Mechanosensitive Hydrolysis of ATP and ADP in Lamina Propria of the Murine Bladder by Membrane-Bound and Soluble Nucleotidases.
Mechanosensitive Hydrolysis of ATP and ADP in Lamina Propria of the Murine Bladder by Membrane-Bound and Soluble Nucleotidases.
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DOI：
10.3389/fphys.2022.918100
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发表时间：
2022
影响因子：
4
通讯作者：
中科院分区：
文献类型：
--
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先前的研究表明，尿路上皮释放的5′-三磷酸腺苷（ATP）在膀胱机械传导中具有突出的作用。尿路上皮ATP通过激活在固有层（LP）中的许多细胞类型（包括传入神经元）中表达的嘌呤能受体来调节排尿周期，并且可能对尿路上皮和逼尿肌之间的直接机械敏感信号传导也很重要。ATP的兴奋作用被酶水解终止，随后产生生物活性代谢物。我们研究了可能的机械敏感性机制ATP水解的LP通过确定1，N 6 -乙烯基-ATP（eATP）的降解在非扩张（空）或扩张（全）小鼠（C57 BL/6 J）逼尿肌无膀胱模型的前腔侧，使用HPLC。eATP和eADP的水解是更大的接触LP的膨胀比nonexpanded膀胱，而水解的eAMP在填充过程中保持不变，这表明在LP中的eATP水解的一些步骤是机械敏感的。eATP和eADP也在与无逼尿肌膀胱的LP接触的腔外溶液（ELS）中被分解代谢，但在添加底物之前从器官腔中除去。这两种嘌呤的降解是更大的ELS从膨胀比从nondiffilled的准备工作，这表明存在的机械敏感性释放可溶性核苷酸酶的LP。Ca ~（2+）和Mg ~（2+）对释放酶活性的影响不同。常见的核苷酸酶抑制剂ARL 67156、POM-1、PSB 06126和ENPP 1 Inhibitor C，但碱性磷酸酶抑制剂（-）-p-bromotetramisole草酸盐不抑制膀胱扩张期间释放的酶。通过Wes免疫检测在组织匀浆和来自膨胀制剂的浓缩ELS中鉴定膜结合核苷酸酶。在尿中核苷酸酶的相对分布为ENTPD 1>> ENPP 1> ENTPD 2 = ENTPD 3> ENPP 3 = NT 5E>> ENTPD 8 = TNAP，在浓缩ELS中为ENTPD 1>> ENTPD 3>> ENPP 3> ENPP 1 = ENTPD 2 = NT 5E>> ENTPD 8 = TNAP，表明在膀胱充盈过程中LP中可能发生膜结合核苷酸酶的调节性胞外域脱落。ATP和ADP的膜结合和可溶性核苷酸酶在LP的机械敏感性降解减少了兴奋性嘌呤的LP在膀胱充盈结束。这可能是一种保护机制，以防止过度兴奋的膀胱。膀胱壁中兴奋性和抑制性嘌呤的适当比例由扩张相关的嘌呤释放和嘌呤代谢决定。
Prior studies suggest that urothelium-released adenosine 5′-triphosphate (ATP) has a prominent role in bladder mechanotransduction. Urothelial ATP regulates the micturition cycle through activation of purinergic receptors that are expressed in many cell types in the lamina propria (LP), including afferent neurons, and might also be important for direct mechanosensitive signaling between urothelium and detrusor. The excitatory action of ATP is terminated by enzymatic hydrolysis, which subsequently produces bioactive metabolites. We examined possible mechanosensitive mechanisms of ATP hydrolysis in the LP by determining the degradation of 1,N 6 -etheno-ATP (eATP) at the anti-luminal side of nondistended (empty) or distended (full) murine (C57BL/6J) detrusor-free bladder model, using HPLC. The hydrolysis of eATP and eADP was greater in contact with LP of distended than of nondistended bladders whereas the hydrolysis of eAMP remained unchanged during filling, suggesting that some steps of eATP hydrolysis in the LP are mechanosensitive. eATP and eADP were also catabolized in extraluminal solutions (ELS) that were in contact with the LP of detrusor-free bladders, but removed from the organ chambers prior to addition of substrate. The degradation of both purines was greater in ELS from distended than from nondistended preparations, suggesting the presence of mechanosensitive release of soluble nucleotidases in the LP. The released enzyme activities were affected differently by Ca2+ and Mg2+. The common nucleotidase inhibitors ARL67156, POM-1, PSB06126, and ENPP1 Inhibitor C, but not the alkaline phosphatase inhibitor (-)-p-bromotetramisole oxalate, inhibited the enzymes released during bladder distention. Membrane-bound nucleotidases were identified in tissue homogenates and in concentrated ELS from distended preparations by Wes immunodetection. The relative distribution of nucleotidases was ENTPD1 >> ENPP1 > ENTPD2 = ENTPD3 > ENPP3 = NT5E >> ENTPD8 = TNAP in urothelium and ENTPD1 >> ENTPD3 >> ENPP3 > ENPP1 = ENTPD2 = NT5E >> ENTPD8 = TNAP in concentrated ELS, suggesting that regulated ectodomain shedding of membrane-bound nucleotidases possibly occurs in the LP during bladder filling. Mechanosensitive degradation of ATP and ADP by membrane-bound and soluble nucleotidases in the LP diminishes the availability of excitatory purines in the LP at the end of bladder filling. This might be a safeguard mechanism to prevent over-excitability of the bladder. Proper proportions of excitatory and inhibitory purines in the bladder wall are determined by distention-associated purine release and purine metabolism.
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影响因子：
19
作者：
Alcedo KP;Bowser JL;Snider NT
通讯作者：
Snider NT
影响因子：
2
作者：
Dunton CL;Purves JT;Hughes FM Jr;Jin H;Nagatomi J
通讯作者：
Nagatomi J
影响因子：
5.5
作者：
Ferguson, DR;Kennedy, I;Burton, TJ
通讯作者：
Burton, TJ
DOI：
10.1016/j.jbc.2021.101526
发表时间：
2022-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Borza R;Salgado-Polo F;Moolenaar WH;Perrakis A
通讯作者：
Perrakis A
影响因子：
2.9
作者：
Bobalova, J;Bobal, P;Mutafova-Yambolieva, VN
通讯作者：
Mutafova-Yambolieva, VN