Expression of cytochrome P450 aromatase transcripts in buffalo (Bubalus bubalis)-ejaculated spermatozoa and its relationship with sperm motility.
The cytochrome P450 aromatase (aromP450) deficient mice are infertile due to an impairment of spermatogenesis associated with a decrease in sperm motility and inability to fertilize oocytes. The sperm analysis showed decreased sperm motility in humans, having Cyp19 gene mutations. Further, in human, it was hypothesized that aromatase could be used as marker of sperm quality, particularly in the acquisition of its motility. However, there is no information regarding the expression of aromP450 in spermatozoa of farm animals including cattle and buffalo. In the present study, the expression of aromP450 in ejaculated buffalo spermatozoa and its relationship with sperm motility of ejaculated spermatozoa was studied by RT-PCR using total RNA isolated from buffalo-ejaculated spermatozoa. The results showed that conventional RT-PCR could not amplify aromatase transcript, while a nested PCR detected the presence of P450arom mRNA in buffalo-ejaculated spermatozoa. RT reaction followed by nested PCR was performed to compare the expression of aromatase transcripts in buffalo-ejaculated spermatozoa of two category semen graded on the basis of mass motility and motile and non-motile spermatozoa separated by swim-up. A higher (P<0.01) expression of aromP450 transcript was found in spermatozoa obtained from the good quality semen (higher mass motility) to that in spermatozoa of poor quality semen (low mass motility). Similarly, higher (P<0.01) expression of aromP450 mRNA was observed in the motile spermatozoa as compared to non-motile spermatozoa separated from good quality semen by swim-up. It is concluded that the present study demonstrates a positive relation between aromatase transcript and mass motility of buffalo-ejaculated spermatozoa, which could be a putative marker for the quality of semen in farm animals, particularly the acquisition of sperm motility.
Tiwari
Ashutosh
A
Molecular Endocrinology Lab., Division of Animal Biochemistry, National Dairy Research Institute, Karnal 132001, Haryana, India.
Singh
Dheer
D
Kumar
O Suneel
OS
Sharma
M K
MK
D000818:Animals; D001141:Aromatase / Q000235:genetics*; D002020:Buffaloes / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D015636:Magnesium Chloride / Q000008:administration & dosage; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa / Q000201:enzymology*
D012333:RNA, Messenger; D015636:Magnesium Chloride; D001141:Aromatase