Expression of caspase-3, caspase-9, GDF-9 and IGF-1 genes in ovine oocytes cultured with optimum and elevated doses of Amphiregulin, Neuregulin-1 and Tumour necrosis factor-α in-vitro

RAMESH HONDARAVALLI SOMASHETTY; NANDI SUMANTA; GIRISH KUMAR VENKATESH

doi:10.56093/ijans.v92i9.115614

Authors

RAMESH HONDARAVALLI SOMASHETTY Karnataka Veterinary, Animal & Fisheries Sciences University, Bidar, Veterinary College, Hebbal, Bengaluru
NANDI SUMANTA NIANP-ICAR, Adugodi, Bengaluru
GIRISH KUMAR VENKATESH Karnataka Veterinary, Animal & Fisheries Sciences University, Bidar, Veterinary College, Hebbal, Bengaluru

https://doi.org/10.56093/ijans.v92i9.115614

Keywords:

Caspase-3, Caspase-9, GDF-9, IGF-1, Oocytes

Abstract

The present study was conducted to evaluate the expression of caspase-3, caspase-9, growth differentiation factor-9 (GDF-9) and insulin growth factor-1 (IGF-1) genes in oocytes cultured in vitro with optimum and elevated doses of amphiregulin (50 ng and 150 ng), neuregulin-1 (25 ng and 150 ng) and tumor necrosis factor-α (25 ng and 150 ng) during in vitro maturation of oocytes based on the results of effects of AREG or NRG-1 or TNF-α concentration on maturation of oocytes which caused significant effect, were selected for gene expression studies along with the basal/control level. Total RNA was extracted from in vitro matured oocytes using Trizol method and Real-time reverse transcription polymerase chain reaction was used to evaluate the expression of genes. There was an upregulation of caspase-3 at 150 ng of TNF-α, caspase-9 at 50 ng of AREG, GDF-9 at 150 ng of AREG and IGF-1 at 150 ng of AREG and 25 ng of TNF-α. These results suggested that AREG at elevated dose and TNF-α at optimum dose enhanced the expression of GDF-9 and IGF-1, while the presence of elevated dose of TNF-α and optimum dose of AREG activated caspase-3 and caspase-9, respectively in oocytes cultured in vitro.

Downloads

Download data is not yet available.

Author Biographies

RAMESH HONDARAVALLI SOMASHETTY, Karnataka Veterinary, Animal & Fisheries Sciences University, Bidar, Veterinary College, Hebbal, Bengaluru

Department of Veterinary Biochemistry, Veterinary college, andÂ Assistant Professor.
NANDI SUMANTA, NIANP-ICAR, Adugodi, Bengaluru

NIANP-ICAR, Principal Scientist.
GIRISH KUMAR VENKATESH, Karnataka Veterinary, Animal & Fisheries Sciences University, Bidar, Veterinary College, Hebbal, Bengaluru

Department of Veterinary Biochemistry and Professor & Head

References

Bari M A, Kabir M E, Sarker M B, Ahnakhan and Moniruzzaman M. 2011. Morphometric analysis of ovarian follicles of Black Bengal goats during winter and summer season. Bangladesh Journal of Animal Sciences 40(1&2): 51–55. DOI: https://doi.org/10.3329/bjas.v40i1-2.10791

Dragovic R A, Ritter L J, Schulz S J, Amato F, Thompson J G, Armstrong D T and Gilchrist R B. 2007. Oocytes-secreted factor activation of smad 2/3 signaling enables initiation of mouse cumulus cell expansion. Biology of Reproduction 76: 848–57. DOI: https://doi.org/10.1095/biolreprod.106.057471

Fenwick M A and Hurst P R. 2002. Immunohistochemical localization of active caspase-3 in the mouse ovary: Growth and atresia of small follicles. Reproduction 124: 659–65. DOI: https://doi.org/10.1530/rep.0.1240659

Field S L, Dasgupta T, Cummings M and Orsi N M. 2014. Cytokines in ovarian folliculogenesis, oocytes maturation and luteinisation. Molecular Reproduction and Development 81: 284–314. DOI: https://doi.org/10.1002/mrd.22285

