Vascular Endothelial Growth Factor Attenuates Neurodegenerative Changes in the NSC-34 Motor Neuron Cell Line Induced by Cerebrospinal Fluid of Sporadic Amyotrophic Lateral Sclerosis Patients

Abstract

Background: Motor neuron disease or amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the selective death of motor neurons in the spinal cord as well as motor cortex. Recently, vascular endothelial growth factor (VEGF) has been identified as a neurotrophic factor in animal models of familial ALS and other neurological diseases. Objective: The present study was designed to investigate the neuroprotective role of VEGF in the more prevalent sporadic form of ALS. Methods: We studied the effect of VEGF on the NSC-34 cell line exposed to cerebrospinal fluid (CSF) from sporadic ALS patients (ALS-CSF) in terms of lactate dehydrogenase (LDH) assay as well as choline acetyltransferase (ChAT) and phosphorylated neurofilament expression by immunocytochemistry and confocal microscopy. NSC-34 cells were exposed to CSF from patients with definite ALS and compared to controls. LDH activity was assessed in the growth media, prior to and 24 h after the addition of VEGF to the cells. At similar time points, the cells were fixed and processed for immunocytochemistry to evaluate ChAT and phosphorylated neurofilament expression. Results: Exposure to ALS-CSF caused morphological changes of NSC-34 cells like reduced differentiation and aggregation of phosphorylated neurofilaments. Enhanced LDH activity and reduced ChAT immunoreactivity were also observed. Addition of VEGF to NSC-34 cells exposed to ALS-CSF was protective in terms of reduced LDH activity and restoration of ChAT expression. Conclusion: The present study confirms that VEGF exerts a neuroprotective effect on the NSC-34 cell line by attenuating the degenerative changes induced by ALS-CSF. It thus has therapeutic potential in sporadic ALS.

Brain-Derived Neurotrophic Factor Facilitates Functional Recovery from ALS-Cerebral Spinal Fluid-Induced Neurodegenerative Changes in the NSC-34 Motor Neuron Cell Line

“…These patients were assessed for their functional status using the modified Norris scoring system and the ALS Functional Rating Scale (ALSFRS) [34]. The patients' mean age and disease duration matched well with our earlier studies [19,20]. A neurologist (A.N.)…”

“…Kage et al [16] found its levels to decline in the spinal cord tissue of ALS patients, whereas Petri et al [17] showed that both FGF-2 and FGFR1 were preserved, even at the end stage of the disease. VEGF has recently emerged as a key neurotrophic factor for the growth, survival, and plasticity of motor neurons [18,19,20]. …”

The Neuroprotection Exerted by Memantine, Minocycline and Lithium, against Neurotoxicity of CSF from Patients with Amyotrophic Lateral Sclerosis, Is Antagonized by Riluzole

“…Additionally, ALS/CSF only increases the cytosolic calcium concentration in motor neurons [24]. Furthermore, in NSC34, a hybrid cell line of neuroblastoma and motor neuron, ALS/CSF also exerts cytotoxic actions; however, this effect is only seen in differentiated cells [35,36]. Thus, it seems reasonable to conclude that the cytotoxic effect elicited by ALS/CSF on our MCN cultures is mostly exerted on cortical motor neurons.…”

Advantages and Pitfalls in Experimental Models Of ALS

“…the involvement of MAP kinases (Guo & Bhat, 2007), the interaction between bcl2-A1 and pro-caspase-3 (Iaccarino et al, 2011) or the involvement of kynurenine pathway implicated in the regulatory mechanisms of the immune response . In addition, different therapeutic agents have been tested in culture, such as the vascular endothelial growth factor (VEGF) (Kulshreshtha et al, 2011) or the semi-synthetic tetracycline called minocycline (Guo & Bhat, 2007), known for their neuroprotective effects in neurodegeneration models (Orsucci et al, 2009;Storkebaum et al, 2004). Similarly, other cell lines have been transfected with WT or mutant (G93A) hSOD1 and used for the same purpose: N18TG2 neuroblastoma, the non-neuronal Madin-Darby Canine Kidney (MDCK) (Raimondi et al, 2006) or the mouse embryonic fibroblast NIH3T3 cell lines (Shinder et al, 2001).…”