Cell calcium最新文献_第4页

CPVT1 point mutations in RyR2 S5 and S6 segments and their Ca2+ signaling consequence RyR2 S5和S6片段CPVT1点突变及其Ca2+信号转导作用。

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-09-14 DOI: 10.1016/j.ceca.2025.103081

Xiao-hua Zhang , Grace Ellen Donch , Naohiro Yamaguchi , Martin Morad

Precise activation of cardiac ryanodine receptor (RyR2) by small influx of Ca²⁺ during the action potential triggers the release of SR Ca²⁺ that activates contraction, a process known as Ca²⁺-induced Ca²⁺ release (CICR). Missense mutations in RyR2 often cause aberrant and unregulated Ca²⁺ releases that are associated with catecholaminergic polymorphic ventricular tachycardia (CPVT), often lethal arrhythmias. Here using CRISPR/Cas9 gene editing in human induced pluripotent stem cells (hiPSCs), we extended our previous studies to include two new arrhythmogenic mutations one, R4822H, located in S5-S6 transmembrane luminal loop near RyR2 selective filter and the other, L4865V, located on S6 segment. TIRF-imaging of voltage-clamped mutant myocytes showed that I_Ca and caffeine-triggered cytosolic Ca²⁺ rise (Fura-2 signal) or ER-GCaMP6 SR Ca²⁺ release signals were significantly suppressed in R4822H but not in L4865V myocytes. Spontaneous Ca²⁺ transients, however, persisted in both mutant lines activating both Fura-2 and ER-GCaMP6 Ca²⁺ transients in L4865V cells, but only Fura-2 Ca²⁺ transients in R4822H mutant. Spontaneous Ca²⁺ sparks igniting frequencies were similar in both mutants, but spark durations were significantly shorter. Although both of these mutations are located at S5 and S6 transmembrane regions of RyR2, their phenotypes diverge markedly. L4865V mutant does not show suppressed E-C coupling function, while R4822H mutant has completely suppressed CICR suggesting that the spontaneous beating in R4822H mutant results from remodeling of dormant Ca²⁺ signaling pathway expressed in hiPSCCMs.

在动作电位期间，通过Ca2+的少量内流精确激活心脏ryanodine受体（RyR2），触发SR Ca2+的释放，激活收缩，这一过程称为Ca2+诱导的Ca2+释放（CICR）。RyR2的错义突变经常引起与儿茶酚胺能多态性室性心动过速（CPVT）相关的异常和不调节的Ca2+释放，通常是致命的心律失常。在这里，我们利用CRISPR/Cas9基因编辑人类诱导多能干细胞（hiPSCs），扩展了我们之前的研究，包括两个新的心律失常突变，一个是位于S5-S6跨膜腔袢靠近RyR2选择过滤器的R4822H，另一个是位于S6段的L4865V。电压箝位突变型肌细胞的tirf成像显示，ICa和咖啡因触发的胞质Ca2+升高（Fura-2信号）或ER-GCaMP6 SR Ca2+释放信号在R4822H中被显著抑制，而在L4865V肌细胞中没有。然而，自发的Ca2+瞬态在两个突变系中持续存在，在L4865V细胞中激活Fura-2和ER-GCaMP6 Ca2+瞬态，但在R4822H突变株中仅激活Fura-2 Ca2+瞬态。自发Ca2+火花点燃频率在两个突变体中相似，但火花持续时间明显短。虽然这两个突变都位于RyR2的S5和S6跨膜区，但它们的表型明显不同。L4865V突变体不表现出抑制E-C偶联功能，而R4822H突变体完全抑制了CICR，这表明R4822H突变体的自发振荡是由hipsccm中表达的休眠Ca2+信号通路的重塑引起的。

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引用次数: 0

SERCA3, ubiquitous but specific calcium pumps? SERCA3，普遍存在但特定的钙泵？

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-09-05 DOI: 10.1016/j.ceca.2025.103079

Maëliss Toth , Shaymaa Alhabib , Boris Manoury , Régis Bobe , Véronique Leblais

The calcium ion (Ca²⁺) is an important second messenger for the organism, participating in the regulation of various physiological responses in all cell types. In view of its crucial role, maintaining calcium homeostasis is important. This is why many players tightly regulate calcium homeostasis. These include a type of transporter historically located in the sarcoplasmic or endoplasmic reticulum (SR or ER), called Sarco-Endoplasmic Reticulum Ca²⁺-ATPase or SERCA calcium pumps (Toyoshima et al., 2000). Existence of these pumps was first demonstrated in the 1960s in rabbit skeletal muscle (Ebashi and Lipmann, 1962). In the mid-80s, only two families of these transporters, SERCA1 and SERCA2, were described in the skeletal muscle and the cardiovascular system, respectively. However, the existence of a third family, named SERCA3, was subsequently revealed. In this review, we present an overview of the current knowledge of the SERCA3. We firstly present the structure of this pump from its gene to the protein and its catalytic properties, highlighting its specific features compared to other isoforms. We then focus on the pathophysiological settings by describing its functional role established in several organs and pointing out the studies assuming its implication in different diseases such as obesity or cancers.

钙离子（Ca2+）是机体重要的第二信使，参与调节各种细胞类型的各种生理反应。鉴于其至关重要的作用，维持钙稳态是很重要的。这就是为什么许多球员严格调节钙稳态的原因。其中包括一种历史上位于肌浆或内质网（SR或ER）的转运体，称为肌内质网Ca2+- atp酶或SERCA钙泵（Toyoshima等，2000）。这些泵的存在于20世纪60年代首次在兔骨骼肌中得到证实（Ebashi和Lipmann， 1962）。在80年代中期，只有两个转运蛋白家族，SERCA1和SERCA2，分别在骨骼肌和心血管系统中被描述。然而，随后发现了第三个家族，名为SERCA3。在这篇综述中，我们对SERCA3的现有知识进行了概述。我们首先介绍了该泵的结构，从其基因到蛋白质及其催化性能，突出了其与其他同工异构体相比的特定特征。然后，我们通过描述其在几个器官中建立的功能作用，并指出其在不同疾病（如肥胖或癌症）中的含义的研究，重点关注病理生理设置。

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引用次数: 0

Dantrolene normalizes heightened Ca2+ influx in activated T cells from the familial Alzheimer's disease TgF344-AD rats 丹曲林使家族性阿尔茨海默病TgF344-AD大鼠激活T细胞中升高的Ca2+内流正常化

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-08-27 DOI: 10.1016/j.ceca.2025.103072

Navdeep K. Uppal , Anthony Valenzuela , Pamela J. Lein , Alla F. Fomina

Aim

Dysregulation of the peripheral immune response contributes to Alzheimer's disease pathogenesis. Dantrolene, a negative allosteric modulator of ryanodine receptor types 1 and 3, reduces neurodegeneration in Alzheimer's disease animal models by an unclear mechanism. Given that Alzheimer's disease causative mutations in amyloid precursor and presenilin proteins interfere with intracellular Ca²⁺ signaling in neurons, we tested the hypotheses that these mutations may impair Ca²⁺ signaling in T lymphocytes and that dantrolene can repair this defect.

Methods

We explored cytosolic Ca²⁺ dynamics and effects of dantrolene sodium in resting and activated splenic T cells derived from adult transgenic TgF344-AD rats expressing mutant human "Swedish" amyloid precursor (APPsw) and presenilin 1 lacking exon 9 (PS1Δ9) proteins, and in control wild-type rats.

Results

We found no differences in the cytosolic Ca²⁺ signaling between resting T cells from TgF344-AD and control rats. In contrast, amplitudes of caffeine-triggered calcium transients and store-operated Ca²⁺ entry were significantly larger in activated TgF344-AD rat T cells relative to wild-type rat T cells. Preincubation with dantrolene sodium reduced the amplitude and the rate of Ca²⁺ influx in activated TgF344-AD rat T cells in the absence of store refilling and after dissipation of inner mitochondrial membrane potential, indicating that it does not involve Ca²⁺ release via ryanodine receptors or mitochondrial Ca²⁺ uptake.

Conclusions

Expression of Alzheimer's disease risk genes upregulates the store-operated Ca²⁺ entry in T cells, which may alter peripheral immune responses and exacerbate Alzheimer's disease pathogenesis. We speculate that dantrolene's neuroprotective effect in Alzheimer's disease animal models may be due to its normalization of the peripheral T cells' Ca²⁺ signaling and functions.

目的外周免疫反应失调参与阿尔茨海默病的发病机制。丹曲林是一种ryanodine受体1型和3型的负变构调节剂，通过一种尚不清楚的机制减少阿尔茨海默病动物模型的神经变性。鉴于阿尔茨海默病的淀粉样蛋白前体和早老素蛋白的致病突变干扰神经元细胞内Ca2+信号，我们测试了这些突变可能损害T淋巴细胞中的Ca2+信号和丹trolene可以修复这种缺陷的假设。方法我们研究了在静止和激活的脾脏T细胞中，由表达突变人类“瑞典”淀粉样蛋白前体（APPsw）和早老素1缺乏外显子9 （PS1Δ9）蛋白的TgF344-AD成年转基因大鼠和对照野生型大鼠衍生的胞浆Ca2+动力学和dantrolene钠的影响。结果我们发现TgF344-AD的静止T细胞和对照大鼠的胞质Ca2+信号没有差异。相比之下，在激活的TgF344-AD大鼠T细胞中，咖啡因触发的钙瞬态和储存操作的Ca2+进入的振幅明显大于野生型大鼠T细胞。在没有储存再填充和线粒体内膜电位耗散的情况下，丹trolene钠预孵育降低了激活的TgF344-AD大鼠T细胞中Ca2+内流的幅度和速率，表明它不涉及通过ryanodine受体或线粒体Ca2+摄取释放Ca2+。结论阿尔茨海默病风险基因的表达上调T细胞储存操作的Ca2+进入，可能改变外周免疫反应，加剧阿尔茨海默病的发病机制。我们推测丹曲林在阿尔茨海默病动物模型中的神经保护作用可能是由于其使外周T细胞的Ca2+信号和功能正常化。

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引用次数: 0

CNO induced Ca2+ store and glutamate-dependent nonspecific Ca2+ signalling in DREADD-free brain slices CNO诱导Ca2+储存和谷氨酸依赖的非特异性Ca2+信号在无dreadd的脑切片中

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-08-26 DOI: 10.1016/j.ceca.2025.103070

Xiao-Yu Zhang , Xi Wu , Rui-yun Bi , Shan Zhang , Ye-Hua Gan

DREADD (design receptors exclusively activated by designer drugs) is a widely used powerful tool designed to study specific cellular functions. However, off-target effects of chemogenetic activators, including clozapine N-oxide (CNO) and deschloroclozapine (DCZ), have been reported. In our study, we demonstrated the direct off-target effects of CNO and DCZ on basal Ca²⁺ levels in the locus coeruleus nucleus in both neurons and astrocytes by combining viral microinjection, Ca²⁺ imaging and electrophysiology. We observed that CNO induced a Ca²⁺ store-dependent increase in basal Ca²⁺ in both DREADD-absent neurons and astrocytes; interestingly, CNO directly increased the frequency of spontaneous presynaptic glutamate release. Furthermore, ionotropic glutamate receptors contributed to the CNO-induced increase in Ca²⁺ in both DREADD-free neurons and astrocytes. Importantly, IP3R2-KO diminished CNO-induced Ca²⁺ raise in astrocytes but not neurons. Our results revealed direct Ca²⁺ store- and glutamate-dependent off-target effects of DREADD agonists during their cellular action, which may help elucidate the mechanisms underlying the off-target effects of chemogenetic tools.

设计药物特异性激活的设计受体（DREADD）是一种被广泛应用于研究特定细胞功能的强大工具。然而，化学发生活化剂的脱靶效应，包括氯氮平n -氧化物（CNO）和去氯氯氮平（DCZ），已被报道。在我们的研究中，我们通过结合病毒显微注射、Ca2+成像和电生理学，证明了CNO和DCZ对神经元和星形胶质细胞蓝斑核基础Ca2+水平的直接脱靶效应。我们观察到，CNO诱导Ca2+储存依赖性的基础Ca2+增加，在缺乏症的神经元和星形胶质细胞；有趣的是，CNO直接增加自发性突触前谷氨酸释放的频率。此外，嗜离子性谷氨酸受体有助于cno诱导的无dreadd神经元和星形胶质细胞中Ca2+的增加。重要的是，IP3R2-KO减少了星形胶质细胞中cno诱导的Ca2+升高，而不是神经元。我们的研究结果揭示了DREADD激动剂在细胞作用过程中直接的Ca2+储存和谷氨酸依赖的脱靶效应，这可能有助于阐明化学发生工具脱靶效应的机制。

