Abstract
The Golgi apparatus (GA) appears disrupted in motor neurons of amyotrophic lateral sclerosis (ALS). Here, mouse motor neuron-like NSC-34 cell lines stably expressing human superoxide dismutase 1 (hSOD1)wt and mutant hSOD1G93A, as an ALS cell model, were constructed. The number of cells with disrupted GA increased from 14% to 34%. Furthermore, NSC-34/hSOD1G93A cells showed lower levels of proliferation and differentiation. GA disruption was not caused by apoptosis as determined by several techniques including caspase-3 activation. Similarly, spinal cords from ALS patients did not show caspase-3 activation. Therefore, NSC-34/hSOD1G93A cells are a suitable cell model to study GA dysfunction in ALS.
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Acknowledgments
We thank Dr. Francis Barr, Max-Planck-Institut für Biochemie, Martinsried, Germany, for polyclonal antibodies FBA 34 and FBA42; Prof. Neil Cashman, Centre for Research in Neurodegenerative Diseases, University of Toronto, Canada, for the NSC-34 cells; Prof. Don Cleveland, Neuroscience and Cellular and Molecular Medicine, University of California of San Diego, for SOD1 coding plasmids; Dr. Paula Alves, Dr. Helena Vieira and Marlene Carmo, Laboratory of Animal Cell Technology, ITQB, for help with FACS analysis; Prof. Mamede de Carvalho, Faculdade de Medicina de Lisboa, Portugal, and Prof. Michael Swash, The Royal London Hospital, for fruitful discussion; Cell Imaging Service (Instituto Gulbenkian de Ciência, Oeiras, Portugal) for the use of the confocal microscope. CG and AP were recipients of PhD fellowships from Fundação para a Ciência e a Tecnologia (FCT), Portugal. This work was funded by projects POCTI/BCI/38631/2001 and POCTI/CBO/43952/2002 from FCT and project STREP LSH-CT-2004-503228 from the European Commission to JC, and projects AG-10124 and AG-17586 to JQT.
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Gomes, C., Palma, A.S., Almeida, R. et al. Establishment of a cell model of ALS disease: Golgi apparatus disruption occurs independently from apoptosis. Biotechnol Lett 30, 603–610 (2008). https://doi.org/10.1007/s10529-007-9595-z
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DOI: https://doi.org/10.1007/s10529-007-9595-z