Data di Pubblicazione:
2012
Citazione:
Gene regulation of the receptor GPR17, a key player in oligodendroglial differentiation / D. Lecca, A. Fratangeli, S. Ceruti, P. Rosa, M.P. Abbracchio. ((Intervento presentato al convegno Opportunity and challenges in the pharmacological modulation of adult neural stem cells tenutosi a Novara nel 2012.
Abstract:
The P2Y-like receptor GPR17 is a G protein-coupled receptor regulating oligodendrogliogenesis. Under physiological conditions, it is expressed in early oligodendrocyte precursor cells (OPCs), representing multipotent slowly-proliferating precursors in the adult brain. We have demonstrated that the activation of GPR17 by its endogenous agonists (i.e. uracil nucleotides and cysteinyl-leukotrienes) promotes OPC maturation. GPR17 is necessary to address OPC differentiation to mature oligodendrocytes, but its expression has to be turned off before terminal maturation. However, the molecular mechanisms at the basis of GPR17 gene regulation are still unknown.
To identify candidate factors involved in induction/repression of GPR17 we have developed a reporter assay. First, we cloned two putative promoter regions of the human GPR17 gene into a reporter vector upstream to a gene encoding for an engineered firefly luciferase and we have transfected these constructs in oligodendroglial cells such as Oli-neu, an immortalized cell line that can be differentiated in vitro to mature oligodendrocytes. Then, we evaluated the bioluminescence produced in response to an array of stimuli, including nucleotides, hormones and growth factors. Results demonstrated that treatment with forskolin induced a 2-fold increase of GPR17 promoter activity in the murine oligodendroglial cell line Oli-neu, suggesting regulation via cAMP-responsive elements. Moreover, incubation of cells with conditioned medium from cortical neurons was able to significantly induce GPR17 promoter activity, even if we still don’t know which is the neuronal factor responsible of this effect. This increase of GPR17 expression after exposure to conditioned medium has been confirmed with both real time PCR and immunocytochemistry.
Understanding how the expression of GPR17 is finely regulated will not only advance basic knowledge in oligodendrogliogenesis, but will also help developing new pharmacological strategies to stimulate the reparative potential of the quiescent OPCs that are still present in the adult brain.
Tipologia IRIS:
14 - Intervento a convegno non pubblicato
Elenco autori:
D. Lecca, A. Fratangeli, S. Ceruti, P. Rosa, M.P. Abbracchio
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