Artificial Neural Network to Predict Varicocele Impact on Male Fertility through Testicular Endocannabinoid Gene Expression Profiles

Figure 3

Morphological analysis and correlation with fertility outcomes. A total of 30 animals (CTR n = 6; LAPARO n = 5; VAR n = 19) were subjected to the morphological evaluation. Panel (a): (A-C) example photomicrographs of left testes sections stained with H&E. (1-4) Tables reporting the mean number of pups recorded in experimental animals displaying different Johnsen scores. (A) Animals with a Johnsen score of 10 showed a complete spermatogenesis with many spermatozoa (black arrow head), and the seminiferous epithelium (E) organized in a regular thickness (white line); polyhedral Leydig cells with a round nucleus and eosinophilic cytoplasm were visible in interstitial compartments (white arrow head). (B) Samples with a Johnsen score of 7 displayed a mild impairment of the spermatogenic process with many spermatids (black arrow head) and some disorganization foci (black arrows) in the germinal epithelium; Leydig cells with smaller nuclei and accumulated connective tissue were also detectable in some interstitia (white arrow head). (C) Animals with a Johnsen score of 4 reported a compromised spermatogenesis with only few spermatocytes (black arrow heads) present and a damaged seminipherous epithelium with many disorganization foci (black arrows); interstitial compartments were not always conserved and also displayed uneven Leydig cells with not clearly identifiable cellular edges (white arrow head). Scale bars = 100 μm. (1) A conserved reproductive outcome was recorded in LAPARO group which showed, as CTR group, a complete spermatogenesis. (2) A conserved spermatogenesis was also recorded in four VAR animals showing a quite variable mean number of pups. (3) A mild impairment of the spermatogenic process was recorded in four VAR animals with a mean number of pups ranging from 4 to 8. (4) The majority of VAR animals (11) displayed a compromised spermatogenesis with a fertility outcome ranging from 0.5 to 10 new-borns. Panel (b): regression analysis of the mean Johnsen score values recorded in left testes versus the mean reproductive outcome; R² = 0,465.

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