title: Involvement of the Notch pathway in terminal astrocytic differentiation: role of PKA.
abstract: The Notch pathway is a highly conserved signaling system essential for modulating neurogenesis and promoting astrogenesis. Similarly, the cAMP signaling cascade can promote astrocytic commitment in several cell culture models, such as the C6 glioma cell line. These cells have the capacity to differentiate into oligodendrocytes or astrocytes, characteristics that allow their use as a glial progenitor model. In this context, we explore here the plausible involvement of cAMP in Notch-dependent signal transactions. The exposure of C6 cells to a non-hydrolysable cAMP analogue resulted in a sustained augmentation of Notch activity, as detected by nuclear translocation of its intracellular domain portion (NICD) and transcriptional activity. The cAMP effect is mediated through the activation of the γ-secretase complex, responsible for Notch cleavage and is sensitive to inhibitors of the cAMP-dependent protein kinase, PKA. As expected, Notch cleavage and nuclear translocation resulted in the up-regulation of the mRNA levels of one of its target genes, the transcription factor Hair and enhancer of split 5. Moreover, the glutamate uptake activity, as well as the expression of astrocytic markers such as glial fibrillary acidic protein, S100β protein and GLAST was also enhanced in cAMP-exposed cells. Our results clearly suggest that during the process of C6 astrocytic differentiation, cAMP activates the PKA/γ-secretase/NICD/RBPJ(κ) pathway and Notch1 expression, leading to transcriptional activation of the genes responsible for glial progenitor cell fate decision.
authors.forename: Carla
authors.initials: C
authors.lastname: Angulo-Rojo
authors.affiliation: †Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México.
authors.forename: Rebeca
authors.initials: R
authors.lastname: Manning-Cela
authors.affiliation: ‡Department of Pharmacological Sciences, State University of New York at Stony Brook, NY, U.S.A.
authors.forename: Adán
authors.initials: A
authors.lastname: Aguirre
authors.affiliation: *Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México.
authors.forename: Arturo
authors.initials: A
authors.lastname: Ortega
authors.affiliation: *Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México.
authors.forename: Esther
authors.initials: E
authors.lastname: López-Bayghen
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000187:drug effects; D001254:Astrocytoma / Q000473:pathology; D003994:Bucladesine / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D005576:Colforsin / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism*; D007546:Isoquinolines / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D051880:Receptors, Notch / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology*
chemical_list: D000276:Adjuvants, Immunologic; D005904:Glial Fibrillary Acidic Protein; D007546:Isoquinolines; D047428:Protein Kinase Inhibitors; D051880:Receptors, Notch; D013449:Sulfonamides; D005576:Colforsin; D003994:Bucladesine; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide