ZFIN Image: El Maï et al., 2023, Fig. 8

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Fig. 8

Gut-specific tert expression extends the healthspan of naturally aged zebrafish.

Expression of tert transgene in the gut of WT fish delays local aging phenotypes such as proliferation, senescence and tissue degeneration. This leads to beneficial systemic impact improving early aging phenotypes such as proliferation capacity. a, Representative immunofluorescence images of proliferation staining (left, PCNA) and quantification (right) in the gut, testes and kidney marrow of 27-month-old WT zebrafish expressing (WT + Cre; n = 7 fish for the gut and kidney marrow and n = 6 fish for the testes) or not expressing (WT no Cre; n = 8 fish for the gut and kidney marrow and n = 6 fish for the testes) tert transgene in the gut. b, Representative images of SA-β-Gal staining of gut, testes and kidney marrow sections of 24-month-old WT zebrafish expressing (WT + Cre) or not expressing (WT no Cre) the tert transgene in the gut (left). RT–qPCR analysis of senescence-associated genes ink4a/b (p15/16) and cdkn1a (p21) in the gut, testes and kidney marrow of either 9- (n = 6 fish) or 27-month-old WT zebrafish expressing (WT + Cre; n = 8 fish) or not expressing (WT no Cre; n = 8 fish) tert mRNA in the gut (right). c, Representative H&E-stained sections of the gut, testes and kidney marrow of 27-month-old WT zebrafish expressing (WT + Cre) or not expressing (WT no Cre) the tert transgene in the gut (left) and respective quantifications of the width of the gut lamina propria (right; n = 7 fish for WT no Cre and n = 6 fish for WT + Cre) and mature spermatid area (right; n = 7 fish for WT no Cre and WT + Cre). The yellow arrows indicate the width of the lamina propria quantified on the left. The dashed lines delineate the mature area of the spermatids. d, Survival curve of WT no Cre (n = 42 fish; similar to the WT curve in Fig. Fig.7)7) and WT + Cre (n = 36 fish) zebrafish. All data are presented as the mean ± s.e.m. *P < 0.05, **P < 0.01, ***P < 0.001, using a two-tailed unpaired t-test for a,c or a one-way ANOVA and post hoc Tukey tests for b; **P < 0.01, a using two-tailed Mann–Whitney U-test). All RT–qPCR graphs represent the mean ± s.e.m. mRNA fold increase after normalization by rps11 gene expression levels. Scale bar, 20 µm.

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