Measurement by quantitative PCR of changes in HPRT, PGK-1, PGK-2, APRT, MTase, and Zfy gene transcripts during mouse spermatogenesis.
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.
Singer-Sam
J
J
Division of Biology, Beckman Research Institute, City of Hope, Duarte, CA 91010.
Robinson
M O
MO
Bellvé
A R
AR
Simon
M I
MI
Riggs
A D
AD
D000228:Adenine Phosphoribosyltransferase / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D005796:Genes; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008297:Male; D008667:Metalloproteins / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010735:Phosphoglycerate Kinase / Q000235:genetics; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis / Q000235:genetics; D013091:Spermatogenesis / Q000235:genetics*; D014158:Transcription, Genetic; D015032:Zinc
D004268:DNA-Binding Proteins; D008667:Metalloproteins; D012313:RNA; D004248:DNA (Cytosine-5-)-Methyltransferases; D000228:Adenine Phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase; D010735:Phosphoglycerate Kinase; D015032:Zinc