Activity and intracellular localization of senescence-associated β-galactosidase in aging Xenopus oocytes and eggs.

Senescence-associated β-galactosidase (SA-β-gal) serves as a marker of senescence in aging somatic cells. However, little is known about SA-β-gal dynamics in aging gamete cells. To address this issue, here we investigated activity and intracellular localization of SA-β-gal in freshly obtained and aging oocytes and eggs of the African clawed frog Xenopus laevis. Data base mining revealed the presence of several homologous β-galactosidase sequences in the annotated Xenopus genome. Some of them were predicted to contain an N-terminal signal peptide sequence, suggesting enzyme translocation to cellular organelles. Biochemical and microscopic analyses confirmed SA-β-gal localization in the particulate and cytosolic fractions of oocytes and eggs. SA-β-gal activity was found to reside predominantly within a fraction of dense cytoplasmic granules that were extensively stained with the lysosome-specific dye LysoTracker Green DND-26 and had an average size of 8.9 ± 5.6 μm. These features identify the SA-β-gal-containing granules as a subpopulation of yolk platelets, specialized late endosomes or lysosomes that accumulate and store processed protein in frog oocytes. Further analysis revealed an increase of SA-β-gal activity in Xenopus eggs, but not in oocytes, aged in vitro over 48 h. Our data suggest that endosomal acidification during egg aging may be responsible for this increase.