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引用次数: 0

Silencing CALB1 enhances prostate cancer radiosensitivity via calcium-mediated mitochondrial dysfunction and cellular senescence 沉默CALB1可通过钙介导的线粒体功能障碍和细胞衰老增强前列腺癌的放射敏感性

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-08-26 DOI: 10.1016/j.ceca.2025.103071

Chen Gong , Senmao Li , Ye An , Chadanfeng Yang , Zhiyong Tan , Wujie Chen , Dihao Lv , Haichao Wu , Haifeng Wang , Shi Fu , Haihao Li , Yanjie Kong , Yinglong Huang , Mingxia Ding

Background

Prostate cancer remains a leading cause of cancer-related deaths in men, with radioresistance limiting treatment efficacy. This study investigates the role of Calbindin 1 (CALB1), a calcium-binding protein regulated by miR-186–5p, in prostate cancer progression and radiation response.

Methods

CALB1 expression was analyzed using GEO and TCGA datasets, and the regulatory relationship with miR-186–5p was validated. Functional studies including CALB1 knockdown, calcium chelation, and mitochondrial rescue interventions were conducted in prostate cancer cells, spheroids, and xenograft models, assessing proliferation, senescence, calcium homeostasis, and radiation response.

Results

We identified CALB1 as a target of downregulated miR-186–5p in prostate cancer. CALB1 silencing inhibited prostate cancer growth by inducing cellular senescence through calcium dysregulation, mitochondrial dysfunction, and oxidative stress. CALB1 depletion significantly enhanced radiosensitivity both in vitro and in vivo, with calcium chelation or mitochondrial interventions partially rescuing these effects.

Conclusions

CALB1 regulates prostate cancer progression and radiation response by maintaining calcium homeostasis. Its depletion triggers calcium overload and mitochondrial dysfunction, enhancing radiation sensitivity and identifying CALB1 as a potential therapeutic target.

前列腺癌仍然是男性癌症相关死亡的主要原因，放射耐药限制了治疗效果。本研究探讨了钙结合蛋白1 (CALB1)，一种由miR-186-5p调节的钙结合蛋白，在前列腺癌进展和放射反应中的作用。方法利用GEO和TCGA数据集分析scalb1的表达，验证其与miR-186-5p的调控关系。在前列腺癌细胞、球状体和异种移植模型中进行了功能研究，包括CALB1敲除、钙螯合和线粒体拯救干预，评估了增殖、衰老、钙稳态和辐射反应。我们发现CALB1是前列腺癌中miR-186-5p下调的靶标。CALB1沉默通过钙失调、线粒体功能障碍和氧化应激诱导细胞衰老，从而抑制前列腺癌的生长。CALB1缺失显著增强了体外和体内的放射敏感性，钙螯合或线粒体干预部分挽救了这些作用。结论scalb1通过维持钙稳态调节前列腺癌的进展和放射反应。它的消耗引发钙超载和线粒体功能障碍，增强辐射敏感性，并将CALB1确定为潜在的治疗靶点。

