{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,28]],"date-time":"2025-09-28T20:32:48Z","timestamp":1759091568640,"version":"3.37.3"},"reference-count":19,"publisher":"Rockefeller University Press","issue":"1","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2016,1,1]]},"abstract":"<jats:p>The divalent cation chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N\u2032,N\u2032-tetraacetic acid (BAPTA), often used to buffer physiological changes in cytosolic Ca2+, also binds Zn2+ with high affinity. In a recently published method (Lamboley et al. 2015. J. Gen. Physiol. http:\/\/dx.doi.org\/10.1085\/jgp.201411250), the absorbance shift of BAPTA at 292 nm was successfully used to determine the total calcium concentrations of various skeletal muscle tissues. In the present study, we show that endogenous Zn2+ in rat skeletal muscle tissue can be unknowingly measured as \u201cCa2+,\u201d unless appropriate measures are taken to eliminate Zn2+ interference. We analyzed two rat skeletal muscle tissues, soleus and plantaris, for total calcium and zinc using either inductively coupled plasma mass spectrometry (ICP-MS) or the BAPTA method described above. ICP-MS analysis showed that total zinc contents in soleus and plantaris were large enough to affect the determination of total calcium by the BAPTA method (calcium = 1.72 \u00b1 0.31 and 1.96 \u00b1 0.14, and zinc = 0.528 \u00b1 0.04 and 0.192 \u00b1 0.01; mean \u00b1 standard error of the mean [SEM]; n = 5; mmole\/kg, respectively). We next analyzed total calcium using BAPTA but included the Zn2+-specific chelator N,N,N\u2032,N\u2032-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) that buffers Zn2+ without affecting Ca2+\/BAPTA binding. We found that estimated concentrations of total calcium ([CaT]WM) in soleus and plantaris were reduced after TPEN addition ([CaT]WM = 3.71 \u00b1 0.62 and 3.57 \u00b1 0.64 without TPEN and 3.39 \u00b1 0.64 and 3.42 \u00b1 0.62 with TPEN; mean \u00b1 SEM; n = 3; mmole\/kg, respectively). Thus, we show that a straightforward correction can be applied to the BAPTA method to improve the accuracy of the determination of total calcium that should be applicable to most any tissue studied. In addition, we show that using TPEN in combination with the BAPTA method allows one to make reasonable estimates of total zinc concentration that are in agreement with the direct determination of zinc concentration by ICP-MS.<\/jats:p>","DOI":"10.1085\/jgp.201511493","type":"journal-article","created":{"date-parts":[[2015,12,28]],"date-time":"2015-12-28T14:28:15Z","timestamp":1451312895000},"page":"95-102","update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":4,"title":["Zinc flexes its muscle: Correcting a novel analysis of calcium for zinc interference uncovers a method to measure zinc"],"prefix":"10.1085","volume":"147","author":[{"given":"Cheng","family":"Qian","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Neuroscience Program, Ohio University, Athens, OH 45701"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert A.","family":"Colvin","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Neuroscience Program, Ohio University, Athens, OH 45701"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"291","published-online":{"date-parts":[[2015,12,28]]},"reference":[{"key":"2023072623434392900_bib1","doi-asserted-by":"crossref","first-page":"2719","DOI":"10.1016\/S0021-9258(18)89421-2","article-title":"Cytosolic Ca2+ homeostasis in Ehrlich and Yoshida carcinomas. A new, membrane-permeant chelator of heavy metals reveals that these ascites tumor cell lines have normal cytosolic free Ca2+","volume":"260","author":"Arslan","year":"1985","journal-title":"J. Biol. Chem."},{"key":"2023072623434392900_bib2","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1042\/bj3220793","article-title":"Study of the interactions of cadmium and zinc ions with cellular calcium homoeostasis using 19F-NMR spectroscopy","volume":"322","author":"Benters","year":"1997","journal-title":"Biochem. J."},{"key":"2023072623434392900_bib3","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1021\/cb500043a","article-title":"Measuring picomolar intracellular exchangeable zinc in PC-12 cells using a ratiometric fluorescence biosensor","volume":"1","author":"Bozym","year":"2006","journal-title":"ACS Chem. Biol."},{"key":"2023072623434392900_bib4","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1111\/j.1748-1716.2009.01983.x","article-title":"Ageing influences myonuclear domain size differently in fast and slow skeletal muscle of rats","volume":"197","author":"Brooks","year":"2009","journal-title":"Acta Physiol. (Oxf.)."},{"key":"2023072623434392900_bib5","doi-asserted-by":"publisher","first-page":"C726","DOI":"10.1152\/ajpcell.00541.2007","article-title":"Insights into Zn2+ homeostasis in neurons from experimental and modeling studies","volume":"294","author":"Colvin","year":"2008","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"2023072623434392900_bib6","doi-asserted-by":"publisher","first-page":"306","DOI":"10.1039\/b926662c","article-title":"Cytosolic zinc buffering and muffling: Their role in intracellular zinc homeostasis","volume":"2","author":"Colvin","year":"2010","journal-title":"Metallomics."