Frank L A, Sutton-mcdowall M L, Brown H M, Russell D L, Gilchrist R B and Thompson J G. 2014. Hyperglycaemic conditions perturb mouse oocytes in vitro developmental competence via beta-O-linked glycosylation of heat shock protein 90. Human Reproduction 29: 1292–1303. DOI: https://doi.org/10.1093/humrep/deu066

Gilchrist R B, Lane M. and Thompson J G. 2008. Oocytes secreted factors: regulators of cumulus cell function and oocytes quality. Human Reproduction Update 14(2): 159–77. Gode F, Gulekli B, Dogan E, Korhan P, Dogan S, Bige O, Cimrin D and Atabey N. 2011. Influence of follicular fluid GDF9 and BMP15 on embryo quality. Fertility and Sterility 95: 2274–78. DOI: https://doi.org/10.1016/j.fertnstert.2011.03.045

Huang Z and Wells D. 2010. The human oocytes and cumulus cells relationship: New insights from the cumulus cell transcriptome. Basic Science of Reproductive Medicine 16(10): 715–25. DOI: https://doi.org/10.1093/molehr/gaq031

Jamnongjit M and Hammes S R. 2006. Ovarian steroids: The good, the bad and the signals that raise them. Cell Cycle 5: 1178–83. DOI: https://doi.org/10.4161/cc.5.11.2803

Li Y I, Li R Q, Ou, S B, Zhang N F, Ren L, Wei LN, Zhang Q X and Yang D Z. 2014. Increased GDF9 and BMP15 mRNA levels in cumulus granulosa cells correlate with oocytes maturation, fertilization, and embryo quality in humans. Reproductive Biology and Endocrinology 12: 81. DOI: https://doi.org/10.1186/1477-7827-12-81

Lin Z L, Li, Y H, Xu Y N, Wang Q L, Namgoong S, Cui X S and Kim N H. 2014. Effects of growth differentiation factor 9 and bone morphogenetic protein 15 on the in vitro maturation of porcine oocytes. Reproduction in Domestic Animals 49: 219–27. DOI: https://doi.org/10.1111/rda.12254

Livak K J and Schmittgen T D. 2001. Analysis of relative gene expression data using real time quantitative PCR and the 2ΔΔCT method. Methods 25(4): 402–08. DOI: https://doi.org/10.1006/meth.2001.1262

Mao J, Smith M F, Rucker E B, Wu G M, Mccauley T C, Cantley T C, Prather R S, Didion B A and Day B N. 2004. Effect of epidermal growth factor and insulin-like growth factor I on porcine preantral follicular growth, antrum formation, and stimulation of granulosal cell proliferation and suppression of apoptosis in vitro. Journal of Animal Science 82(7): 1967–75. DOI: https://doi.org/10.2527/2004.8271967x

Milan blaha, Lucie N, Katerina V K, Petr V and Prochazka R. 2015. Gene expression analysis of pig cumulus oocytes complexes stimulated in vitro with follicle stimulating hormone or epidermal growth factor like peptides. Reproductive Biology 13: 113. DOI: https://doi.org/10.1186/s12958-015-0112-2

Nandi S, Ravidranath B M, Gupta P S P and Sarma P V. 2002. Timing of sequential changes in cumulus cells and first polar body extrusion during in vitro maturation of buffalo oocytes. Theriogenology 57: 1151–59. DOI: https://doi.org/10.1016/S0093-691X(01)00709-9

Nandi S, Girishkumar V, Gupta P S P, Ramesh H S and Manjunatha B M. 2006. Effect of ovine follicular fluid peptide on follicle, oocytes, and somatic cell culture in buffalo (Bubalus bubalis). Animal Reproduction 3: 61–69. DOI: https://doi.org/10.1111/j.1440-169X.2007.00901.x

