Retinoic Acid-Differentiated Neuroblastoma SH-SY5Y Is an Accessible In Vitro Model to Study Native Human Acid-Sensing Ion Channels 1a (ASIC1a)

Abstract

Human neuroblastoma SH-SY5Y is a prominent neurobiological tool used for studying neuropathophysiological processes. We investigated acid-sensing (ASIC) and transient receptor potential vanilloid-1 (TRPV1) and ankyrin-1 (TRPA1) ion channels present in untreated and differentiated neuroblastoma SH-SY5Y to propose a new means for their study in neuronal-like cells. Using a quantitative real-time PCR and a whole-cell patch-clamp technique, ion channel expression profiles, functionality, and the pharmacological actions of their ligands were characterized. A low-level expression of ASIC1a and ASIC2 was detected in untreated cells. The treatment with 10 μM of retinoic acid (RA) for 6 days resulted in neuronal differentiation that was accompanied by a remarkable increase in ASIC1a expression, while ASIC2 expression remained almost unaltered. In response to acid stimuli, differentiated cells showed prominent ASIC-like currents. Detailed kinetic and pharmacological characterization suggests that homomeric ASIC1a is a dominant isoform among the present ASIC channels. RA-treatment also reduced the expression of TRPV1 and TRPA1, and minor electrophysiological responses to their agonists were found in untreated cells. Neuroblastoma SH-SY5Y treated with RA can serve as a model system to study the effects of different ligands on native human ASIC1a in neuronal-like cells. This approach can improve the characterization of modulators for the development of new neuroprotective and analgesic drugs.

Lignans as Pharmacological Agents in Disorders Related to Oxidative Stress and Inflammation: Chemical Synthesis Approaches and Biological Activities

“…Sevanol in vitro dose-dependently inhibited human and rat ASIC3 channels and, although with less efficiency, rat ASIC1a channels, heterologously expressed in oocytes of Xenopus laevis . In the model of neuronal-like cells, differentiated from the SH-SY5Y cell line by retinoic acid, sevanol showed an inhibitory effect on native human ASIC1a [ 118 ]. In Complete Freund’s Adjuvant (CFA)-induced thermal hyperalgesia test in vivo, sevanol showed an anti-inflammatory effect by significantly increasing the withdrawal latency on a hot plate and reducing edema of the inflamed hind paw [ 119 , 120 , 121 ].…”

Hypoxia Induces DPSC Differentiation versus a Neurogenic Phenotype by the Paracrine Mechanism

“…To verify the ability of DPSCs to induce neuronal differentiation by an autocrine/paracrine mechanism mediated by growth factor secretion, we verified the ability of DPSC-CM (5H, 16H) to differentiate SH-SY5Y neuroblastoma cells into cells possessing a more mature, neuron-like phenotype. As demonstrated by several authors, SH-SY5Y cells, in their undifferentiated form, express immature neuronal markers and lack mature neuronal markers, but following treatment with differentiation-inducing agents, SH-SY5Y cells become morphologically more like primary neurons [60].…”

“…These abilities make DPSCs an efficient cellular model for the study of neurodegenerative diseases and could contribute to autologous and allogenic cell therapy for CNS injury [36,60,[69][70][71][72].…”

“…In SH-SY5Y exposed for 10 days with 16H CM, the neuronal marker expression of NFH and β3-Tubulin reached values of 96.1% and 92.8%, respectively (Figure 4B). ronal markers, but following treatment with differentiation-inducing agents, SH-SY5Y cells become morphologically more like primary neurons [60].…”

Mechanisms of Action of the Peptide Toxins Targeting Human and Rodent Acid-Sensing Ion Channels and Relevance to Their In Vivo Analgesic Effects

“…A higher membrane targeting of the ASIC1a subunit was observed in acutely resected human cortical tissue (from patients undergoing surgical treatment of intractable epilepsy) compared to mice, possibly linked to a more efficient trafficking due to an amino acid difference at position 285 between mASIC1a and hASIC1a [ 213 ]. A dominant expression of ASIC1 compared to ASIC2 was also reported from human central neuronal cell line cultures [ 214 ], associated with a dominant ASIC1a current (strongly inhibited by PcTx1) and a native hASIC1a current was also recorded from neuroblastoma differentiated into neuronal-like phenotype, associated with the co-expression of ASIC2 but not ASIC3, leading to some Mamb-sensitive heterotrimeric ASIC1a/2a [ 215 ].…”