Calcium dysregulation, mitochondrial pathology and protein aggregation in a culture model of amyotrophic lateral sclerosis: Mechanistic relationship and differential sensitivity to intervention

Mitochondrial dysfunction in familial amyotrophic lateral sclerosis

“…Cytosolic ATP levels were significantly reduced in neuroblastoma cells expressing mutant SOD1 G37R (Coussee et al 2011) and decreased more rapidly and to a greater extent in rotenonetreated SOD1 G93A neuronal cells . Impairment of intracellular Ca 2+ homeostasis has been reported in cells expressing mutant SOD1 G93A or SOD1 G37R (Carri et al 1997;Coussee et al 2011;Tradewell et al 2011) and in motor neurons from mutant SOD1 G93A transgenic mice Kruman et al 1999). Also isolated mitochondria from transgenic mutant SOD1 G93A and SOD1 G85R mice showed a significant decrease in Ca 2+ loading capacity (Damiano et al 2006;Nguyen et al 2009;Vila et al 2003).…”

In Vivo and In Vitro Models to Study Amyotrophic Lateral Sclerosis

“…Indeed, at the E12-13 embryonic developmental stage, the spinal cord of mice embryos is easy to access and remove, and motor neurons, astrocytes and microglia can be extracted, purified and maintain in culture with high viability yields (Gingras, et al, 2007b;Sanelli, et al, 2007;Schnaar and Schaffner, 1981). These cells can be dissociated from the spinal cord and cultured together to facilitate motor neuron survival (Tradewell, et al, 2011;Tradewell and Durham, 2010). Motor neurons will be easy to identify because of their large cell body (>20µm in diameter) and dendritic trees, and can be stained for expression of the transcription factor Hb9 and choline acetyltransferase, specific molecular markers of motor neuron (Fig 3) (Gingras, et al, 2007b;Tradewell, et al, 2011).…”

“…These cells can be dissociated from the spinal cord and cultured together to facilitate motor neuron survival (Tradewell, et al, 2011;Tradewell and Durham, 2010). Motor neurons will be easy to identify because of their large cell body (>20µm in diameter) and dendritic trees, and can be stained for expression of the transcription factor Hb9 and choline acetyltransferase, specific molecular markers of motor neuron (Fig 3) (Gingras, et al, 2007b;Tradewell, et al, 2011). Cultures of purified motor neurons without the trophic support of glial cells are difficult to maintain for more than 2 weeks (Bar, 2000;Lunn, et al, 2009).…”

“…Cultures of purified motor neurons without the trophic support of glial cells are difficult to maintain for more than 2 weeks (Bar, 2000;Lunn, et al, 2009). In order to perform studies on a chronic disease like ALS, especially while looking for long-term cell survival effects, dissociated spinal cord cell cultures constitute a better choice as they can be maintain in culture for up to 7 weeks (Tradewell, et al, 2011). Purified motor neurons can also be cultured on an astrocyte feeder layer for several weeks on which they display characteristics (neurite pattern) closer to adult motor neurons (Bar, 2000).…”

“…Purified motor neurons can also be cultured on an astrocyte feeder layer for several weeks on which they display characteristics (neurite pattern) closer to adult motor neurons (Bar, 2000). To reproduce an ALS phenotype, motor neurons can be induced to express multiple copies of the gene of interest, for example, mutated human G93A SOD1, by microinjection of the vectors into the cells identified by their specific morphology (Tradewell, et al, 2011;Tradewell and Durham, 2010). Motor neurons can also be purified and cultured separately, to allow their adenoviral transduction with G93A SOD1 (Lunn, et al, 2009), and then, to enhance their survival, they can be plated on a glial cell feeder layer treated with arabinofuranosyl cytidine (AraC), to prevent cell proliferation (De Paola, et al, 2008).…”