Expression of functional NR1/NR2B-type NMDA receptors in neuronally differentiated SK-N-SH human cell line

Marina Pizzi, Flora Boroni, Andrea Bianchetti, Costantinos Moraitis, Ilenia Sarnico, Marina Benarese, Francesca Goffi, Alessandra Valero, PierFranco F. Spano

Research output: Contribution to journalArticlepeer-review

Abstract

The present study demonstrates that human SK-N-SH neuroblastoma cells, differentiated by retinoic acid (RA), express functional NMDA receptors and become vulnerable to glutamate toxicity. During exposure to RA, SK-N-SH cells switched from non-neuronal to neuronal phenotype by showing antigenic changes typical of postmitotic neurons together with markers specific for cholinergic cells. Neuronally differentiated cells displayed positive immunoreactivity to the vesicular acetylcholine transporter and active acetylcholine release in response to depolarizing stimuli. The differentiation correlated with the expression of NMDA receptors. RT-PCR and immunoblotting analysis identified NMDA receptor subunits NR1 and NR2B, in RA-differentiated cultures. The NR1 protein immunolocalized to the neuronal cell population and assembled with the NR2B subunit to form functional N-methyl-D-aspartate (NMDA) receptors. Glutamate or NMDA application, concentration-dependently increased the intracellular Ca2+ levels and acetylcholine release in differentiated cultures, but not in undifferentiated SK-N-SH cells. Moreover, differentiated cultures became vulnerable to NMDA receptor-mediated excitotoxicity. The glutamate effects were enhanced by glycine application and were prevented by the NMDA receptor blocker MK 801, as well as by the NR2B selective antagonist ifenprodil. These data suggest that SK-N-SH cells differentiated by brief treatment with RA may represent an unlimited source of neuron-like cells suitable for studying molecular events associated with activation of human NR1/NR2B receptors.

Original languageEnglish
Pages (from-to)2342-2350
Number of pages9
JournalEuropean Journal of Neuroscience
Volume16
Issue number12
DOIs
Publication statusPublished - 2002

Keywords

  • Acetylcholine release
  • Excitotoxicity
  • Ifenprodil
  • Neuroblastoma
  • NR1
  • NR2B

ASJC Scopus subject areas

  • Neuroscience(all)

Fingerprint Dive into the research topics of 'Expression of functional NR1/NR2B-type NMDA receptors in neuronally differentiated SK-N-SH human cell line'. Together they form a unique fingerprint.

Cite this