{"title":"Silencing CALB1 enhances prostate cancer radiosensitivity via calcium-mediated mitochondrial dysfunction and cellular senescence","authors":"Chen Gong , Senmao Li , Ye An , Chadanfeng Yang , Zhiyong Tan , Wujie Chen , Dihao Lv , Haichao Wu , Haifeng Wang , Shi Fu , Haihao Li , Yanjie Kong , Yinglong Huang , Mingxia Ding","doi":"10.1016/j.ceca.2025.103071","DOIUrl":"10.1016/j.ceca.2025.103071","url":null,"abstract":"<div><h3>Background</h3><div>Prostate cancer remains a leading cause of cancer-related deaths in men, with radioresistance limiting treatment efficacy. This study investigates the role of Calbindin 1 (CALB1), a calcium-binding protein regulated by miR-186–5p, in prostate cancer progression and radiation response.</div></div><div><h3>Methods</h3><div>CALB1 expression was analyzed using GEO and TCGA datasets, and the regulatory relationship with miR-186–5p was validated. Functional studies including CALB1 knockdown, calcium chelation, and mitochondrial rescue interventions were conducted in prostate cancer cells, spheroids, and xenograft models, assessing proliferation, senescence, calcium homeostasis, and radiation response.</div></div><div><h3>Results</h3><div>We identified CALB1 as a target of downregulated miR-186–5p in prostate cancer. CALB1 silencing inhibited prostate cancer growth by inducing cellular senescence through calcium dysregulation, mitochondrial dysfunction, and oxidative stress. CALB1 depletion significantly enhanced radiosensitivity both in vitro and in vivo, with calcium chelation or mitochondrial interventions partially rescuing these effects.</div></div><div><h3>Conclusions</h3><div>CALB1 regulates prostate cancer progression and radiation response by maintaining calcium homeostasis. Its depletion triggers calcium overload and mitochondrial dysfunction, enhancing radiation sensitivity and identifying CALB1 as a potential therapeutic target.</div></div>","PeriodicalId":9678,"journal":{"name":"Cell calcium","volume":"132 ","pages":"Article 103071"},"PeriodicalIF":4.0,"publicationDate":"2025-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144926570","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Regulation of K+-dependent Na+/Ca2+-exchanger subtype 4, NCKX4, by palmitoylation 棕榈酰化对K+依赖性Na+/Ca2+交换器亚型4 （NCKX4）的调控

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-08-14 DOI: 10.1016/j.ceca.2025.103069

By Maryam Al-Khannaq , Jonathan Lytton

Mammalian K⁺-dependent Na⁺/Ca²⁺ exchangers (NCKX), encoded by the SLC24 gene family, are crucial for maintaining Ca²⁺ homeostasis. NCKX4, widely expressed in the brain and sensory neurons, plays a key role in neuronal satiety and enamel formation. Despite its importance, the regulatory mechanisms of NCKX4 remain largely unexplored. This study investigates how palmitoylation, a post-translational modification affecting membrane proteins, regulates NCKX4 and influences its cellular localization and function.

Using Acyl-RAC and palmitate-based click-chemistry, we found that approximately 14% of NCKX4 is palmitoylated at steady-state in both endogenous and transfected systems. The level of this modification is highly dynamic, being regulated by inhibitors of palmitoylation (2-bromopalmitate) and depalmitoylation (palmostatin B), resulting in greater than a two-fold decrease or increase, respectively. Site-directed mutagenesis of six cysteine residues revealed two key sites (Cys118 and Cys425) critical for NCKX4 palmitoylation.

The subcellular distribution of palmitoylated NCKX4 was examined via proximity ligation and click-chemistry. NCKX4 was found across multiple membrane compartments, with a higher fraction localizing to the plasma membrane when palmitoylation was inhibited by 2-bromopalmitate. However, a Ca²⁺ imaging assay in HEK293T cells showed no significant change in aggregate cellular NCKX4-mediated Ca²⁺ transport upon modulation of palmitoylation status. These data suggest palmitoylation promotes internalization of the NCKX4 protein while also activating it, counter-acting effects that result in unchanged NCKX4-mediated cellular Ca²⁺ transport activity.

In summary, NCKX4 is subject to dynamic palmitoylation, which influences both distribution across cellular compartments and intrinsic Ca²⁺ transport activity. These findings contribute to our understanding of the regulation and functional roles of NCKX4 in cellular physiology.

哺乳动物K+依赖性Na+/Ca2+交换器（NCKX）由SLC24基因家族编码，对维持Ca2+稳态至关重要。NCKX4广泛表达于脑和感觉神经元，在神经元饱腹感和牙釉质形成中起关键作用。尽管它很重要，但NCKX4的调控机制在很大程度上仍未被探索。本研究探讨了影响膜蛋白的翻译后修饰棕榈酰化（palmitoylation）如何调控NCKX4并影响其细胞定位和功能。使用酰基rac和棕榈酸盐为基础的点击化学，我们发现大约14%的NCKX4在内源性和转染系统中处于稳态棕榈酰化。这种修饰的水平是高度动态的，受棕榈酰化（2-溴铝酸酯）和去棕榈酰化（棕榈抑素B）抑制剂的调节，分别导致大于两倍的减少或增加。6个半胱氨酸残基的定点突变揭示了NCKX4棕榈酰化的两个关键位点（Cys118和Cys425）。通过近端结扎和点击化学检测棕榈酰化NCKX4的亚细胞分布。NCKX4跨越多个膜区室，当2-溴铝酸盐抑制棕榈酰化时，NCKX4在质膜上的定位比例更高。然而，HEK293T细胞中的Ca2+成像分析显示，在棕榈酰化状态的调节下，聚集细胞nckx4介导的Ca2+运输没有显著变化。这些数据表明，棕榈酰化促进了NCKX4蛋白的内化，同时也激活了它，抵消了NCKX4介导的细胞Ca2+运输活性不变的影响。总之，NCKX4受动态棕榈酰化的影响，这影响了细胞间室的分布和内在的Ca2+运输活性。这些发现有助于我们理解NCKX4在细胞生理学中的调控和功能作用。

