Klamt 2013 Abstract IOC79

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Klamt F (2013) Differences in basal mitochondrial respiratory parameters and reactive species generation between the cholinergic, dopaminergic and tumoral phenotypes of SH-SY5Y human neuroblastoma cell line. Mitochondr Physiol Network 18.07.

Link:

Bristot IJ, Lopes FM, de Medeiros LM, Lisboa da Motta L, de Oliveira Porciuncula L, Klamt F (2013)

Event: IOC79

The molecular mechanisms underlying the neuronal lost in several neurodegenerative diseases are not completely understood. Part of this difficulty is related to the lack of reliable experimental models. Aiming to establish better in vitro models for neuroscience research, our group previously established the experimental conditions to differentiate the human neuroblastoma SH-SY5Y cell line into dopaminergic and cholinergic neuron-like cells with the use of retinoic acid (RA) and brain derived neurotrophic factor (BDNF), respectively. Since the mitochondrial dysfunction and bioenergetics dysregulation are associated with neurodegenerative diseases and cancer, we aimed here to characterize the differences in the abovementioned parameters between the cholinergic, dopaminergic and tumoral phenotypes of SH-SY5Y human neuroblastoma cell line. Exponentially growing SH-SY5Y cells were maintained with DMEM/F12 medium plus 10% of fetal bovine serum (FBS). Dopaminergic differentiation was triggered by the combination of 10 μM RA plus 1% of FBS for 7 days. Cholinergic differentiation was induced by the addition of BDNF at the 4th day of RA-treatment. Oroboros Oxygraph-2k was used for high-resolution respirometry, where O2 consumption, basal/maximum respiration rates, respiratory reserve and residual O2 consumption were evaluated. Reactive species generation were determined using DCF-DA oxidation and Amplex Red® fluorescence. Mitotracker dye was used to analyze differences in mitochondrial morphology between phenotypes. Our data demonstrates relevant differences between these phenotypes, which could help to establish then as in vitro cell model to be used for molecular and cellular studies in Parkinson/Alzheimer disease and in tumors of the central nervous system.

Keywords: tumor cell metabolism, lung cancer, in vitro neuronal model, neuronal differentiation

O2k-Network Lab: BR Porto Alegre Klamt F


Labels: MiParea: Respiration, mt-Medicine  Pathology: Alzheimer's, Neurodegenerative, Parkinson's 

Organism: Human  Tissue;cell: Neuroblastoma  Preparation: Intact cells 


Coupling state: ROUTINE 

HRR: Oxygraph-2k 



References

Ivi Juliana Bristot1,2, Fernanda Martins Lopes1,2, Liana Marengo de Medeiros1,2, Leonardo Lisbôa da Motta1,2, Lisiane de Oliveira Porciúncula1, Fábio Klamt1,2

  1. Laboratory of Cellular Biochemistry (24), Department of Biochemistry, ICBS / Federal University of Rio Grande do Sul (UFRGS)
  2. National Institutes for Sciences & Technology Translational Medicine (MCT/CNPq INCT-TM);

Support

MCT/CNPq Universal (470306/2011-4), PqG/FAPERGS (2414-2551/12-8), PRONEX/FAPERGS (1000274), PRONEN/FAPERGS (1120325), and MCT/CNPq INCT-TM (573671/2008-7).