Human sperm express a functional androgen receptor: effects on PI3K/AKT pathway.
BACKGROUND
Results from mice lacking the androgen receptor (AR) showed that it is critical for the proper development and function of the testes. The aim of this study was to investigate whether a functional AR is present in human sperm.


METHODS
The expression of AR and its effects on sperm were evaluated by RT-PCR, Western Blot, Immunocytochemistry, PI3Kinase and DNA laddering assays.


RESULTS
We showed in human sperm that AR is located at the head region. Dihydrotestosterone (DHT), in a dose-dependent manner, leads to the rapid phosphorylation of the AR on tyrosine, serine and threonine residues and this effect was reduced by the AR antagonist hydroxyflutamide (OH-Flut). The effects of AR were evaluated on the phosphoinositide-3 kinase/protein kinase B (PI3K/AKT) pathway. Specifically, 0.1 and 1 nM DHT stimulated PI3K activity, whereas 10 nM DHT decreased PI3K activity and levels of p-AKT S473 and p-AKT T308, p-BCL2, and enhanced phosphatase and tensin homologue (PTEN) phosphorylation. In addition, 10 nM DHT was able to induce the cleavage of caspases 8, 9 and 3 and cause DNA laddering, and these effects were reversed either by casodex or OHFlut. By using wortmannin, a specific PI3K inhibitor, the cleavage of caspase 3 was reproduced, confirming that in sperm the PI3K/AKT pathway is involved in caspase activation.


CONCLUSIONS
Human sperm express a functional AR that have the ability to modulate the PI3K/AKT pathway, on the basis of androgen concentration.
Department of Pharmaco-Biology, Faculty of Pharmacy, University of Calabria, 87036 Arcavacata di Rende, Cosenza, Italy.
Saveria
S
Aquila
Emilia
E
Middea
Stefania
S
Catalano
Stefania
S
Marsico
Marilena
M
Lanzino
Ivan
I
Casaburi
Ines
I
Barone
Rosalinda
R
Bruno
Silvia
S
Zupo
Sebastiano
S
Andò
D000728:Androgens / Q000494:pharmacology; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002470:Cell Survival; D013196:Dihydrotestosterone / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006801:Humans; D008297:Male; D051059:PTEN Phosphohydrolase / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011944:Receptors, Androgen / Q000096:biosynthesis*; D013094:Spermatozoa / Q000378:metabolism*; D013739:Testosterone / Q000494:pharmacology; D019061:src-Family Kinases / Q000378:metabolism
D000728:Androgens; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D011944:Receptors, Androgen; D013196:Dihydrotestosterone; D013739:Testosterone; D019869:Phosphatidylinositol 3-Kinases; D019061:src-Family Kinases; D051057:Proto-Oncogene Proteins c-akt; D051059:PTEN Phosphohydrolase; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9