The aim of this study was to investigate the effects of nerve growth factor (NGF) on the in vitro culture of goat preantral follicles. Ovarian cortex fragments were cultured in α-MEM+ supplemented with 0, 1, 10, 50, 100 or 200 ng/ml NGF for 1 or 7 days. Small fragments of noncultured ovarian tissue as well as those cultured for 1 or 7 days were processed for histology and transmission electron microscopy. The results showed that after 1 or 7 days of culture at all concentrations of NGF, except at 1 ng/ml after 1 day of culture, there was a significant reduction in the percentage of normal follicles compared to noncultured tissues. At higher NGF concentrations (100 and 200 ng/ml) after 7 days of culture, there was a significant reduction in the percentage of normal follicles compared to tissues cultured in α-MEM+ alone or at the other concentrations of NGF. It is important to note that ultrastructural and fluorescent analyses confirmed only the integrity of follicles cultured with 1 ng/ml of NGF after 7 days. In contrast to noncultured control tissues, the percentage of developing follicles was significantly increased at all concentrations of NGF after 1 or 7 days of culture. We observed that follicular diameter was greater at 1 and 10 ng/ml NGF after culture for 7 days than at the other concentrations but was similar to follicles cultured in α-MEM+ alone. In conclusion, NGF improved the survival of goat preantral follicles cultured in vitro in a dose-dependent manner.

1.
Abir, R., B. Fisch, S. Jin, M. Barnnet, A. Ben-Haroush, C. Felz, G. Kessler-Icekson, D. Feldberg, S. Nitke (2005) Presence of NGF and its receptors in ovaries from human fetuses and adults. Mol Hum Reprod 11: 229–236.
2.
Abir, R., B. Fisch, A.H. Raz, S. Nitke, Z. Ben Rafael (1998) Preservation of fertility in women undergoing chemotherapy – current approach and future prospects. J Assist Reprod Genet 15: 469–478.
3.
Anderson, R.A., L.L.L. Robinson, J. Brooks, N. Spears (2002) Neurotropins and their receptors are expressed in the human fetal ovary. J Clin Endocrinol Metab 87: 890–897.
4.
Anesetti, G., P. Lombide, H. D’Albora, S.R. Ojeda (2001) Intrinsic neurons in the human ovary. Cell Tissue Res 306: 231–237.
5.
Barker, P.A. (2004) p75NTR is positively promiscuous: novel partners and new insights. Neuron 42: 529–533.
6.
Barrett, G.L. (2000) The p75 neurotrophin receptor and neuronal apoptosis. Prog Neurobiol 61: 205–229.
7.
Bhakar, A.L., J.L. Howell, C.E. Paul, A.H. Salehi, E.B. Becker, F. Said, A. Bonni, P.A. Barker (2003) Apoptosis induced by p75NTR overexpression requires Jun kinase-dependent phosphorylation of Bad. J Neurosci 23: 11373–11381.
8.
Botchkarev, V.A., N.V. Botchkareva, K.M. Albers, L. Chen, P. Welker, R. Paus (2000) A role for p75 neurotrophin receptor in the control of apoptosis-driven hair follicle regression. FASEB J 14: 1931–1942.
9.
Braw-Tal, R., S. Yossefi (1997) Studies in vivo and in vitro on the initiation of follicle growth in the bovine ovary. J Reprod Fertil 109: 165–171.
10.
Cantarella, G., L. Lempereur, M. Presta, D. Ribatti, G. Lombardo, F. Lazarovici, G. Zappalà, C. Pafumi, R. Bernardini (2002) Nerve growth factor-endothelial cell interaction leads to angiogenesis in vitro and in vivo. FASEB J 21: 21–29.
11.
Chaves, R.N., F.S. Martins, M.V.A. Saraiva, J.J.H. Celestino, C.A.P. Lopes, J.C. Correia, I.B. Lima-Verde, M.H.T. Matos, S.N. Báo, K.P.O. Name, C.C. Campello, J.R.V. Silva, J.R. Figueiredo (2008) Chilling ovarian fragments during transportation improves viability and growth of goat preantral follicles cultured in vitro. Reprod Fertil Dev 20: 640–647.
12.
Dees, W.L., J.K. Hiney, T.D. Schultea, A. Meyerhofer, M. Danilchik, G.A. Dissen, S.R. Ojeda (1995) The primate ovary contains a population of catecholaminergic neuron-like cells expressing nerve growth factor receptors. Endocrinology 136: 5760–5768.
13.
Dissen, G.A., C. Garcia-Rudaz, S.R. Ojeda (2009a) Role of neurotrophic factors in early ovarian development. Semin Reprod Med 27: 24–31.
14.
Dissen, G.A., C. Garcia-Rudaz, A. Paredes, C. Mayer, A. Mayerhofer, S.R. Ojeda (2009b) Excessive ovarian production of nerve growth factor facilitates development of cystic ovarian morphology in mice and is a feature of polycystic ovarian syndrome in humans. Endocrinology 150: 2906–2914.
15.
Dissen, G.A., D.F. Hill, M.E. Costa, W.L. Dees, H.E. Lara, S.R. Ojeda (1996a) A role for trkA nerve growth factor receptors in mammalian ovulation. Endocrinology 137: 198–209.
16.
Dissen, G.A., D.F. Hill, M.E. Costa, Y.J. Ma, S.R. Ojeda (1991) Nerve growth factor in the peripubertal rat ovary. Mol Endocrinol 5: 1642–1650.
17.
Dissen, G.A., A.N. Hirshfield, S. Malamd, S.R. Ojeda (1995) Expression of neurotropins and their receptors in the mammalian ovary is developmentally regulated: changes at the time of folliculogensis. Endocrinology 136: 4681–4692.
18.
Dissen, G.A., H.E. Lara, V. Leyton, A. Paredes, D.F. Hill, M.E. Costa, A. Martinez-Serrano, S.R. Ojeda (2000a) Intraovarian excess of nerve growth factor increases androgen secretion and disrupts estrous cyclicity in the rat. Endocrinology 141: 1073–1082.
19.
Dissen, G.A., A. Mayerhofer, S.R. Ojeda (1996b) Participation of nerve growth factor in the regulation of ovarian function. Zygote 4: 309–312.
20.
Dissen, G.A., J.A. Parrott, M.K. Skinner, D.F. Hill, M.E. Costa, S.R. Ojeda (2000b) Direct effects of nerve growth factor on theca cells from antral ovarian follicles. Endocrinology 141: 4736–4750.
21.
Dissen, G.A., C. Romero, A.N. Hirshfield, S.R. Ojeda (2001) Nerve growth factor is required for early follicular development in the mammalian ovary. Endocrinology 142: 2078–2086.
22.
Dissen, G.A., C. Romero, A. Paredes, S.R. Ojeda (2002) Neurotrophic control of ovarian development. Microsc Res Tech 59: 509–515.
23.
Fortune, J.E., S. Kito, S.A. Wandji, V. Srsen (1998) Activation of bovine and baboon primordial follicles in vitro. Theriogenology 49: 441–449.
24.
Harsum, S., J.D. Clarke, P. Martin (2001) A reciprocal relationship between cutaneous nerves and repairing skin wounds in the developing chick embryo. Dev Biol 238: 27–29.
25.
Hartshorne, G.M. (1997) In vitro culture of ovarian follicles. Rev Reprod 2: 94–104.
26.
Hovatta, O., R. Silye, R. Anir, T. Krausz, R.M.L. Winston (1997) Extracellular matrix improves survival of both stored and fresh human primordial and primary ovarian follicles in long-term culture. Hum Reprod 12: 1032–1036.
27.
Hoyle, G.W., R.M. Graham, J.B. Finkelstein, K.P.T. Nguyen, D. Gozal, M. Friedman (1998) Hyperinnervation of the airways in transgenic mice overexpressing nerve growth factor. Am J Respir Cell Mol Biol 18: 149–157.
28.
Lara, H.E., W.L. Dees, J.K. Hiney, G.A. Dissen, C. Rivier, S.R. Ojeda (1991) Functional recovery of the developing rat ovary after transplantation: contribution of the extrinsic innervations. Endocrinology 129: 1849–1860.
29.
Lara, H.E., D.F. Hill, K.H. Katz, S.R. Ojeda (1990a) The gene encoding nerve growth factor is expressed in the immature rat ovary: effect of denervation and hormonal treatment. Endocrinology 126: 357–363.
30.
Lara, H.E., J.K. McDonald, S.R. Ojeda (1990b) Involvement of nerve growth factor in female sexual development. Endocrinology 126: 364–375.
31.
Levanti, M.B., A. Germana, F. Abbate, G. Montalbano, J.A. Vega, G. Germana (2005) TrkA and p75NTR in the ovary of adult cow and pig. J Anat 207: 93–96.
32.
Lopes, C.A.P., R.R. Dos Santos, J.J. Celestino, M.A. Melo, R.N. Chaves, C.C. Campello, J.R. Silva, S.N. Báo, K. Jewgenow, J.R. Figueiredo (2009) Short-term preservation of canine preantral follicles: effects of temperature, medium and time. Anim Reprod Sci 115: 201–214.
33.
Lucci, C.M., C.A. Amorim, S.N. Báo, J.R. Figueiredo, A.P.R. Rodrigues, J.R. Silva, P.B.D. Gonçalves (1999) Effect of the interval of serial sections of ovarian tissue in the tissue chopper on the number of isolated caprine preantral follicles. Anim Reprod Sci 56: 39–49.
34.
Lucci, C.M., R.V. Silva, C.A. Carvalho, J.R. Figueiredo, S.N. Báo (2001) Light microscopical and ultrastrutural characterization of goat preantral follicles. Small Rumin Res 41: 61–69.
35.
