Vitamin B12-impaired metabolism produces apoptosis and Parkinson phenotype in rats expressing the transcobalamin-oleosin chimera in substantia nigra
2009
Orozco-Barrios , Carlos Enrique (Centro de Investigación y de Estudios Avanzados del Instituto Politécnico NacionalUniversité Nancy 2, MexicoNancy(France). Department of Physiology, Biophysics and NeuroscienceFaculté de Médecine) | Battalia-Hsu , Shyue-Fang (Université Nancy 2, Nancy(France). Faculté de Médecine) | Arango-Rodriguez , Martha Ligia (Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Mexico(Mexique). Department of Physiology, Biophysics and Neuroscience) | Ayala-Davila , José (Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Mexico(Mexique). Department of Physiology, Biophysics and Neuroscience) | Chery , Celine (Université Nancy 2, Nancy(France). Faculté de Médecine) | Alberto , Jean-Marc (Université Nancy 2, Nancy(France). Faculté de Médecine) | Schroeder , Henry (INRA , Vandoeuvre-Les-Nancy (France). USC 0340 USC INRA / INPL / ENSAIA / Univ. Nancy 1 : Animal et Fonctionnalités des Produits Animaux) | Daval , Jean-Luc (Université Nancy 2, Nancy(France). Faculté de Médecine) | Martinez-Fong , Daniel (Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Mexico(Mexique). Department of Physiology, Biophysics and Neuroscience) | Guenant , Jean-Louis (Université Nancy 2, Nancy(France). Faculté de Médecine)
Background: Vitamin B12 is indispensable for proper brain functioning and cytosolic synthesis of S-adenosylmethionine. Whether its deficiency produces effects on viability and apoptosis of neurons remains unknown. There is a particular interest in investigating these effects in Parkinson disease where Levodopa treatment is known to increase the consumption of Sadenosylmethionine. To cause deprivation of vitamin B12, we have recently developed a cell model that produces decreased synthesis of S-adenosylmethionine by anchoring transcobalamin (TCII) to the reticulum through its fusion with Oleosin (OLEO). [br/] Methodology: Gene constructs including transcobalamin-oleosin (TCII-OLEO) and control constructs, green fluorescent protein-transcobalamin-oleosin (GFP-TCII-OLEO), oleosin-transcobalamin (OLEO-TCII), TCII and OLEO were used for expression in N1E-115 cells (mouse neuroblastoma) and in substantia nigra of adult rats, using a targeted transfection with a Neurotensin polyplex system. We studied the viability and the apoptosis in the transfected cells and targeted tissue. The turning behavior was evaluated in the rats transfected with the different plasmids. [br/] Principal Findings: The transfection of N1E-115 cells by the TCII-OLEO-expressing plasmid significantly affected cell viability and increased immunoreactivity of cleaved Caspase-3. No change in propidium iodide uptake (used as a necrosis marker) was observed. The transfected rats lost neurons immunoreactive to tyrosine hydroxylase. The expression of TCII-OLEO was observed in cells immunoreactive to tyrosine hydroxylase of the substantia nigra, with a superimposed expression of cleaved Caspase-3. These cellular and tissular effects were not observed with the control plasmids. Rats transfected with TCII-OLEO expressing plasmid presented with a significantly higher number of turns, compared with those transfected with the other plasmids. [br/] Conclusions/Significance: In conclusion, the TCII-OLEO transfection was responsible for apoptosis in N1E-115 cells and rat substantia nigra and for Parkinson-like phenotype. This suggests evaluating whether vitamin B12 deficit could aggravate the PD in patients under Levodopa therapy by impairing S-adenosylmethionine synthesis in substantia nigra.
Mostrar más [+] Menos [-]Información bibliográfica
Este registro bibliográfico ha sido proporcionado por Institut national de la recherche agronomique