Lysophosphatidic acid-induced membrane ruffling and brain-derived neurotrophic factor gene expression are mediated by ATP release in primary microglia
Lysophosphatidic acid-induced membrane ruffling and brain-derived neurotrophic factor gene expression are mediated by ATP release in primary microglia
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DOI：
10.1111/j.1471-4159.2008.05599.x
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发表时间：
2008-10-01
影响因子：
4.7
通讯作者：
Ueda, Hiroshi
中科院分区：
文献类型：
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作者：
Fujita, Ryousuke;Ma, Yan;Ueda, Hiroshi
我们研究了溶血磷脂酸（LPA）对小胶质细胞的影响，这可能在神经病理性疼痛的发生和维持中起重要作用。扫描电子显微镜检测到LPA引起膜皱褶，并增加大鼠小胶质细胞原代培养物中脑源性神经营养因子（BDNF）的表达，该细胞表达LPA（3），但不表达LPA（1）或LPA（2）受体。这些作用被G α（q/11）-反义寡脱氧核苷酸（AS-ODN）、磷脂酶C（PLC）抑制剂U 73122和特异性降解ATP和ADP的腺苷三磷酸双磷酸酶抑制。当使用荧光素酶生物发光测定法测量ATP释放时，LPA显示出以LPA（3）和PLC底物可逆的方式增加ATP释放。然而，LPA诱导的ATP释放也被G α（q/11）AS-ODN阻断，但不被百日咳毒素阻断。这些结果表明，LPA通过LPA（3）受体、G α（q/11）和PLC诱导原代培养的大鼠小胶质细胞释放ATP，释放的ATP或异位转化的ADP可能通过P2 Y（12）受体和G α（i/o）激活引起膜皱褶，并通过P2 X（4）受体激活BDNF表达。
We examined the effects of lysophosphatidic acid (LPA) on microglia, which may play an important role in the development and maintenance of neuropathic pain. LPA caused membrane ruffling as detected by scanning electron microscopy, and increased the expression of brain-derived neurotrophic factor (BDNF) in a primary culture of rat microglia, which express LPA(3), but not LPA(1) or LPA(2) receptors. These actions were inhibited by a G alpha(q/11)-antisense oligodeoxynucleotide (AS-ODN), U73122, an inhibitor of phospholipase C (PLC), and apyrase, which specifically degrades ATP and ADP. When ATP release was measured using a luciferin-luciferase bioluminescence assay, LPA was shown to increase it in an LPA(3) and PLC inhibitor-reversible manner. However, LPA-induced ATP release was also blocked by the G alpha(q/11) AS-ODN, but not by pertussis toxin. These results suggest that LPA induces the release of ATP from rat primary cultured microglia via the LPA(3) receptor, G alpha(q/11) and PLC, and that the released ATP or ectopically converted ADP may in turn cause membrane ruffling via P2Y(12) receptors and G alpha(i/o) activation, and BDNF expression via activation of P2X(4) receptors.