Neuroprotective properties of RT10, a fraction isolated from Parawixia bistriata spider venom, against excitotoxicity injury in neuron-glia cultures

Abstract

Background: L- Glutamate ( L- Glu), the major excitatory neurotransmitter in the mammalian Central Nervous System (CNS), is essential to cognitive functions. However, when L- Glu is accumulated in large concentrations at the synaptic cleft, it can induce excitotoxicity that results in secondary damage implicated in many neurological disorders. Current therapies for the treatment of neurological disorders are ineffective and have side effects associated with their use; therefore, there is a need to develop novel treatments. In this regard, previous studies have shown that neuroactive compounds obtained from the venom of the spider Parawixia bistriata have neuroprotective effects in vitro and in vivo. In this sense, this work aimed to evaluate potential neuroprotective effects of fraction RT10, obtained from this spider venom, on primary cultures of neuron and glial cells subjected to glutamate excitotoxicity insults. Methods: Primary cultures of neurons and glia were obtained from the cerebral tissue of 1-day-old postnatal Wistar rats. After 7 days in vitro (DIV), the cultures were incubated with fraction RT10 (0.002; 0.02; 0.2 and 2 µg/µL) or riluzole (100 µM) for 3-hours before application of 5 mM L- Glu. After 12 hours, the resazurin sodium salt (RSS) test was applied to measure metabolic activity and proliferation of living cells, whereas immunocytochemistry for MAP2 was performed to measure neuronal survival. In addition, the cells were immunolabeled with NeuN and GFAP in baseline conditions. Results: In the RSS tests, we observed that pre-incubation with RT10 before the excitotoxic insults from L -Glu resulted in neuroprotection, shown by a 10% reduction in the cell death level. RT10 was more effective than riluzole, which resulted in a cell-death reduction of 5%. Moreover, qualitative analysis of neuronal morphology (by MAP2 staining, expressed as fluorescence intensity (FI), an indirect measure of neuronal survival) indicate that RT10 reduced the toxic effects of L -Glu, as shown by a 38 % increase in MAP2 fluorescence when compared to L -Glu insult. On the other hand, the riluzole treatment resulted in 17% increase of MAP2 fluorescence; therefore, the neuroprotection from RT10 was more efficacious. Conclusion: RT10 fraction exhibits neuroprotective effects against L -Glu excitotoxicity in neuron-glia cultured in vitro .

The synergic effects of presynaptic calcium channel antagonists purified from spiders on memory elimination of glutamate-induced excitotoxicity in the rat hippocampus trisynaptic circuit

“…Therefore, the concurrent use of N- and P/Q-type VGCC blockers shows a synergic effect. Additionally, our results are supported by some previously published studies in which NMDA antagonists were used against excitotoxicity ( Rodrigues et al, 2001 ; Fontana et al, 2003 ; Rodrigues et al, 2004 ; Primini et al, 2019 ). In some other studies, the Parawixin protein family extracted from the Parawixia bistriata spider was examined.…”

The synergic effects of presynaptic calcium channel antagonists purified from spiders on memory elimination of glutamate-induced excitotoxicity in the rat hippocampus trisynaptic circuit

“…Therefore, the concurrent use of N- and P/Q-type VGCC blockers shows a synergic effect. Additionally, our results are supported by some previously published studies in which NMDA antagonists were used against excitotoxicity ( Rodrigues et al, 2001 ; Fontana et al, 2003 ; Rodrigues et al, 2004 ; Primini et al, 2019 ). In some other studies, the Parawixin protein family extracted from the Parawixia bistriata spider was examined.…”

Scutellariae Radix Ameliorates Prenatal Stress Induced Anxiety-like and Depression-like Behavior in the Offspring via Reversing HPA Axis Hyperfunction and Ameliorating Neurodevelopmental Dysfunction

