Man'ko V V, Bychkova S V, Klevets' M Iu
Ukr Biokhim Zh (1999). 2004 Jan-Feb;76(1):65-71.
The presence of two types of well-characterised Ca2+ release channels, namely IP3-receptors (Ins(1,4,5)P3Rs) and ryanodine-receptors (RyRs), was detected in the salivary glands secretory cells of Chironomus plumosus L. For this aim different blockators and activators of these Ca2+ -transport systems were used. The conditions for permeabilization of these cells by saponine were experimentally chosen for their more intensive action. It was shown that IP3 decreased calcium content in saponine-treated gland tissue by (41.14 +/- 11.75)%. The effect of IP3 was not observed under condition of heparin and eosin Y presence in the incubation medium, but heparin alone did not cause any action on calcium content in saponine-treated gland tissue. The observed effects of IP3 are supposed to be the evidences of Ins (1,4,5)P3Rs presence in the intracellular membrane of this object. It was also shown that calcium content in intact gland tissue increased by (67.12 +/- 22.60)% in presence of heparin (500 mkg/ml) in the incubation medium. This effect of heparin was also observed with presence of verapamil (100 mkM) and eosin Y (5, 20 mkM) in incubation medium. So, this effect is not connected with function of voltage-gated Ca2+ -channels and Ca2+ -pumps. Ryanodine in concentration of 5nM decreased calcium content in saponine-treated gland tissue by (35.18 +/- 3.87)% but it caused the increase of calcium content at high concentration (500 nM) by (40.72 +/- 12.52)%. It improved the presence of RyRs in intracellular membrane of secretory cells of this object. Besides, these channels, perhaps, belong to "non-sensitive" to caffeine, because caffeine did not affect calcium content in the gland tissue neither in presence nor with absence of eosin Y.
在摇蚊唾液腺分泌细胞中检测到两种特征明确的Ca2+释放通道,即肌醇三磷酸受体(Ins(1,4,5)P3Rs)和兰尼碱受体(RyRs)。为此,使用了这些Ca2+转运系统的不同阻断剂和激活剂。通过实验选择了皂苷使这些细胞透化的条件,以使其作用更强。结果表明,肌醇三磷酸使皂苷处理的腺体组织中的钙含量降低了(41.14±11.75)%。在孵育培养基中存在肝素和伊红Y的情况下未观察到肌醇三磷酸的作用,但单独的肝素对皂苷处理的腺体组织中的钙含量没有任何影响。观察到的肌醇三磷酸的作用被认为是该对象细胞内膜中存在Ins (1,4,5)P3Rs的证据。还表明,在孵育培养基中存在肝素(500μg/ml)时,完整腺体组织中的钙含量增加了(67.12±22.60)%。在孵育培养基中存在维拉帕米(100μM)和伊红Y(5、20μM)时也观察到了肝素的这种作用。因此,这种作用与电压门控Ca2+通道和Ca2+泵的功能无关。5nM浓度的兰尼碱使皂苷处理的腺体组织中的钙含量降低了(35.18±3.87)%,但在高浓度(500 nM)时使钙含量增加了(40.72±12.52)%。这证明了该对象分泌细胞的内膜中存在兰尼碱受体。此外,这些通道可能属于对咖啡因“不敏感”的类型,因为无论有无伊红Y,咖啡因都不会影响腺体组织中的钙含量。