cDNA copies of the testis-specific lactate dehydrogenase (LDH-C) mRNA are present in spermatogenic cells in mice, but processed pseudogenes are not derived from mRNAs that are expressed in haploid and late meiotic spermatogenic cells.
Retroposons are a class of genes created by reverse transcribing a processed mRNA and inserting the DNA copy into genomic DNA in germ-line cells. The present study concerns the question: Are retroposons created in meiotic and haploid spermatogenic cells? We demonstrate that polymerase chain reaction amplifies cytoplasmic DNAs with the expected intronless-structure of endogenous reverse transcriptase copies of the processed lactate dehydrogenase C mRNA encoding the testis-specific isoform of lactate dehydrogenase. Quantification of cytoplasmic LDH-C mRNA and endogenous cDNA by competitive RT-PCR and PCR, respectively, indicates that the level of LDH-C cDNA is lower by a factor of about 10(7) than the level of LDH-C mRNA in the cytoplasmic nucleic acids extracted from the testes of 14-day-old mice, and that about 1 in 10(5) meiotic cells contains an endogenous cDNA copy of LDH-C mRNA. A review of the literature reveals that a large number of genes including the LDH-C gene, whose expression is restricted to spermatogenic cells, are always single copy. Collectively, these observations suggest that reverse transcriptase cDNA copies of mRNAs are present in meiotic and haploid spermatogenic cells, but these cDNAs are not integrated into genomic DNA.
Zhong
X
X
Department of Biology, University of Massachusetts Boston, 100 Morrissey Blvd., Boston, MA 02125-3393, USA.
Kleene
K C
KC
D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000235:genetics*; D008297:Male; D008540:Meiosis; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011544:Pseudogenes; D012194:RNA-Directed DNA Polymerase / Q000235:genetics; D018626:Retroelements; D013094:Spermatozoa / Q000502:physiology*; D013737:Testis / Q000201:enzymology*
D018076:DNA, Complementary; D007527:Isoenzymes; D018626:Retroelements; D007770:L-Lactate Dehydrogenase; D012194:RNA-Directed DNA Polymerase