RNA extraction and leptin receptor mRNA detection in bull ejaculated spermatozoa

ABSTRACT

Leptin, known as a potential satiety factor, plays an important role in both metabolism and reproduction. The presence of leptin in human seminal plasma and human spermatozoa has been shown; recently, leptin receptors [Ob-R] have been localized in human spermatozoa, thus suggesting a possible action of this hormone even on these cells. Our aim was to detect leptin receptor mRNA in bull ejaculated spermatozoa by reverse transcriptase-polymerase chain reaction [RT-PCR]. Total RNA was isolated from bull ejaculated spermatozoa and purified by different methods. Our results have revealed that sodium dodecylsulphate [SDS] and SDS/citric acid extraction methods are superior to guanidinium isothiocyanate in terms of yield and reproducibility of RNA recovery. The mRNA for Ob-Rb was detected in all samples examined. We conclude that Ob-R mRNA is present in bull spermatozoa where seminal plasma leptin can exert its effects