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IGF1 stimulates differentiation of primary follicles and their growth in ovarian explants of zebrafish (Danio rerio) cultured in vitro

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Abstract

The present study is an attempt to elucidate the involvement of insulin-like growth factor (IGF1) in the differentiation and growth of primary follicles in ovarian explant cultures of zebrafish. Ovaries from adult females were cultured in triplicate sets/treatment group for 15 days at 22°C in the laboratory. Culture medium was supplemented with either insulin (1 ng/mL) or IGF1 (1 ng/mL) or insulin + IGF1 (Experiment 1) or 0.1 or 1.0 or 10 ng/mL of IGF1 (Experiment 2). Ovaries cultured in medium alone served as controls and those fixed at the beginning of the culture as initial controls. Experiments were repeated. On the 16th day ovarian explants were fixed in Bouin’s fluid and processed for paraffin embedding, sections (3 µm) were cut and stained with hematoxylin-eosin. Follicles were classified into 6 stages and atretic follicles (AF). Previtellogenic, vitellogenic and total follicle number was calculated. At the start of the culture, ovaries contained all stages of growing and degenerating follicles. In in vitro cultured control ovaries, vitellogenic follicles underwent atresia, while, primary follicles remained unaffected. Insulin or insulin + IGF1 treated ovaries did not differ significantly while IGF1 exposed ovarian explants had greater (P < 0.05) number of primary follicles compared to controls. IGF1 also caused an increase in the number and growth of primary follicles in a dose dependent manner although; cultures were not supplemented with gonadotrophic hormones. Results suggest that locally derived intra-ovarian IGF1 may have a role in the differentiation and growth of primary follicles in zebrafish ovary.

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References

  • Berishvili G, D’Cotta H, Baroiller JF, Segner H and Reinecke M 2006 Differential expression of IGF-I mRNA and peptide in the male and female gonad during early development of a bony fish, the tilapia Oreochromis niloticus. Gen. Comp. Endocrinol. 46 204–210

    Article  Google Scholar 

  • Campbell RC 1989 Statistics for Biologists, III Edition Textbook. Cambridge University Press, Cambridge

  • Campbell B, Dickey J, Beckman B, Young G, Pierce A, Fukada H and Swanson P 2006 Previtellogenic oocyte growth in salmon: relationships among body growth, plasma insulin-like growth factor-1, estradiol-17beta, follicle-stimulating hormone and expression of ovarian genes for insulin-like growth factors, steroidogenic acute regulatory protein and receptors for gonadotropins, growth hormone, and somatolactin. Biol. Reprod. 75 34–44

    Article  CAS  PubMed  Google Scholar 

  • Clelland E and Peng C 2009 Endocrine/paracrine control of zebrafish ovarian development. Mol. Cell Endocrinol. 312 42–52

    Article  CAS  PubMed  Google Scholar 

  • Das D, Khan PP and Maitra S 2013 Participation of PI3-kinase/Akt signaling in insulin stimulation of p34cdc2 activation in zebrafish oocyte: Phosphodiesterase 3 as a potential downstream target. Mol. Cell Endocrinol. 374 46–55

    Article  CAS  PubMed  Google Scholar 

  • Das D, Pal S and Maitra S 2016a Releasing prophase arrest in zebrafish oocyte: synergism between maturational steroid and Igf1. Reproduction 151 59–72

    Article  CAS  PubMed  Google Scholar 

  • Das D, Nath P, Pal S, Hajra S, Ghosh P and Maitra S 2016b Expression of two insulin receptor subtypes, insra and insrb, in zebrafish (Danio rerio) ovary and involvement of insulin action in ovarian function. Gen. Comp. Endocrinol. 239 21–31

    Article  CAS  PubMed  Google Scholar 

  • Forsgren KL and Young G 2012 Stage-Specific Effects of Androgens and Estradiol-17beta on the Development of Late Primary and Early Secondary Ovarian Follicles of Coho Salmon (Oncorhynchus kisutch) In-vitro. Biol. Reprod. 87 64 1–14

    Google Scholar 

  • Garcia-Lopez A, Sanchez-Amaya MI, Tyler CR and Prat E 2011 Mechanisms of oocyte development in European sea bass (Dicentrarchus labrax, 2L.): Investigations via application of unilateral ovariectomy. Reproduction 142 243–253

