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The tether function of the anoctamins 蛋白拮抗剂的拴系功能
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-20 DOI: 10.1016/j.ceca.2024.102875
Wei-Yin Lin, Woo Young Chung, Shmuel Muallem

The core functions of the anoctamins are Cl channel activity and phosphatidylserine (and perhaps other lipids) scrambling. These functions have been extensively studied in various tissues and cells. However, another function of the anoctamins that is less recognized and minimally explored is as tethers at membrane contact sites. This short review aims to examine evidence supporting the localization of the anoctamins at membrane contact sites, their tether properties, and their functions as tethers.

阳离子维生素的核心功能是 Cl- 通道活性和磷脂酰丝氨酸(或许还有其他脂质)扰乱。这些功能已在各种组织和细胞中得到广泛研究。然而,阳起素的另一种功能是作为膜接触位点的拴系物,但这一功能较少被认识和探索。这篇简短的综述旨在研究支持阳起糖蛋白在膜接触位点定位的证据、它们的系链特性及其作为系链的功能。
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引用次数: 0
The physiological roles of anoctamin2/TMEM16B and anoctamin1/TMEM16A in chemical senses 鹅膏蕈素2/TMEM16B和鹅膏蕈素1/TMEM16A在化学感官中的生理作用
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-18 DOI: 10.1016/j.ceca.2024.102889
Michele Dibattista , Simone Pifferi , Andres Hernandez-Clavijo , Anna Menini

Chemical senses allow animals to detect and discriminate a vast array of molecules. The olfactory system is responsible of the detection of small volatile molecules, while water dissolved molecules are detected by taste buds in the oral cavity. Moreover, many animals respond to signaling molecules such as pheromones and other semiochemicals through the vomeronasal organ. The peripheral organs dedicated to chemical detection convert chemical signals into perceivable information through the employment of diverse receptor types and the activation of multiple ion channels. Two ion channels, TMEM16B, also known as anoctamin2 (ANO2) and TMEM16A, or anoctamin1 (ANO1), encoding for Ca2+-activated Cl¯ channels, have been recently described playing critical roles in various cell types. This review aims to discuss the main properties of TMEM16A and TMEM16B-mediated currents and their physiological roles in chemical senses. In olfactory sensory neurons, TMEM16B contributes to amplify the odorant response, to modulate firing, response kinetics and adaptation. TMEM16A and TMEM16B shape the pattern of action potentials in vomeronasal sensory neurons increasing the interspike interval. In type I taste bud cells, TMEM16A is activated during paracrine signaling mediated by ATP. This review aims to shed light on the regulation of diverse signaling mechanisms and neuronal excitability mediated by Ca-activated Cl¯ channels, hinting at potential new roles for TMEM16A and TMEM16B in the chemical senses.

化学感官使动物能够检测和分辨大量分子。嗅觉系统负责检测挥发性小分子,而口腔中的味蕾则检测溶于水的分子。此外,许多动物还通过绒毛器官对信息素等信号分子和其他半化学物质做出反应。专门检测化学物质的外周器官通过利用不同类型的受体和激活多种离子通道,将化学信号转化为可感知的信息。两种离子通道,即 TMEM16B(又称 anoctamin2 (ANO2))和 TMEM16A(又称 anoctamin1 (ANO1)),编码 Ca2+ 激活的 Cl¯ 通道,最近被描述为在各种细胞类型中发挥关键作用。本综述旨在讨论 TMEM16A 和 TMEM16B 介导的电流的主要特性及其在化学感官中的生理作用。在嗅觉神经元中,TMEM16B 有助于放大气味反应、调节发射、反应动力学和适应。TMEM16A 和 TMEM16B 塑造了绒毛感觉神经元的动作电位模式,增加了间隔时间。在 I 型味蕾细胞中,TMEM16A 在 ATP 介导的旁分泌信号中被激活。本综述旨在阐明钙激活Cl'通道介导的多种信号机制和神经元兴奋性的调控,提示TMEM16A和TMEM16B在化学感官中的潜在新作用。
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引用次数: 0
VSI: The anoctamins: Structure and function VSI:营养素:结构与功能
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-17 DOI: 10.1016/j.ceca.2024.102888
Karl Kunzelmann, Jiraporn Ousingsawat, Rainer Schreiber

