title: Production of monoclonal antibodies against recombinant human zona pellucida glycoproteins: utility in immunolocalization of respective zona proteins in ovarian follicles.
abstract: The zona pellucida (ZP) glycoproteins play an important role in oocyte development and gamete biology. To analyze their expression in follicles during various developmental stages, murine monoclonal antibodies (MAbs) were generated against the baculovirus-expressed recombinant human ZP2, ZP3 and ZP4. A panel of MAbs specific for the respective zona protein in ELISA and Western blot, and devoid of cross-reaction with other zona proteins was selected. Immunohistochemistry has shown that ZP2 MAb, MA-1620, did not react with oocytes in resting primordial follicles but showed reactivity with degenerating oocytes in primordial follicles undergoing atresia, and with oocytes in growing and antral follicles. Three MAbs against ZP3 did not react with oocytes in primordial follicles, but reacted only with oocytes in growing and antral follicles. Out of four MAbs against ZP4, three MAbs reacted with oocytes in primordial, growing and antral follicles. No reactivity of these MAbs with other ovarian cell types and other tissues studied (endometrium, uterine cervix, fallopian tubes and kidney) was detected except for a strong reactivity of ZP2 MA-1620 with epithelial cells of the uterine ectocervix or endometrium in some samples investigated. Altogether, these studies document generation of MAbs exhibiting high specificity for human zona proteins, which will be useful reagents to study their immunobiology.
authors.affiliation: Laboratory of Development, Differentiation and Cancer, The University of Tennessee Graduate School of Medicine, 1924 Alcoa Highway, Knoxville, TN 37920, USA. buko@utk.edu
authors.forename: Antonin
authors.initials: A
authors.lastname: Bukovsky
authors.forename: Satish K
authors.initials: SK
authors.lastname: Gupta
authors.forename: Pankaj
authors.initials: P
authors.lastname: Bansal
authors.forename: Sanchita
authors.initials: S
authors.lastname: Chakravarty
authors.forename: Manjari
authors.initials: M
authors.lastname: Chaudhary
authors.forename: Marta
authors.initials: M
authors.lastname: Svetlikova
authors.forename: Rachel S
authors.initials: RS
authors.lastname: White
authors.forename: Pleas
authors.initials: P
authors.lastname: Copas
authors.forename: Nirmala B
authors.initials: NB
authors.lastname: Upadhyaya
authors.forename: Stuart E
authors.initials: SE
authors.lastname: Van Meter
authors.forename: Michael R
authors.initials: MR
authors.lastname: Caudle
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D004527:Egg Proteins / Q000032:analysis* / Q000276:immunology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology*; D008875:Middle Aged; D006080:Ovarian Follicle / Q000737:chemistry* / Q000166:cytology; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D015044:Zona Pellucida / Q000737:chemistry*; D000072216:Zona Pellucida Glycoproteins
chemical_list: D000911:Antibodies, Monoclonal; D004527:Egg Proteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; C000606515:ZP2 protein, human; C000606517:ZP3 protein, human; D000072216:Zona Pellucida Glycoproteins