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Expression profiling of primary cultured buffalo granulosa cells from different follicular size in comparison with their in vivo counterpart

Published online by Cambridge University Press:  10 March 2020

Ahmed S.A. Sosa ,
Sally Ibrahim ,
Karima Gh. M. Mahmoud Open the ORCID record for Karima Gh. M. Mahmoud [Opens in a new window] ,
Mohamed M. Ayoub ,
Mohamed S.S. Abdo  and
Mahmoud F. Nawito
Ahmed S.A. Sosa
Affiliation:
Department of Animal Reproduction and AI, Veterinary Research Division, National Research Centre, Dokki, Giza, Egypt
Sally Ibrahim
Affiliation:
Department of Animal Reproduction and AI, Veterinary Research Division, National Research Centre, Dokki, Giza, Egypt
Karima Gh. M. Mahmoud*
Affiliation:
Department of Animal Reproduction and AI, Veterinary Research Division, National Research Centre, Dokki, Giza, Egypt
Mohamed M. Ayoub
Affiliation:
Department of Theriogenology, Cairo University, Cairo, Egypt
Mohamed S.S. Abdo
Affiliation:
Department of Theriogenology, Cairo University, Cairo, Egypt
Mahmoud F. Nawito
Affiliation:
Department of Animal Reproduction and AI, Veterinary Research Division, National Research Centre, Dokki, Giza, Egypt
*
Address for correspondence: Karima Gh. M. Mahmoud. Department of Animal Reproduction and AI, Veterinary Research Division, National Research Centre, Dokki, Giza, Egypt, Tel: +2 01001827716. E-mail: karimamahmoud@yahoo.com
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Summary

This study aimed to: (i) characterize cultured granulosa cells (GCs) from different follicle sizes morphologically and molecularly; and (ii) select a suitable model according to follicular size that maintained GC function during culture. Buffalo ovaries were collected from a slaughterhouse and follicles were classified morphologically into: first group ≤ 4 mm, second group 5–8 mm, third group 9–15 mm and fourth group 16–20 mm diameter. GC pellets were divided into two portions. The first portion served as the control fresh pellet, and the secondwas used for 1 week for GC culture. Total RNA was isolated, and qRT-PCR was performed to test for follicle-stimulating hormone receptor (FSHR), cytochrome P450 19 (CYP19), luteinizing hormone/choriogonadotropin receptor (LHCGR), proliferating cell nuclear antigen (PCNA), apoptosis-related cysteine peptidase (CASP3), anti-Müllerian hormone (AMH), and phospholipase A2 group III (PLA2G3) mRNAs. Estradiol (E2) and progesterone (P4) levels in the culture supernatant and in follicular fluids were measured using enzyme-linked immunosorbent assay (ELISA). Basic DMEM-F12 medium maintained the morphological appearance of cultured GCs. The relative abundance of FSHR, CYP19, and LHCGR mRNAs was 0.001 ≤ P ≤ 0.01 and decreased at the end of culture compared with the fresh pellet. There was a fine balance between expression patterns of the proliferation marker gene (PCNA) and the proapoptotic marker gene (CASP3). AMH mRNA was significantly increased (P < 0.001) in cultured GCs from small follicles, while cultured GCs from other three categories (5–8 mm, 9–15 mm and 16–20 mm) showed a clear reduction (P < 0.001). Interestingly, the relative abundance of PLA2G3 mRNA was significantly (P < 0.001) increased in all cultured GCs. E2 and P4 concentrations were significantly (P < 0.001) decreased in all cultured groups. Primary cultured GCs from small follicles could be a good model for better understanding follicular development in Egyptian buffaloes.

Keywords

BuffaloGene expressionPrimary GCs cultureSteroidogenesis
Type
Research Article
Information
Zygote , Volume 28 , Issue 3 , June 2020 , pp. 233 - 240
Copyright
© Cambridge University Press 2020