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引用次数: 0

NAADP-mediated calcium release promotes angiopoietin 2 secretion by regulating Rab46-dependent Weibel-Palade body trafficking naadp介导的钙释放通过调节rab46依赖性Weibel-Palade体运输促进血管生成素2的分泌

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-08-14 DOI: 10.1016/j.ceca.2025.103068

Ryan D. Murray , Melissa Rose , Katarina T. Miteva , David J. Beech , Lynn McKeown

Angiopoietin2 (Ang2), a regulator of angiogenesis, is stored with other pro-inflammatory and pro-thrombotic mediators, in endothelial-specific vesicles called Weibel-Palade bodies (WPBs). Acute stimulation of endothelial cells with histamine, delays Ang2 secretion by activating Rab46-specific trafficking of Ang2-containing WPBs to the microtubule organising centre (MTOC), where they persist until Ca²⁺ binds to the EF-hand of Rab46, enabling detachment. Here, using Ca²⁺ imaging and high-resolution light microscopy, we pharmacologically investigated the contribution of endolysosomal two-pore channel proteins (TPC) to the Ca²⁺ signal necessary for WPB detachment and Ang2 secretion. We show an increase in the histamine-evoked clustering of Rab46 (and thus WPBs) at the MTOC in the presence of TPC inhibitors Ned-19 and tetrandrine, and a decrease in the presence of a TPC2 agonist, TPC2-A1-N. Histamine-evoked secretion of Ang2 was decreased by pharmacological inhibition of TPC channels but potentiated in the presence of TPC2-A1-N. These data suggest that histamine-mediated Ca²⁺ release via TPC2 channels is necessary for the Rab46-dependent detachment of Ang2-positive WPBs from the MTOC and thus Ang2 secretion.

Summary

Ca²⁺ binding to the EF-hand of Rab46 in endothelial cells has previously been reported but the molecular mechanisms and functional relevance are unclear. Here, the authors show that Ca²⁺ released from TPC channels regulates the detachment of Rab46-positive WPBs from the MTOC, which thereby promotes secretion of Ang2.

血管生成素2 （Ang2）是一种血管生成调节剂，与其他促炎和促血栓介质一起储存在内皮特异性囊泡中，称为韦贝尔-帕拉德小体（WPBs）。用组胺急性刺激内皮细胞，通过激活含有Ang2的WPBs到微管组织中心（MTOC）的Rab46特异性运输，延迟Ang2的分泌，在那里它们持续存在，直到Ca 2 +与Rab46的EF-hand结合，使其脱离。在这里，我们使用Ca 2 +成像和高分辨率光学显微镜，从药理学上研究了内溶酶体两孔通道蛋白（TPC）对WPB脱离和Ang2分泌所必需的Ca 2 +信号的贡献。我们发现，在TPC2抑制剂Ned-19和粉防己碱存在的情况下，MTOC处组胺诱发的Rab46聚类（以及WPBs）增加，而TPC2激动剂TPC2- a1 - n存在的情况下，组胺诱发的Rab46聚类（以及WPBs）减少。组胺诱发的Ang2分泌因药物抑制TPC通道而减少，但在TPC2-A1-N存在时增强。这些数据表明，组胺介导的Ca +通过TPC2通道释放是rab46依赖性的Ang2阳性WPBs脱离MTOC和Ang2分泌所必需的。ca 2 +在内皮细胞中与Rab46的EF-hand结合已有报道，但其分子机制和功能相关性尚不清楚。在这里，作者证明了从TPC通道释放的Ca 2 +可以调节rab46阳性WPBs从MTOC上脱离，从而促进Ang2的分泌。

