Abstract
Because no isoform-specific blocker of NCX has ever been synthesized, a more selective strategy to identify the role of each antiporter isoform in the brain was represented by the generation of knockout and knockin mice for the different isoforms of the antiporter.
Experiments performed in NCX2 and NCX3 knockout mice provided evidence that these two isoforms participate in spatial learning and memory consolidation, although in an opposite manner. These new data from ncx2−/− and ncx3−/− mice may open new experimental avenues for the development of effective therapeutic compounds that, by selectively inhibiting or activating these molecular targets, could treat patients affected by cognitive impairment including Alzheimer’s, Parkinson’s, Huntington’s diseases, and infarct dementia.
More importantly, knockout and knockin mice also provided new relevant information on the role played by NCX in maintaining the intracellular Na+ and Ca2+ homeostasis and in protecting neurons during brain ischemia. In particular, both ncx2−/− and ncx3−/− mice showed an increased neuronal vulnerability after the ischemic insult induced by transient middle cerebral artery occlusion.
As the ubiquitous deletion of NCX1 brings about to an early death of embryos because of a lack of heartbeat, this strategy could not be successfully pursued. However, information on the role of NCX1 in normal and ischemic brain could be obtained by developing conditional knockout mice lacking NCX1 in the brain. Preliminarily results obtained in these conditional mice suggest that also NCX1 protects neurons from ischemic cell death.
Overall, the use of genetic-modified mice for NCX1, NCX2, and NCX3 represents a fruitful strategy to characterize the physiological role exerted by NCX in CNS and to identify the isoforms of the antiporter as potential molecular targets for therapeutic intervention in cerebral ischemia.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
L. Annunziato, G. Pignataro, G.F. Di Renzo, Pharmacology of brain Na+/Ca2+ exchanger: from molecular biology to therapeutic perspectives. Pharmacol. Rev. 56, 633–654 (2004)
L. Annunziato, G. Pignataro, F. Boscia, R. Sirabella, L. Formisano, M. Saggese, O. Cuomo, R. Gala, A. Secondo, D. Viggiano, P. Molinaro, V. Valsecchi, A. Tortiglione, A. Adornetto, A. Scorziello, M. Cataldi, GF. Di Renzo ncx1, ncx2, and ncx3 gene product expression and function in neuronal anoxia and brain ischemia. Ann. N. Y. Acad. Sci. 1099, 413-426 (2007)
F. Boscia, R. Gala, A. Pannaccione, A. Secondo, A. Scorziello, G. Di Renzo, L. Annunziato, NCX1 expression and functional activity increase in microglia invading the infarct core. Stroke 40, 3608–3617 (2009)
F. Boscia, C. D’Avanzo, A. Pannaccione, A. Secondo, A. Casamassa, L. Formisano, N. Guida, L. Annunziato, Silencing or knocking out the Na+/Ca2+ exchanger-3 (NCX3) impairs oligodendrocyte differentiation. Cell. Death. Differ. 19, 562–572 (2011)
A. Canitano, M. Papa, F. Boscia, P. Castaldo, S. Sellitti, M. Taglialatela, L. Annunziato, Brain distribution of the Na+/Ca2+ exchanger-encoding genes NCX1, NCX2, and NCX3 and their related proteins in the central nervous system. Ann. N. Y. Acad. Sci. 976, 394–404 (2002)
C.H. Cho, S.S. Kim, M.J. Jeong, C.O. Lee, H.S. Shin, The Na+ -Ca2+ exchanger is essential for embryonic heart development in mice. Mol. Cells 10, 712–722 (2000)