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References

Andersen, CL, Jensen, JL and Ørntoft, TF (2004) Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res 64, 5245–50.CrossRefGoogle ScholarPubMed
Araújo, VR, Gastal, MO, Figueiredo, JR and Gastal, EL (2014) In vitro culture of bovine preantral follicles: a review. Reprod Biol Endocrinol 12, 78.CrossRefGoogle ScholarPubMed
Baufeld, A and Vanselow, J (2018) A tissue culture model of estrogen-producing primary bovine granulosa cells. JoVE 139, e58208.Google Scholar
Baumgarten, SC and Stocco, C (2018) Granulosa cells. In Encyclopedia of Reproduction vol. 2 (ed. Skinner, M.K.), pp. 8–13. Academic Press: Elsevier.CrossRefGoogle Scholar
Bertevello, P, Teixeira-Gomes, A-P, Seyer, A, Vitorino Carvalho, A, Labas, V, Blache, M-C, Banliat, C, Cordeiro, L, Duranthon, V and Papillier, P (2018) Lipid identification and transcriptional analysis of controlling enzymes in bovine ovarian follicle. Int J Mol Sci 19, 3261.CrossRefGoogle ScholarPubMed
Bhide, P and Homburg, R (2016) Anti-Müllerian hormone and polycystic ovary syndrome. Best Pract Res CL OB 37, 38–45.CrossRefGoogle ScholarPubMed
Boone, DL and Tsang, BK (1998) Caspase-3 in the rat ovary: localization and possible role in follicular atresia and luteal regression. Biol Reprod 58, 1533–9.CrossRefGoogle ScholarPubMed
Bravo, R and Macdonald-Bravo, H (1987) Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J Cell Biol 105, 1549–54.CrossRefGoogle ScholarPubMed
Campbell, BK, Onions, V, Kendall, N, Guo, L and Scaramuzzi, R (2010) The effect of monosaccharide sugars and pyruvate on the differentiation and metabolism of sheep granulosa cells in vitro. Reproduction, 140, 541–50.CrossRefGoogle ScholarPubMed
Diouf, MN, Sayasith, K, Lefebvre, R, Silversides, DW, Sirois, J and Lussier, JG (2006) Expression of phospholipase A2 group IVA (PLA2G4A) is upregulated by human chorionic gonadotropin in bovine granulosa cells of ovulatory follicles. Biol Reprod 74, 1096–103.CrossRefGoogle ScholarPubMed
Doyle, L, Walker, C and Donadeu, F (2010) VEGF modulates the effects of gonadotropins in granulosa cells. Domest Anim Endocrinol 38, 127–37.CrossRefGoogle ScholarPubMed
El-Salam, MHA and El-Shibiny, S (2011) A comprehensive review on the composition and properties of buffalo milk. Dairy Sci Technol 91, 663.CrossRefGoogle Scholar
FAOSTAT (2015) http://faostat.fao.orgGoogle Scholar
Fitzpatrick, F and Soberman, R (2001) Regulated formation of eicosanoids. J Clin Invest 107, 1347–51.CrossRefGoogle ScholarPubMed
Fuentes, L, Hernández, M, Fernández-Avilés, FJ, Crespo, MSN and Nieto, ML (2002) Cooperation between secretory phospholipase A2 and TNF-receptor superfamily signaling: implications for the inflammatory response in atherogenesis. Circ Res 91, 681–8.CrossRefGoogle ScholarPubMed
Gougeon, A (2010) Human ovarian follicular development: from activation of resting follicles to preovulatory maturation. In Annales d’Endocrinologie vol. 71. Elsevier, pp. 132–43.CrossRefGoogle ScholarPubMed
Gutierrez, C, Glazyrin, A, Robertson, G, Campbell, B, Gong, J, Bramley, T and Webb, R (1997) Ultra-structural characteristics of bovine granulosa cells associated with maintenance of oestradiol production in vitro. Mol Cell Endocrinol 134, 51–8.CrossRefGoogle ScholarPubMed
Hatzirodos, N, Irving-Rodgers, HF, Hummitzsch, K, Harland, ML, Morris, SE and Rodgers, RJ (2014) Transcriptome profiling of granulosa cells of bovine ovarian follicles during growth from small to large antral sizes. BMC Genomics 15, 24.CrossRefGoogle ScholarPubMed
Hatzirodos, N, Hummitzsch, K, Irving-Rodgers, HF and Rodgers, RJ (2015) Transcriptome comparisons identify new cell markers for theca interna and granulosa cells from small and large antral ovarian follicles. PLoS One 10, e0119800.CrossRefGoogle ScholarPubMed
Hatzirodos, N, Glister, C, Hummitzsch, K, Irving-Rodgers, HF, Knight, PG and Rodgers, RJ (2017) Transcriptomal profiling of bovine ovarian granulosa and theca interna cells in primary culture in comparison with their in vivo counterparts. PLoS One 12, e0173391.CrossRefGoogle ScholarPubMed
Hernandez-Medrano, J, Campbell, B and Webb, R (2012) Nutritional influences on folliculogenesis. Reprod Domest Anim 47, 274–82.CrossRefGoogle ScholarPubMed
Hunzicker-Dunn, M and Maizels, ET (2006) FSH signaling pathways in immature granulosa cells that regulate target gene expression: branching out from protein kinase A. Cell Signal 18, 1351–9.CrossRefGoogle ScholarPubMed
Kereilwe, O and Kadokawa, H (2019) Bovine gonadotrophs express anti-Müllerian hormone (AMH): comparison of AMH mRNA and protein expression levels between old Holsteins and young and old Japanese black females. Reprod Fertil Dev 31, 810–19.CrossRefGoogle Scholar
Koskela, S (2013) Granulosa Cell Anti-Müllerian Hormone Secretion in Ovarian Development and Disease, University of Oulu. e-bookGoogle Scholar
Kruip, TA and Dieleman, S (1982) Macroscopic classification of bovine follicles and its validation by micromorphological and steroid biochemical procedures. Reprod Nutr Dev 22, 465–73.CrossRefGoogle ScholarPubMed
Lappas, M and Rice, G (2004) Phospholipase A2 isozymes in pregnancy and parturition. Prostaglandin Leukot Essent Fatty Acids 70, 87–100.CrossRefGoogle ScholarPubMed
Law, NC, Weck, J, Kyriss, B, Nilson, JH and Hunzicker-Dunn, M (2013) Lhcgr expression in granulosa cells: roles for PKA-phosphorylated β-catenin, TCF3, and FOXO1. Mol Endocrinol 27, 1295–310.CrossRefGoogle ScholarPubMed
Matti, N, Irving-Rodgers, HF, Hatzirodos, N, Sullivan, TR and Rodgers, RJ (2010) Differential expression of focimatrix and steroidogenic enzymes before size deviation during waves of follicular development in bovine ovarian follicles. Mol Cell Endocrinol 321, 207–14.CrossRefGoogle ScholarPubMed
Mohammed, BT and Donadeu, FX (2018) Bovine granulosa cell culture. In Epithelial Cell Culture pp. 79–87. Springer.CrossRefGoogle Scholar
Monga, R, Sharma, I, Datta, T and Singh, D (2011) Characterization of serum-free buffalo granulosa cell culture and analysis of genes involved in terminal differentiation from FSH-to LH-responsive phenotype. Domest Anim Endocrinol 41, 195–206.CrossRefGoogle ScholarPubMed
Mora, JM, Fenwick, MA, Castle, L, Baithun, M, Ryder, TA, Mobberley, M, Carzaniga, R, Franks, S and Hardy, K (2012) Characterization and significance of adhesion and junction-related proteins in mouse ovarian follicles. Biol Reprod 86, 153, 1–14.CrossRefGoogle ScholarPubMed
Munakata, Y, Kawahara-Miki, R, Shiratsuki, S, Tasaki, H, Itami, N, Shirasuna, K, Kuwayama, T and Iwata, H (2016) Gene expression patterns in granulosa cells and oocytes at various stages of follicle development as well as in in vitro grown oocyte and granulosa cell complexes. J Reprod Dev 62, 359–66.CrossRefGoogle ScholarPubMed
Oktay, K, Schenken, RS and Nelson, JF (1995) Proliferating cell nuclear antigen marks the initiation of follicular growth in the rat. Biol Reprod 53, 295–301.CrossRefGoogle ScholarPubMed
Palma, GA, Arganaraz, ME, Barrera, AD, Rodler, D, Mutto, AA and Sinowatz, F (2012) Biology and biotechnology of follicle development. Sci World J 2012, 938138.CrossRefGoogle ScholarPubMed
Park, Y, Maizels, ET, Feiger, ZJ, Alam, H, Peters, CA, Woodruff, TK, Unterman, TG, Lee, EJ, Jameson, JL and Hunzicker-Dunn, M (2005) Induction of cyclin D2 in rat granulosa cells requires FSH-dependent relief from FOXO1 repression coupled with positive signals from Smad. J Biol Chem 280, 9135–48.CrossRefGoogle ScholarPubMed