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引用次数: 0

Histamine 1 receptors and reverse-mode Na+/Ca2+ exchanger drive extracellular Na+-dependent intracellular Ca2+ oscillations in human cerebrovascular endothelial cells 组胺1受体和反向模式Na+/Ca2+交换驱动细胞外Na+依赖的细胞内Ca2+振荡在人脑血管内皮细胞

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-08-12 DOI: 10.1016/j.ceca.2025.103067

Valentina Brunetti , Roberto Berra-Romani , Nayeli Coyotl-Santiago , Yair Esquitin-Gonzalez , Giorgia Chinigò , Gerardo Rosario Biella , Francesco Moccia , Giorgia Scarpellino

Cerebrovascular endothelial cells represent the core component of the blood-brain barrier, (BBB) which plays a critical role in regulating the local ionic microenvironment around the synapses. Therefore, cerebrovascular endothelial cells experience dramatic changes in the extracellular concentrations of potassium and sodium ions during intense neuronal firing or pathological conditions, such as spreading depression.

Herein, we assessed the mechanisms by which a reduction in extracellular sodium concentration ([Na⁺]_o) triggers complex Ca²⁺ signals in the hCMEC/D3 cell line, which is the most widespread model of human BBB.

We demonstrate that lowering the [Na⁺]_o elicits a variety of Ca²⁺ signals, including monotonic increases in intracellular Ca²⁺ concentration ([Ca²⁺]_i) and repetitive oscillations in [Ca²⁺]_i, which are triggered by the reverse-mode Na⁺/Ca²⁺ exchanger and histamine 1 receptor (H1R). Furthermore, we provide the first evidence that H1R may play a critical role in translating a reduction in [Na⁺]_o into the activation of phospholipase C and following production of inositol triphosphate (InsP₃), thereby inducing the rhythmic activation of InsP₃ receptors on the endoplasmic reticulum (ER) and progressive depletion of the ER Ca²⁺ pool. The fall in the ER Ca²⁺ concentration leads to quick Store-Operated Ca²⁺ Entry activation, which maintains the intracellular Ca²⁺ oscillations by rapidly refilling the ER Ca²⁺ store. The endothelial Ca²⁺ oscillations induced by the reduction in [Na⁺]_o may then lead to nitric oxide release.

These findings, therefore, shed novel light on the mechanisms whereby G_q protein coupled receptors (G_qPCRs) can shape endothelial Ca²⁺ signaling and Ca²⁺-dependent events at the human neurovascular unit.

脑血管内皮细胞是血脑屏障（BBB）的核心组成部分，在调节突触周围局部离子微环境中起着关键作用。因此，在强烈的神经元放电或病理状态（如扩张性抑郁）时，脑血管内皮细胞的胞外钾和钠离子浓度会发生剧烈变化。在此，我们评估了细胞外钠浓度（[Na+]o）的降低触发hCMEC/D3细胞系中复杂Ca2+信号的机制，hCMEC/D3细胞系是人类血脑屏障最广泛的模型。我们证明降低[Na+]o引起多种Ca2+信号，包括细胞内Ca2+浓度（[Ca2+]i）的单调增加和[Ca2+]i的重复振荡，这是由反向模式Na+/Ca2+交换器和组胺1受体（H1R）触发的。此外，我们提供了第一个证据，证明H1R可能在将[Na+]o的减少转化为磷脂酶C的激活和随后的肌醇三磷酸（InsP3）的产生中发挥关键作用，从而诱导内质网（ER）上的InsP3受体的节律性激活和ER Ca2+池的逐渐耗尽。内质网Ca2+浓度的下降导致快速存储操作的Ca2+进入激活，通过快速重新填充内质网Ca2+存储来维持细胞内Ca2+振荡。由[Na+]o的减少引起的内皮Ca2+振荡可能导致一氧化氮释放。因此，这些发现揭示了Gq蛋白偶联受体（gqpcr）在人类神经血管单元中塑造内皮Ca2+信号和Ca2+依赖事件的机制。

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引用次数: 0

Chemical but not mechanical stimulation reduce TRPA1 channel lateral mobility 化学而非机械刺激可降低TRPA1通道的横向流动性

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-07-31 DOI: 10.1016/j.ceca.2025.103059