Plasma membrane localized anoctamin 1, 2 and 6 (TMEM16A, B, F) have been examined in great detail with respect to structure and function, but much less is known about the other seven intracellular members of this exciting family of proteins. This is probably due to their limited accessibility in intracellular membranous compartments, such as the endoplasmic reticulum (ER) or endosomes. However, these so-called intracellular anoctamins are also found in the plasma membrane (PM) which adds to the confusion regarding their cellular role. Probably all intracellular anoctamins except of ANO8 operate as intracellular phospholipid (PL) scramblases, allowing for Ca2+-activated, passive transport of phospholipids like phosphatidylserine between both membrane leaflets. Probably all of them also conduct ions, which is probably part of their physiological function. In this brief overview, we summarize key findings on the biological functions of ANO3, 4, 5, 7, 8, 9 and 10 (TMEM16C, D, E, G, H, J, K) that are gradually coming to light. Compartmentalized regulation of intracellular Ca2+ signals, tethering of the ER to specific PM contact sites, and control of intracellular vesicular trafficking appear to be some of the functions of intracellular anoctamins, while loss of function and abnormal expression are the cause for various diseases.

人们已经对质膜定位的鹅膏蕈素 1、2 和 6(TMEM16A、B、F)的结构和功能进行了详细的研究,但对这一令人兴奋的蛋白质家族的其他七个细胞内成员的了解却少得多。这可能是由于它们在细胞内膜隔室(如内质网(ER)或内体)中的可及性有限。然而,这些所谓的细胞内营养素也存在于质膜(PM)中,这就使人们对它们在细胞中的作用更加困惑。除 ANO8 外,可能所有细胞内抗外激素都是细胞内磷脂(PL)扰乱酶,允许在 Ca2+ 激活下在两片膜之间被动运输磷脂(如磷脂酰丝氨酸)。它们可能都能传导离子,这可能也是其生理功能的一部分。在这篇简短的综述中,我们总结了有关 ANO3、4、5、7、8、9 和 10(TMEM16C、D、E、G、H、J、K)生物功能的主要发现,这些发现正逐渐被人们所了解。细胞内Ca2+信号的区系化调节、ER与特定PM接触点的拴系以及细胞内囊泡贩运的控制似乎是细胞内anoctamins的部分功能,而功能缺失和异常表达则是导致各种疾病的原因。
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引用次数: 0
The anoctamins: Structure and function 营养素结构与功能
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-15 DOI: 10.1016/j.ceca.2024.102885
Rainer Schreiber, Jiraporn Ousingsawat, Karl Kunzelmann

When activated by increase in intracellular Ca2+, anoctamins (TMEM16 proteins) operate as phospholipid scramblases and as ion channels. Anoctamin 1 (ANO1) is the Ca2+-activated epithelial anion-selective channel that is coexpressed together with the abundant scramblase ANO6 and additional intracellular anoctamins. In salivary and pancreatic glands, ANO1 is tightly packed in the apical membrane and secretes Cl. Epithelia of airways and gut use cystic fibrosis transmembrane conductance regulator (CFTR) as an apical Cl exit pathway while ANO1 supports Cl secretion mainly by facilitating activation of luminal CFTR and basolateral K+ channels. Under healthy conditions ANO1 modulates intracellular Ca2+ signals by tethering the endoplasmic reticulum, and except of glands its direct secretory contribution as Cl channel might be small, compared to CFTR. In the kidneys ANO1 supports proximal tubular acid secretion and protein reabsorption and probably helps to excrete HCO3in the collecting duct epithelium. However, under pathological conditions as in polycystic kidney disease, ANO1 is strongly upregulated and may cause enhanced proliferation and cyst growth. Under pathological condition, ANO1 and ANO6 are upregulated and operate as secretory channel/phospholipid scramblases, partly by supporting Ca2+-dependent processes. Much less is known about the role of other epithelial anoctamins whose potential functions are discussed in this review.

