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RNA isolated from spermatozoa or washed semen samples may facilitate the detection of mutations and deletions in testis-expressed AZF-linked genes. Spermatozoa (or spermatids) may express a reverse transcriptase, the role of which is unknown. Ejaculate spermatozoa may be a very useful tool in the identification of genes linked to an infertile phenotype.
We have circumstantial evidence for the presence of an RT in ejaculate sperm based on the detection of transcripts for ORF2 of LINE 1 encoding such an enzyme. Both SINES and LINES belong to the retroposon class of repeat elements, which are thought to proliferate via an intermediate RNA that is converted to DNA by an RNA-dependent DNA polymerase (reverse transcriptase). High levels of transcribed repetitive sequences are present in human spermatozoa, including medium reiteration repeats (MERs) and short and long nuclear interspersed repeats (SINES and LINES). We have further evidence for a complex population of transcripts based on screening a human testis cDNA library with a heterologous spermatozoal probe. To date, gene-specific RT-PCR or in situ hybridization has detected transcripts for β-actin, heat shock proteins (HSP) 70 and 90, protamines (PRM) 1 and 2, transition protein (TNp) 2, HLA II, β-integrins, and, most recently, phosphodiesterase subtypes.
Human ejaculate spermatozoa contain multiple mRNA species carried over from earlier stages in spermatogenesis.
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