title: NMDA receptor modulation by a conditioned medium derived from rat cerebellar granule cells.
abstract: Our previous studies have shown that the response to the excitotoxic action of glutamate by cultured cerebellar granule cells depends upon the cell density or the volume of medium in which they have been grown: the higher the cell density or the lower the volume, the higher the response to glutamate. We have hypothesized that this variable response is due to the formation in culture of a glutamate-sensitizing activity GSA more abundantly in conditioned medium derived from high-density or low-volume cultures than that present in low-density or high volume cultures and capable of restoring sensitivity in previously resistant granule cells. In order to elucidate the mechanism of action of glutamate-sensitizing activity, we measured the extent and function of NMDA receptors in low- and high-volume cultures and assessed the effect of glutamate-sensitizing activity on the same receptors. We found that under high-volume conditions the extent of MK-801 binding, the amount of NMDA receptor type 1, the currents evoked in whole cells after an NMDA pulse and the response of cultured cells to this ligand were markedly reduced compared with low-volume cultures. Addition of glutamate-sensitizing activity to high-volume cultures increased their glutamate sensitivity, the NMDA-evoked currents, the extent of MK-801 binding and the amount of NMDA receptor type 1 protein present. The corresponding mRNA transcripts, on the contrary, were unchanged in high-volume, low-volume and high-volume GSA-treated cultures.
authors.affiliation: Istituto di Neurobiologia, CNR, Rome, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Dus
authors.forename: N
authors.initials: N
authors.lastname: Canu
authors.forename: C
authors.initials: C
authors.lastname: Zona
authors.forename: M T
authors.initials: MT
authors.lastname: Ciotti
authors.forename: P
authors.initials: P
authors.lastname: Calissano
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology*; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D016291:Dizocilpine Maleate / Q000494:pharmacology; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D018698:Glutamic Acid / Q000378:metabolism; D016202:N-Methylaspartate / Q000633:toxicity; D009474:Neurons / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D018408:Patch-Clamp Techniques; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism*; D014316:Tritium
chemical_list: D017077:Culture Media, Conditioned; D018690:Excitatory Amino Acid Agonists; D018691:Excitatory Amino Acid Antagonists; D012333:RNA, Messenger; D016194:Receptors, N-Methyl-D-Aspartate; D014316:Tritium; D018698:Glutamic Acid; D016202:N-Methylaspartate; D016291:Dizocilpine Maleate