Leptin Chronic Effect on Differentiation, Ion Currents and Protein Expression in N1E-115 Neuroblastoma Cells

Rebeca Isabel Vergara-Reyes, Patricia Cervantes-Acosta, Antonio Hernández-Beltrán, Manuel Barrientos-Morales and Belisario Domínguez-Mancera

Background and Objective: Arcuate nucleus (ARC), a component of appetite-regulatory factors, contains populations of both orexigenic and anorexigenic neurons and one of the fundamental components of its system is leptin. Studies have evidenced the critical neurotrophic role in the development of ARC. To determine such effects on neuron development, N1E-115 neuroblastoma cells were used as an ARC model. Materials and Methods: N1E-115 neuroblastoma cells were treated with leptin [10 nM] for 24, 48 and 72 hrs. Dimethyl sulfoxide (DMSO) 1.5% was used as a known drug that promotes neurite expression. Cells percentage (%) that developed neurites was evaluated by bright field microscopy. Patch-clamp electrophysiology was used to analyze membrane ion currents, RT-PCR for quantifying changes in mRNA expression of anorexic peptides, proopiomelanocortin (POMC) and cocaine and amphetamine-related transcript (CART), in addition to principal Na_v, Ca_v ion channel subunits. Results: N1E-115 cells treated with leptin show neurite expression after 24 hrs of treatment, similar effects were obtained with DMSO. Leptin (time-dependent) increases the inward current in comparison with the control value at 72 hrs. Outward currents were not affected by leptin. Leptin and DMSO increased Na⁺ and Ca²⁺ current without changes in the kinetic properties. Lastly, leptin promotes an increase in mRNA level expression of transcripts to POMC, CART, Na_v1.2 and Ca_v1.3. Conclusion: Leptin chronic treatment promotes neurite expression, Up-regulation of Na⁺ and Ca²⁺ ion channels determining neuronal excitability, besides increasing the mRNA level expression of anorexic peptides POMC and CART in neuroblastoma N1E-115.

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