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Expression of the neurosecretory process in pc12 cells is governed by rest
Author(s) -
D’Alessandro Rosalba,
Klajn Andrijana,
Stucchi Laura,
Podini Paola,
Malosio Maria Luisa,
Meldolesi Jacopo
Publication year - 2008
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2008.05259.x
Subject(s) - exocytosis , transfection , vesicle , microbiology and biotechnology , repressor , biology , gene silencing , transcription factor , transcription (linguistics) , gene , secretion , endocrinology , genetics , linguistics , philosophy , membrane
The neurosecretory process is acquired during differentiation and can be lost en block by differentiated cells. To investigate the role of REST/NRSF, a transcription repressor, in the maintenance of the process we studied two PC12 clones, one wt and one defective, expressing low and high levels of endogenous RE‐1 silencing transcription (factor) (REST), respectively. Stable transfection of constructs demonstrated that REST represses 10 genes coding for proteins of neurosecretory vesicles and their exocytosis, eight including and two lacking the REST‐binding sequence, RE‐1. Of these genes, those of chromogranins were strongly repressed by fewfold increases of REST, those of VAMP2 and syntaxin1a required much higher levels. Moreover, in wt cells transfected with an active construct the dense‐core vesicles, still competent for regulated exocytosis, were much smaller, with lighter cores; in defective cells, the dominant‐negative construct induced the rescue of many vesicle/exocytosis genes but not of those of chromogranins. Small dense‐core vesicles, exocytized upon stimulation, were rescued when the construct‐transfected defective cells were transfected also with chromograninA or treated with trichostatinA, a blocker of histone deacetylases. Our results identify REST, working by direct and indirect mechanisms, as the factor governing the maintenance of the neurosecretory process and the properties of dense‐core vesicles in PC12 cells.