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Nucleic Acids Research, 1990, Vol. 18, No. 5 1255-1259
© 1990

MOLECULAR BIOLOGY

Measurement by quantitative PCR of changes in HPRT, PGK-1, PGK-2, APRT, MTase, and Zfy gene transcripts during mouse spermatogenesis

Judith Singer-Sam, Murray O. Robinson1, Anthony R. Bellvé2, Melvin l. Simon1 and Arthur D. Riggs

Division of Biology, Beckman Research Institute of the City of Hope Duarte, CA 91010 1Caltech Division of Biology Pasadena, CA 91125 2Department of Anatomy and Cell Biology, Urology and the Center for Reproductive Sciences, College of Physicians and Surgeons, Columbia University New York, NY 10032, USA

Received October 12, 1989. Revised February 6, 1990. Accepted February 6, 1990.

A reverse transcriptase-polymerase chain reaction assay (RT-PCR) was used quantitatively to measure accumulated levels of RNA transcripts in total mouse RNAs derived from male germ cells at various spermatogenic stages. RNA levels for two X-linked enzymes, phosphoglycerate kinase (PGK-1) and hypoxanthine phosphoribosyl transferase (HPRT), both decrease during spermatogenesis, although the transcript levels decrease much more rapidly for PGK-1. RNA for the Y-linked ZFY (zinc finger protein) is elevated in all spermatogenic cell fractions tested, being particularly high in leptotene/zygotene spermatocytes and round spermatids. RNA for adenine phosphoribosyltransferase (APRT) Increases 5-fold to a peak during late pachynema. RNA for PGK-2, undetectable in spermatogonial cells, increases at least 50-fold by the round spermatid stage. DNA (cytosine-5-)-methyltransferase (MTase) transcript levels are over an order of magnitude higher throughout spermatogenesis than in non-dividing liver cells.


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