当细胞内 Ca2+ 增加而激活时,外胚层蛋白(TMEM16 蛋白)可作为磷脂扰乱酶和离子通道发挥作用。Anoctamin 1(ANO1)是由 Ca2+ 激活的上皮阴离子选择性通道,它与丰富的扰乱酶 ANO6 和其他细胞内 anoctamins 共同表达。在唾液腺和胰腺中,ANO1 紧贴顶端膜并分泌 Cl-。气道和肠道上皮细胞利用囊性纤维化跨膜传导调节器(CFTR)作为顶端 Cl- 出口途径,而 ANO1 主要通过促进腔内 CFTR 和基底侧 K+ 通道的激活来支持 Cl- 分泌。在健康条件下,ANO1 通过拴系内质网来调节细胞内 Ca2+ 信号,除腺体外,与 CFTR 相比,ANO1 作为 Cl- 通道的直接分泌贡献可能较小。在肾脏中,ANO1 支持近端肾小管的酸分泌和蛋白质重吸收,并可能帮助集合管上皮细胞排泄 HCO3。然而,在多囊肾等病理情况下,ANO1 会强烈上调,并可能导致增殖和囊肿生长。在病理情况下,ANO1 和 ANO6 会上调,并作为分泌通道/磷脂扰乱酶发挥作用,部分是通过支持 Ca2+ 依赖性过程。本综述将讨论其他上皮营养素的潜在功能。
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引用次数: 0
pH regulating mechanisms of astrocytes: A critical component in physiology and disease of the brain 星形胶质细胞的 pH 值调节机制:大脑生理和疾病的关键组成部分
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-08 DOI: 10.1016/j.ceca.2024.102882
Shefeeq M. Theparambil , Gulnaz Begum , Christine R. Rose

Strict homeostatic control of pH in both intra- and extracellular compartments of the brain is fundamentally important, primarily due to the profound impact of free protons ([H+]) on neuronal activity and overall brain function. Astrocytes, crucial players in the homeostasis of various ions in the brain, actively regulate their intracellular [H+] (pHi) through multiple membrane transporters and carbonic anhydrases. The activation of astroglial pHi regulating mechanisms also leads to corresponding alterations in the acid-base status of the extracellular fluid. Notably, astrocyte pH regulators are modulated by various neuronal signals, suggesting their pivotal role in regulating brain acid-base balance in both health and disease. This review presents the mechanisms involved in pH regulation in astrocytes and discusses their potential impact on extracellular pH under physiological conditions and in brain disorders. Targeting astrocytic pH regulatory mechanisms represents a promising therapeutic approach for modulating brain acid-base balance in diseases, offering a potential critical contribution to neuroprotection.

对大脑细胞内和细胞外的 pH 值进行严格的平衡控制至关重要,这主要是由于游离质子([H+])对神经元活动和大脑整体功能有着深远的影响。星形胶质细胞是大脑中各种离子平衡的关键角色,它们通过多种膜转运体和碳酸酐酶积极调节细胞内的[H+](pHi)。星形胶质细胞 pHi 调节机制的激活也会导致细胞外液酸碱状态的相应改变。值得注意的是,星形胶质细胞 pH 调节器受各种神经元信号的调节,这表明它们在调节健康和疾病状态下的大脑酸碱平衡中发挥着关键作用。本综述介绍了星形胶质细胞 pH 值调节的相关机制,并讨论了它们在生理条件下和脑部疾病中对细胞外 pH 值的潜在影响。针对星形胶质细胞的 pH 调节机制是调节疾病中脑酸碱平衡的一种很有前景的治疗方法,为神经保护做出了潜在的重要贡献。
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引用次数: 0
Simultaneous TIRF imaging of subplasmalemmal Ca2+ dynamics and granule fusions in insulin-secreting INS-1 cells reveals coexistent synchronized and asynchronous release 对分泌胰岛素的 INS-1 细胞浆膜下 Ca2+ 动态和颗粒融合进行同步 TIRF 成像,发现同步和非同步释放同时存在
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-08 DOI: 10.1016/j.ceca.2024.102883
Charlotte Suckert , Carolin Zosel , Michael Schaefer

