonal antibodies (Mabs) were produced and two (Mab 1 and Mab 2) were selected for further characterization. Periodate oxidation of Western blots of oviductal fluid did not affect the binding of Mab 1 or Mab 2, thus suggesting that these antibodies recognized protein rather than carbohydrate epitopes on SOG. The specificity of Mab 1 was determined by Western blot analysis of tissues obtained from estrous rabbits and from the male rabbit reproductive tract. SOG was identified in tissue extracts of both the oviductal ampulla and isthmus. Cervix was the only non-oviductal tissue with which Mab 1 cross-reacted. Mab 1 was used to isolated SOG from whole oviductal fluid by immuno-affinity chromatography. Affinity-purified SOG and Mab 1 were used to develop a quantitative, SOG-specific, competitive enzyme-linked immunosorbent assay. This assay was used to quantify SOG in rabbit oviductal fluid collected during estrus and pseudopregnancy. SOG secretion during pseudopregnancy was resolved into two transient episodes of increased secretion. Maximum SOG secretion (X = 1039 +/- 199 micrograms/day) occurred within 48 h of the induction of pseudopregnancy. A second period of enhanced SOG secretion (X = 308 +/- 46 micrograms/day) occurred during the fifth and sixth days of pseudopregnancy. Baseline SOG secretion occurred during estrus at approximately 60% of maximum postovulatory secretion.
Erickson-Lawrence
M F
MF
Department of Anatomy, University of Virginia School of Medicine, Charlottesville 22908.
Turner
T T
TT
Ross
P
P
Thomas
T S
TS
Oliphant
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D015153:Blotting, Western; D002846:Chromatography, Affinity; D051152:Clusterin; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005187:Fallopian Tubes / Q000032:analysis*; D005260:Female; D006023:Glycoproteins / Q000032:analysis*; D006825:Hybridomas / Q000032:analysis; D007132:Immunoglobulin Isotypes / Q000032:analysis; D051379:Mice; D018832:Molecular Chaperones; D009928:Organ Specificity; D010084:Oxidation-Reduction; D010504:Periodic Acid; D011555:Pseudopregnancy / Q000378:metabolism; D011817:Rabbits
D000911:Antibodies, Monoclonal; C495243:Clu protein, mouse; D051152:Clusterin; D006023:Glycoproteins; D007132:Immunoglobulin Isotypes; D018832:Molecular Chaperones; D010504:Periodic Acid; C009288:metaperiodate