    Article  CAS  PubMed  Google Scholar 

  • Geva E, Jaff RB 2000 Role of vascular endothelial growth factor in ovarian physiology and pathology. Fertil. Steril. 74 429–38

    Article  CAS  PubMed  Google Scholar 

  • Good NE, Winget GD, Winter W, Connolly TN, Izawa E and Singh RMM 1966 Hydrogen ion buffers for biological research. Biochemistry 5 2

    Article  Google Scholar 

  • Irwin DA 2011 The regulation and function of the ovarian-derived insulin-like growth factor system in zebrafish (Danio rerio), Thesis. University of Guelph, Canada

  • Irwin DA and Kraak GVD 2012 Regulation and actions of insulin-like growth factors in the ovary of zebrafish (Danio rerio). Gen. Comp. Endocrinol. 177 187–194

    Article  CAS  PubMed  Google Scholar 

  • Johnston TK 2013 In vitro model of the fathead minnow hypothalamic-pituitary gonadal axis for toxicological evaluation, Thesis, University of Illinois at Urbana-Champaign

  • Juengel JL, Haydon LJ, Mester B, Thomson JB, Beaumont M and Eckery DC 2010 The role of IGFs in the regulation of ovarian follicular growth in the brushtail possum (Trichosurus vulpecula). Reproduction 140 295–303

    Article  CAS  PubMed  Google Scholar 

  • Kagawa H, Moriyama S and Kawauchi H 1995 Immunocytochemical localization of IGF-I in the ovary of the red seabream, Pagrus major. Gen. Comp. Endocrinol. 99 307–315

    Article  CAS  PubMed  Google Scholar 

  • Kortner TM, Rocha E, Silva P, Castro LFC and Arukwe A 2008 Genomic approach in evaluating the role of androgens on the growth of Atlantic cod (Gadus morhua) previtellogenic oocytes. Comp. Biochem. Physiol. D 3 205–218

    Google Scholar 

  • Kortner TM, Rocha E and Arukwe A 2009 Androgenic modulation of early growth of Atlantic Cod (Gadus morhua L.) previtellogenic oocytes and zona radiata-related. J. Toxicol Environ. Health Sci. A 72 184–195

    Article  CAS  Google Scholar 

  • Leibovitz A 1963 The Growth and Maintenance of tissue-cell cultures in free gas exchange with the atmosphere. Am. J. Hygeine 78 173–180

    CAS  Google Scholar 

  • Li CW and Ge W 2011 Spatiotemporal Expression of bone morphogenetic protein family ligands and receptors in the zebrafish ovary: A potential paracrine-signaling mechanism for oocyte-follicle cell communication. Biol. Reprod. 85 97–98

    Google Scholar 

  • Li CW and Ge W. 2013 Regulation of the activin-inhibin-follistatin system by bone morphogenetic proteins in the zebrafish ovary. Biol. Reprod. 89 55–110

    PubMed  Google Scholar 

  • Li J, Chu L, Sun X, Liu Y, and Cheng CHK 2015 IGFs Mediate the action of LH on oocyte maturation in zebrafish. Mol. Endocrinol. 29 373–383

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu L and Ge W 2007 Growth differentiation factor 9 and its spatiotemporal expression and regulation in the zebrafish ovary. Biol. Reprod. 76 294–302

    Article  PubMed  Google Scholar 

  • Liu L, Korzh V, Balasubramaniyan NV, Ekker M and Ge R 2002 Platelet derived growth factor A (pdgf-a) expression during zebrafish embryonic development. Dev. Genes Evol. 212 298–301

    Article  CAS  PubMed  Google Scholar 

  • Lokman PM, George KAN, Divers SL, Algie M and Young G 2007 11 Ketotestosterone and IGF-I increase the size of previtellogenic oocytes from short finned eel, Anguilla australis, in vitro. Reproduction 133 955–967

    Article  CAS  PubMed  Google Scholar 

  • Lubzens E, Young G, Bobe J and Cerd J 2010 Oogenesis in teleosts: How fish eggs are formed. Gen. Comp. Endocrinol. 165 367–389

    Article  CAS  PubMed  Google Scholar 

  • Lyrakou S, Hultén MA and Hartshorne GM 2002 Growth factors promote meiosis in mouse fetal ovaries in vitro. Mol. Hum. Reprod. 8 906–911

