Ratio-metric measurement of intracellular calcium in visceral muscles via selective expression of a yellow cameleon calcium sensor
Ratio-metric measurement of intracellular calcium in visceral muscles via selective expression of a yellow cameleon calcium sensor
复制标题
通过黄色cameleon钙传感器的选择性表达对内脏肌细胞内钙进行比率测量
DOI：
10.1016/j.snb.2022.131756
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Shinsuke Nakayama
中科院分区：
文献类型：
--
作者：
Chiho Takai;Naoko Iwata;Kazunori Kanemaru;Kenji F. Tanaka;Yao Yu;Satoshi Iino;Shinsuke Nakayama
内脏肌肉根据它们的位置和功能表现出不同的收缩模式。大多数内脏肌肉由平滑肌组成，细胞内钙离子浓度([Ca~(2+)]i)的增加会引起收缩，就像横纹肌一样。为了严格评估各种内脏运动的机制，测量[Ca~(2+)]i很重要。然而，由于传感方法的局限性，很难比较不同器官和个体之间的[Ca~(2+)]i。因此，在这项研究中，我们使用转基因小鼠测量了内脏肌肉中的[Ca~(2+)]i，转基因小鼠在骨骼肌和平滑肌肉中都选择性地表达了一种基因编码的比率钙指示剂(YC-Nano50)。我们分别获取了YC-Nano50的CFP和YFP荧光图像，以评估反映[Ca~(2+)]i的荧光共振能量转移(FRET)。在静息状态下，在离体肌中测得的YFP/CFP比值低于骨骼肌，分别对应于[Ca~(2+)]i在约20-40nM和50-80nM的范围内。在内脏平滑肌中，膀胱的YFP/CFP比值最高，胃(胃窦)最低。在由骨骼肌组成的食道中，YFP/CFP比值高于其他内脏肌肉，与腹壁和横隔肌相当。此外，即使在运动过程中，我们也能够测量肌片和从肠道分离的完整节段中的钙瞬变和振荡。这种方法能够阐明钙信号在内脏肌不同功能中的生物学意义。
Visceral muscles exhibit various patterns of contractions depending on their locations and functions. Most visceral muscles are comprised of smooth muscle, and an increase in the intracellular Ca2+concentration ([Ca2+]i) induces contraction, as in striated muscle. In order to critically assess the mechanisms underlying a variety of visceral movements, it is important to measure [Ca2+]i. However, due to limitations of sensing methods, it is difficult to compare [Ca2+]ibetween different organs and individuals. In this study, we thus measured [Ca2+]iin visceral muscles using transgenic mice that selective expressed a genetically encoded ratiometric Ca2+indicator (YC-Nano50) in both skeletal and smooth muscles. We acquired CFP and YFP fluorescence images of YC-Nano50 separately, to evaluate the fluorescence resonance energy transfer (FRET), which reflects [Ca2+]i. In the resting condition, the YFP/CFP ratio measured in isolated muscularis was lower in smooth muscle than in skeletal muscle, corresponding to [Ca2+]iin the range of approximately 20–40 nM and 50–80 nM, respectively. Among visceral smooth muscles, the YFP/CFP ratio was highest in the urinary bladder, and lowest in the stomach (antrum). In the esophagus, comprised of skeletal muscle, the YFP/CFP ratio was higher than in other visceral smooth muscles, and was comparable to that in abdominal wall and diaphragm skeletal muscles. In addition, we were able to measure Ca2+transients and oscillations in muscle sheets and intact segments isolated from the intestine even during movement. This method is able to elucidate the biological significance of Ca2+signaling in diverse functions of visceral muscles.
登录
查看更多内容
脊椎动物肌肉的细胞内 pH 调节。
影响因子：
18.2
作者：
C. Aickin
通讯作者：
C. Aickin
调节 [Ca2]i 平滑肌的细胞机制。
影响因子：
18.2
作者：
C. Breemen;K. Saida
通讯作者：
K. Saida
钙与牛蛙骨骼肌小白蛋白结合的稳态特性：Mg2、pH、离子强度和温度的影响。
影响因子：
2.7
作者：
Y. Ogawa;Masaru Tanokura
通讯作者：
Masaru Tanokura
豚鼠盲肠绦虫平滑肌中的细胞内游离镁：脱钠后镁和 pH 值的伴随分析
DOI：
--
发表时间：
1994
期刊：
The Journal of General Physiology
影响因子：
--
作者：
S. Nakayama;H. Nomura;T. Tomita
通讯作者：
T. Tomita
使用 Yellow Cameleon-Nano140 融合 α-肌动蛋白同时观察年轻心肌细胞的肌节动态和局部 Ca2+ 浓度
DOI：
--
发表时间：
2017
期刊：
影响因子：
--
作者：
塚本精一;藤井輝之;大山廣太郎;下澤東吾;小比類巻生;石渡信一;福田紀男
通讯作者：
福田紀男