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通过19F核磁共振对心动周期中的钙离子瞬变、镁离子和pH值进行测量。

Ca2+ transient, Mg2+, and pH measurements in the cardiac cycle by 19F NMR.

作者信息

Kirschenlohr H L, Metcalfe J C, Morris P G, Rodrigo G C, Smith G A

机构信息

Department of Biochemistry, University of Cambridge, United Kingdom.

出版信息

Proc Natl Acad Sci U S A. 1988 Dec;85(23):9017-21. doi: 10.1073/pnas.85.23.9017.

Abstract

19F NMR indicators have been used to measure the free cytosolic cation concentrations ([Mn+]i, where M is the atomic symbol and n is the value of the charge) of Ca2+, H+, and Mg2+ in perfused ferret hearts. The [Ca2+]i transient, cytosolic pH (pHi), and [Mg2+]i have also been followed at 16 phases in the cardiac cycle in hearts paced at 1.25 Hz at 30 degrees C. The initial [Ca2+]i rose rapidly after a 50-ms delay, was maximal at greater than 1.5 microM after 150 ms, and declined thereafter to the initial concentration. In contrast, no significant changes in pHi (pH 7.03 +/- 0.08) or [Mg2+]i (1.2 +/- 0.1 mM) were detected in the cycle. A decrease in developed pressure when the [Ca2+]i indicator (but not the pHi or [Mg2+]i indicator) was loaded into hearts was substantially reversed by the addition of 50 microM ZnCl2 to the perfusion medium. The Zn2+ was taken up into the myoplasm and displaced Ca2+ bound to the indicator, a symmetrically substituted difluoro derivative of 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (5FBAPTA), as evidenced by the appearance of the Zn-5FBAPTA resonance. The decrease in developed pressure caused by 5FBAPTA, therefore, may be due to its Ca2+ buffering effect on the myoplasm. By coloading hearts with the [Ca2+]i and pHi indicators, simultaneous measurement of several [Mn+]i was demonstrated, which should provide a useful addition to the methods available to monitor cardiac function and pharmacology.

摘要

19F核磁共振指示剂已被用于测量灌注雪貂心脏中Ca2+、H+和Mg2+的游离胞质阳离子浓度([Mn+]i,其中M为原子符号,n为电荷值)。在30℃下以1.25Hz起搏的心脏的心动周期的16个阶段,还对[Ca2+]i瞬变、胞质pH(pHi)和[Mg2+]i进行了跟踪。初始[Ca2+]i在延迟50毫秒后迅速上升,在150毫秒后大于1.5微摩尔时达到最大值,此后下降至初始浓度。相比之下,在该周期中未检测到pHi(pH 7.03±0.08)或[Mg2+]i(1.2±0.1毫摩尔)有显著变化。当将[Ca2+]i指示剂(而非pHi或[Mg2+]i指示剂)加载到心脏中时,所产生压力的降低通过向灌注培养基中添加50微摩尔ZnCl2而基本得到逆转。Zn2+被摄取到肌浆中,并取代了与指示剂结合的Ca2+,该指示剂是1,2-双(邻氨基苯氧基)乙烷-N,N,N',N'-四乙酸(5FBAPTA)的对称取代二氟衍生物,Zn-5FBAPTA共振的出现证明了这一点。因此,5FBAPTA引起的所产生压力的降低可能是由于其对肌浆的Ca2+缓冲作用。通过将[Ca2+]i和pHi指示剂共同加载到心脏中,证明了可以同时测量几种[Mn+]i,这应为监测心脏功能和药理学的现有方法提供有益补充。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b5c8/282653/a6def8fe0e2a/pnas00302-0257-a.jpg

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