原代大脑皮层培养物中兴奋性氨基酸受体刺激的磷酸肌醇代谢

Excitatory amino acid receptor-stimulated phosphoinositide turnover in primary cerebrocortical cultures.

作者信息

Birrell G J, Marcoux F W

机构信息

Parke-Davis Pharmaceutical Research Division, Warner-Lambert Company, Ann Arbor, MI 48106.

出版信息

Br J Pharmacol. 1993 Jun;109(2):379-85. doi: 10.1111/j.1476-5381.1993.tb13580.x.

DOI:10.1111/j.1476-5381.1993.tb13580.x

PMID:8395285

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2175676/

Abstract

Characterization of excitatory amino acid-induced accumulation of [3H]-phosphoinositides was carried out in primary cerebrocortical cultures isolated from foetal rats. 2. All of the excitatory amino acid receptor agonists examined caused concentration-dependent enhancement of phosphoinositide (PI) formation. The most potent excitatory amino acid receptor agonists were quisqualate, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid ((1S,3R)-ACPD), ibotenate and glutamate with mean EC50 values of 0.9 +/- 0.4 microM, 15 +/- 5 microM, 15 +/- 3 microM and 41 +/- 8 microM respectively. 3. The selective ionotropic receptor antagonists kynurenic acid (1 mM), 2,3-dihydroxy-6-nitro-7-sulphamoyl-benzo(F)quinoxaline (NBQX, 10 microM) and (+/-)-4-(3-phosphonopropyl)-2 piperazinecarboxylic acid (CPP, 100 microM), failed to block responses to quisqualate, (1S,3R)-ACPD or glutamate. D,L-2-Amino-3-phosphonopropionate (D,L-AP3) did not block 1S,3R-ACPD or quisqualate-induced PI turnover, but had an additive effect with quisqualate or (1S,3R)-ACPD. 4. Exposure of cultures to agonists in the absence of added extracellular calcium reduced the maximal quisqualate response by approximately 45%, revealing a two-component concentration-response curve. Concentration-response curves to ibotenate and glutamate became flattened by omission of extracellular calcium, whereas (1S,3R)-ACPD-stimulated PI turnover was unaffected. 5. Pretreatment of cultures with pertussis toxin markedly inhibited PI responses evoked by (1S,3R)-ACPD. 6. These results suggest that excitatory amino acid-stimulated PI turnover in cerebrocortical cultures is independent of ionotropic receptor activation and is mediated via specific G-protein-linked metabotropic receptors. The partial dependence of the responses to quisqualate, ibotenate and glutamate on the presence of extracellular calcium suggests that the effects of these agonists may be mediated by more than one receptor subtype.

摘要

在从胎鼠分离的原代大脑皮质培养物中进行了兴奋性氨基酸诱导的[3H] - 磷酸肌醇积累的特性研究。2. 所检测的所有兴奋性氨基酸受体激动剂均引起磷酸肌醇（PI）形成的浓度依赖性增强。最有效的兴奋性氨基酸受体激动剂是喹啉酸、（1S，3R）-1-氨基环戊烷-1,3-二羧酸（（1S，3R）-ACPD）、鹅膏蕈氨酸和谷氨酸，其平均EC50值分别为0.9±0.4微摩尔、15±5微摩尔、15±3微摩尔和41±8微摩尔。3. 选择性离子型受体拮抗剂犬尿烯酸（1毫摩尔）、2,3-二羟基-6-硝基-7-氨磺酰基苯并（F）喹喔啉（NBQX，10微摩尔）和（±）-4-（3-膦酰基丙基）-2-哌嗪羧酸（CPP，100微摩尔）未能阻断对喹啉酸、（1S，3R）-ACPD或谷氨酸的反应。D，L-2-氨基-3-膦酰基丙酸（D，L-AP3）未阻断（1S，3R）-ACPD或喹啉酸诱导的PI周转，但与喹啉酸或（1S，3R）-ACPD有相加作用。4. 在无细胞外钙添加的情况下将培养物暴露于激动剂可使喹啉酸的最大反应降低约45%，显示出双组分浓度 - 反应曲线。省略细胞外钙会使鹅膏蕈氨酸和谷氨酸的浓度 - 反应曲线变平，而（1S，3R）-ACPD刺激的PI周转不受影响。5. 用百日咳毒素预处理培养物可显著抑制（1S，3R）-ACPD引起的PI反应。6. 这些结果表明，大脑皮质培养物中兴奋性氨基酸刺激的PI周转独立于离子型受体激活，并通过特定的G蛋白偶联代谢型受体介导。对喹啉酸、鹅膏蕈氨酸和谷氨酸反应对细胞外钙存在的部分依赖性表明，这些激动剂的作用可能由不止一种受体亚型介导。