Bombesin stimulation of gastrin release from canine gastrin cells in primary culture.
Bombesin stimulation of gastrin release from canine gastrin cells in primary culture.
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铃蟾肽刺激原代培养的犬胃泌素细胞释放胃泌素。
DOI：
10.1152/ajpgi.1987.252.3.g413
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发表时间：
1987
期刊：
影响因子：
--
通讯作者：
Walsh,JH
中科院分区：
文献类型：
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作者：
Giraud,AS;Soll,AH;Cuttitta,F;Walsh,JH
本文报道了犬胃窦胃泌素(G)细胞的分离和短期培养方法。组织用酶分散，淋洗浓缩G细胞，培养40h，G细胞占12%，含生长抑素或5-羟色胺的细胞不到2%。蛙皮素(0.001-100 pm)能以线性方式刺激细胞培养2小时内释放胃泌素。蛙素特异性单抗2A11呈剂量依赖性地阻断蛙皮素的刺激作用。生长抑素(0.001-1,000 nM)抑制蛙皮素刺激的胃泌素释放。抗生长抑素抗体(单抗S8)可阻断外源性生长抑素的抑制作用，但不改变蛙皮素刺激或基础胃泌素的释放。P物质(SP)类似物Spantide(1 nM-1微米)不能抑制蛙皮素刺激的胃泌素释放。受体后，Forsklin激活腺苷环化酶，佛波酯β-TPA激活蛋白激酶C，导致胃泌素释放。钙离子载体A23187也以剂量依赖的方式释放胃泌素。这种方法允许对胃窦G细胞进行浓缩和短期培养；这些细胞具有刺激性蛙黄素和抑制生长抑素受体，这表明这些多肽对胃窦G细胞有直接作用。此外，G细胞被cAMP和钙/磷脂酰肌醇依赖的机制激活。
A method is described for the isolation and short-term culture of canine antral gastrin (G) cells. Tissue was dispersed by enzymes and G cells enriched by elutriation and cultured for 40 h. These cultures contained 12% G cells and less than 2% somatostatin- or serotonin-containing cells. Bombesin (0.001–100 pM) potently stimulated gastrin release from cell cultures in a linear fashion over 2 h. The bombesin-specific monoclonal antibody 2A11 dose-dependently blocked bombesin stimulation. Somatostatin (0.001–1,000 nM) inhibited bombesin-stimulated gastrin release. Antibody to somatostatin (Mab S8) prevented the inhibition by exogenous somatostatin but did not alter bombesin-stimulated or basal gastrin release. The substance P (SP) analogue spantide (1 nM-1 microM) did not inhibit bombesin-stimulated gastrin release. Postreceptor activation of adenylate cyclase by forskolin and of protein kinase C by the phorbol ester, beta-TPA, caused gastrin release. The calcium ionophore A23187 also released gastrin in a dose-dependent fashion. This methodology allows enrichment and short-term culture of antral G cells; these cells have stimulatory bombesin and inhibitory somatostatin receptors, suggesting that these peptides have a direct action on antral G cells. Furthermore, G cells are activated by cAMP and calcium/phosphatidylinositol-dependent mechanisms.