Alicia Sampieri , Alexander Asanov , Aaron Pavel Rodríguez-Hernández , Ileana Tobías-Juárez , Daniel Martínez-Flores , Luis Vaca

The transient Receptor Potential Ankyrin 1 (TRPA1) is a member from the TRP superfamily of ion channels. TRPA1 channels are calcium-permeable nonselective cation channels, which are highly conserved throughout the animal kingdom. Mammals have only one member (TRPA1), while zebrafish has two (TRPA1a and TRPA1b). TRPA1 channels are activated by a plethora of stimuli, including noxious cold, mechanical stimulation, calcium, pH, reactive oxygen, and carbonyl species. In the present study we characterize the modulation of TRPA1b channel lateral mobility by Allyl isothiocyanate (AITC) and mechanical stimulation. We show that only AITC stimulation alters channel diffusion at the plasma membrane.

瞬时受体电位锚蛋白1 （TRPA1）是离子通道TRP超家族的成员。TRPA1通道是钙渗透性非选择性阳离子通道，在动物界高度保守。哺乳动物只有一个成员（TRPA1），而斑马鱼有两个成员（TRPA1a和TRPA1b）。TRPA1通道可被多种刺激激活，包括毒冷、机械刺激、钙、pH、活性氧和羰基物质。在本研究中，我们描述了异硫氰酸烯丙酯（AITC）和机械刺激对TRPA1b通道横向迁移的调节。我们发现只有AITC刺激改变了质膜上的通道扩散。

引用次数: 0

Calmodulin enhancement of mitochondrial calcium uniporter function in isolated mitochondria 钙调素增强离体线粒体钙转运蛋白功能

IF 4 2区生物学 Q2 CELL BIOLOGY

Cell calcium

Pub Date : 2025-07-19 DOI: 10.1016/j.ceca.2025.103056

Sara A. Garcia , Anne M. Neumaier , Michael Kohlhaas , Anton Xu , Alexander Nickel , Katharina J. Ermer , Luzia Enzner , Christoph Maack , Vasco Sequeira , Christopher N. Johnson

Mitochondrial calcium (Ca²⁺) uptake and factors that regulate this process have been an area of immense interest given the roles in cellular energetics. Here, we have investigated the ability of the Ca²⁺ sensing protein Calmodulin (CaM) to modify the function of the Mitochondrial Ca²⁺ Uniporter (MCU). Our data leveraged recombinantly produced CaM and mitochondria isolated from healthy and MCU impaired/diseased mice (Barth syndrome model). We found CaM enhanced Ca²⁺ uptake in both the absence and presence of CaMKII inhibition (KN93 as well as AIP). Mitochondria lacking function MCU (Barth syndrome model) validated that MCU was responsible for Ca²⁺ uptake in our experiments. Control experiments demonstrate that the observed CaM enhancement does not arise from CaM Ca²⁺ buffering. Fitting the Ca²⁺fluorescence data supported a monophasic decay process where the presence of CaM yielded enhanced kinetic rates of Ca²⁺ uptake. This CaM enhancement effect persisted in the presence of PTP impairment (cyclosporin), and subtle modification to the CaM protein sequence (D131E) revealed that an intact CaM-C domain Ca²⁺ binding was required for enhancement of MCU function.

线粒体钙（Ca2+）摄取和调节这一过程的因素一直是一个非常感兴趣的领域，因为它在细胞能量学中的作用。在这里，我们研究了Ca2+传感蛋白钙调蛋白（CaM）改变线粒体Ca2+单转运蛋白（MCU）功能的能力。我们的数据利用了从健康和MCU受损/患病小鼠（Barth综合征模型）中分离的重组产生的CaM和线粒体。我们发现CaM在CaMKII抑制（KN93和AIP）缺失和存在的情况下都能增强Ca2+摄取。线粒体缺乏功能MCU （Barth综合征模型）在我们的实验中证实了MCU负责Ca2+摄取。对照实验表明，观察到的CaM增强不是由CaM Ca2+缓冲引起的。拟合Ca2+荧光数据支持单相衰减过程，其中CaM的存在产生增强的Ca2+摄取的动力学速率。这种CaM增强效应在PTP损伤（环孢素）存在时持续存在，并且对CaM蛋白序列（D131E）的细微修饰表明，增强MCU功能需要完整的CaM- c结构域Ca2+结合。

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