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ETUDE DES EFFETS DE LA STIMULATION ELECTRIQUE A HAUTE FREQUENCE DANS UN MODELE CELLULAIRE IN VITRO

Abstract : A system for electrical stimulation on cells lines in vitro was optimized for studying cellular and molecular mechanism of high frequency electrical stimulation. Two cells lines (GH3 and PC12) were analysed at the transcriptomic and proteomic level as well as for the impact on hormonal secretion and neurotransmitter release. We compared PRL secretion of GH3 cell by HFS, LFS and dopamine as well as catecolamine (DA, AD and NA) secretion of PC12 cell treated by HFS, LFS and 6-OHDA. Proteomic synthesis of stimulated cells were analysed by 35S methionine incorporation and by SELDI-TOF-MS. At last we have investigated modifications of gene expression of stimulated cells using DNA nylon microarray based on long oligonucleotides and radioactive detection. First experiment demonstrated that HFS induced a significant decrease of the amount of prolactin to a level comparable to the level obtained with dopamine, one well known inhibitor of prolactin. Similarly, the amount of catecholamines detected in the cell culture medium was significantly down regulated. Transcriptomic data manifested the existence of characteristic expression profiles for high frequency electrical stimulation. About 100 discriminatory genes were individualized upon HFS involved in the proteomic synthesis, calcium signalisation and cellular energetic. We confirmed the impact of HFS on protein synthesis with SELDI-TOF technique and methionine incorporation.One original HFS mechanism has been validated, involving protein synthesis neutralisation in the reacted inactivation of neuronal structures. Implication of calcium signalisation and PKC pathway were also suggested.
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https://tel.archives-ouvertes.fr/tel-00009865
Contributor : Rong Xia <>
Submitted on : Monday, August 1, 2005 - 4:12:33 AM
Last modification on : Friday, November 6, 2020 - 4:08:15 AM
Long-term archiving on: : Friday, November 25, 2016 - 9:20:17 AM

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Rong Xia. ETUDE DES EFFETS DE LA STIMULATION ELECTRIQUE A HAUTE FREQUENCE DANS UN MODELE CELLULAIRE IN VITRO. Biologie cellulaire. Université Joseph-Fourier - Grenoble I, 2005. Français. ⟨tel-00009865⟩

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