Abstract
The female reproductive system is important as the site for development and fertilization of an oocyte, for implantation and development of an embryo, and for growth and delivery of the fetus. It also produces protein and steroid hormones that help maintain a female’s health. Although the female phenotype is the default pathway for the development of the urogenital system, many processes can become disrupted during and after development which may originate from developmental problems. Improper development can be the underlying cause of structural malformations, sub- or infertility, hormonal abnormalities, endometriosis, carcinogenesis, or other detrimental outcomes. Our research programs examine the normal physiology and function of the female reproductive system and how it can become damaged due to pathologies or environmental/therapeutic exposures, with a focus on the ovary, ovarian follicles, and ovarian hormones. This chapter will describe detailed protocols of an in vitro organ culture system and methods to analyze changes in follicle formation, follicle development, and ovarian physiology. These methods can also be applied to the study of other aspects of female reproduction.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kezele P. and Skinner M. K. (2003) Regulation of ovarian primordial follicle assembly and development by estrogen and progesterone: endocrine model of follicle assembly. Endocrinology 144, 3329–37.
Chen Y., Jefferson W. N., Newbold R. R., Padilla-Banks E. and Pepling M. E. (2007) Estradiol, progesterone, and genistein inhibit oocyte nest breakdown and primordial follicle assembly in the neonatal mouse ovary in vitro and in vivo. Endocrinology 148, 3580–90.
Skinner M. K. (2005) Regulation of primordial follicle assembly and development. Hum Reprod Update 11 (5), 461–71.
Kezele P. R., Nilsson E. E. and Skinner M. K. (2002) Insulin but not insulin-like growth factor-1 promotes the primordial to primary follicle transition. Mol Cell Endocrinol 192, 37–43.
Nilsson E. E., Kezele P. and Skinner M. K. (2002) Leukemia inhibitory factor (LIF) promotes the primordial to primary follicle transition in rat ovaries. Mol Cell Endocrinol 188, 65–73.
Gigli I., Cushman R. A., Wahl C. M. and Fortune J. E. (2005) Evidence for a role for anti-Mullerian hormone in the suppression of follicle activation in mouse ovaries and bovine ovarian cortex grafted beneath the chick chorioallantoic membrane. Mol Reprod Dev 71, 480–88.
Fortune J. E., Kito S. and Byrd D. D. (1999) Activation of primordial follicles in vitro. J Reprod Fertil Suppl 54, 439–48.
Desmeules P. and Devine P. J. (2005) Characterizing the ovotoxicity of cyclophosphamide metabolites on cultured mouse ovaries. Toxicol Sci 90, 500–09.
Devine P. J., Sipes I. G. and Hoyer P. B. (2004) Initiation of delayed ovotoxicity by in vitro and in vivo exposures of rat ovaries to 4-vinylcyclohexene diepoxide. Reprod Toxicol 19, 71–77.
Rajapaksa K. S., Sipes I. G. and Hoyer P. B. (2007) involvement of microsomal epoxide hydrolase enzyme in ovotoxicity caused by 7,12-dimethylbenz[a]anthracene. Toxicol Sci 96, 327–34.
Rajapaksa K. S., Cannady E. A., Sipes I. G. and Hoyer P. B. (2007) Involvement of CYP 2E1 enzyme in ovotoxicity caused by 4-vinylcyclohexene and its metabolites. Toxicol Appl Pharmacol 221, 215–21.
Hirshfield A. N. (1991) Development of follicles in the mammalian ovary. Int Rev Cytol 124, 43–101.
Parrott J. A. and Skinner M. K. (1999) Kit-ligand/stem cell factor induces primordial follicle development and initiates folliculogenesis. Endocrinology 140, 4262–71.
Cooke P. S., Young P. F. and Cunha G. R. (1987) A new model system for studying androgen-induced growth and morphogenesis in vitro: the bulbourethral gland. Endocrinology 121, 2161–70.
Pedersen T. and Peters H. (1968) Proposal for a classification of oocytes and follicles in the mouse ovary. J Reprod Fertil 17, 555–57.
Brooks H. L., Ageloff S., Kwon T. H., Brandt W., Terris J. M., Seth A., et al. (2003) cDNA array identification of genes regulated in rat renal medulla in response to vasopressin infusion. Am J Physiol Renal Physiol 284, F218–28.
McReynolds M. R., Taylor-Garcia K. M., Greer K. A., Hoying J. B. and Brooks H. L. (2005) Renal medullary gene expression in aquaporin-1 null mice. Am J Physiol Renal Physiol 288, F315–21.
Schwartz M. A., Stone A. L., Greer K. A., Hoying J. B. and Williams S. K. (2005) Gene expression in tissue associated with extracellular matrix modified ePTFE. J Biomed Mater Res A 73, 30–38.
Kezele P. R., Ague J. M., Nilsson E. and Skinner M. K. (2005) Alterations in the ovarian transcriptome during primordial follicle assembly and development. Biol Reprod 72, 241–55.
Nilsson E., Rogers N. and Skinner M. K. (2007) Actions of anti-Mullerian hormone on the ovarian transcriptome to inhibit primordial to primary follicle transition. Reproduction 134, 209–21.
Lutfalla G. and Uze G. (2006) Performing quantitative reverse-transcribed polymerase chain reaction experiments. Methods Enzymol 410, 386–400.
Hu X. M., Christian P. J., Sipes I. G. and Hoyer P. B. (2001) Expression and redistribution of cellular bad, bax and bcl-xl protein is associated with VCD-induced ovotoxicity in rats. Biol Reprod 65, 1489–95.
Devine P. J., Sipes I. G., Skinner M. K. and Hoyer P. B. (2002) Characterization of a rat in vitro ovarian culture system to study the ovarian toxicant 4-vinylcyclohexene diepoxide. Toxicol Appl Pharmacol 184, 107–15.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Devine, P.J., Hoyer, P.B., Keating, A.F. (2009). Current Methods in Investigating the Development of the Female Reproductive System. In: Vaillancourt, C., Lafond, J. (eds) Human Embryogenesis. Methods in Molecular Biology, vol 550. Humana Press. https://doi.org/10.1007/978-1-60327-009-0_8
Download citation
DOI: https://doi.org/10.1007/978-1-60327-009-0_8
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60327-008-3
Online ISBN: 978-1-60327-009-0
eBook Packages: Springer Protocols