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Quantitative assessment of (Ca2+)i levels in rat skeletal muscle in vivo.

机译:体内大鼠骨骼肌中(Ca2 +)i水平的定量评估。

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Intracellular free Ca2+ concentration ([Ca2+]i) plays an essential role in physiological regulatory processes and common pathological conditions. Better understanding of these phenomena is still hampered by problems encountered in the quantitative assessment of [Ca2+]i changes, especially in blood-perfused organs. This study demonstrates that the ratiometric fluorescence technique can be adapted for quantitative in vivo [Ca2+]i determinations. The rat spinotrapezius muscle was topically loaded with indo 1-AM and imaged by a cooled digital camera. Ratio images were calculated in small regions (100 micrometers x 100 micrometers) practically devoid of large vessels in the resting state, after 30 min of ischemia, 20 min of reperfusion, or ionomycin or manganate treatments. When we assumed an average [Ca2+]i of 100 nM in the resting blood-perfused muscle, ischemia increased [Ca2+]i to approximately 200 nM. During reperfusion [Ca2+]i decreased to approximately 140 nM. Ionomycin induced an increase in [Ca2+]i to well above 750 nM. Manganate reduced Ca2+-dependent fluorescence to virtually zero. Our main conclusion is that changes in [Ca2+]i can be monitored and quantitatively determined in vivo.
机译:细胞内游离Ca2 +浓度([Ca2 +] i)在生理调节过程和常见病理状况中起着至关重要的作用。对[Ca2 +] i变化的定量评估(尤其是在血液灌注器官中)遇到的问题仍然无法更好地理解这些现象。这项研究表明比例荧光技术可适用于定量体内[Ca2 +] i测定。大鼠脊柱前肌局部装有印度1-AM,并通过冷却的数码相机成像。在缺血30分钟,再灌注20分钟或离子霉素或锰酸盐处理后,在静止状态几乎没有大血管的小区域(100微米x 100微米)中计算比率图像。当我们假设静息血液灌注的肌肉中的平均[Ca2 +] i为100 nM时,缺血会使[Ca2 +] i升高至大约200 nM。在再灌注过程中,[Ca2 +] i降至约140 nM。碘霉素诱导[Ca 2+] i增加至远高于750 nM。锰酸盐将依赖于Ca2 +的荧光降低到几乎为零。我们的主要结论是[Ca2 +] i的变化可以在体内进行监测和定量测定。

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