Knock-down of pantothenate kinase 2 severely affects the development of the nervous and vascular system in zebrafish, providing new insights into PKAN disease

Zizioli, D., Tiso, N., Guglielmi, A., Saraceno, C., Busolin, G., Giuliani, R., Khatri, D., Monti, E., Borsani, G., Argenton, F., Finazzi, D.
Neurobiology of disease   85: 35-48 (Journal)
Registered Authors
Argenton, Francesco, Borsani, Giuseppe, Busolin, Giorgia, Tiso, Natascia
Coenzyme A, Morpholino, NBIA, Neurodegeneration, Pank2, Pantethine, Zebrafish
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Animals, Genetically Modified
  • COS Cells
  • Cardiovascular System/enzymology*
  • Cardiovascular System/growth & development*
  • Cardiovascular System/pathology
  • Cell Line, Tumor
  • Disease Models, Animal
  • Gene Knockdown Techniques
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Nervous System/enzymology*
  • Nervous System/growth & development*
  • Nervous System/pathology
  • Pantothenate Kinase-Associated Neurodegeneration/pathology
  • Pantothenate Kinase-Associated Neurodegeneration/physiopathology
  • Phenotype
  • Phosphotransferases (Alcohol Group Acceptor)/genetics
  • Phosphotransferases (Alcohol Group Acceptor)/metabolism*
  • Sequence Homology, Amino Acid
  • Zebrafish
26476142 Full text @ Neurobiol. Dis.
Pantothenate Kinase Associated Neurodegeneration (PKAN) is an autosomal recessive disorder with mutations in the pantothenate kinase 2 gene (PANK2), encoding an essential enzyme for Coenzyme A (CoA) biosynthesis. The molecular connection between defects in this enzyme and the neurodegenerative phenotype observed in PKAN patients is still poorly understood. We exploited the zebrafish model to study the role played by the pank2 gene during embryonic development and get new insight into PKAN pathogenesis. The zebrafish orthologue of hPANK2 lies on chromosome 13, is a maternal gene expressed in all development stages and, in adult animals, is highly abundant in CNS, dorsal aorta and caudal vein. The injection of a splice-inhibiting morpholino induced a clear phenotype with perturbed brain morphology and hydrocephalus; edema was present in the heart region and caudal plexus, where hemorrhages with reduction of blood circulation velocity were detected. We characterized the CNS phenotype by studying the expression pattern of wnt1 and neurog1 neural markers and by use of the Tg(neurod:EGFP/sox10:dsRed) transgenic line. The results evidenced that downregulation of pank2 severely impairs neuronal development, particularly in the anterior part of CNS (telencephalon). Whole-mount in situ hybridization analysis of the endothelial markers cadherin-5and fli1a, and use of Tg(fli1a:EGFP/gata1a:dsRed) transgenic line, confirmed the essential role of pank2 in the formation of the vascular system. The specificity of the morpholino-induced phenotype was proved by the restoration of a normal development in a high percentage of embryos co-injected with pank2 mRNA. Also, addition of pantethine or CoA, but not of vitamin B5, to pank2 morpholino-injected embryos rescued the phenotype with high efficiency. The zebrafish model indicates the relevance of pank2 activity and CoA homeostasis for normal neuronal development and functioning and provides evidence of an unsuspected role for this enzyme and its product in vascular development.
Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes