Modulation of germ cell apoptosis with a nitric oxide synthase inhibitor in a murine model of congenital cryptorchidism.
OBJECTIVE
Apoptosis has been implicated in testicular germ cell loss in experimental models of cryptorchidism. Nitric oxide synthase (NOS) has been shown to have a role in apoptosis in many cell types. The Hoxa 11 knockout mouse has congenital bilateral cryptorchidism and is uniformly sterile. We examined the time course of apoptosis in this model and attempted to attenuate this response in vivo by inhibition of NOS.


METHODS
The offspring of heterozygous Hoxa 11 knockout mice were genotyped by polymerase chain reaction. Homozygous knockout mice treated with the NOS inhibitor Nomega-nitro-L-arginine methyl ester (L-NAME) and untreated controls were sacrificed at weekly intervals at 3 to 13 weeks of age. Spermatogenesis was evaluated with hematoxylin and eosin staining. Germ cell apoptosis was assessed with a TUNEL assay and DNA staining. Co-localization of NOS activity was measured with a polyclonal antibody to endothelial NOS.


RESULTS
Impaired spermatogenesis was observed in Hoxa 11 knockout mice. Testis/body weight ratios were decreased in this group at weeks 6 and 7, while body weights were unchanged. Germ cell apoptosis was significantly higher in the knockout group compared to wild-type controls. Co-localization was observed between endothelial NOS activity and apoptotic cells, while mice treated with L-NAME demonstrated improved spermatogenesis and attenuated apoptosis.


CONCLUSIONS
Apoptosis and NOS reactivity appeared to co-localize in the seminiferous tubules in the Hoxa 11 knockout mouse model. Treatment with the NOS inhibitor L-NAME attenuated apoptosis and improved spermatogenesis. This finding suggests that early treatment might serve as an adjunct to early surgical intervention to reduce testicular atrophy, although any impact on long-term fertility remains to be determined.
DeFoor
William R
WR
Division of Pediatric Urology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio 45229-3039, USA.
Kuan
Chia-Yi
CY
Pinkerton
Malinda
M
Sheldon
Curtis A
CA
Lewis
Alfor G
AG
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D003456:Cryptorchidism / Q000201:enzymology* / Q000473:pathology*; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005854:Germ Cells / Q000166:cytology* / Q000187:drug effects*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors*
D004791:Enzyme Inhibitors; D019001:Nitric Oxide Synthase; D019331:NG-Nitroarginine Methyl Ester