Key result
Increasing extracellular calcium concentration from 1.2 mM to 2.0 mM enhanced the amplitude of cytosolic calcium transients by 70% in neonatal rat ventricular myocytes.
Why the study?
Does supraphysiological extracellular calcium concentration alter myocardial calcium handling in neonatal rat ventricular myocytes?
Does supraphysiological extracellular calcium concentration alter myocardial calcium handling in neonatal rat ventricular myocytes?
Absolute Event Rate: 0.528% vs 0.313%
p-value: p=<0.001
Using physiological extracellular calcium concentrations (1.2 mM) is critical for in vitro experiments with neonatal rat cardiomyocytes to prevent artifactual alterations in calcium handling and transporter function.
Supraphysiological calcium may artifactually boost transients in neonatal myocytes; leaves open physiological concentrations for accurate in vitro models.
Calcium ions play an important role in several cell functions, from fertilization to cell death. The cytosolic Ca(2+) concentration is much lower than the extracellular concentration ([Ca(2+)](o)). The latter may markedly affect Ca(2+) fluxes across the cell membrane and thus the cellular Ca(2+) load. Thus, when working with preparations in vitro, it is important to keep [Ca(2+)](o) close to the in vivo value. In this study, we determined the calcemia in immature rats, for which values are currently unavailable, and investigated how supraphysiological [Ca(2+)](o) affects myocardial Ca(2+) handling. Blood ionized [Ca(2+)] was similar in neonatal (2-5 days old) and adults Wistar rats (1.28 ± 0.03 and 1.31 ± 0.03 mmol/l; n=6 and 5, respectively, P>0.37), and lower than the [Ca(2+)](o) range often used in experiments with neonatal myocardial preparations. Cytosolic Ca(2+) transients, measured with indo-1 in neonatal ventricular myocytes, were enhanced by an increase in [Ca(2+)](o) from 1.2 to 2 mM, which also increased the Ca(2+) content in the sarcoplasmic reticulum (SR), and changed the pattern of competition between the main transporters that remove Ca(2+) from the cytosol (SR Ca(2+)-ATPase and Na(+)/Ca(2+) exchanger). These observations stress the importance of using physiological [Ca(2+)](o) values for reliability of results. It is expected that the present calcemia data, reported for the first time in immature rats, may contribute to the refinement of in vitro experiments with neonatal rat preparations.
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Bassani et al. (2012) studied Healthy Wistar rats (neonatal and adult) (n=20). Supraphysiological extracellular calcium concentration ([Ca2+]o) vs. Physiological extracellular calcium concentration (1.2 mM) was evaluated on Twitch Ca2+ transient amplitude (µM) (p=<0.001). Increasing extracellular calcium concentration from 1.2 mM to 2.0 mM enhanced the amplitude of cytosolic calcium transients by 70% in neonatal rat ventricular myocytes.
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