Autoregulation in PC12 cells via P2Y receptors: Evidence for non-exocytotic nucleotide release from neuroendocrine cells.
Autoregulation in PC12 cells via P2Y receptors: Evidence for non-exocytotic nucleotide release from neuroendocrine cells.
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DOI：
10.1007/s11302-007-9062-2
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发表时间：
2007-09
影响因子：
3.5
通讯作者：
Boehm S
中科院分区：
文献类型：
--
作者：
Hussl S;Kubista H;Boehm S
核苷酸不仅从神经元释放，而且从各种其他类型的细胞释放，包括成纤维细胞、上皮细胞、内皮细胞和神经胶质细胞。虽然从非神经细胞释放ATP通常是Ca 2+非依赖性的，并且大多数是非囊泡的，但神经元ATP释放通常被认为是通过胞吐作用发生的。为了评估从神经内分泌细胞释放核苷酸是否可能涉及非囊泡组分，使用P2 Y12受体的自分泌/旁分泌激活作为从PC 12细胞释放核苷酸的生物传感器。编码肉毒杆菌毒素C1轻链的质粒的表达导致在PC 12膜的免疫印迹中检测到的突触融合蛋白1的减少。与此同时，自发以及去极化诱发的释放先前纳入[3 H]去甲肾上腺素从转染的细胞显着减少相比，从未转染的细胞释放，从而表明，胞吐受损。在表达肉毒杆菌毒素C1轻链的PC 12细胞中，ADP降低环AMP合成的程度与未转染细胞相同。同样，由于P2 Y12受体的阻断或由于腺苷三磷酸双磷酸酶降解细胞外核苷酸而引起的环AMP合成的增强在未转染的和肉毒杆菌毒素C1轻链表达细胞之间没有不同。然而，去极化诱发释放内源性核苷酸引起的环AMP合成的抑制被取消或大大减少在表达肉毒杆菌毒素C1轻链的细胞中。总之，这些结果表明，从神经内分泌细胞的自发核苷酸释放可能发生独立的囊泡胞吐，而去极化诱发的核苷酸释放主要依赖于胞吐机制。
Nucleotides are released not only from neurons, but also from various other types of cells including fibroblasts, epithelial, endothelial and glial cells. While ATP release from non-neural cells is frequently Ca2+ independent and mostly non-vesicular, neuronal ATP release is generally believed to occur via exocytosis. To evaluate whether nucleotide release from neuroendocrine cells might involve a non-vesicular component, the autocrine/paracrine activation of P2Y12 receptors was used as a biosensor for nucleotide release from PC12 cells. Expression of a plasmid coding for the botulinum toxin C1 light chain led to a decrease in syntaxin 1 detected in immunoblots of PC12 membranes. In parallel, spontaneous as well as depolarization-evoked release of previously incorporated [3H]noradrenaline from transfected cells was significantly reduced in comparison with the release from untransfected cells, thus indicating that exocytosis was impaired. In PC12 cells expressing the botulinum toxin C1 light chain, ADP reduced cyclic AMP synthesis to the same extent as in non-transfected cells. Likewise, the enhancement of cyclic AMP synthesis either due to the blockade of P2Y12 receptors or due to the degradation of extracellular neucleotides by apyrase was not different between non-transfected and botulinum toxin C1 light chain expressing cells. However, the inhibition of cyclic AMP synthesis caused by depolarization-evoked release of endogenous nucleotides was either abolished or greatly reduced in cells expressing the botulinum toxin C1 light chain. Together, these results show that spontaneous nucleotide release from neuroendocrine cells may occur independently of vesicle exocytosis, whereas depolarization-evoked nucleotide release relies predominantly on exocytotic mechanisms.
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影响因子：
4.8
作者：
Gerasimovskaya, EV;Ahmad, S;Stenmark, KR
通讯作者：
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影响因子：
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DOI：
10.1152/ajprenal.00293.2000
发表时间：
2002-02-01
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