Abstract
Our previous studies using puromycin aminonucleoside (PAN) established that podocyte damage leads to glomerular growth arrest during development and glomerulosclerosis later in life. This study examined the potential benefit of maintaining podocyte-derived VEGF in podocyte defense and survival after PAN injury using conditional transgenic podocytes and mice, in which human VEGF-A (hVEGF) transgene expression is controlled by tetracycline responsive element (TRE) promoter and reverse tetracycline transactivator (rtTA) in podocytes. In vitro experiments used primary cultured podocytes harvested from mice carrying podocin-rtTA and TRE-hVEGF transgenes, in which hVEGF can be induced selectively. Induction of VEGF in PAN-exposed podocytes resulted in preservation of intrinsic VEGF, α-actinin-4 and synaptopodin, antiapoptotic marker Bcl-xL/Bax, as well as attenuation in apoptotic marker cleaved/total caspase-3. In vivo, compared with genotype controls, PAN-sensitive neonatal mice with physiologically relevant levels of podocyte-derived VEGF showed significantly larger glomeruli. Furthermore, PAN-induced up-regulation of desmin, down-regulation of synaptopodin and nephrin, and disruption of glomerular morphology were significantly attenuated in VEGF-induced transgenic mice. Our data indicate that podocyte-derived VEGF provides self-preservation functions, which can rescue the cell after injury and preempt subsequent deterioration of the glomerulus in developing mice.
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Abbreviations
- Ctrl:
-
genotype control
- Dox:
-
doxycycline
- Flt1:
-
fms-like tyrosine kinase 1
- hVEGF:
-
human VEGF-A
- mVEGF:
-
mouse VEGF-A
- PAN:
-
puromycin aminonucleoside
- rhVEGF:
-
recombinant human VEGF165
- rtTA:
-
reverse tetracycline transactivator
- sFlt1:
-
soluble fms-like tyrosine kinase 1
- Tg:
-
double-transgenic for podocin-rtTA and TRE-hVEGF
- TRE:
-
tetracycline responsive element
- WT1:
-
Wilms' tumor 1
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Supported by National Institute of Diabetes and Digestive and Kidney Diseases (DK037868 and DK044757) and American Heart Association (09BGIA2261364).
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Ma, J., Matsusaka, T., Yang, HC. et al. Induction of Podocyte-Derived VEGF Ameliorates Podocyte Injury and Subsequent Abnormal Glomerular Development Caused by Puromycin Aminonucleoside. Pediatr Res 70, 83–89 (2011). https://doi.org/10.1203/PDR.0b013e31821bdf1c
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DOI: https://doi.org/10.1203/PDR.0b013e31821bdf1c
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