Fig. 5.

TUNEL assay reveals increased levels of apoptosis in testes with Pmca4 deletion, compared to WT, with no effect of the diet. (a) Quantification of TUNEL results in WT and Pmca4^−/− testes from control and walnut-diet. No significant difference is evident between genotype or diet-type in the interstitial space; however, significant differences are seen between genotypes for spermatogonia (Gonia) and spermatocytes. The increase in spermatids did not reach significance. Implementation of the walnut-enriched diet reduced the number of apoptotic cells in Pmca4^−/− but not significantly. (Two-way ANOVA: N = 3, ±SEM). (b-c) Histological staining of testis tissue is consistent with apoptotic activity in Pmca4^−/−, evident through increased tubule disorganization with spaces between germ cells. Scale bar = 50 μm. (d) No statistically significant difference was detected in testicular weight between genotypes or treatment.