SILENCING OF PNPLA6, THE NEUROPATHY TARGET ESTERASE (NTE) CODIFYING GENE, ALTERS HUMAN NEURODIFFERENTIATION IN VITRO

ABSTRACT: Neuropathy Target Esterase (NTE) is a protein involved in the development of a polyneuropathy caused by exposure to certain organophosphorus compounds. In vivo and in vitro studies have also associated NTE with embryonic development since NTE null mice embryos are non-viable, and silencing the NTE-codifying gene (Pnpla6) in mouse embryonic stem cells strongly alters the differentiation of vascular and nervous systems. In this paper, human embryonal carcinoma stem cells (hNT2) are used as in vitro neurodifferentiation model to determine whether PNPLA6 gene silencing is able to alter the differentiation into a neuronal phenotype. In controls, the PNPLA6 mRNA levels increased in parallel with other neuroectodermal markers during the neurodifferentiation. PNPLA6 gene silencing with specific interference RNA led to a 97% maximum decrease in gene expression by day 3 after transfection, with a maximum of 50% reduction in NTE enzymatic activity by day 4. Microarray analyses of the PNPLA6-silenced cells showed alterations in several developmental processes, mainly neurogenesis and epithelium tube morphogenesis. PNPLA6 silencing also led to a reduction in electrical activity and an altered neuronal phenotype. This work is the first proof supporting the hypothesis that NTE protein plays a critical role in human early neurodevelopment using a human cell differentiation model. PNPA6 gene silencing in undifferentiatied hNT2 cells was performed, together with a Control (non-gene silencing cells) and a Negative Control (non-specific RNA silencer). Three biological replicates.

ORGANISM(S): Homo sapiens

SUBMITTER: Marco Fabbri 

PROVIDER: E-GEOD-55761 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress