Localization of species conserved zona pellucida antigens in mammalian ovaries.
The mammalian zona pellucida (ZP) consists of three glycoproteins (ZP1, ZP2 and ZP3), which are variably conserved among species at the genomic and amino acid levels. In order to evaluate the expression of ZP during ovarian development, a population of antibodies was selected that recognize species conserved antigenic domains of the three ZP proteins. Domain specific antibodies were selected from sera of rabbits immunized with all three native pig ZP proteins by elution of antibodies bound to each of the three human ZP recombinant proteins expressed from cDNAs, using the baculovirus expression system in insect cells. Immunoblot analysis was used to characterize the specificity of the antibodies and immunohistochemistry was used to evaluate the stage specific expression of ZP proteins during ovarian follicular development of the mouse, baboon and human. This study demonstrates that the conserved domains of all three ZP proteins are localized in the oocyte extracellular ZP matrix as well as in a subset of granulosa cells. However, this expression does vary among species with respect to the stage and cell type during early stages of ovarian follicular development. These antibodies should serve as excellent markers for evaluating early stages of human ovarian follicular development and in the development of contraceptive agents.
Cariño
Cecilia
C
Department of Reproductive Biology, The National Institute of Medical Sciences and Nutrition Salvador Zubirán, Mexico City, Mexico.
Prasad
Sarvamangala
S
Skinner
Sheri
S
Dunbar
Bohn
B
Chirinos
Mayel
M
Schwoebel
Eric
E
Larrea
Fernando
F
Dunbar
Bonnie
B
D000818:Animals; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis* / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004527:Egg Proteins / Q000235:genetics / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000737:chemistry; D005260:Female; D006107:Granulosa Cells / Q000737:chemistry; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology*; D006080:Ovarian Follicle / Q000502:physiology; D010053:Ovary / Q000737:chemistry*; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins / Q000276:immunology; D013045:Species Specificity; D018411:Spodoptera / Q000378:metabolism; D013552:Swine; D014162:Transfection; D015044:Zona Pellucida / Q000737:chemistry / Q000648:ultrastructure; D000072216:Zona Pellucida Glycoproteins
D000941:Antigens; D018076:DNA, Complementary; D004527:Egg Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; C000606514:ZP1 protein, human; C000606515:ZP2 protein, human; C000606517:ZP3 protein, human; D000072216:Zona Pellucida Glycoproteins; C000606519:Zp1 protein, mouse; C000606520:Zp2 protein, mouse; C000606521:Zp3 protein, mouse