O. Cuomo, R. Gala, G. Pignataro, F. Boscia, A. Secondo, A. Scorziello, A. Pannaccione, D. Viggiano, A. Adornetto, P. Molinaro, X.F. Li, J. Lytton, G. Di Renzo, L. Annunziato, A critical role for the potassium-dependent sodium-calcium exchanger NCKX2 in protection against focal ischemic brain damage. J. Neurosci. 28, 2053–2063 (2008)
S.M. Finkbeiner, Glial calcium. Glia 9, 83–104 (1993)
A. Fontan-Lozano, R. Romero-Granados, Y. del-Pozo-Martin, I. Suarez-Pereira, J.M. Delgado-Garcia, J.M. Penninger, A.M. Carrion, Lack of DREAM protein enhances learning and memory and slows brain aging. Curr. Biol. 19, 54–60 (2009)
W.F. Goldman, P.J. Yarowsky, M. Juhaszova, B.K. Krueger, M.P. Blaustein, Sodium/calcium exchange in rat cortical astrocytes. J. Neurosci. 14, 5834–5843 (1994)
R. Gomez-Villafuertes, B. Torres, J. Barrio, M. Savignac, N. Gabellini, F. Rizzato, B. Pintado, A. Gutierrez-Adan, B. Mellstrom, E. Carafoli, J.R. Naranjo, Downstream regulatory element antagonist modulator regulates Ca2+ homeostasis and viability in cerebellar neurons. J. Neurosci. 25, 10822–10830 (2005)
H. Hasegawa, M. Muraoka, K. Matsui, A. Kojima, Discovery of a novel potent Na+/Ca2+ exchanger inhibitor: design, synthesis and structure-activity relationships of 3,4-dihydro-2(1H)-quinazolinone derivatives. Bioorg. Med. Chem. Lett. 13, 3471–3475 (2003)
S.A. Henderson, J.I. Goldhaber, J.M. So, T. Han, C. Motter, A. Ngo, C. Chantawansri, M.R. Ritter, M. Friedlander, D.A. Nicoll, J.S. Frank, M.C. Jordan, K.P. Roos, R.S. Ross, K.D. Philipson, Functional adult myocardium in the absence of Na+-Ca2+ exchange: cardiac-specific knockout of NCX1. Circ. Res. 95, 604–611 (2004)
A. Holgado, L. Beauge, Effects of external monovalent cations on Na+-Ca2+ exchange in cultured rat glial cells. Ann. N. Y. Acad. Sci. 779, 279–281 (1996)
T. Iwamoto, S. Kita, YM-244769, a novel Na+/Ca2+ exchange inhibitor that preferentially inhibits NCX3, efficiently protects against hypoxia/reoxygenation-induced SH-SY5Y neuronal cell damage. Mol. Pharmacol. 70, 2075–2083 (2006)
T. Iwamoto, Y. Inoue, K. Ito, T. Sakaue, S. Kita, T. Katsuragi, The exchanger inhibitory peptide region-dependent inhibition of Na+/Ca2+ exchange by SN-6 [2-[4-(4-nitrobenzyloxy)benzyl]thiazolidine-4-carboxylic acid ethyl ester], a novel benzyloxyphenyl derivative. Mol. Pharmacol. 66, 45–55 (2004)
D. Jeon, Y.M. Yang, M.J. Jeong, K.D. Philipson, H. Rhim, H.S. Shin, Enhanced learning and memory in mice lacking Na+/Ca2+ exchanger 2. Neuron 38, 965–976 (2003)
D. Jeon, K. Chu, K.H. Jung, M. Kim, B.W. Yoon, C.J. Lee, U. Oh, H.S. Shin, Na+/Ca2+ exchanger 2 is neuroprotective by exporting Ca2+ during a transient focal cerebral ischemia in the mouse. Cell Calcium 43, 482–491 (2008)
M. Juhaszova, H. Shimizu, M.L. Borin, R.K. Yip, E.M. Santiago, G.E. Lindenmayer, M.P. Blaustein, Localization of the Na+-Ca2+ exchanger in vascular smooth muscle, and in neurons and astrocytes. Ann. N. Y. Acad. Sci. 779, 318–335 (1996)
S.V. Koushik, J. Wang, R. Rogers, D. Moskophidis, N.A. Lambert, T.L. Creazzo, S.J. Conway, Targeted inactivation of the sodium-calcium exchanger (Ncx1) results in the lack of a heartbeat and abnormal myofibrillar organization. FASEB J. 15, 1209–1211 (2001)
Z. Li, S. Matsuoka, L.V. Hryshko, D.A. Nicoll, M.M. Bersohn, E.P. Burke, R.P. Lifton, K.D. Philipson, Cloning of the NCX2 isoform of the plasma membrane Na+-Ca2+ exchanger. J. Biol. Chem. 269, 17434–17439 (1994)
X.F. Li, L. Kiedrowski, F. Tremblay, F.R. Fernandez, M. Perizzolo, R.J. Winkfein, R.W. Turner, J.S. Bains, D.E. Rancourt, J. Lytton, Importance of K+-dependent Na+/Ca2+-exchanger 2, NCKX2, in motor learning and memory. J. Biol. Chem. 281, 6273–6282 (2006)
