title: The effects of immunizing sheep with different BMP15 or GDF9 peptide sequences on ovarian follicular activity and ovulation rate.
abstract: The aims of these studies were to determine the abilities of antisera against different regions of ovine bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) to inhibit ovarian follicular activity, estrus (mating), and ovulation in sheep. The 9-15-mer peptides were conjugated to keyhole limpet hemocyanin (KLH) and used to generate antibodies against the flexible N-terminal regions of the mature protein as well as against regions in which dimerization of the protein or interaction with a type 1 BMP or a type 2 TGFB or BMP receptor was predicted to occur. Ewes (n = 10 per treatment group) were vaccinated with KLH or the KLH-BMP15 (n = 9 different peptides) or KLH-GDF9 (n = 10) peptides in Freund adjuvant at five consecutive monthly intervals. Overall, antisera generated against peptides that corresponded to amino acid residues 1-15 of the N-terminus of the BMP15 or GDF9 mature protein or GDF9 amino acid residues 21-34 were the most potent at inhibiting ovulation following primary and single booster vaccination. Several other BMP15 (8/9) or GDF9 (6/10) treatment groups, but not KLH alone, also produced significant reductions in the numbers of animals that ovulated, although 2, 3 or 4 booster vaccinations were required. Anovulation was commonly associated with the inhibition of normal ovarian follicular development and anestrus. The in vitro neutralization studies with IgG from the BMP15 or GDF9 immunized ewes showed that the mean inhibition of BMP15 plus GDF9 stimulation of (3)H-thymidine uptake by rat granulosa cells was approximately 70% for animals without corpora lutea (CL), whereas for animals with one to three CL or more than three CL, the inhibition was 24%-33% or 27%-42%, respectively. In summary, these data suggest that reagents that block the biological actions of BMP15 or GDF9 at their N-termini have potential as contraceptives or sterilizing agents.
authors.affiliation: AgResearch, Wallaceville Animal Research Centre, Upper Hutt 6007, New Zealand. Kenneth.mcnatty@vuw.ac.nz
authors.forename: Kenneth P
authors.initials: KP
authors.lastname: McNatty
authors.forename: Norma L
authors.initials: NL
authors.lastname: Hudson
authors.forename: Lynda
authors.initials: L
authors.lastname: Whiting
authors.forename: Karen L
authors.initials: KL
authors.lastname: Reader
authors.forename: Stan
authors.initials: S
authors.lastname: Lun
authors.forename: Andrea
authors.initials: A
authors.lastname: Western
authors.forename: Derek A
authors.initials: DA
authors.lastname: Heath
authors.forename: Peter
authors.initials: P
authors.lastname: Smith
authors.forename: Lloyd G
authors.initials: LG
authors.lastname: Moore
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Juengel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D055430:Bone Morphogenetic Protein 15; D002478:Cells, Cultured; D003269:Contraception, Immunologic; D003338:Corpus Luteum / Q000033:anatomy & histology / Q000187:drug effects; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D055429:Growth Differentiation Factor 9; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000276:immunology*; D008297:Male; D008969:Molecular Sequence Data; D006080:Ovarian Follicle / Q000033:anatomy & histology / Q000276:immunology / Q000502:physiology*; D010060:Ovulation / Q000276:immunology / Q000502:physiology*; D010446:Peptide Fragments / Q000276:immunology / Q000494:pharmacology; D051381:Rats; D017386:Sequence Homology, Amino Acid; D012756:Sheep; D014611:Vaccination
chemical_list: C525904:Bmp15 protein, rat; D055430:Bone Morphogenetic Protein 15; C525899:Gdf9 protein, rat; D055429:Growth Differentiation Factor 9; D036341:Intercellular Signaling Peptides and Proteins; D010446:Peptide Fragments