Biosynthesis of the Canine Zona Pellucida Requires the Integrated Participation of Both Oocytes and Granulosa Cells

Daniel G. Blackmore, Lucan R. Baillie, Janet E. Holt, Lynda Dierkx, R. John Aitken, Eileen A. McLaughlin

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

In the dog, attempts to localize the expression of zona pellucida (ZP) proteins during folliculogenesis have failed to demonstrate conclusively whether any or all of the zona proteins are synthesized in the oocyte or the granulosa cells. Probing of paraformaldehyde-fixed prepubertal canine ovarian tissue sections with a panel of fluorescently conjugated lectins localized the expression of glycoproteins during folliculogenesis. We confirm that six lectins (PSA, s-WGA, ECL, GSL-II, LEL, and STL) consistently labeled the ZP and adjacent granulosa cells of the developing follicle and that canine ZP expresses β-gal(1,4)glcNAc, β-gal(1,3)galNac, α-mannose, and terminal sialic acid residues in a developmentally specific manner. Riboprobes for canine ZPA and ZPC genes were produced and used for in situ hybridization studies of mRNA expression in canine folliculogenesis. In addition, we isolated a partial cDNA transcript from total ovarian RNA for the canine ZPB gene having a high degree of sequence identity with the felid and porcine ZPB homologues. Subsequently, the ZPA gene transcripts were localized to the cytoplasm of oocytes in primordial, primary, and early secondary follicles. We then localized expression of ZPB and ZPC gene transcripts to the granulosa cells of growing follicles, but not in squamous granulosa cells of primordial follicles or oocytes. These observations indicate that in the juvenile canine ovary, the oocyte is responsible for synthesis of the ZPA protein and directing synthesis of the ZPB and ZPC proteins by the granulosa cells and that ZP gene transcription occurs in a sequential manner during folliculogenesis.

Original languageEnglish
Pages (from-to)661-668
Number of pages8
JournalBiology of Reproduction
Volume71
Issue number2
DOIs
Publication statusPublished - 1 Aug 2004
Externally publishedYes

Fingerprint

Zona Pellucida
Granulosa Cells
Oocytes
Canidae
Genes
Lectins
Proteins
Herpes Zoster
N-Acetylneuraminic Acid
Mannose
In Situ Hybridization
Ovary
Glycoproteins
Cytoplasm
Swine
Complementary DNA
Epithelial Cells
Dogs
RNA
Messenger RNA

Cite this

Blackmore, D. G., Baillie, L. R., Holt, J. E., Dierkx, L., Aitken, R. J., & McLaughlin, E. A. (2004). Biosynthesis of the Canine Zona Pellucida Requires the Integrated Participation of Both Oocytes and Granulosa Cells. Biology of Reproduction, 71(2), 661-668. https://doi.org/10.1095/biolreprod.104.028779
Blackmore, Daniel G. ; Baillie, Lucan R. ; Holt, Janet E. ; Dierkx, Lynda ; Aitken, R. John ; McLaughlin, Eileen A. / Biosynthesis of the Canine Zona Pellucida Requires the Integrated Participation of Both Oocytes and Granulosa Cells. In: Biology of Reproduction. 2004 ; Vol. 71, No. 2. pp. 661-668.
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Biosynthesis of the Canine Zona Pellucida Requires the Integrated Participation of Both Oocytes and Granulosa Cells. / Blackmore, Daniel G.; Baillie, Lucan R.; Holt, Janet E.; Dierkx, Lynda; Aitken, R. John; McLaughlin, Eileen A.

In: Biology of Reproduction, Vol. 71, No. 2, 01.08.2004, p. 661-668.

Research output: Contribution to journalArticle

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T1 - Biosynthesis of the Canine Zona Pellucida Requires the Integrated Participation of Both Oocytes and Granulosa Cells

AU - Blackmore, Daniel G.

AU - Baillie, Lucan R.

AU - Holt, Janet E.

AU - Dierkx, Lynda

AU - Aitken, R. John

AU - McLaughlin, Eileen A.

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AB - In the dog, attempts to localize the expression of zona pellucida (ZP) proteins during folliculogenesis have failed to demonstrate conclusively whether any or all of the zona proteins are synthesized in the oocyte or the granulosa cells. Probing of paraformaldehyde-fixed prepubertal canine ovarian tissue sections with a panel of fluorescently conjugated lectins localized the expression of glycoproteins during folliculogenesis. We confirm that six lectins (PSA, s-WGA, ECL, GSL-II, LEL, and STL) consistently labeled the ZP and adjacent granulosa cells of the developing follicle and that canine ZP expresses β-gal(1,4)glcNAc, β-gal(1,3)galNac, α-mannose, and terminal sialic acid residues in a developmentally specific manner. Riboprobes for canine ZPA and ZPC genes were produced and used for in situ hybridization studies of mRNA expression in canine folliculogenesis. In addition, we isolated a partial cDNA transcript from total ovarian RNA for the canine ZPB gene having a high degree of sequence identity with the felid and porcine ZPB homologues. Subsequently, the ZPA gene transcripts were localized to the cytoplasm of oocytes in primordial, primary, and early secondary follicles. We then localized expression of ZPB and ZPC gene transcripts to the granulosa cells of growing follicles, but not in squamous granulosa cells of primordial follicles or oocytes. These observations indicate that in the juvenile canine ovary, the oocyte is responsible for synthesis of the ZPA protein and directing synthesis of the ZPB and ZPC proteins by the granulosa cells and that ZP gene transcription occurs in a sequential manner during folliculogenesis.

KW - Follicle

KW - Granulosa cells

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KW - Ovary

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