Role of myosin Va in purinergic vesicular neurotransmission in the gut.
Role of myosin Va in purinergic vesicular neurotransmission in the gut.
复制标题
DOI：
10.1152/ajpgi.00330.2011
复制
发表时间：
2012-03-15
期刊：
影响因子：
--
通讯作者：
Goyal RK
中科院分区：
文献类型：
--
作者：
Chaudhury A;He XD;Goyal RK
我们检验了这样的假设：肌球蛋白 Va 通过将嘌呤能囊泡转运至静脉曲张膜进行胞吐作用，在嘌呤能囊泡神经传递中发挥关键作用。研究在野生型 (WT) 和肌球蛋白 Va 缺乏的稀、棕色、九孔雀 (DBA) 小鼠中进行。在小鼠窦肌条上进行细胞内微电极记录。在屏蔽氮能连接电位后，在非肾上腺素能非胆碱能条件下记录嘌呤能抑制连接电位（pIJP）。 DBA 小鼠表现出 pIJP 降低，但对 P2Y1 受体激动剂 MRS-2365 的超极化反应正常。为了研究 DBA 小鼠嘌呤能传递减少的机制，通过超速离心和蔗糖缓冲纯化从整个肠道组织匀浆中获得分离的静脉曲张，进行了研究。通过对囊泡 ATP 转运蛋白（溶质载体蛋白 SLC17A9）进行免疫染色，在组织切片和孤立的静脉曲张中鉴定出嘌呤能静脉曲张。 WT 和 DBA 小鼠的静脉曲张相似。与 WT 静脉曲张相比，DBA 静脉曲张中的肌球蛋白 Va 显着降低。邻近连接实验显示肌球蛋白Va与SLC17A9密切相关。通过静脉曲张的 FM1-43 染色检查囊泡胞吐作用，结果表明与 WT 静脉曲张相比，DBA 静脉曲张在 KCl 刺激后胞吐作用受损。这些研究表明，SLC17A9 可以识别含有 ATP 的嘌呤能静脉曲张。肌球蛋白 Va 与 SLC17A9 染色的囊泡结合，并可能将其转运至静脉曲张膜进行胞吐作用。在肌球蛋白 Va 缺乏的小鼠中，嘌呤能抑制性神经传递受损。
We examined the hypothesis that myosin Va, by transporting purinergic vesicles to the varicosity membrane for exocytosis, plays a key role in purinergic vesicular neurotransmission. Studies were performed in wild-type (WT) and myosin Va-deficient dilute, brown, nonagouti (DBA) mice. Intracellular microelectrode recordings were made in mouse antral muscle strips. Purinergic inhibitory junction potential (pIJP) was recorded under nonadrenergic noncholinergic conditions after masking the nitrergic junction potentials. DBA mice showed reduced pIJP but normal hyperpolarizing response to P2Y1 receptor agonist MRS-2365. To investigate the mechanism of reduced purinergic transmission in DBA mice, studies were performed in isolated varicosities obtained from homogenates of whole gut tissues by ultracentrifugation and sucrose cushion purification. Purinergic varicosities were identified in tissue sections and in isolated varicosities by immunostaining for the vesicular ATP transporter, the solute carrier protein SLC17A9. The varicosities were similar in WT and DBA mice. Myosin Va was markedly reduced in DBA varicosities compared with the WT varicosities. Proximity ligation assay showed that myosin Va was closely associated with SLC17A9. Vesicular exoendocytosis was examined by FM1–43 staining of varicosities, which showed that exoendocytosis after KCl stimulation was impaired in DBA varicosities compared with WT varicosities. These studies show that SLC17A9 identifies ATP-containing purinergic varicosities. Myosin Va associates with SLC17A9-stained vesicles and possibly transports them to varicosity membrane for exocytosis. In myosin Va-deficient mice, purinergic inhibitory neurotransmission is impaired.
登录
查看更多内容
DOI：
10.3410/b1-46
发表时间：
2009-06-09
期刊：
F1000 biology reports
影响因子：
--
作者：
Burnstock G
通讯作者：
Burnstock G
影响因子：
25
作者：
Lewis, Tommy L., Jr.;Mao, Tianyi;Svoboda, Karel;Arnold, Don B.
通讯作者：
Arnold, Don B.
DOI：
10.1146/annurev-cellbio-100809-151502
发表时间：
2011
影响因子：
11.3
作者：
Hartman MA;Finan D;Sivaramakrishnan S;Spudich JA
通讯作者：
Spudich JA
影响因子：
64.5
作者：
CHENEY, RE;OSHEA, MK;MOOSEKER, MS
通讯作者：
MOOSEKER, MS
DOI：
10.1152/ajpgi.00164.2011
发表时间：
2011-09-01
影响因子：
4.5
作者：
Chaudhury, Arun;He, Xue-Dao;Goyal, Raj K.
通讯作者：
Goyal, Raj K.