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Investigation of DNA Loop Domains using Fluorescent in Situ Hybridization (FISH) and Epifluorescence Microscopy

Published online by Cambridge University Press:  02 July 2020

Angela V. Klaus ,
Siobhan McCarthy ,
John McCarrey  and
W. Steven Ward
Angela V. Klaus
Affiliation:
Division of Urology, Robert Wood Johnson Medical School, New Brunswick, NJ08903
Siobhan McCarthy
Affiliation:
Division of Urology, Robert Wood Johnson Medical School, New Brunswick, NJ08903
John McCarrey
Affiliation:
Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, TX, 78228
W. Steven Ward
Affiliation:
Division of Urology, Robert Wood Johnson Medical School, New Brunswick, NJ08903
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Extract

The average sperm nucleus is only 8 micrometers long, but contains 1 meter of DNA. We have previously described how the sperm cell is able to package this DNA into the small volume of the sperm head. The only DNA structure that exists in both somatic cells and sperm nuclei is the organization of DNA loop domains that are attached at their bases to the nuclear skeleton, the nuclear matrix. We compared the structure of one gene in sperm nuclei to that of spermatogenic and adult cells.

Individual loop domains of the 5S rRNA gene cluster were visualized by fluorescent in situ hybridization (FISH). Nuclei were isolated from hamster spermatozoa, spermatogonia, pachytene spermatocytes, round spermatids, and adult liver cells. They were then extracted to remove the protamines or histones, fixed, hybridized to a biotinylated 5S rDNA probe, then viewed by epifluorescence microscopy.

Type
Cytochemistry (Light and Electron Histochemistry)
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 1116 - 1117
Copyright
Copyright © Microscopy Society of America

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References

References:

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2.Nadel, B., De, Lara J., Ward, WS.J. of Andrology. 16(6): 517–22, 1995 Nov-Dec.Google Scholar