Springer Protocols: Abstract: Epigenetic Control in Male Germ Cells: A Transillumination-Assisted Microdissection Method for the Analysis of Developmentally Regulated Events

Epigenetic Control in Male Germ Cells: A Transillumination-Assisted Microdissection Method for the Analysis of Developmentally Regulated Events

By: Durga Prasad Mishra³, Paolo Sassone-Corsi³

Abstract

Full Text

Download PDF (206K)

Germ cells constitute the vehicles of genetic information and thereby of inheritance through generations. The epigenetic control mechanisms that govern maintenance and reprogramming of the germline in diverse organisms have gained increasing interest as they reveal essential regulatory pathways imphcated in health and disease. We describe a step-by-step transillumination-assisted microdissec tion method currently used in our laboratory to routinely characterize various spermatogenic cell types. This methodology is ideal to analyze nuclear chromatin condensation and allows further evaluation of epigenetic modifications in male germ cells.

Affiliation(s): (3) Department of Pharmacology, School of Medicine, University of California,Irvine, Irvine, CA

Book Title: Germline Stem Cells

Series: Methods in Molecular Biology | Volume: 450 | Pub. Date: Apr-01-2008 | Page Range: 119-126 | DOI: 10.1007/978-1-60327-214-8_8

Subject: Cell Biology

Key Words: Chromatin - epigenetic reprogramming - germ cells - meiosis - spermatogenesis

References

Download References

1.	1. Holliday, R. (1989) DNA methylation and epigenetic mechanisms. Cell Biophys. 15, 15–20.

2.	2. Sassone-Corsi, P. (2002) Unique chromatin remodeling and transcriptional regulation in sper matogenesis. Science. 296, 2176–2178.

3.	3.DeBaun, M.R., Niemitz, E.L., and Feinberg, A.R (2003) Association of in vitro fertilization with Beckwith—Wiedemann syndrome and epigenetic alterations of LITl and H19. Am. J. Hum. Genet. 72, 156–160.

4.	4. Brinster, R.L. (2002) Germline stem cell transplantation and transgenesis. Science. 296, 2174–2176.

5.	5. Kimmins, S., and Sassone-Corsi, P. (2005) Chromatin remodelling and epigenetic features of germ cells. Nature. 31, 583–589.

6.	6. Russell, L.D., Ettlin, R.A., SinhaHikim, A.R, and Clegg, E.D. (1990) Mammalian sperma togenesis, in Histological and histopathological evaluation of the testis, Cache River Press, St. Louis, MO, pp. 1–40.

7.	7. Perey, B., Clermont, Y., and Leblond, C.P (1961) The wave of the seminiferous epithelium in the rat. Am. J. Anat. 108, 47–77.

8.	8.Parvinen, M., and Vanha-Perttula, T. (1972) Identification and enzyme quantitation of the stages of the seminiferous epithelial wave in the rat. Anat. Rec. 174, 435–450.

9.	9.Toppari, J., Eerola, E., and Parvinen, M. (1985) Flow cytometric DNA analysis of defined stages of rat seminiferous epithelial cycle during in vitro differentiation. J. Androl. 6, 325–333.

10.	10. Toppari, J., and Parvinen, M. (1985) In vitro differentiation of rat seminiferous tubular seg ments from defined stages of the epithelial cycle morphologic and immunolocalization analy sis. J. Androl. 6, 334–343.

11.	11. Leblond, C.P, and Clermont, Y. (1952) Definition of the stages of the cycle of the seminiferous epithelium in the rat. Ann. N. Y. Acad. Sci. 55, 548–573.

12.	12. Parvinen, M. (1982) Regulation of the seminiferous epithelium. Endocr Rev. 3, 404–417.

13.	13. Toppari, J., Bishop, PC, Parker, J.W., and diZerega, G.S. (1988) DNA flow cytometric analy sis of mouse seminiferous epithelium. Cytometry. 9, 456–462.

14.	14. Kangasniemi, M., Kaipia, A., Mali, P., Toppari, J., Huhtaniemi, I., and Parvinen, M. (1990) Modulation of basal and FSH-dependent cyclic AMP production in rat seminiferous tubules staged by an improved transillumination technique. Anat. Rec. 227, 62–76.

15.	15. Parvinen, M., Toppari, J., and Lähdetie J. (1993) Transillumination-phase contrast microscopic techniques for evaluation of male germ cell toxicity and mutagenicity, in Methods in Reproductive Toxicology (Chapin, R.E., and Heindel, J., eds.). Academic Press, Orlando, Fl, pp. 142–165.

16.	16. Parvinen, M., and Hecht, N.B. (1981) Identification of living spermatogenic cells of the mouse by transillumination-phase contrast microscopic technique for “in situ” analyses of DNA polymerase activities. Histochemistry. 71, 567–579.

17.	17. Kotaja, N., Kimmins, S., Brancorsini, S., et al. (2004) Preparation, isolation and characteriza tion of stage-specific spermatogenic cells for cellular and molecular analysis. Nat. Methods. 3, 249–254.

18.	18. Kotaja, N., Lin, H., Parvinen, M., and Sassone-Corsi, P.(2006) Interplay of PIWI/Argonaute protein MIWI and kinesin KIF17b in chromatoid bodies of male germ cells. J. Cell Sci. 119, 2819–2825.

19.	19. Kotaja, N., Bhattacharyya, S.N., Jaskiewicz, L., et al. (2006) The chromatoid body of male germ cells: similarity with processing bodies and presence of Dicer and microRNA pathway components. Proc. Natl. Acad. Sci. U. S. A. 103, 2647–2652.

20.	20. Martianov, I., Fimia, G.M., Dierich, A., Parvinen, M., Sassone-Corsi, P., and Davidson, I. (2001) Late arrest of spermiogenesis and germ cell apoptosis in mice lacking the TBP-like TLF/TRF2 gene. Mol. Cell. 7, 509–515.

21.	21. Martianov, I., Brancorsini, S., Gansmuller, A., Parvinen, M., Davidson, I., and Sassone-Corsi, P. (2002) Distinct functions of TBP and TLF/TRF2 during spermatogenesis: requirement of TLF for heterochromatic chromocenter formation in haploid round spermatids. Development. 129, 945–955.

22.	22. Henriksen, K., and Parvinen, M. (1998) Stage-specific apoptosis of male germ cells in the rat: mechanisms of cell death studied by supravital squash preparations. Tissue Cell. 30, 692–701.

23.	23. Ventelä, S., Toppari, J., and Parvinen, M. (2003) Intercellular organelle traffic through cyto plasmic bridges in early spermatids of the rat: mechanisms of haploid gene product sharing. Mol Biol. Cell. 14, 2768–2780.

24.	24. Kimmins, S., Crosio, C, Kotaja, N., et al. (2007) Differential functions of the Aurora-B and Aurora-C kinases in mammalian spermatogenesis. Mol. Endocrinol. 21, 726–739.

Comments (Loading...)

Post comment/View all comments