Inducible overexpression of endothelial proNGF as a mouse model to study microvascular dysfunction

Biochim Biophys Acta Mol Basis Dis. 2018 Mar;1864(3):746-757. doi: 10.1016/j.bbadis.2017.12.023. Epub 2017 Dec 16.


Impaired maturation of nerve growth factor precursor (proNGF) and its accumulation has been reported in several neurodegenerative diseases, myocardial infarction and diabetes. To elucidate the direct impact of proNGF accumulation identified the need to create a transgenic model that can express fully mutated cleavage-resistant proNGF. Using Cre-Lox technology, we developed an inducible endothelial-specific proNGF transgenic mouse (proNGFLoxp) that overexpresses GFP-conjugated cleavage-resistant proNGF123 when crossed with VE-cadherin-CreERT2 (Cre). Expression of proNGF, inflammatory mediators, NGF and VEGF was evaluated by PCR, Western blot and immunohistochemistry. EC-proNGF overexpression was confirmed using colocalization of anti-proNGF within retinal vasculature. EC-proNGF did not cause retinal neurotoxicity or marked glial activation at 4-weeks. Microvascular preparation from Cre-proNGF mice showed significant imbalance of proNGF/NGF ratio, enhanced expression of TNF-α and p75NTR, and tendency to impair TrkA phosphorylation compared to controls. EC-proNGF overexpression triggered mRNA expression of p75NTR and inflammatory mediators in both retina and renal cortex compared to controls. EC-proNGF expression induced vascular permeability including breakdown of BRB and albuminuria in the kidney without affecting VEGF level at 4-weeks. Histopathological changes were assessed after 8-weeks and the results showed that EC-proNGF triggered formation of occluded (acellular) capillaries, hall mark of retinal ischemia. EC-proNGF resulted in glomerular enlargement and kidney fibrosis, hall mark of renal dysfunction. We have successfully created an inducible mouse model that can dissect the contribution of autocrine direct action of cleavage-resistant proNGF on systemic microvascular abnormalities in both retina and kidney, major targets for microvascular complication.

Keywords: Acellular capillary; Apoptosis; Barrier dysfunction; Diabetic retinopathy; Inflammation; Nerve growth factor; Neurodegeneration; Neurotrophin.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Disease Models, Animal
  • Endothelium, Vascular / metabolism*
  • Endothelium, Vascular / pathology
  • Endothelium, Vascular / physiopathology*
  • Gene Expression Regulation
  • Kidney Diseases / genetics
  • Kidney Diseases / metabolism
  • Kidney Diseases / pathology
  • Kidney Diseases / physiopathology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microvessels / physiopathology*
  • Nerve Growth Factor / genetics*
  • Nerve Growth Factor / metabolism
  • Protein Precursors / genetics
  • Protein Processing, Post-Translational
  • Retina / metabolism
  • Retina / pathology
  • Retina / physiopathology
  • Retinal Diseases / genetics
  • Retinal Diseases / metabolism
  • Retinal Diseases / pathology
  • Retinal Diseases / physiopathology
  • Retinal Vessels / metabolism
  • Retinal Vessels / pathology
  • Retinal Vessels / physiopathology


  • Protein Precursors
  • Nerve Growth Factor