Detection of mRNA transcripts of cyclic nucleotide phosphodiesterase subtypes in ejaculated human spermatozoa

Multiple types and subtypes, including splice variants, of cyclic nucleotid e phosphodiesterases (PDE) have been shown to be expressed in various tissu es and organs. They control the intracellular level of cyclic nucleotides a nd are involved in hormonal signalling. In human spermatozoa, PDE play an i mportant role in the regulation of motility, capacitation and acrosome reac tion. The aim of this study was to investigate which transcripts of the dif ferent PDE types and subtypes could be found in human spermatozoa using rev erse transcription-polymerase chain reaction (RT-PCR). Ejaculated spermatoz oa from 10 single semen samples as well as another three semen sample pools were separated by swim-up and were investigated by RT-PCR. We obtained PCR products of the PDE types/subtypes 1A/B/C. 2, 3A/B, 4A/B/C, 5, and 8 with different intensities. Control PCR for leukocyte contamination were negativ e and contamination by other somatic cells was excluded by the spermatozoa preparation protocol, immunohistochemistry and visual examination. These re sults demonstrated for the first time that human ejaculated spermatozoa con tain an extended pattern of PDE mRNA transcripts.