Modulation of muscarinic cholinoceptor-stimulated inositol 1,4,5-trisphosphate accumulation by N-methyl-D-aspartate in neonatal rat cerebral cortex

• Published in: Neuropharmacology Vol. 33; no. 1; p. 15
• Main Authors: Challis, R A, Mistry, R, Gray, D W, Nahorski, S R
• Format: Journal Article
• Language: English
• Published: England 01-01-1994
• Subjects: 2-Amino-5-phosphonovalerate - pharmacology; Animals; Animals, Newborn - metabolism; Carbachol - pharmacology; Cerebral Cortex - drug effects; Cerebral Cortex - metabolism; Dizocilpine Maleate - pharmacology; Female; Glycine - pharmacology; In Vitro Techniques; Inositol 1,4,5-Trisphosphate - metabolism; Magnesium - pharmacology; Male; N-Methylaspartate - pharmacology; Rats; Rats, Wistar; Receptors, Muscarinic - drug effects
• ISSN: 0028-3908
• DOI: 10.1016/0028-3908(94)90092-2

The mechanisms by which N-methyl-D-aspartate (NMDA) receptor activation can modulate muscarinic receptor-stimulated phosphoinositide turnover have been studied in neonatal rat cerebral cortex slices. A maximally effective concentration of carbachol (1 mM) caused a large stimulation of both total [3H]inositol phosphate ([3H]InsPx) accumulation (30-40-fold over basal levels after 15 min in the presence of 5 mM LiCl) and inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] mass accumulation (consisting of a rapid peak increase of about 8-10-fold within 15 sec followed by a sustained plateau rise of 4-5-fold which persisted for > 10 min). Low concentrations of NMDA enhanced carbachol-stimulated [3H]InsPx and Ins(1,4,5)P3 accumulations with a maximal effect being observed at 10 microM NMDA. However, at higher concentrations of NMDA (30-300 microM) a dramatic inhibition of these indices of phosphoinositide turnover was observed. Time-course studies demonstrated that NMDA (100 microM) caused a significant enhancement of the initial increases in [3H]InsPx and Ins(1,4,5)P3 accumulations stimulated by carbachol, with the profound inhibitory effects becoming evident at longer incubation times. The modulatory effects of NMDA were antagonized by D-2-amino-5-phosphonopentanoate and MK-801. Reducing extracellular calcium concentration ([Ca2+]e) to the low micromolar range decreased basal Ins(1,4,5)P3 accumulation and attenuated the response to carbachol. Under these conditions NMDA (10-100 microM) caused only a potentiation of agonist-stimulated Ins(1,4,5)P3 accumulation. Under control conditions ([Ca2+]e = 1.3 mM), addition of MK-801 (1 microM) 10 min after carbachol + 100 microM NMDA challenge failed to reverse the inhibitory effect of NMDA on carbachol-stimulated [3H]InsPx accumulation. Furthermore, pre-incubation of cerebral cortex slices with 100 microM NMDA for 15 min (followed by extensive washing of slices to remove NMDA) dramatically decreased [3H]inositol incorporation into the cellular inositol phospholipid fraction and decreased basal and carbachol-stimulated Ins(1,4,5)P3 mass accumulations. We conclude that the enhancement of agonist-stimulated phosphoinositide turnover seen at concentrations of NMDA up to 10 microM may be due to Ca2+ entry and Ca2+ facilitation of phosphoinositide-specific phospholipase C activity. In contrast, the inhibitory effect of high concentrations of NMDA on agonist-stimulated phosphoinositide turnover may be due to progressive, irreversible and, at least in part, Ca(2+)-dependent damage to the cell populations in the slice preparation responding to muscarinic-receptor stimulation.