ZFIN ID: ZDB-FIG-170907-7
Sugimoto et al., 2017 - Dissection of zebrafish shha function using site-specific targeting with a Cre-dependent genetic switch.. eLIFE   6 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Anatomical Terms:
Stage: Adult
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage: Adult

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; shhact/ct (control, left) and tcf21:DsRed; tcf21:CreER; shhact/ct hearts (right). (D) Immunofluorescence images of the subepicardial (top) and trabecular areas (bottom) of heart sections obtained from 4-HT–treated 7 dpi shhact/ct (control) or tcf21:CreER; shhact/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 shhact/ct (control) or tcf21:CreER; shhact/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.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
actb2 ae1Tg/+ (EKW) amputation: cardiac ventricle Adult regenerating tissue cardiac muscle cell RTPCR
f1Tg/+ (EKW) standard conditions Adult cardiac muscle cell RTPCR
pd37Tg/+ (EKW) standard conditions Adult epicardium RTPCR
pd37Tg/+ (EKW) amputation: cardiac ventricle Adult epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+; pd42Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
aldh1a2 pd37Tg/+ (EKW) standard conditions Adult Not Detected epicardium RTPCR
pd37Tg/+ (EKW) amputation: cardiac ventricle Adult epicardium RTPCR
Adult regenerating tissue epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+; pd42Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
dhh pd37Tg/+ (EKW) standard conditions Adult epicardium RTPCR
pd37Tg/+ (EKW) amputation: cardiac ventricle Adult epicardium RTPCR
Adult regenerating tissue epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+; pd42Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
DsRed2 ae1Tg/+; pd37Tg/+ (EKW) amputation: cardiac ventricle Adult epicardium IHC
EGFP ae1Tg/+; pd37Tg/+ (EKW) amputation: cardiac ventricle Adult cardiac muscle cell IHC
gli1 ae1Tg/+ (EKW) amputation: cardiac ventricle Adult regenerating tissue cardiac muscle cell RTPCR
f1Tg/+ (EKW) standard conditions Adult cardiac muscle cell RTPCR
ihhb pd37Tg/+ (EKW) standard conditions Adult epicardium RTPCR
pd37Tg/+ (EKW) amputation: cardiac ventricle Adult epicardium RTPCR
Adult regenerating tissue epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+; pd42Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
ptch1 ae1Tg/+ (EKW) amputation: cardiac ventricle Adult regenerating tissue cardiac muscle cell RTPCR
f1Tg/+ (EKW) standard conditions Adult cardiac muscle cell RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+; pd42Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
shha pd37Tg/+ (EKW) standard conditions Adult Not Detected epicardium RTPCR
pd37Tg/+ (EKW) amputation: cardiac ventricle Adult epicardium RTPCR
Adult regenerating tissue epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium RTPCR
shhavcc8Gt/vcc8Gt; pd37Tg/+; pd42Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult Not Detected epicardium RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
pd37Tg/+ (EKW) amputation: cardiac ventricle Adult epicardium aldh1a2 expression increased amount, abnormal
Adult epicardium dhh expression increased amount, abnormal
Adult epicardium ihhb expression increased amount, abnormal
Adult epicardium shha expression increased amount, abnormal
shhavcc8Gt/vcc8Gt; pd37Tg/+; pd42Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult epicardium shha expression absent, abnormal
shhavcc8Gt/vcc8Gt; pd42Tg/+ (EKW) amputation: cardiac ventricle, chemical treatment by environment: tamoxifen Adult regenerating tissue cardiac muscle cell proliferation decreased occurrence, abnormal
Acknowledgments:
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