PUBLICATION

Deficiency of a transmembrane prolyl 4-hydroxylase in the zebrafish leads to basement membrane defects and compromised kidney function

Authors
Hyvarinen, J., Parikka, M., Sormunen, R., Ramet, M., Tryggvason, K., Kivirikko, K.I., Myllyharju, J., and Koivunen, P.
ID
ZDB-PUB-101027-9
Date
2010
Source
The Journal of biological chemistry   285(53): 42023-42032 (Journal)
Registered Authors
Keywords
Basement membrane, Enzyme catalysis, Eye, Hypoxia, Kidney, HIF, Prolyl 4-hydroxylase, Zebrafish
MeSH Terms
  • Animals
  • Basement Membrane/metabolism*
  • Catalysis
  • Gene Expression Regulation, Enzymologic*
  • Heart/physiology
  • Hypoxia
  • In Situ Hybridization
  • Kidney/metabolism*
  • Microscopy, Electron, Transmission/methods
  • Models, Biological
  • Podocytes
  • Procollagen-Proline Dioxygenase/metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Zebrafish
PubMed
20952382 Full text @ J. Biol. Chem.
Abstract
Prolyl 4-hydroxylases (P4Hs) catalyze the hydroxylation of collagens and hypoxia inducible factor (HIF)-α subunits. We studied the zebrafish homologue of the recently characterized human transmembrane P4H (P4H-TM) that can hydroxylate HIF-α, but not collagens, in vitro and influence HIF-α levels in cellulo. The zebrafish P4H-TM mRNA had its highest expression in the eye and brain and lower levels in other tissues, including the kidney. Morpholino knock-down of P4H-TM in embryos resulted in a reduction in the size of the eye and head and morphological alterations in the head from 2 days post fertilization (dpf) onwards. In addition, pericardial edema, regarded as a sign of kidney dysfunction, developed from 3 dpf onwards. The phenotype was dependent on the P4H-TM catalytic activity, as similar results were obtained with morpholinos targeting either translation initiation or catalytic residues of the enzyme. Structural and functional analyses of the morphant pronephric kidneys revealed fragmented glomerular basement membranes (BMs), disorganized podocyte foot processes and severely compromised pronephric kidney function leading to proteinuria. The opacity of the eye lens was reduced due to the presence of extra nuclei and deposits, and the structure of the lens capsule BM was altered. Our data suggest that P4H-TM catalytic activity is required for the proper development of the glomerular and lens capsule BMs. Many HIF target genes were induced in the P4H-TM-deficient morphants, but the observed phenotype is not likely to be mediated at least solely via the HIF pathway, and thus P4H-TM probably has additional, as yet unknown, substrates.
Genes / Markers
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Human Disease / Model
Sequence Targeting Reagents
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Mapping