Poole, DH, Ocón-Grove, OM and Johnson, AL (2016) Anti-Müllerian hormone (AMH) receptor type II expression and AMH activity in bovine granulosa cells. Theriogenology 86, 1353–60.CrossRefGoogle ScholarPubMed
Portela, VM, Zamberlam, G and Price, CA (2010) Cell plating density alters the ratio of estrogenic to progestagenic enzyme gene expression in cultured granulosa cells. Fertil Steril 93, 2050–5.CrossRefGoogle ScholarPubMed
Robker, RL and Richards, JS (1998) Hormone-induced proliferation and differentiation of granulosa cells: a coordinated balance of the cell cycle regulators cyclin D2 and p27Kip1. Mol Endocrinol 12, 924–40.CrossRefGoogle ScholarPubMed
Rozen, S and Skaletsky, H (2000) Primer3 on the WWW for general users and for biologist programmers. In Bioinformatics Methods and Protocols pp. 365–86. Springer.Google Scholar
Sadeu, J and Smitz, J (2008) Growth differentiation factor-9 and anti-Müllerian hormone expression in cultured human follicles from frozen–thawed ovarian tissue. Reprod Biomed Online 17, 537–48.CrossRefGoogle ScholarPubMed
Salmon, NA, Handyside, AH and Joyce, IM (2004) Oocyte regulation of anti-Müllerian hormone expression in granulosa cells during ovarian follicle development in mice. Dev Biol 266, 201–8.CrossRefGoogle ScholarPubMed
Salvetti, NR, Panzani, CG, Gimeno, EJ, Neme, LG, Alfaro, NS and Ortega, HH (2009) An imbalance between apoptosis and proliferation contributes to follicular persistence in polycystic ovaries in rats. Reprod Biol Endocrinol 7, 68.CrossRefGoogle ScholarPubMed
Sartori, R, Fricke, PM, Ferreira, JC, Ginther, O and Wiltbank, MC (2001) Follicular deviation and acquisition of ovulatory capacity in bovine follicles. Biol Reprod 65, 1403–9.CrossRefGoogle ScholarPubMed
Seifer, DB and Merhi, Z (2014) Is AMH a regulator of follicular atresia? J Assist Reprod Genet 31, 1403–7.CrossRefGoogle ScholarPubMed
Skinner, MK (2005) Regulation of primordial follicle assembly and development. Hum Reprod Update 11, 461–71.CrossRefGoogle ScholarPubMed
Strober, W (2001) Trypan blue test of cell viability. Curr Protoc Immunol 21, A.3B.1–2.CrossRefGoogle Scholar
Taru Sharma, G, Dubey, P and Sai Kumar, G (2011) Localization and expression of follicle-stimulating hormone receptor gene in buffalo (Bubalus bubalis) pre-antral follicles. Reprod Domest Anim 46, 114–20.CrossRefGoogle Scholar
Teh, A, Izzati, U, Mori, K, Fuke, N, Hirai, T, Kitahara, G and Yamaguchi, R (2018) Histological and immunohistochemical evaluation of granulosa cells during different stages of folliculogenesis in bovine ovaries. Reprod Domest Anim 53, 569–81.CrossRefGoogle ScholarPubMed
Visser, JA, de Jong, FH, Laven, JS and Themmen, AP (2006) Anti-Müllerian hormone: a new marker for ovarian function. Reproduction 131, 1–9.CrossRefGoogle ScholarPubMed
Wandji, S-A, Sršeň, V, Voss, A, Eppig, J and Fortune, J (1996) Initiation of in vitro growth of bovine primordial follicles. Biol Reprod 55, 942–8.CrossRefGoogle ScholarPubMed
Wandji, S, Sršeň, V, Nathanielsz, P, Eppig, J and Fortune, J (1997) Initiation of growth of baboon primordial follicles in vitro. Hum Reprod 12, 1993–2001.CrossRefGoogle ScholarPubMed
Wei, S, Shen, X, Gong, Z, Deng, Y, Lai, L and Liang, H (2017) FSHR and LHR expression and signaling as well as maturation and apoptosis of cumulus–oocyte complexes following treatment with FSH receptor binding inhibitor in sheep. Cell Physiol Biochem 43, 660–9.CrossRefGoogle Scholar
Yadav, M, Agrawal, H, Pandey, M, Singh, D and Onteru, SK (2018) Three-dimensional culture of buffalo granulosa cells in hanging drop mimics the preovulatory follicle stage. J Cell Physiol 233, 1959–70.CrossRefGoogle ScholarPubMed
Zahid, N (2014) Role of anti-Müllerian hormone (AMH) in polycystic ovary syndrome (PCOS). A mini review. Reprod Syst Sex Disord 3, 143.Google Scholar