The basal and glucose-induced insulin secretion from pancreatic beta cells is a tightly regulated process that is triggered in a Ca2+-dependent fashion and further positively modulated by substances that raise intracellular levels of adenosine 3′,5′-cyclic monophosphate (cAMP) or by certain antidiabetic drugs. In a previous study, we have temporally resolved the subplasmalemmal [Ca2+]i dynamics in beta cells that are characterized by trains of sharply delimited spikes, reaching peak values up to 5 µM. Applying total internal reflection fluorescence (TIRF) microscopy and synaptopHluorin to visualize fusion events of individual granules, we found that several fusion events can coincide within 50 to 150 ms. To test whether subplasmalemmal [Ca2+]i microdomains around single or clustered Ca2+ channels may cause a synchronized release of insulin-containing vesicles, we applied simultaneous dual-color TIRF microscopy and monitored Ca2+ fluctuations and exocytotic events in INS-1 cells at high frame rates. The results indicate that fusions can be triggered by subplasmalemmal Ca2+ spiking. This, however, does account for a minority of fusion events. About 90 %-95 % of fusion events either happen between Ca2+ spikes or incidentally overlap with subplasmalemmal Ca2+ spikes. We conclude that only a fraction of exocytotic events in glucose-induced and tolbutamide- or forskolin-enhanced insulin release from INS-1 cells is tightly coupled to Ca2+ microdomains around voltage-gated Ca2+ channels.

胰岛β细胞的基础胰岛素分泌和葡萄糖诱导的胰岛素分泌是一个受到严格调控的过程,它以 Ca2+ 依赖性方式触发,并进一步受到能提高细胞内腺苷-3′,5′-环磷酸(cAMP)水平的物质或某些抗糖尿病药物的积极调节。在之前的一项研究中,我们对β细胞浆膜下[Ca2+]i的动态进行了时间解析,其特征是一连串界限分明的尖峰,峰值可达5 µM。应用全内反射荧光(TIRF)显微镜和突触荧光素来观察单个颗粒的融合事件,我们发现在 50 到 150 毫秒内可以同时发生几个融合事件。为了检验单个或聚集的 Ca2+ 通道周围的浆膜下[Ca2+]i 微域是否会导致含胰岛素的囊泡同步释放,我们同时应用了双色 TIRF 显微镜,并以高帧频监测 INS-1 细胞中的 Ca2+ 波动和外泌事件。结果表明,融合可由浆膜下 Ca2+ 尖峰触发。不过,这只占融合事件的少数。大约 90%-95% 的融合事件要么发生在 Ca2+ 尖峰之间,要么偶然与浆膜下 Ca2+ 尖峰重叠。我们的结论是,在葡萄糖诱导和甲苯磺丁酰胺或福斯可林增强的 INS-1 细胞胰岛素释放过程中,只有一小部分外渗事件与电压门控 Ca2+ 通道周围的 Ca2+ 微域紧密耦合。
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引用次数: 0
A novel probe to monitor lysosome-mitochondria contact sites opens up a new path to study neurodegenerative diseases 监测溶酶体-线粒体接触点的新型探针为研究神经退行性疾病开辟了新途径
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-08 DOI: 10.1016/j.ceca.2024.102887
Mai Makino , Shuhei Nakamura
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引用次数: 0
TRPM7 in neurodevelopment and therapeutic prospects for neurodegenerative disease 神经发育中的 TRPM7 和神经退行性疾病的治疗前景
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-06 DOI: 10.1016/j.ceca.2024.102886
Zhengwei Luo , Xinyang Zhang , Andrea Fleig , Daniel Romo , Kenneth G. Hull , F. David Horgen , Hong-Shuo Sun , Zhong-Ping Feng