B. Linck, Z. Qiu, Z. He, Q. Tong, D.W. Hilgemann, K.D. Philipson, Functional comparison of the three isoforms of the Na+/Ca2+ exchanger (NCX1, NCX2, NCX3). Am. J. Physiol. 274, C415–C423 (1998)
J. Luo, Y. Wang, X. Chen, H. Chen, D.B. Kintner, G.E. Shull, K.D. Philipson, D. Sun, Increased tolerance to ischemic neuronal damage by knockdown of Na+-Ca2+ exchanger isoform 1. Ann. N. Y. Acad. Sci. 1099, 292–305 (2007)
T. Matsuda, N. Arakawa, K. Takuma, Y. Kishida, Y. Kawasaki, M. Sakaue, K. Takahashi, T. Takahashi, T. Suzuki, T. Ota, A. Hamano-Takahashi, M. Onishi, Y. Tanaka, K. Kameo, A. Baba, SEA0400, a novel and selective inhibitor of the Na+-Ca2+ exchanger, attenuates reperfusion injury in the in vitro and in vivo cerebral ischemic models. J. Pharmacol. Exp. Ther. 298, 249–256 (2001)
P. Molinaro, O. Cuomo, G. Pignataro, F. Boscia, R. Sirabella, A. Pannaccione, A. Secondo, A. Scorziello, A. Adornetto, R. Gala, D. Viggiano, S. Sokolow, A. Herchuelz, S. Schurmans, G. Di Renzo, L. Annunziato, Targeted disruption of Na+/Ca2+ exchanger 3 (NCX3) gene leads to a worsening of ischemic brain damage. J. Neurosci. 28, 1179–1184 (2008)
P. Molinaro, D. Viggiano, R. Nistico, R. Sirabella, A. Secondo, F. Boscia, A. Pannaccione, A. Scorziello, B. Mehdawy, S. Sokolow, A. Herchuelz, G.F. Di Renzo, L. Annunziato, Na+-Ca2+ exchanger (NCX3) knock-out mice display an impairment in hippocampal long-term potentiation and spatial learning and memory. J. Neurosci. 31, 7312–7321 (2011)
P. Molinaro, M. Cantile, O. Cuomo, A. Secondo, A. Pannaccione, P. Ambrosino, G. Pignataro, F. Fiorino, B. Severino, E. Gatta, M.J. Sisalli, M. Milanese, A. Scorziello, G. Bonanno, M. Robello, V. Santagada, G. Caliendo, G. Di Renzo, L. Annunziato, Neurounina-1, a Novel Compound that Increases Na+/Ca2+ Exchanger Activity, Effectively Protects Against Stroke Damage. Mol Pharmacol. doi:10.1124/mol.112.080986 (2012)
D.A. Nicoll, S. Longoni, K.D. Philipson, Molecular cloning and functional expression of the cardiac sarcolemmal Na+-Ca2+ exchanger. Science 250, 562–565 (1990)
D.A. Nicoll, B.D. Quednau, Z. Qui, Y.R. Xia, A.J. Lusis, K.D. Philipson, Cloning of a third mammalian Na+-Ca2+ exchanger, NCX3. J. Biol. Chem. 271, 24914–24921 (1996)
M. Papa, A. Canitano, F. Boscia, P. Castaldo, S. Sellitti, H. Porzig, M. Taglialatela, L. Annunziato, Differential expression of the Na+-Ca2+ exchanger transcripts and proteins in rat brain regions. J. Comp. Neurol. 461, 31–48 (2003)
G. Pignataro, R. Gala, O. Cuomo, A. Tortiglione, L. Giaccio, P. Castaldo, R. Sirabella, C. Matrone, A. Canitano, S. Amoroso, G. Di Renzo, L. Annunziato, Two sodium/calcium exchanger gene products, NCX1 and NCX3, play a major role in the development of permanent focal cerebral ischemia. Stroke 35, 2566–2570 (2004)
A.J. Pintado, C.J. Herrero, A.G. Garcia, C. Montiel, The novel Na+/Ca2+ exchange inhibitor KB-R7943 also blocks native and expressed neuronal nicotinic receptors. Br. J. Pharmacol. 130, 1893–1902 (2000)
H. Reuter, S.A. Henderson, T. Han, T. Matsuda, A. Baba, R.S. Ross, J.I. Goldhaber, K.D. Philipson, Knockout mice for pharmacological screening: testing the specificity of Na+-Ca2+ exchange inhibitors. Circ. Res. 91, 90–92 (2002a)
H. Reuter, S.A. Henderson, T. Han, R.S. Ross, J.I. Goldhaber, K.D. Philipson, The Na+-Ca2+ exchanger is essential for the action of cardiac glycosides. Circ. Res. 90, 305–308 (2002b)
A. Secondo, R.I. Staiano, A. Scorziello, R. Sirabella, F. Boscia, A. Adornetto, V. Valsecchi, P. Molinaro, L.M. Canzoniero, G. Di Renzo, L. Annunziato, BHK cells transfected with NCX3 are more resistant to hypoxia followed by reoxygenation than those transfected with NCX1 and NCX2: Possible relationship with mitochondrial membrane potential. Cell Calcium 42, 521–535 (2007)
A. Secondo, A. Pannaccione, P. Molinaro, P. Ambrosino, P. Lippiello, A. Esposito, M. Cantile, P.R. Khatri, D. Melisi, G. Di Renzo, L. Annunziato, Molecular pharmacology of the amiloride analog 3-amino-6-chloro-5-[(4-chloro-benzyl)amino]-n-[[(2,4-dimethylbenzyl)-amino] iminomethyl]-pyrazinecarboxamide (CB-DMB) as a pan inhibitor of the Na+-Ca2+ exchanger isoforms NCX1, NCX2, and NCX3 in stably transfected cells. J. Pharmacol. Exp. Ther. 331, 212–221 (2009)
A.J. Silva, R. Paylor, J.M. Wehner, S. Tonegawa, Impaired spatial learning in alpha-calcium-calmodulin kinase II mutant mice. Science 257, 206–211 (1992a)
A.J. Silva, C.F. Stevens, S. Tonegawa, Y. Wang, Deficient hippocampal long-term potentiation in alpha-calcium-calmodulin kinase II mutant mice. Science 257, 201–206 (1992b)
S. Sokolow, M. Manto, P. Gailly, J. Molgo, C. Vandebrouck, J.M. Vanderwinden, A. Herchuelz, S. Schurmans, Impaired neuromuscular transmission and skeletal muscle fiber necrosis in mice lacking Na/Ca exchanger 3. J. Clin. Invest. 113, 265–273 (2004)
K. Takuma, T. Matsuda, H. Hashimoto, S. Asano, A. Baba, Cultured rat astrocytes possess Na+-Ca2+ exchanger. Glia 12, 336–342 (1994)
M. Tsoi, K.H. Rhee, D. Bungard, X.F. Li, S.L. Lee, R.N. Auer, J. Lytton, Molecular cloning of a novel potassium-dependent sodium-calcium exchanger from rat brain. J. Biol. Chem. 273, 4155–4162 (1998)
K. Wakimoto, K. Kobayashi, O.M. Kuro, A. Yao, T. Iwamoto, N. Yanaka, S. Kita, A. Nishida, S. Azuma, Y. Toyoda, K. Omori, H. Imahie, T. Oka, S. Kudoh, O. Kohmoto, Y. Yazaki, M. Shigekawa, Y. Imai, Y. Nabeshima, I. Komuro, Targeted disruption of Na+/Ca2+ exchanger gene leads to cardiomyocyte apoptosis and defects in heartbeat. J. Biol. Chem. 275, 36991–36998 (2000)
T. Watano, Y. Harada, K. Harada, N. Nishimura, Effect of Na+/Ca2+ exchange inhibitor, KB-R7943 on ouabain-induced arrhythmias in guinea-pigs. Br. J. Pharmacol. 127, 1846–1850 (1999)
L.J. Wu, B. Mellstrom, H. Wang, M. Ren, S. Domingo, S.S. Kim, X.Y. Li, T. Chen, J.R. Naranjo, M. Zhuo, DREAM (downstream regulatory element antagonist modulator) contributes to synaptic depression and contextual fear memory. Mol. Brain 3, 3 (2010)
Acknowledgments
This work was supported by COFIN2008; Ricerca-Sanitaria RF-FSL352059 Ricerca finalizzata 2006; Ricerca-Oncologica 2006; Progetto-Strategico2007; Progetto Ordinario 2007; Ricerca finalizzata 2009; Ricerca-Sanitaria progetto Ordinario by Ministero della Salute 2008 all to LA
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this chapter
Cite this chapter
Molinaro, P. et al. (2013). Genetically Modified Mice as a Strategy to Unravel the Role Played by the Na+/Ca2+ Exchanger in Brain Ischemia and in Spatial Learning and Memory Deficits. In: Annunziato, L. (eds) Sodium Calcium Exchange: A Growing Spectrum of Pathophysiological Implications. Advances in Experimental Medicine and Biology, vol 961. Springer, Boston, MA. https://doi.org/10.1007/978-1-4614-4756-6_18
Download citation
DOI: https://doi.org/10.1007/978-1-4614-4756-6_18
Published:
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4614-4755-9
Online ISBN: 978-1-4614-4756-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)