    Article  CAS  PubMed  Google Scholar 

  • Maures T, Chan SJ, Xu B, Sun H, Ding J and Duan C 2002 Structural, Biochemical, and Expression Analysis of Two Distinct Insulin-Like Growth Factor I Receptors and their Ligands in Zebrafish. Endocrinology 143 1858–1871

    Article  CAS  PubMed  Google Scholar 

  • Miura T, Yamauchi K, Takahashi H and Nagahama Y 1991 Hormonal induction of all stages of spermatogenesis in vitro in the male Japanese eel (Anguilla japonica). PNAS. 88 5774–5778

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Miura T, Miura C, Konda Y and Yamauchi K 2002 Spermatogenesis-preventing substance in Japanese eel. Development 129 2689–2697

    CAS  PubMed  Google Scholar 

  • Miura T and Miura C 2008 Progestin is an essential factor for the initiation of the meiosis in spermatogenesis and early oogenesis in fish. Cybium 32 130–132

    Google Scholar 

  • Mukherjee D, Majumdar S, Moulick SR, Pal P and Mallick B 2014 Signalling Pathways in Insulin and IGF-1 Mediated Oocyte Maturation in Lower vertebrates. Indian J. Biochem. Biophys. 51 520–526

    CAS  PubMed  Google Scholar 

  • Nagahama Y, Yoshikuni M, Yamashita M, Tokumoto T and Katsu Y 1995 Regulation of oocyte growth and maturation in fish. Curr. Top. Dev. Biol. 30 103–145

    Article  CAS  PubMed  Google Scholar 

  • Nagahama Y and Yamashita M 2008 Regulation of oocyte maturation in fish. Dev. Growth Differ. 50 195–219

    Article  Google Scholar 

  • Nakamura S, Kobayashi K, Nishimura T and Tanaka M 2011 Ovarian Germline Stem Cells in the Teleost Fish, Medaka (Oryzias latipes). Int. J. Biol. Sci. 7 403–409

    Article  PubMed  PubMed Central  Google Scholar 

  • Nelson SN and Kraak GVD 2010a The role of insulin-like growth factor (IGF) system in zebrafish (Danio rerio) ovarian development. Gen. Comp. Endocrinol. 168 103–110

    Article  CAS  PubMed  Google Scholar 

  • Nelson SN and Kraak GVD 2010b Characterization and regulation of the insulin-like growth factor (IGF) system in the zebrafish (Danio rerio) ovary. Gen. Comp. Endocrinol. 168 111–120

    Article  CAS  PubMed  Google Scholar 

  • Pang Y and Ge W 2002 Epidermal growth factor and TGFα promote zebrafish oocyte maturation in vitro: potential role of the ovarian activin regulatory system. Endocrinology 143 47–54

    Article  CAS  PubMed  Google Scholar 

  • Peng C, Clelland E and Tan Q 2009 Potential role of bone morphogenetic protein-15 in zebrafish follicle development and oocyte maturation. Comp. Biochem. Physiol. A. 153 83–87

    Article  Google Scholar 

  • Perrot V, Moiseeva EB, Gozes Y, Chan SJ and Funkenstein B 2000 Insulin-like growth factor receptors and their ligands in gonads of a hermaphroditic species, the gilthead seabream (Sparus aurata): expression and cellular localization. Biol. Reprod. 6 229–241

    Article  Google Scholar 

  • Rajapurohit SV and Pancharatna K 2007 Development and hatching success of zebrafish (Danio rerio) under laboratory conditions. J. Adv. Zool. 28 21–25

    Google Scholar 

  • Reinecke M 2010 Insulin-like Growth Factors and Fish Reproduction. Biol. Reprod. 82 656 –661

    Article  CAS  PubMed  Google Scholar 

  • Roith DL, Kavsan VM, Koval AP and Roberts CT 1993 Phylogeny of the insulin-like growth factors (IGFs) and receptors: a molecular approach. Mol. Reprod. Dev. 35 332–338

    Article  Google Scholar 

  • Sanchez F and Smitz J 2012 Molecular control of oogenesis. Biochimica. Et. Biophysica. Act. 1822 1896–1912

    Article  CAS  Google Scholar 

  • Schlueter PJ, Peng G, Westerfield M and Duan C 2007 Insulin-like growth factor signaling regulates zebrafish embryonic growth and development by promoting cell survival and cell cycle progression. Cell Death Differ. 14 1095–1105

