FIGURE

Fig. 5

ID

ZDB-FIG-170907-7

Publication

Sugimoto et al., 2017 - Dissection of zebrafish shha function using site-specific targeting with a Cre-dependent genetic switch.

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Fig. 5

Epicardial shha expression promotes subepicardial cardiomyocyte proliferation during heart regeneration.

(A) Schematic of the experiment. (B) Semi-qRT-PCR analysis of shha in purified tcf21:DsRed2⁺ epicardial cells obtained from uninjured and injured (7 dpi) tcf21:DsRed hearts. Injury was confirmed by the induction of raldh2 expression. (C) Semi-qRT-PCR analysis of Hh pathway genes using purified tcf21:DsRed2⁺ epicardial cells obtained from 4-HT–treated 7 dpi tcf21:DsRed; shha^ct/ct (control, left) and tcf21:DsRed; tcf21:CreER; shha^ct/ct hearts (right). (D) Immunofluorescence images of the subepicardial (top) and trabecular areas (bottom) of heart sections obtained from 4-HT–treated 7 dpi shha^ct/ct (control) or tcf21:CreER; shha^ct/ct hearts. Brackets, subepicardial areas. Dotted lines, approximate amputation plane. Arrows indicate proliferating cardiomyocytes. (E) Quantification of cardiomyocyte proliferation in the subepicardial and trabecular areas of the heart sections obtained from 4-HT–treated 7 dpi shha^ct/ct (control) or tcf21:CreER; shha^ct/ct hearts shown in D (n = 6 each). The data are presented as the mean ± SEM (**p<0.01, Mann–Whitney U test). N.S., not significant (p=0.3367). (F) Image of heart sections obtained from 7 dpi gata4:EGFP; tcf21;DsRed2 fish. Subepicardial cardiomyocytes (green, arrows) associate with epicardial cells (magenta, arrowheads). (G) Semi-qRT-PCR analysis of shha pathway genes using purified subepicardial cardiomyocytes obtained from 7 dpi gata4:EGFP ventricles. Cardiomyocytes purified from uninjured cmlc2:EGFP ventricles were used as negative controls. Scale bar, 50 μm.

Expression Data

Genes:	actb2 aldh1a2 dhh DsRed2 EGFP gli1 ihhb ptch1 shha
Fish:	ae1Tg/+ (EKW) f1Tg/+ (EKW) pd37Tg/+ (EKW) ae1Tg/+; pd37Tg/+ (EKW) shha^{vcc8Gt/vcc8Gt}; pd37Tg/+ (EKW) shha^{vcc8Gt/vcc8Gt}; pd37Tg/+; pd42Tg/+ (EKW)
Conditions:	amputation: cardiac ventricle, chemical treatment by environment: tamoxifen amputation: cardiac ventricle
Anatomical Terms:	cardiac muscle cell epicardium regenerating tissue cardiac muscle cell regenerating tissue epicardium
Stage:	Adult

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	pd37Tg/+ (EKW) shha^{vcc8Gt/vcc8Gt}; pd37Tg/+; pd42Tg/+ (EKW) shha^{vcc8Gt/vcc8Gt}; pd42Tg/+ (EKW)
Conditions:	amputation: cardiac ventricle, chemical treatment by environment: tamoxifen amputation: cardiac ventricle
Observed In:	epicardium regenerating tissue cardiac muscle cell proliferation
Stage:	Adult

Phenotype Detail

Acknowledgments

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