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Molecular characterization of tumor necrosis alpha-induced protein 6 and its human chorionic gonadotropin-dependent induction in theca and mural granulosa cells of equine preovulatory follicles.
Abstract: The preovulatory rise in gonadotropins causes an expansion of the cumulus-oocyte complex, a process requiring the induction of several genes. The objectives of this study were to clone the equine tumor necrosis factor alpha-induced protein 6 (TNFAIP6), and investigate its regulation in equine follicles during human chorionic gonadotropin (hCG)-induced ovulation. The isolation of the equine TNFAIP6 cDNA revealed that it contains an open reading frame of 834 bp (including the stop codon), encoding a predicted 277 amino acid protein that is highly similar (91-93% identity) to known mammalian homologs. The regulation of TNFAIP6 mRNA was studied in equine follicles isolated during estrus between 0 and 39 h post-hCG and in corpora lutea (CL) obtained on day 8 of the estrous cycle. Results from semi-quantitative RT-PCR/Southern blot showed that levels of TNFAIP6 mRNA were low in follicles obtained at 0 h, increased at 12 h, returned to basal levels at 24 h, and increased again at 36 h post-hCG (P<0.05). Levels of TNFAIP6 transcripts were relatively moderate in CL, but low in non-ovarian tissues tested. Analyses performed with isolated preparations of theca and granulosa cells indicated that TNFAIP6 mRNA was regulated in both layers, with a maximal induction obtained 33-36 h post-hCG (P<0.05). Immunohistochemical staining of sections of equine follicles isolated at 0 and 33 h post-hCG confirmed the induction of TNFAIP6 protein in both cell types after hCG treatment. Thus, the present study describes for the first time the gonadotropin-dependent regulation of follicular TNFAIP6 during the ovulation in a monoovulatory species. The biphasic induction of TNFAIP6 in equine theca and granulosa cells differs from the pattern observed in rodents, suggesting a distinct control of gene expression in this monoovulatory species.
Publication Date: 2007-01-25 PubMed ID: 17244740DOI: 10.1530/rep.1.01200Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research presents a comprehensive study of the equine tumor necrosis factor alpha-induced protein 6 (TNFAIP6) in equine preovulatory follicles, along with its regulatory effects during human chorionic gonadotropin (hCG)-induced ovulation.
Molecular Characterization of TNFAIP6
- The research aimed to clone the horses’ TNFAIP6 and explore its regulation during hCG-induced ovulation. Isolation of the equine TNFAIP6 cDNA revealed it comprises an 834 base pairs open reading frame, including the stop codon. This encodes a predicted 277 amino acid protein highly similar to known mammalian homologs, with an identity match of 91-93%.
Regulation of TNFAIP6 mRNA in Equine Follicles
- The study also explored the regulation of TNFAIP6 mRNA in equine follicles during the estrus, specifically in the period between 0 to 39 hours post-hCG. Examination of corpora lutea (CL) obtained on the estrous cycle’s eighth day was also performed.
- Analysis revealed that the levels of TNFAIP6 mRNA were low in follicles at 0 hours, then increased at 12 hours, returned to basal levels at 24 hours, and again increased at 36 hours post-hCG.
- In contrast, the levels of TNFAIP6 transcripts in CL were moderately high but low in non-ovarian tissues tested. Similar trends were exhibited in both the theca and granulosa cells.
Induction in Theca and Granulosa Cells
- The study found that the hCG treatment resulted in the induction of TNFAIP6 protein in both the theca and granulosa cells, which was confirmed by immunohistochemical staining.
- It was noted that the biphasic induction of TNFAIP6 differs from the patterns observed in rodents, suggesting distinct gene expression control in horses, a monoovulatory species.
Conclusion
- Overall, this is the first study to describe the gonadotropin-dependent regulation of follicular TNFAIP6 during ovulation in a monoovulatory species. The findings contribute to the understanding of this protein’s role in ovulation, with potential implications for fertility and reproductive biology studies.
Cite This Article
APA
Sayasith K, Doré M, Sirois J.
(2007).
Molecular characterization of tumor necrosis alpha-induced protein 6 and its human chorionic gonadotropin-dependent induction in theca and mural granulosa cells of equine preovulatory follicles.
Reproduction, 133(1), 135-145.
https://doi.org/10.1530/rep.1.01200 Publication
Researcher Affiliations
- Centre de Recherche en Reproduction Animale, Université de Montréal, Canada. k.sayasith@umontreal.ca
MeSH Terms
- Amino Acid Sequence
- Animals
- Base Sequence
- Blotting, Southern
- Cattle
- Chorionic Gonadotropin / pharmacology
- Corpus Luteum / chemistry
- Female
- Follicular Phase
- Granulosa Cells / chemistry
- Horses / metabolism
- Humans
- Hyaluronan Receptors / analysis
- Hyaluronan Receptors / genetics
- Immunohistochemistry
- Mice
- Molecular Sequence Data
- Ovulation Induction
- RNA, Messenger
- Reverse Transcriptase Polymerase Chain Reaction
- Sequence Homology
- Theca Cells / chemistry
Citations
This article has been cited 3 times.- Benoit G, Warma A, Lussier JG, Ndiaye K. Gonadotropin regulation of ankyrin-repeat and SOCS-box protein 9 (ASB9) in ovarian follicles and identification of binding partners. PLoS One 2019;14(2):e0212571.
- Schuermann Y, Rovani MT, Gasperin B, Ferreira R, Ferst J, Madogwe E, Gonçalves PB, Bordignon V, Duggavathi R. ERK1/2-dependent gene expression in the bovine ovulating follicle. Sci Rep 2018 Nov 1;8(1):16170.
- Nagyova E. The Biological Role of Hyaluronan-Rich Oocyte-Cumulus Extracellular Matrix in Female Reproduction. Int J Mol Sci 2018 Jan 18;19(1).
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