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Zan Pan, Aihong Liu, Natalia Karagianni, Mark I. Rosenblatt; Vegf Promotes Trigeminal Neuron Growth Through Multiple VEGF Receptor Types. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2005.
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© ARVO (1962-2015); The Authors (2016-present)
Our previous data demonstrated that vascular endothelial growth factor (VEGF) can regulate the growth of corneal nerves. We sought to identify the VEGF receptor sub-types (VEGFR1, VEGFR2, or neuropilin-1(NPR1)) which mediate the potent effect of VEGF on isolated trigeminal neurons in vitro.
Thy1-YFP mice were sacrificed and trigeminal ganglia (TG) isolated following craniotomy. Isolated neurons were isolated from TG by limited enzymatic treatment with papain and collagenase/dispase followed by Percoll gradient centrifugation. Neurons were seeded on poly-D-lysine coated culture plates and incubated in Neurobasal A media containing 1% B27 supplement. Two hours after plating, neurons were treated for 1 hour with either VEGFR2 kinase inhibitors (SU1498 and Ki8751), specific neutralizing antibodies for VEGFR1, VEGFR2 or NPR1 or random IgG. Subsequently, 50ng/ml VEGF was added to each pre-treated well to evaluate for VEGF-dependent neuronal growth. Neurons were imaged by fluorescence microscopy at multiple time points. Images were analyzed using quantitative neuronal tracing software (Neurolucida).
Primary cultured trigeminal neurons extended dendrites 24 hr after plating. By 72 hours, 50 ng/ml VEGF increased dendrite numbers, branches, length and tortuosity by 3-fold. SU1498 (5uM) or Ki 8751 (5nM) completely suppressed dendrite elongation in the presence of VEGF. Neutralizing antibodies for VEGFR1, VEGFR2, and NPR1 inhibited dendrite growth in a dose-dependent manner with nearly complete inhibition achieved by 10ug/ml anti-VEGFR1, 2.5 ng/ml anti-VEGFR2, and 2ug/ml anti-NPR1. Random IgG did not affect dendrite growth.
VEGF promotion of trigeminal neuron growth is mediated through multiple receptors, including VEGFR receptors type 1, 2 and NPR 1. VEGF represents a potential drug target to promote restoration of corneal innervation following corneal injury. Further study is needed to identify opportunities to optimize VEGF-mediated neurogenesis while minimizing its possible angiogenic effects.
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