Silencing of the hydra serine protease inhibitor Kazal1 gene mimics the human SPINK1 pancreatic phenotype.
In hydra, the endodermal epithelial cells carry out the digestive function together with the gland cells that produce zymogens and express the evolutionarily conserved gene Kazal1. To assess the hydra Kazal1 function, we silenced gene expression through double-stranded RNA feeding. A progressive Kazal1 silencing affected homeostatic conditions as evidenced by the low budding rate and the induced animal death. Concomitantly, a dramatic disorganization followed by a massive death of gland cells was observed, whereas the cytoplasm of digestive cells became highly vacuolated. The presence of mitochondria and late endosomes within those vacuoles assigned them as autophagosomes. The enhanced Kazal1 expression in regenerating tips was strongly diminished in Kazal1(-) hydra, and the amputation stress led to an immediate disorganization of the gland cells, vacuolization of the digestive cells and death after prolonged silencing. This first cellular phenotype resulting from a gene knock-down in cnidarians suggests that the Kazal1 serine-protease-inhibitor activity is required to prevent excessive autophagy in intact hydra and to exert a cytoprotective function to survive the amputation stress. Interestingly, these functions parallel the pancreatic autophagy phenotype observed upon mutation within the Kazal domain of the SPINK1 and SPINK3 genes in human and mice, respectively.
Chera
Simona
S
Department of Zoology and Animal Biology, University of Geneva, Sciences III, 30 Quai Ernest Ansermet, CH-1211 Geneva 4, Switzerland.
de Rosa
Renaud
R
Miljkovic-Licina
Marijana
M
Dobretz
Kevin
K
Ghila
Luiza
L
Kaloulis
Kostas
K
Galliot
Brigitte
B
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D020868:Gene Silencing; D006801:Humans; D006829:Hydra / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D010179:Pancreas / Q000378:metabolism*; D010641:Phenotype; D012330:RNA, Double-Stranded / Q000235:genetics; D016415:Sequence Alignment; D015842:Serine Proteinase Inhibitors / Q000235:genetics*; D014359:Trypsin Inhibitor, Kazal Pancreatic
D002352:Carrier Proteins; D012330:RNA, Double-Stranded; C486642:SPINK1 protein, human; D015842:Serine Proteinase Inhibitors; D014359:Trypsin Inhibitor, Kazal Pancreatic