Noma N, Kawashima I, Fan H Y, Fujita Y, Kawai T, Tomoda Y, Mihara T, Richards J S and Shimada M. 2011. LH-Induced Neuregulin 1 (NRG-1) type III transcripts control granulosa cell differentiation and oocytes maturation. Molecular Endocrinology 25(1): 104–16. DOI: https://doi.org/10.1210/me.2010-0225

Orisaka M, Tajima K, Tsang B K and Kotsuji F. 2009. Oocytes granulosa-theca cell interactions during preantral follicular development. Journal of Ovarian Research 2: 1–9. DOI: https://doi.org/10.1186/1757-2215-2-9

Park J Y, Su Y Q, Ariga M, Law E, Jin S L and Conti M. 2004. EGF-like growth factors as mediators of LH action in the ovulatory follicle. Science 303: 682–84. DOI: https://doi.org/10.1126/science.1092463

Prange-kiel J, Kreutzkamm C, Wehrenbergu and Runegm. 2001. Role of tumor necrosis factor in preovulatory follicles of swine. Biology of Reproduction 65: 928–35. DOI: https://doi.org/10.1095/biolreprod65.3.928

Ramesh H S, Sumanta N and Venkatesh G K. 2021. Effect of different concentrations of Amphiregulin/Neuregulin/Tumour necrosis factor-α on ovine oocytes maturation in vitro. The Indian Journal of Veterinary Sciences and Biotechnology 17(4): 00–5.

Richani D, Ritter L J, Thompson J G and Gilchrist R B. 2013. Mode of oocytes maturation affects EGF-like peptide function and oocytes competence. Molecular Human Reproduction 19: 500–09. DOI: https://doi.org/10.1093/molehr/gat028

Robles R Y, Morita K K, Mann G I, Perez S, Yang T, Matikainen D H, Sherr J L and Tilly. 2000. The aryl hydrocarbon receptor, a basic helix-loop-helix transcription factor of the PAS gene family is required for normal ovarian germ cell dynamics in the mouse. Endocrinology 141: 450–53. DOI: https://doi.org/10.1210/endo.141.1.7374

Sudiman J, Ritter L J, Feil D K, Wang X, Chan K, Mottershead D G, Robertson D M, Thompson J G and Gilchrist R B. 2014. Effects of differing oocytes-secreted factors during mouse in vitro maturation on subsequent embryo and fetal development. Journal of Assisted Reproduction and Genetics 31: 295–306. DOI: https://doi.org/10.1007/s10815-013-0152-5

Sugimura S, Ritter L J, Sutton-mcdowall M L, Mottershead D G, Thompson J G and Gilchrist. R B. 2014. Amphiregulin co-operates with bone morphogenetic protein 15 to increase bovine oocytes developmental competence: Effects on gap junction mediated metabolite supply. Molecular Human Reproduction 20: 499–513. DOI: https://doi.org/10.1093/molehr/gau013

Sugimura S, Ritter L J, Rose R, Thompson J G, Smitz J, Mottershead D G and Gilchrist R B. 2015. Promotion of EGF receptor signaling improves the quality of low developmental competence oocytes. Developmental Biology 403(2): 139–49. DOI: https://doi.org/10.1016/j.ydbio.2015.05.008

Thornton B and Basu C. 2011. Real-Time PCR (q-PCR) primer design using free online software. Biochemistry and Molecular Biology Education 39: 145–54. DOI: https://doi.org/10.1002/bmb.20461

Yamamoto Y, Kuwahar A, Taniguchi Y, Yamasaki M, Tanaka Y, Mukai Y, Yamashita M and Matsuzaki T. 2015. Tumor necrosis factor alpha inhibits ovulation and induces granulosa cell death in rat ovaries. Reproductive Medicine and Biology 14: 107–15. DOI: https://doi.org/10.1007/s12522-014-0201-5

Yan C, Wang P, Demayo J, Elvin J A, Carino C, Prasad S V, Skinner S S, Dunbar B S, Dube J L, Celeste A J and Matzuk M M. 2001. Synergistic roles of bone morphogenetic protein 15 and growth differentiation factor 9 in ovarian function. Molecular Endocrinology 15: 854–66. DOI: https://doi.org/10.1210/mend.15.6.0662