},{"key":"2023072623434392900_bib7","doi-asserted-by":"publisher","first-page":"1111","DOI":"10.1039\/C5MT00084J","article-title":"Understanding metal homeostasis in primary cultured neurons. Studies using single neuron subcellular and quantitative metallomics","volume":"7","author":"Colvin","year":"2015","journal-title":"Metallomics."},{"key":"2023072623434392900_bib8","doi-asserted-by":"publisher","first-page":"838","DOI":"10.1016\/S0006-291X(89)80042-7","article-title":"Zinc forms complexes with higher kinetical stability than calcium, 5-F-BAPTA as a good example","volume":"165","author":"Csermely","year":"1989","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2023072623434392900_bib9","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1002\/biof.1114","article-title":"Zinc signals and immune function","volume":"40","author":"Haase","year":"2014","journal-title":"Biofactors."},{"key":"2023072623434392900_bib10","doi-asserted-by":"crossref","first-page":"8148","DOI":"10.1016\/S0021-9258(18)34309-6","article-title":"Divalent cation-nucleotide complex at the exchangeable nucleotide binding site of tubulin","volume":"257","author":"Jemiolo","year":"1982","journal-title":"J. Biol. Chem."},{"key":"2023072623434392900_bib11","doi-asserted-by":"publisher","first-page":"2125","DOI":"10.1073\/pnas.75.5.2125","article-title":"Isolation of a low molecular weight Ca2+ carrier from calf heart inner mitochondrial membrane","volume":"75","author":"Jeng","year":"1978","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072623434392900_bib12","doi-asserted-by":"publisher","first-page":"244","DOI":"10.1039\/C4MT00094C","article-title":"Relationship between the architecture of zinc coordination and zinc binding affinity in proteins\u2014insights into zinc regulation","volume":"7","author":"Kocha\u0144czyk","year":"2015","journal-title":"Metallomics."},{"key":"2023072623434392900_bib13","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1085\/jgp.201411250","article-title":"New method for determining total calcium content in tissue applied to skeletal muscle with and without calsequestrin","volume":"145","author":"Lamboley","year":"2015","journal-title":"J. Gen. Physiol."},{"key":"2023072623434392900_bib14","doi-asserted-by":"publisher","first-page":"202","DOI":"10.1039\/C4MT00230J","article-title":"Analyzing free zinc(II) ion concentrations in cell biology with fluorescent chelating molecules","volume":"7","author":"Maret","year":"2015","journal-title":"Metallomics."},{"key":"2023072623434392900_bib15","doi-asserted-by":"publisher","first-page":"393","DOI":"10.1016\/j.ceca.2006.04.008","article-title":"Determining zinc with commonly used calcium and zinc fluorescent indicators, a question on calcium signals","volume":"40","author":"Martin","year":"2006","journal-title":"Cell Calcium."},{"key":"2023072623434392900_bib16","doi-asserted-by":"publisher","first-page":"2396","DOI":"10.1021\/bi00552a018","article-title":"New calcium indicators and buffers with high selectivity against magnesium and protons: design, synthesis, and properties of prototype structures","volume":"19","author":"Tsien","year":"1980","journal-title":"Biochemistry."},{"key":"2023072623434392900_bib17","doi-asserted-by":"publisher","first-page":"634","DOI":"10.1093\/cvr\/cvq352","article-title":"Intracellular free zinc during cardiac excitation\u2013contraction cycle: calcium and redox dependencies","volume":"89","author":"Tuncay","year":"2011","journal-title":"Cardiovasc. Res."},{"key":"2023072623434392900_bib18","doi-asserted-by":"publisher","first-page":"539","DOI":"10.1016\/0041-008X(84)90190-X","article-title":"In vitro cadmium-DNA interactions: Cooperativity of cadmium binding and competitive antagonism by calcium, magnesium, and zinc","volume":"75","author":"Waalkes","year":"1984","journal-title":"Toxicol. Appl. Pharmacol."},{"key":"2023072623434392900_bib19","doi-asserted-by":"publisher","first-page":"17599","DOI":"10.1074\/jbc.M115.661280","article-title":"Intracellular zinc modulates cardiac ryanodine receptor-mediated calcium release","volume":"290","author":"Woodier","year":"2015","journal-title":"J. Biol. Chem."}],"container-title":["Journal of General Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/147\/1\/95\/1795152\/jgp_201511493.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/147\/1\/95\/1795152\/jgp_201511493.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,26]],"date-time":"2023-07-26T23:44:03Z","timestamp":1690415043000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jgp\/article\/147\/1\/95\/43474\/Zinc-flexes-its-muscle-Correcting-a-novel-analysis"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,12,28]]},"references-count":19,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2016,1,1]]}},"URL":"https:\/\/doi.org\/10.1085\/jgp.201511493","relation":{},"ISSN":["1540-7748","0022-1295"],"issn-type":[{"type":"electronic","value":"1540-7748"},{"type":"print","value":"0022-1295"}],"subject":[],"published":{"date-parts":[[2015,12,28]]}}}