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Progranulin (PGRN) functions in neuronal cultures Guo, Aobo

Abstract

Null mutations in the progranulin gene (PGRN) have been identified as a major cause of frontotemporal lobe dementia with ubiquitinated inclusions (FTLD-U). In this disorder, the ubiquitinated aggregated protein inclusions of a normally nuclear-located RNA processing protein called TAR DNA binding protein (TDP-43) accumulate in the cytoplasm of neurons. To determine whether aspects of this clinical pathology can be established in primary cultures of mouse cortical neurons, PGRN levels were knocked down in cultures using lentiviral vectors to introduce siRNA constructs. Similar to observations in the brains of FTLD-U patients, TDP-43 levels were markedly decreased in the nucleus of PGRN knockdown neurons relative to cytoplasmic levels in comparison to control neurons and significantly recovered by over-expressed PGRN, although the TDP-43 cleavage was not obvious in this model. Meanwhile, depletion of PGRN elevated levels of activated caspase-3, and the PGRN-deficient neurons demonstrated enhanced vulnerability to sub-lethal doses of NMDA and H₂O₂. These results show that it is possible to recapitulate key features of the PGRN null mutation in a culture dish within a short period of time and suggest that the seeds of this form of frontotemporal dementia may be sown early in life.

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