Neurodevelopment, a complex and highly regulated process, plays a foundational role in shaping the structure and function of the nervous system. The transient receptor potential melastatin 7 (TRPM7), a divalent cation channel with an α-kinase domain, mediates a wide range of cellular functions, including proliferation, migration, cell adhesion, and survival, all of which are essential processes in neurodevelopment. The global knockout of either TRPM7 or TRPM7-kinase is embryonically lethal, highlighting the crucial role of TRPM7 in development in vivo. Subsequent research further revealed that TRPM7 is indeed involved in various key processes throughout neurodevelopment, from maintaining pluripotency during embryogenesis to regulating gastrulation, neural tube closure, axonal outgrowth, synaptic density, and learning and memory. Moreover, a discrepancy in TRPM7 expression and/or function has been associated with neuropathological conditions, including ischemic stroke, Alzheimer's disease, and Parkinson's disease. Understanding the mechanisms of proper neurodevelopment may provide us with the knowledge required to develop therapeutic interventions that can overcome the challenges of regeneration in CNS injuries and neurodegenerative diseases. Considering that ion channels are the third-largest class targeted for drug development, TRPM7′s dual roles in development and degeneration emphasize its therapeutic potential. This review provides a comprehensive overview of the current literature on TRPM7 in various aspects of neurodevelopment. It also discusses the links between neurodevelopment and neurodegeneration, and highlights TRPM7 as a potential therapeutic target for neurodegenerative disorders, with a focus on repair and regeneration.

神经发育是一个复杂而高度调控的过程,在塑造神经系统的结构和功能方面起着基础性作用。瞬时受体电位美司他丁 7(TRPM7)是一种具有α激酶结构域的二价阳离子通道,它介导多种细胞功能,包括增殖、迁移、细胞粘附和存活,所有这些都是神经发育的基本过程。全面敲除TRPM7或TRPM7-激酶会导致胚胎死亡,这突显了TRPM7在体内发育过程中的关键作用。随后的研究进一步发现,TRPM7 确实参与了整个神经发育过程中的各种关键过程,从维持胚胎发生过程中的多能性到调节胃形成、神经管闭合、轴突生长、突触密度以及学习和记忆。此外,TRPM7 表达和/或功能的差异还与神经病理学疾病有关,包括缺血性中风、阿尔茨海默病和帕金森病。了解神经正常发育的机制可能会为我们提供开发治疗干预措施所需的知识,从而克服中枢神经系统损伤和神经退行性疾病再生所面临的挑战。考虑到离子通道是药物开发的第三大目标类别,TRPM7 在发育和退化中的双重作用凸显了其治疗潜力。本综述全面概述了目前有关 TRPM7 在神经发育各方面作用的文献。它还讨论了神经发育和神经退行性变之间的联系,并强调 TRPM7 是神经退行性疾病的潜在治疗靶点,重点关注修复和再生。
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引用次数: 0
PDX1, a transcription factor essential for organ differentiation, regulates SERCA-dependent Ca2+ homeostasis in sensory neurons 器官分化所必需的转录因子 PDX1 可调节感觉神经元中依赖 SERCA 的 Ca2+ 稳态
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-04-02 DOI: 10.1016/j.ceca.2024.102884
Jami L. Saloman , Ariel Y. Epouhe , Catherine F. Ruff , Kathryn M. Albers

Pancreatic and duodenal homeobox 1 (PDX1) is a transcription factor required for the development and differentiation of the pancreas. Previous studies indicated that PDX1 expression was restricted to the gastrointestinal tract. Using a cre-dependent reporter, we observed PDX1-dependent expression of tdtomato (PDX1-tom) in a subpopulation of sensory nerves. Many of these PDX1-tom afferents expressed the neurofilament 200 protein and projected to the skin. Tdtomato-labeled terminals were associated with hair follicles in the form of longitudinal and circumferential lanceolate endings suggesting a role in tactile and proprioceptive perception. To begin to examine the functional significance of PDX1 in afferents, we used Fura-2 imaging to examine calcium (Ca2+) handling under naïve and nerve injury conditions. Neuropathic injury is associated with increased intracellular Ca2+ signaling that in part results from dysregulation of the sarco/endoplasmic reticulum calcium transport ATPase (SERCA). Here we demonstrate that under naïve conditions, PDX1 regulates expression of the SERCA2B isoform in sensory neurons. In response to infraorbital nerve injury, a significant reduction of PDX1 and SERCA2B expression and dysregulation of Ca2+ handling occurs in PDX1-tom trigeminal ganglia neurons. The identification of PDX1 expression in the somatosensory system and its regulation of SERCA2B and Ca2+ handling provide a new mechanism to explain pathological changes in primary afferents that may contribute to pain associated with nerve injury.