    Article  CAS  PubMed  Google Scholar 

  • Schmid AC, Naef E, Kloas W and Reinecke M 1999 IGF-I and IGF-II in the ovary of a bony fish Oreochromis mossambicus, the tilapia: in situ hybridization, immunohistochemical localization, Northern Blot and cDNA sequences. Mol. Cell Endocrinol. 156 141 149

    Google Scholar 

  • Selman K, Wallace RA, Sarka A and Xiaoping Q 1993 Stages of Oocyte Development in the Zebrafish, Brachydanio rerio. J. Morphol. 218 203–224

    Article  Google Scholar 

  • Shoae A. 2012 Insulin-Like Growth Factor System in a Previtellogenic Fish, The New Zealand Short finned Eel, Anguilla australis, University of Otago, New Zealand. Thesis

  • Silva JRV, Figueiredo JR and Hurk RVD 2009 Involvement of growth hormone (GH) and insulin like growth factor (IGF) system in ovarian folliculogenesis. Theriogenology 71(8) 1193–1208

    Article  CAS  PubMed  Google Scholar 

  • Spicer LJ 2004 Proteolytic degradation of insulin-like growth factor binding proteins by ovarian follicles: a control mechanism for selection of dominant follicles. Biol. Reprod. 70 1223–1230

    Article  CAS  PubMed  Google Scholar 

  • Tse ACK and Ge W 2009 Differential regulation of betacellulin and heparin-binding EGF like growth factor in cultured zebrafish ovarian follicle cells by EGF family ligands. Comp. Biochem. Physiol. A 153 13–17

    Article  Google Scholar 

  • Tse ACK and Ge W 2010 Spatial localization of EGF family ligands and receptors in the zebrafish ovarian follicle and their expression profiles during folliculogenesis. Gen. Comp. Endocrinol. 167 397–407

    Article  CAS  PubMed  Google Scholar 

  • Wang Y and Ge W 2004 Developmental profiles of activin bA, bB, and follistatin expression in the zebrafish ovary: Evidence for their differential roles during sexual maturation and ovulatory cycle. Biol. Reprod. 71 205–620

    Google Scholar 

  • Werner H, Weinstein D and Bentov I. 2008. Similarities and differences between insulin and IGF-I: Structures, receptors, and signaling pathways. Arch. Physiol. Biochem. 114 17–22

    Article  CAS  PubMed  Google Scholar 

  • Wilson JM, Bunte RM and Carty AJ 2009 Evaluation of rapid cooling and tricaine methanesulfonate (MS222) as methods of euthanasia in zebrafish (Danio rerio). JAALAS. 48(6) 785–789

    CAS  PubMed  PubMed Central  Google Scholar 

  • Yao K and Ge W 2010 Kit system in the zebrafish ovary: evidence for functional divergence of two isoforms of kit (kita and kitb) and kit ligand (kitlga and kitlgb) during folliculogenesis. Biol. Reprod. 82(6) 1216–26

    Article  CAS  PubMed  Google Scholar 

  • Zhang Z, Lau SM, Zhang L and Ge W 2015 Disruption of follicle stimulating hormone receptor (fshr) but not luteinising hormone receptor (lhcgr) by TALEN leads to failed follicle activation in females followed by sexual reversal to males. Endocrinology 156 3747–3762

    Article  CAS  PubMed  Google Scholar 

  • Zhou R, Tsang AHK, Lau SW and Ge W 2011 Pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptors in the zebrafish ovary: evidence for potentially dual roles of PACAP in controlling final oocyte maturation. Biol. Reprod. 85 615–625

    Article  CAS  PubMed  Google Scholar 

  • Zhou R, Yu SMY and Ge W 2016 Expression and functional characterization of intrafollicular GH IGF system in the zebrafish ovary. Gen. Comp. Endocrinol. 232 32–42

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by UGC-SAP DSA-Phase I [F. 4-18/2015/DSA-I (SAP-II)] grant from New Delhi, India.

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Correspondence to Pancharatna A Katti.

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Corresponding editor: Subhash C Lakhotia

Corresponding editor: Subhash C Lakhotia

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Katti, P.A., Narvekar, S.S., Goundadkar, B.B. et al. IGF1 stimulates differentiation of primary follicles and their growth in ovarian explants of zebrafish (Danio rerio) cultured in vitro . J Biosci 42, 647–656 (2017). https://doi.org/10.1007/s12038-017-9716-3

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