Immunohistochemical Approaches to the Study of Human Fetal Ovarian Development
The development of primordial germ cells into oocytes within primordial follicles involves a complex sequence of proliferation, developmental commitment, entry and arrest in meiosis, and association with surrounding somatic cells. These processes occur over the first few months of development in the human, with multiple stages of development present at any one time point. Immunohistochemistry has been hugely instructive in identifying the various key stages in ovarian development, by allowing simultaneous visualization of different stages of germ cell development, and their spatial arrangement. These studies allow comparison with other species and have identified key differences between human and murine ovarian development as well as giving a basis for functional studies. In this chapter we describe the main methodologies used in immunohistochemistry, using both chromogen and fluorescence approaches, and both single and double antigen detection.
Key wordsImmunohistochemistry Immunofluorescence Germ cell Detection systems Antibodies Antigen retrieval
We are grateful to Anne Saunderson and the staff of the Bruntsfield Suite of the Royal Infirmary of Edinburgh for assistance with patient recruitment and specimen collection and to members of the Anderson Lab and Histology core facility at the MRC Centre for Reproductive Health for assistance in developing these protocols. This work was supported by the Medical Research Council (programme grant G1100357 to R.A.A.) and Medical Research Scotland (research grant 354 FRG to A.J.C.).
This work was supported by the Medical Research Council (programme grant G1100357 to R.A.A.) and Medical Research Scotland (research grant 354 FRG to A.J.C.).
- 2.Wartenburg H (1981) Differentiation and development of the testes. In: Burger H, de Kretser DM (eds) The testis, 2nd edn. Raven, New York, pp 39–81Google Scholar
- 18.Anderson G, Gordon K (1996) Tissue processing, microtomy and paraffin sections. In: Bankroft JD, Stevens A (eds) Theory and practice of histological techniques, 4th edn. Churchill Livingstone, New YorkGoogle Scholar