Short photoperiods evoke testicular apoptosis in white-footed mice (Peromyscus leucopus)

Many small, nontropical mammals stop breeding during winter. Chronic exposure of males to short days (<12.5h light/day) causes the testes to atrophy and both steroidogenesis and gametogenesis to decrease. Male white-footed mice (Peromyscus leucopus) exposed to inhibitory short day lengths provide a natural animal model to study the cellular mechanisms regulating testicular regression. In the present study, the possible role of apoptosis was assessed during naturally occurring, short day-induced gonadal regression in white-footed mice by in situ terminal transferase-mediated end labeling (TUNEL), quantitative DNA 3'-end-labeling autoradiography (laddering) of DNA fragments, and quantification of Fas protein expression, an early initiator of apoptosis. Sexually mature male mice were exposed to short (8 h of light, 16 h of darkness) or long (16 h of light, 8 h of darkness) day lengths for 2, 4, 6, 8, or 10 weeks; gonads were then removed and processed for detection of apoptotic activity. In common with previous studies, the first significant reduction in relative testis mass was observed at week 10 of short day exposure. A 2- to 3-fold increase in apoptotic (TUNEL-positive) germ cells per seminiferous tubule was observed in the testes of mice exposed to short days for 4, 6, 8, or 10 weeks compared with the testes of long day animals. The extent of 3'-end labeling of low mol wt DNA increased with 4-8 weeks of short day exposure. Western blot analysis revealed an up-regulation of the Fas protein in the testes of short day males at 4, 8, and 10 weeks. Fas staining was primarily localized to spermatocytes and spermatids. Plasma testosterone concentrations decreased in short compared with long day animals after 6, 8, or 10 weeks. The increase in TUNEL positive-labeled germ cells, testicular DNA fragmentation, and up-regulation of the Fas protein before short day reductions of testis mass and function suggest that apoptosis is important for the mediation of photoperiod-induced testicular regression in white-footed mice.