Markström, E., E.C. Svensson, R. Shao, B. Svanberg, H. Billig (2002) Survival factors regulating ovarian apoptosis – dependence on follicle differentiation. Reproduction 123: 23–30.
36.
Matos, M.H.T., I.B. Lima-Verde, M.C.A. Luque, J.E. Maia Jr, J.R.V. Silva, J.J.H. Celestino, F.S. Martins, S.N. Báo, C.M. Lucci, J.R. Figueiredo (2007) Essential role of follicle stimulating hormone in the maintenance of caprine preantral follicle viability in vitro. Zygote 15: 173–182.
37.
Mattioli, M., B. Barboni, L. Gioia, P. Lucidi (1999) Nerve growth factor production in sheep antral follicles. Domest Anim Endocrinol 17: 361–371.
38.
Ojeda, S.R., G.A. Dissen, C. Romero (1999) Role of neurotrophic factors in the control of ovarian development. Front Endocrinol 21: 171–178.
39.
Ojeda, S.R., C. Romero, V. Tapia, G.A. Dissen (2000) Neurotrophic and cell-cell dependent control of early follicular development. Mol Cell Endocrinol 163: 67–71.
40.
Paredes, A., C. Romero, G.A. Dissen, T.M. Dechiara, L. Reichardt, A. Cornea, S.R. Ojeda, B. Xu (2004) TrkB receptors are required for growth and oocyte survival in the mammalian ovary. Dev Biol 267: 430–449.
41.
Richards, J.S. (1994) Hormonal control of gene expression in the ovary. Endocr Rev 15: 725–751.
42.
Romero, C., A. Paredes, G.A. Dissen, S.R. Ojeda (2002) Nerve growth factor induces the expression of functional FSH receptors in newly formed follicles of the rat ovary. Endocrinology 143: 1485–1494.
43.
Rosseto, R., I.B. Lima-Verde, M.H.T. Matos, M.V.A. Saraiva, F.S. Martins, L.S. Faustino, V.R. Araújo, C.M. Silva, K.P. Name, S.N. Báo, C.C. Campello, J.R. Figueiredo, H. Blume (2009) Interaction between ascorbic acid and follicle-stimulating hormone maintains follicular viability after long-term in vitro culture of caprine preantral follicles. Domest Anim Endocrinol 37: 112–123.
44.
Salas, C., M. Julio-Pieper, M. Valladares, R. Pommer, M. Vega, C. Mastronardi, B. Kerr, S.R. Ojeda, H.E. Lara, C. Romero (2006) Nerve growth factor-dependent activation of trkA receptors in the human ovary results in synthesis of follicle-stimulating hormone receptors and estrogen secretion. J Clin Endocrinol Metab 91: 2396–2403.
45.
Schotanus, K., W.J. Hage, H. Vanderstichele, R. van den Hurk (1997) Effects of conditioned media from murine granulose cell lines on the growth of isolated bovine preantral follicles. Theriogenology 48: 471–483.
46.
Shi, Z., W. Jim, G. Watanabe, A.K. Suzuki, S. Takahashi, K. Taya (2004) Expression of nerve growth factor (NGF), and its receptors trkA and p75 in ovaries of the cyclic golden hamster (Mesocricetus auratus) and the regulation of their production by luteinizing hormone. J Reprod Dev 50: 605–611.
47.
Silva, J.R., M.A.L Ferreira, S.H.F. Costa, R.R. Santos, F.C.A. Carvalho, A.P.R. Rodrigues, C.M. Lucci, S.N. Báo, J.R. Figueiredo (2002) Degeneration rate of preantral follicles in the ovaries of goats. Small Rumin Res 43: 203–209.
48.
Silva, J.R., T. Tharasanit, M.A. Taverne, G.C. Van Der Weijden, R.R. Santos, J.R. Figueiredo, R. Van Den Hurk (2006) The activin-follistatin system and in vitro early follicle development in goats. J Endocrinol 29: 113–125.
49.
Silva, J.R.V., R. van den Hurk, M.H.T. Matos, R.R. Santos, C. Pessoa, M.O. Moraes, J.R. Figueiredo (2004) Influences of FSH and EGF on primordial follicles during in vitro culture of caprine ovarian cortical tissue. Theriogenology 61: 1691–1704.
50.
Spears, N., M.D. Molinek, L.L.L. Robinson, N. Fulton, H. Cameron, K. Shimoda, E.E. Telfer, R.A. Anderson, D.J. Price (2003) The role of neurotropin receptors in female germ-cell survival in mouse and human. Development 130: 5481–5491.
51.
Steel, R.G.D., J.H. Torrie, D.A. Dickey (1997) Principles and Procedures of Statistics: A Biometrical Approach, ed 3. New York, McGraw-Hill, p 666.
52.
Van den Hurk, R., R. Abir, E.E. Telfer, M.M. Bevers (2000) Primate and bovine immature oocytes and follicles as sources of fertilizable oocytes. Hum Reprod Update 6: 457–474.
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