Role of sodium-calcium exchanger (Ncx1) in embryonic heart development: a transgenic rescue?
Na(+)/Ca(2+) exchanger (Ncx-1) is highly expressed in cardiomyocytes, is thought to be required to maintain a low intracellular Ca(2+) concentration, and may play a role in excitation-contraction coupling. Significantly, targeted deletion of Ncx-1 results in Ncx1-null embryos that do not have a spontaneously beating heart and die in utero. Ultrastructural analysis revealed gross anomalies in the Ncx1-null contractile apparatus, but physiologic analysis showed normal field-stimulated Ca(2+) transients, suggesting that Ncx-1 function may not be critical for Ca(2+) extrusion from the cytosol as previously thought. Using caffeine to empty the intracellular Ca(2+) stores, we show that the sarcoplasmic reticulum is not fully functional within the 9.5-dpc mouse heart, indicating that the sarcoplasmic reticulum is unlikely to account for the unexpected maintenance of intracellular Ca(2+) homeostasis. Using the Ncx1-lacZ reporter, our data indicate restricted expression patterns of Ncx1 and that Ncx1 is highly expressed within the conduction system, suggesting Ncx1 may be required for spontaneous pacemaking activity. To test this hypothesis, we used transgenic mice overexpressing one of the two known adult Ncx1 isoforms under the control of the cardiac-specific a-myosin heavy chain promoter to restore Ncx1 expression within the Ncx1-null hearts. Results indicate that the transgenic re-expression of one Ncx1 isoform was unable to rescue the lethal null mutant phenotype. Furthermore, our in situ results indicate that both known adult Ncx1 isoforms are coexpressed within the embryonic heart, suggesting that effective transgenic rescue may require the presence of both isoforms within the developing heart.
Conway
Simon J
SJ
Institute of Molecular Medicine and Genetics and Department of Cell Biology and Anatomy, Medical College of Georgia, Augusta, Georgia 30912, USA. sconway@mail.mcg.edu
Kruzynska-Frejtag
Agnieszka
A
Wang
Jian
J
Rogers
Rhina
R
Kneer
Paige L
PL
Chen
Hongmei
H
Creazzo
Tony
T
Menick
Donald R
DR
Koushik
Srinagesh V
SV
D000818:Animals; D017931:DNA Primers; D005314:Embryonic and Fetal Development; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006321:Heart / Q000196:embryology / Q000254:growth & development / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009200:Myocardial Contraction / Q000502:physiology; D020033:Protein Isoforms / Q000235:genetics; D019831:Sodium-Calcium Exchanger / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D017931:DNA Primers; D020033:Protein Isoforms; D019831:Sodium-Calcium Exchanger; C110374:sodium-calcium exchanger 1; D001616:beta-Galactosidase