Mechanism of androgen-induced thymic atrophy in the wall lizard, Hemidactylus flaviviridis: an in vitro study.
The present in vitro study demonstrates the effect of androgen on thymocyte apoptosis leading to thymic atrophy in the wall lizard, Hemidactylus flaviviridis. Thymocytes collected from castrated lizards were incubated with varying concentrations of dihydrotestosterone (DHT) to observe its effect on proliferation and apoptosis. DHT treatment reduced the tritiated thymidine incorporation in thymocytes, suggesting that androgen directly inhibits thymocyte proliferation. It also caused apoptosis of thymocytes effectively at 10(-7)M. However, the increased apoptotic action of DHT was indirectly mediated through thymic epithelial cell-rich stromal cell components (TEC). This observation was reaffirmed by in vitro incubation of thymocytes with DHT-pretreated TEC-conditioned medium. However, the DHT-induced TEC-secreted apoptotic factors could induce thymocyte DNA fragmentation only when DHT was added to the conditioned medium. It implies that DHT priming of thymocytes is required for the apoptotic effect of DHT-induced TEC-secreted factor. DHT-induced thymocyte apoptosis was found to be caspase-dependent since it activated the initiator (caspase-9) and effector caspases (caspases-3 and -7) as well as cleaved the enzyme substrate poly(ADP-ribose) polymerase (PARP). Further, the apoptotic effect of DHT was routed through its classical receptors, as non-steroidal antiandrogen flutamide blocked the DHT-induced thymocyte apoptosis. The inhibition of apoptosis by transcription/translation inhibitors further substantiates the genomic pathway of DHT action. It can be concluded that DHT, in addition to inhibiting thymocyte proliferation directly, accelerates caspase-dependent apoptotic process in thymocytes indirectly through TEC via a genomic pathway. Nevertheless, the priming of thymocytes with DHT is required for the apoptotic effect of TEC-secreted factor.
Hareramadas
B
B
Comparative Endocrinology Laboratory, Department of Zoology, University of Delhi (North Campus), Delhi 110 007, India.
Rai
U
U
D000726:Androgen Antagonists / Q000494:pharmacology; D000728:Androgens / Q000494:pharmacology / Q000502:physiology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D020169:Caspases / Q000378:metabolism; D049109:Cell Proliferation; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D013196:Dihydrotestosterone / Q000494:pharmacology; D005485:Flutamide / Q000494:pharmacology; D066298:In Vitro Techniques; D008116:Lizards / Q000502:physiology*; D008297:Male; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013950:Thymus Gland / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*
D000726:Androgen Antagonists; D000728:Androgens; D011500:Protein Synthesis Inhibitors; D013196:Dihydrotestosterone; D003609:Dactinomycin; D005485:Flutamide; D003513:Cycloheximide; D020169:Caspases