TY - JOUR
T1 - The metabotropic P2Y 4 receptor participates in the commitment to differentiation and cell death of human neuroblastoma SH-SY5Y cells
AU - Cavaliere, Fabio
AU - Nestola, Valeria
AU - Amadio, Susanna
AU - D'Ambrosi, Nadia
AU - Angelini, Daniela F.
AU - Sancesario, Giuseppe
AU - Bernardi, Giorgio
AU - Volonté, Cinzia
PY - 2005/2
Y1 - 2005/2
N2 - Extracellular nucleotides exert a variety of biological actions through different subtypes of P2 receptors. Here we characterized in the human neuroblastoma SH-SY5Y cells the simultaneous presence of various P2 receptors, belonging to the P2X ionotropic and P2Y metabotropic families. Western blot analysis detected the P2X 1,2,4,5,6,7 and P2Y 1,2,4,6, but not the P2X 3 and P2Y 12 receptors. We then investigated which biological effects were mediated by the P2Y 4 subtype and its physiological pyrimidine agonist UTP. We found that neuronal differentiation of the SH-SY5Y cells with dibutiryl-cAMP increased the expression of the P2Y 4 protein and that UTP itself was able to positively interfere with neuritogenesis. Moreover, transient transfection and activation of P2Y 4 also facilitated neuritogenesis in SH-SY5Y cells, as detected by morphological phase contrast analysis and confocal examination of neurofilament proteins NFL. This was concurrent with increased transcription of immediate-early genes linked to differentiation such as cdk-5 and NeuroD6, and activity of AP-1 transcription family members such as c-fos, fos-B, and jun-D. Nevertheless, a prolonged activation of the P2Y 4 receptor by UTP also induced cell death, both in naive, differentiated, and P2Y 4- transfected SH-SY5Y cells, as measured by direct count of intact nuclei and cytofluorimetric analysis of damaged DNA. Taken together, our data indicate that the high expression and activation of the P2Y 4 receptor participates in the neuronal differentiation and commitment to death of SH-SY5Y cells.
AB - Extracellular nucleotides exert a variety of biological actions through different subtypes of P2 receptors. Here we characterized in the human neuroblastoma SH-SY5Y cells the simultaneous presence of various P2 receptors, belonging to the P2X ionotropic and P2Y metabotropic families. Western blot analysis detected the P2X 1,2,4,5,6,7 and P2Y 1,2,4,6, but not the P2X 3 and P2Y 12 receptors. We then investigated which biological effects were mediated by the P2Y 4 subtype and its physiological pyrimidine agonist UTP. We found that neuronal differentiation of the SH-SY5Y cells with dibutiryl-cAMP increased the expression of the P2Y 4 protein and that UTP itself was able to positively interfere with neuritogenesis. Moreover, transient transfection and activation of P2Y 4 also facilitated neuritogenesis in SH-SY5Y cells, as detected by morphological phase contrast analysis and confocal examination of neurofilament proteins NFL. This was concurrent with increased transcription of immediate-early genes linked to differentiation such as cdk-5 and NeuroD6, and activity of AP-1 transcription family members such as c-fos, fos-B, and jun-D. Nevertheless, a prolonged activation of the P2Y 4 receptor by UTP also induced cell death, both in naive, differentiated, and P2Y 4- transfected SH-SY5Y cells, as measured by direct count of intact nuclei and cytofluorimetric analysis of damaged DNA. Taken together, our data indicate that the high expression and activation of the P2Y 4 receptor participates in the neuronal differentiation and commitment to death of SH-SY5Y cells.
KW - Dibutiryl-cAMP
KW - Extracellular nucleotides
KW - Immediate-early genes
KW - Neurofilament proteins
KW - UTP
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UR - http://www.scopus.com/inward/citedby.url?scp=11844266620&partnerID=8YFLogxK
U2 - 10.1016/j.nbd.2004.09.001
DO - 10.1016/j.nbd.2004.09.001
M3 - Article
C2 - 15649700
AN - SCOPUS:11844266620
VL - 18
SP - 100
EP - 109
JO - Neurobiology of Disease
JF - Neurobiology of Disease
SN - 0969-9961
IS - 1
ER -