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Am J Physiol Cell Physiol 292: C1942-C1950, 2007. First published January 10, 2007; doi:10.1152/ajpcell.00564.2006
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MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

Changes in regulation of sodium/calcium exchanger of avian ventricular heart cells during embryonic development

Neal Shepherd, Victoria Graham, Bhavya Trevedi, and Tony L. Creazzo

Neonatal/Perinatal Research Institute, Department of Pediatrics/Neonatology Division, Duke University, Durham, North Carolina

Submitted 8 November 2006 ; accepted in final form 4 January 2007

It has been suggested that the sodium/calcium exchanger NCX1 may have a more important physiological role in embryonic and neonatal hearts than in adult hearts. However, in chick heart sarcolemmal vesicles, sodium-dependent calcium transport is reported to be small and, moreover, to be 3–12 times smaller in hearts at embryonic day (ED) 4–5 than at ED18, the opposite of what would be expected of a transporter that is more important in early development. To better assess the role of NCX1 in calcium regulation in the chick embryonic heart, we measured the activity of NCX1 in chick embryonic hearts as extracellular calcium-activated exchanger current (INCX) under controlled ionic conditions. With intracellular calcium concentration ([Ca2+]i) = 47 nM, INCX density increased from 1.34 ± 0.28 pA/pF at ED2 to 3.22 ± 0.55 pA/pF at ED11 (P = 0.006); however, with [Ca2+]i = 481 nM, the increase was small and statistically insignificant, from 4.54 ± 0.77 to 5.88 ± 0.73 pA/pF (P = 0.20, membrane potential = 0 mV, extracellular calcium concentration = 2 mM). Plots of INCX density against [Ca2+]i were well fitted by the Michaelis-Menton equation and extrapolated to identical maximal currents for ED2 and ED11 cells (extracellular calcium concentration = 1, 2, or 4 mM). Thus the increase in INCX at low [Ca2+]i appeared to reflect a developmental change in allosteric regulation of the exchanger by intracellular calcium rather than an increase in the membrane density of NCX1. Supporting this conclusion, RT-PCR demonstrated little change in the amount of mRNA encoding NCX1 expression from ED2 through ED18.

NCX1; chick embryo; allosteric regulation; sodium/calcium exchange current


Address for reprint requests and other correspondence: N. Shepherd, Neonatal/Perinatal Research Institute, Dept. of Pediatrics/Neonatology Division, Duke Univ. Medical Center, DUMC Box 3719, Durham, NC 27710 (e-mail: sheph052{at}duke.edu)






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