An experimental study of the effect of two distinct surgical techniques of orchiopexy on spermatogenesis and testicular damage in cryptorchid testes.

OBJECTIVE: To compare the effect of two different techniques of testicular fixation on testicular function. DESIGN: Experimental study. SETTING: Surgical animal laboratory at an academic medical center. PATIENT(S): Sixteen mature golden hamsters underwent classic transfixation orchiopexy and true dartos pouch orchiopexy. INTERVENTION(S): Classic transfixation orchiopexy (CTO) involved transfixation of the testicular wall at two different points and fixation of the dartos fascia. True dartos pouch orchiopexy (TDPO) involved creating a window in the dartos fascia, passage of the testicle, and closure of the window from both sides of the testicle. MAIN OUTCOME MEASURE(S): Flow cytometric separation of testicular cells into haploid, diploid, and tetraploid fractions for histogram analysis. RESULT(S): A significant decrease in testicular weight was observed in 6 out of 16 animals undergoing CTO. Diploid cells comprised the main cell fraction, and almost no haploid or tetraploid cells were observed, while in the 16 animals undergoing TDPO no change from the control pattern was observed. CONCLUSION(S): This experimental work supports our clinical impression that TDPO should replace CTO as the method of choice for the treatment of an undescended testicle in children.

Use of confocal microscopy for the study of spermatogenesis.

Spermatogenesis consists of spermatogonial proliferation, meiosis and spermatid differentiation. Laser scanning confocal microscopy (LSCM) may be used as an advanced analytical tool to follow spermatogenesis inside the seminiferous tubules without performing histological sections. For this purpose, separated seminiferous tubules are fixed in 0.5% paraformaldehyde, stained for DNA with propidium iodide and analyzed by LSCM. By producing longitudinal optical sections in the layer of spermatogonia, spermatocytes and spermatids, stage-specific changes in their structure may be followed within the tubules by LSCM. Longitudinal z-sections may be obtained to produce three-dimensional images of the seminiferous tubules. In addition, different proteins may be followed during spermatogenesis in a stage specific manner within the tubule by incubation of the fixed seminiferous tubules with appropriate antibodies. As an example of the spermatogenesis studies using described LSCM techniques, detailed examination of spermatogonia, spermatocytes and spermatids during golden hamster spermatogenesis is presented. LSCM analysis of c-kit and SC3 protein expression at different stages of hamster spermatogenesis is demonstrated.