Heat-shock protein 60 is required for blastema formation and maintenance during regeneration.
Zebrafish fin regeneration requires the formation and maintenance of blastema cells. Blastema cells are not derived from stem cells but behave as such, because they are slow-cycling and are thought to provide rapidly proliferating daughter cells that drive regenerative outgrowth. The molecular basis of blastema formation is not understood. Here, we show that heat-shock protein 60 (hsp60) is required for blastema formation and maintenance. We used a chemical mutagenesis screen to identify no blastema (nbl), a zebrafish mutant with an early fin regeneration defect. Fin regeneration failed in nbl due to defective blastema formation. nbl also failed to regenerate hearts. Positional cloning and mutational analyses revealed that nbl results from a V324E missense mutation in hsp60. This mutation reduced hsp60 function in binding and refolding denatured proteins. hsp60 expression is increased during formation of blastema cells, and dysfunction leads to mitochondrial defects and apoptosis in these cells. These data indicate that hsp60 is required for the formation and maintenance of regenerating tissue.
Makino
Shinji
S
Howard Hughes Medical Institute, Department of Cardiology, Children's Hospital, Harvard Medical School, Boston, MA 02115, USA. smakino@enders.tch.harvard.edu
Whitehead
Geoffrey G
GG
Lien
Ching-Ling
CL
Kim
Soo
S
Jhawar
Payal
P
Kono
Akane
A
Kawata
Yasushi
Y
Keating
Mark T
MT
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D015152:Blotting, Northern; D001973:Bromodeoxyuridine; D049109:Cell Proliferation; D018834:Chaperonin 60 / Q000235:genetics* / Q000378:metabolism*; D002874:Chromosome Mapping; D004252:DNA Mutational Analysis; D017931:DNA Primers; D005121:Extremities / Q000502:physiology*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D020125:Mutation, Missense / Q000235:genetics; D012038:Regeneration / Q000502:physiology*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D015027:Zebrafish / Q000502:physiology*
D018834:Chaperonin 60; D017931:DNA Primers; D001973:Bromodeoxyuridine