Published February 1, 2024
| Version v1
Publication
Electrical pulse stimulation parameters modulate N2a neuronal differentiation
Contributors
Others:
- Universidad de Sevilla. Departamento de Biología Celular
- Universidad de Sevilla. Departamento de Bioquímica y Biología Molecular
- Universidad de Sevilla. Departamento de Tecnología Electrónica
- Universidad de Sevilla. BIO132: Citoquímica Ultraestructural
- Universidad de Sevilla. CTS257: Envejecimiento y Neurodegeneración
- Universidad de Sevilla. TIC178: Diseño y Test de Circuitos Integrados de Señal Mixta
- MCIN/ AEI /10.13039/501100011033/ and FEDER PID2021-122529OB-I00
Description
Electrical pulse stimulation has been used to enhance the differentiation or proliferation of neuronal progenitor cells in tissue
engineering and cancer treatment. Therefore, a comprehensive investigation of the effects caused by its parameters is crucial for
improvements in those fields. We propose a study of pulse parameters, to allow the control of N2a cell line fate and behavior. We
have focused on designing an experimental setup that allows for the knowledge and control over the environment and the
stimulation signals applied. To map the effects of the stimulation on N2a cells, their morphology and the cellular and molecular
reactions induced by the pulse stimulation have been analyzed. Immunofluorescence, rt-PCR and western blot analysis have been
carried out for this purpose, as well as cell counting. Our results show that low-amplitude electrical pulse stimulation promotes
proliferation of N2a cells, whilst amplitudes in the range 250 mV/mm–500 mV/mm induce differentiation. Amplitudes higher than
750 mV/mm produce cell damage at low frequencies. For high frequencies, large amplitudes are needed to cause cell death. An
inverse relation has been found between cell density and pulse-induced neuronal differentiation. The best condition for neuronal
differentiation was found to be 500 mV/mm at 100 Hz. These findings have been confirmed by up-regulation of the Neurod1 gene.
Our preliminary study of the molecular effects of electrical pulse stimulation on N2a offers premonitory clues of the PI3K/Akt/GSK3β pathway implications on the neuronal differentiation process through ES. In general, we have successfully mapped the
sensitivity of N2a cells to electrical pulse stimulation parameters.
Additional details
Identifiers
- URL
- https://idus.us.es/handle//11441/154445
- URN
- urn:oai:idus.us.es:11441/154445