胰腺和十二指肠同源框 1(PDX1)是胰腺发育和分化所需的转录因子。以前的研究表明,PDX1 的表达仅限于胃肠道。利用cre依赖性报告基因,我们在感觉神经亚群中观察到了tdtomato(PDX1-tom)的PDX1依赖性表达。其中许多 PDX1-tom 传入神经表达神经丝蛋白 200,并投射到皮肤。Tdtomato标记的末梢以纵向和环向披针形末梢的形式与毛囊相关,这表明它们在触觉和本体感觉中发挥作用。为了开始研究 PDX1 在传入神经中的功能意义,我们使用 Fura-2 成像技术研究了神经损伤条件下的钙离子(Ca2+)处理。神经损伤与细胞内 Ca2+ 信号的增加有关,而 Ca2+ 信号的增加部分源于肌浆/内质网钙离子转运 ATP 酶(SERCA)的失调。我们在此证明,在原始条件下,PDX1 可调节感觉神经元中 SERCA2B 同工酶的表达。在眶下神经损伤后,PDX1 和 SERCA2B 的表达显著减少,PDX1-tom 三叉神经节神经元的 Ca2+ 处理失调。PDX1在躯体感觉系统中的表达及其对SERCA2B和Ca2+处理的调控为解释初级传入神经的病理变化提供了一种新的机制,这种病理变化可能导致与神经损伤相关的疼痛。
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引用次数: 0
Utilization of the genetically encoded calcium indicator Salsa6F in cardiac applications 基因编码钙指示剂 Salsa6F 在心脏领域的应用
IF 4 2区 生物学 Q2 CELL BIOLOGY Pub Date : 2024-03-20 DOI: 10.1016/j.ceca.2024.102873
Karla M. Márquez-Nogueras, Elisa Bovo, Jacy E. Neczypor, Quan Cao, Aleksey V. Zima, Ivana Y. Kuo

Calcium signaling is a critical process required for cellular mechanisms such as cardiomyocyte contraction. The inability of the cell to properly activate or regulate calcium signaling can lead to contractile dysfunction. In isolated cardiomyocytes, calcium signaling has been primarily studied using calcium fluorescent dyes, however these dyes have limited applicability to whole organs. Here, we crossed the Salsa6f mouse which expresses a genetically encoded ratiometric cytosolic calcium indicator with a cardiomyocyte specific inducible cre to temporally-induce expression and studied cytosolic calcium transients in isolated cardiomyocytes and modified Langendorff heart preparations. Isolated cardiomyocytes expressing Salsa6f or Fluo-4AM loaded were compared. We also crossed the Salsa6f mouse with a floxed Polycystin 2 (PC2) mouse to test the feasibility of using the Salsa6f mouse to measure calcium transients in PC2 heterozygous or homozygous knock out mice. Although there are caveats in the applicability of the Salsa6f mouse, there are clear advantages to using the Salsa6f mouse to measure whole heart calcium signals.

钙信号传导是心肌细胞收缩等细胞机制所需的一个关键过程。如果细胞不能正确激活或调节钙信号,就会导致收缩功能障碍。在离体心肌细胞中,主要使用钙荧光染料研究钙信号转导,但这些染料对整个器官的适用性有限。在这里,我们将表达基因编码的比率细胞钙指示剂的 Salsa6f 小鼠与心肌细胞特异性诱导性克隆杂交,以在时间上诱导表达,并研究了离体心肌细胞和改良朗根多夫心脏制备物中的细胞钙瞬态。我们对表达 Salsa6f 或负载 Fluo-4AM 的离体心肌细胞进行了比较。我们还将 Salsa6f 小鼠与杂合多囊胞素 2(PC2)小鼠杂交,以测试使用 Salsa6f 小鼠测量 PC2 杂合或同源基因敲除小鼠钙离子瞬态的可行性。虽然 Salsa6f 小鼠的适用性存在一些注意事项,但使用 Salsa6f 小鼠测量整个心脏钙信号的优势显而易见。
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引用次数: 0
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Cell calcium
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