“…The results demonstrated that SR mitigated the neurogenesis defects observed in the hippocampal DG of offspring mice. Glu serves as the primary excitatory neurotransmitter in the mammalian central nervous system [49]. Previous investigations have demonstrated that stress can induce damage to the central nervous system, leading to an increased concentration of Glu in the synaptic space.…”

Neuroactive venom compounds obtained from Phlogiellus bundokalbo as potential leads for neurodegenerative diseases: insights on their acetylcholinesterase and beta-secretase inhibitory activities in vitro

“…Current therapies for treating neurological diseases are ineffective and have side effects associated with their use; therefore, there is a need to develop novel therapies. In this regard, previous studies have shown that neuroactive compounds obtained from the spider venom with neuroprotective effects in vitro and in vivo [ 73 ].…”

The synergic effects of presynaptic calcium channel antagonists purified from spiders on memory elimination of glutamate-induced excitotoxicity in the rat hippocampus trisynaptic circuit

“…Therefore, the concurrent use of N- and P/Q-type VGCC blockers shows a synergic effect. Additionally, our results are supported by some previously published studies in which NMDA antagonists were used against excitotoxicity ( Rodrigues et al, 2001 ; Fontana et al, 2003 ; Rodrigues et al, 2004 ; Primini et al, 2019 ). In some other studies, the Parawixin protein family extracted from the Parawixia bistriata spider was examined.…”

Scutellariae Radix Ameliorates Prenatal Stress Induced Anxiety-like and Depression-like Behavior in the Offspring via Reversing HPA Axis Hyperfunction and Ameliorating Neurodevelopmental Dysfunction

“…The results demonstrated that SR mitigated the neurogenesis defects observed in the hippocampal DG of offspring mice. Glu serves as the primary excitatory neurotransmitter in the mammalian central nervous system [49]. Previous investigations have demonstrated that stress can induce damage to the central nervous system, leading to an increased concentration of Glu in the synaptic space.…”

Neuroactive venom compounds obtained from Phlogiellus bundokalbo as potential leads for neurodegenerative diseases: insights on their acetylcholinesterase and beta-secretase inhibitory activities in vitro

“…Current therapies for treating neurological diseases are ineffective and have side effects associated with their use; therefore, there is a need to develop novel therapies. In this regard, previous studies have shown that neuroactive compounds obtained from the spider venom with neuroprotective effects in vitro and in vivo [ 73 ].…”

The synergic effects of presynaptic calcium channel antagonists purified from spiders on memory elimination of glutamate-induced excitotoxicity in the rat hippocampus trisynaptic circuit

“…Therefore, the concurrent use of N- and P/Q-type VGCC blockers shows a synergic effect. Additionally, our results are supported by some previously published studies in which NMDA antagonists were used against excitotoxicity ( Rodrigues et al, 2001 ; Fontana et al, 2003 ; Rodrigues et al, 2004 ; Primini et al, 2019 ). In some other studies, the Parawixin protein family extracted from the Parawixia bistriata spider was examined.…”

Scutellariae Radix Ameliorates Prenatal Stress Induced Anxiety-like and Depression-like Behavior in the Offspring via Reversing HPA Axis Hyperfunction and Ameliorating Neurodevelopmental Dysfunction

“…The results demonstrated that SR mitigated the neurogenesis defects observed in the hippocampal DG of offspring mice. Glu serves as the primary excitatory neurotransmitter in the mammalian central nervous system [49]. Previous investigations have demonstrated that stress can induce damage to the central nervous system, leading to an increased concentration of Glu in the synaptic space.…”

Neuroactive venom compounds obtained from Phlogiellus bundokalbo as potential leads for neurodegenerative diseases: insights on their acetylcholinesterase and beta-secretase inhibitory activities in vitro

“…Current therapies for treating neurological diseases are ineffective and have side effects associated with their use; therefore, there is a need to develop novel therapies. In this regard, previous studies have shown that neuroactive compounds obtained from the spider venom with neuroprotective effects in vitro and in vivo [ 73 ].…”