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Role of VEGF and VEGFR2 Receptor in Reversal of ALS-CSF Induced Degeneration of NSC-34 Motor Neuron Cell Line

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Abstract

Vascular endothelial growth factor (VEGF), the well-known angiogenic factor is both neurotrophic and neuroprotective. Altered VEGF signalling is implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS), a fatal degenerative disease of motor neurons. We have shown earlier that VEGF protects NSC-34 motor neuronal cell line, when exposed to cerebrospinal fluid (CSF) from sporadic ALS patients (ALS-CSF). Here, we have investigated the consequences of ALS-CSF and VEGF supplementation on the VEGFR2 receptor and endogenous VEGF expression. ALS-CSF caused significant down-regulation of VEGFR2 as well as the Calbindin-D28K levels, but not endogenous VEGF. Exogenous supplementation restored the depletion of VEGFR2 and Calbindin-D28K with a concomitant up-regulation of endogenous VEGF. The up-regulated caspase 3 in the ALS-CSF group was reinstated to basal levels along with a significant reduction in the number of TUNEL-positive cells. Electron photomicrographs of ALS-CSF-exposed cells divulged presence of cytoplasmic vacuoles alongside severe damage to organelles like mitochondria, endoplasmic reticulum, etc. Substantial recovery of most of the damaged organelles was noted in response to VEGF supplementation. While the enhancement in endogenous VEGF levels highlights the autocrine functions, the up-regulation of VEGFR2 receptor emphasizes the paracrine functions of VEGF in modulating its neuroprotective effect against ALS-CSF. The revival of cellular organellar structure, increased calbindin expression and enhanced survival in response to VEGF supplementation consolidates the opinion that VEGF indeed has a therapeutic potential in sporadic ALS.

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Acknowledgments

This study was partly funded by the National Institute of Mental Health and Neurosciences and Indian Council for Medical Research (5/4-5/7-Neuro/2009-NCD-I dated 16/3/2010). Sailendra K and Lalitha Vishwanath, project trainees, performed the offline quantification of VEGFR2 and calbindin, respectively. Research Associateship for VK was funded by Council for Scientific and Industrial Research (CSIR), Government of India. SS and AMV are CSIR Senior Research Fellows. PSM is a UGC-CSIR Senior Research Fellow. SR is a Senior Research Fellow in the project funded by ICMR.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Neurophysiology, National Institute of Mental Health and Neuro Sciences, Hosur Road, Bangalore, 560 029, India

    K. Vijayalakshmi, R. Sumitha, S. Shruthi, Anu Mary Varghese, Poojashree Mishra, S. Gowri Manohari, T. N. Sathyaprabha, T. R. Raju & Phalguni Anand Alladi

  2. Department of Neurology, National Institute of Mental Health and Neuro Sciences, Hosur Road, Bangalore, 560 029, India

    Piyush Ostwal & A. Nalini

  3. Department of Neuropathology, National Institute of Mental Health and Neuro Sciences, Hosur Road, Bangalore, 560 029, India

    B. C. Sagar

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  1. K. Vijayalakshmi
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Vijayalakshmi, K., Ostwal, P., Sumitha, R. et al. Role of VEGF and VEGFR2 Receptor in Reversal of ALS-CSF Induced Degeneration of NSC-34 Motor Neuron Cell Line. Mol Neurobiol 51, 995–1007 (2015). https://doi.org/10.1007/s12035-014-8757-y

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