Circulation Research. 2002
Published online before print June 13, 2002, doi: 10.1161/01.RES.0000025638.24255.AA
Published online before print June 13, 2002, doi: 10.1161/01.RES.0000025638.24255.AA
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Submitted on April 4, 2002
Revised on May 30, 2002
Accepted on May 31, 2002
Cardiac Gap Junction Channels Show Quantitative Differences in Selectivity
Virginijus Valiunas ;
From the Department of Physiology and Biophysics (V.V., P.R.B.), State University of New York at Stony Brook, Stony Brook, NY; and the Department of Pediatrics (E.C.B.), Section of Hematology/Oncology, University of Chicago, Chicago, Ill. V.V. is on leave from the Institute for Biomedical Research, Kaunas Medical University, Kaunas, Lithuania.
* To whom correspondence should be addressed. E-mail: peter{at}mail.pnb.sunysb.edu.
Several proteins including connexin40 (Cx40) and connexin43 (Cx43) form gap junctions between cells of the heart; they may be found separately or may be coexpressed. These connexins form channels with differing conductance and permeability properties. Cx40 and Cx43 are each required for normal electrical conduction between cells in different regions of the heart. We hypothesized that the major difference between these connexins might be in their selective intercellular passage of small molecules such as second messengers, which can be assessed using biologically inert fluorescent probes. Therefore, we designed experimental paradigms to quantitate the permeability properties of these cardiac connexins using simultaneous measurement of junctional conductance (gj) by the double whole-cell patch-clamp technique and intercellular transfer of Lucifer Yellow (LY) by fluorescence microscopy. These studies were performed in HeLa cells stably transfected with Cx40 or Cx43 or cotransfected with both connexins. We found that homotypic Cx43 channels were about 5 times more permeable to LY than homotypic Cx40 channels (flux of 
1560 versus 300 molecules/channel per second). Channels between heterotypic (Cx40-Cx43) cell pairs and between pairs of coexpressing cells exhibited intermediate LY permeability. The permeability ratio for LY relative to monovalent cation (K+) ranged from 0.0025 for Cx40 to 0.028 for Cx43. These permeability ratios suggest that the connexins are highly selective for solutes in the size and charge range of many second messengers. Moreover, the data indicate that coexpression of connexins does not generate unique permeability characteristics, but rather results in an intermediate permeability for solutes involved in metabolic/biochemical coupling.
Key words: connexin40 • connexin43 • electrophysiology • gap junction • permselectivity
This article has been cited by other articles:
 |
|
 |
|
  V. Valiunas, G. Kanaporis, L. Valiuniene, C. Gordon, H. Z. Wang, L. Li, R. B. Robinson, M. R. Rosen, I. S. Cohen, and P. R. Brink Coupling an HCN2-expressing cell to a myocyte creates a two-cell pacing unit J. Physiol., November 1, 2009; 587(21): 5211 - 5226. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Palacios-Prado, S. Sonntag, V. A. Skeberdis, K. Willecke, and F. F. Bukauskas Gating, permselectivity and pH-dependent modulation of channels formed by connexin57, a major connexin of horizontal cells in the mouse retina J. Physiol., July 1, 2009; 587(13): 3251 - 3269. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Yao, T. Oite, and M. Kitamura Gap junctional intercellular communication in the juxtaglomerular apparatus Am J Physiol Renal Physiol, May 1, 2009; 296(5): F939 - F946. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Akoyev, S. Das, S. Jena, L. Grauer, and D. J. Takemoto Hypoxia-Regulated Activity of PKC{epsilon} in the Lens Invest. Ophthalmol. Vis. Sci., March 1, 2009; 50(3): 1271 - 1282. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. K. Verselis, M. P. Trelles, C. Rubinos, T. A. Bargiello, and M. Srinivas Loop Gating of Connexin Hemichannels Involves Movement of Pore-lining Residues in the First Extracellular Loop Domain J. Biol. Chem., February 13, 2009; 284(7): 4484 - 4493. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Burt, T. K. Nelson, A. M. Simon, and J. S. Fang Connexin 37 profoundly slows cell cycle progression in rat insulinoma cells Am J Physiol Cell Physiol, November 1, 2008; 295(5): C1103 - C1112. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Mese, V. Valiunas, P. R. Brink, and T. W. White Connexin26 deafness associated mutations show altered permeability to large cationic molecules Am J Physiol Cell Physiol, October 1, 2008; 295(4): C966 - C974. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Makino, O. Platoshyn, J. Suarez, J. X.-J. Yuan, and W. H. Dillmann Downregulation of connexin40 is associated with coronary endothelial cell dysfunction in streptozotocin-induced diabetic mice Am J Physiol Cell Physiol, July 1, 2008; 295(1): C221 - C230. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Kanaporis, G. Mese, L. Valiuniene, T. W. White, P. R. Brink, and V. Valiunas Gap Junction Channels Exhibit Connexin-specific Permeability to Cyclic Nucleotides J. Gen. Physiol., March 31, 2008; 131(4): 293 - 305. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. DeRosa, C.-H. Xia, X. Gong, and T. W. White The cataract-inducing S50P mutation in Cx50 dominantly alters the channel gating of wild-type lens connexins J. Cell Sci., December 1, 2007; 120(23): 4107 - 4116. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. W. Yum, J. Zhang, V. Valiunas, G. Kanaporis, P. R. Brink, T. W. White, and S. S. Scherer Human connexin26 and connexin30 form functional heteromeric and heterotypic channels Am J Physiol Cell Physiol, September 1, 2007; 293(3): C1032 - C1048. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Rackauskas, V. K. Verselis, and F. F. Bukauskas Permeability of homotypic and heterotypic gap junction channels formed of cardiac connexins mCx30.2, Cx40, Cx43, and Cx45 Am J Physiol Heart Circ Physiol, September 1, 2007; 293(3): H1729 - H1736. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Swietach, A. Rossini, K. W. Spitzer, and R. D. Vaughan-Jones H+ Ion Activation and Inactivation of the Ventricular Gap Junction: A Basis for Spatial Regulation of Intracellular pH Circ. Res., April 13, 2007; 100(7): 1045 - 1054. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-Z. Wang, P. R. Brink, and G. J. Christ Gap junction channel activity in short-term cultured human detrusor myocyte cell pairs: gating and unitary conductances Am J Physiol Cell Physiol, December 1, 2006; 291(6): C1366 - C1376. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Dong, X. Liu, H. Li, B. M. Vertel, and L. Ebihara Role of the N-terminus in permeability of chicken connexin45.6 gap junctional channels J. Physiol., November 1, 2006; 576(3): 787 - 799. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. F. Ek-Vitorin, T. J. King, N. S. Heyman, P. D. Lampe, and J. M. Burt Selectivity of Connexin 43 Channels Is Regulated Through Protein Kinase C-Dependent Phosphorylation Circ. Res., June 23, 2006; 98(12): 1498 - 1505. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. E. Ebong, S. Kim, and N. DePaola Flow regulates intercellular communication in HAEC by assembling functional Cx40 and Cx37 gap junctional channels Am J Physiol Heart Circ Physiol, May 1, 2006; 290(5): H2015 - H2023. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. F. Ek-Vitorin and J. M. Burt Quantification of gap junction selectivity Am J Physiol Cell Physiol, December 1, 2005; 289(6): C1535 - C1546. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V Valiunas, Y. Y Polosina, H Miller, I. A Potapova, L Valiuniene, S Doronin, R. T Mathias, R. B Robinson, M. R Rosen, I. S Cohen, et al. Connexin-specific cell-to-cell transfer of short interfering RNA by gap junctions J. Physiol., October 15, 2005; 568(2): 459 - 468. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Vogalis, C. C. Hegg, and M. T. Lucero Electrical Coupling in Sustentacular Cells of the Mouse Olfactory Epithelium J Neurophysiol, August 1, 2005; 94(2): 1001 - 1012. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. E. Isakson and B. R. Duling Heterocellular Contact at the Myoendothelial Junction Influences Gap Junction Organization Circ. Res., July 8, 2005; 97(1): 44 - 51. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Thimm, A. Mechler, H. Lin, S. Rhee, and R. Lal Calcium-dependent Open/Closed Conformations and Interfacial Energy Maps of Reconstituted Hemichannels J. Biol. Chem., March 18, 2005; 280(11): 10646 - 10654. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Valiunas, R. Mui, E. McLachlan, G. Valdimarsson, P. R. Brink, and T. W. White Biophysical characterization of zebrafish connexin35 hemichannels Am J Physiol Cell Physiol, December 1, 2004; 287(6): C1596 - C1604. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Swietach and R. D. Vaughan-Jones Novel method for measuring junctional proton permeation in isolated ventricular myocyte cell pairs Am J Physiol Heart Circ Physiol, November 1, 2004; 287(5): H2352 - H2363. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Kansui, K. Fujii, K. Nakamura, K. Goto, H. Oniki, I. Abe, Y. Shibata, and M. Iida Angiotensin II receptor blockade corrects altered expression of gap junctions in vascular endothelial cells from hypertensive rats Am J Physiol Heart Circ Physiol, July 1, 2004; 287(1): H216 - H224. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Bao, L. Reuss, and G. A. Altenberg Regulation of Purified and Reconstituted Connexin 43 Hemichannels by Protein Kinase C-mediated Phosphorylation of Serine 368 J. Biol. Chem., May 7, 2004; 279(19): 20058 - 20066. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. P Moreno Biophysical properties of homomeric and heteromultimeric channels formed by cardiac connexins Cardiovasc Res, May 1, 2004; 62(2): 276 - 286. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. E. Saffitz and A. G. Kleber Effects of Mechanical Forces and Mediators of Hypertrophy on Remodeling of Gap Junctions in the Heart Circ. Res., March 19, 2004; 94(5): 585 - 591. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Valiunas, S. Doronin, L. Valiuniene, I. Potapova, J. Zuckerman, B. Walcott, R. B. Robinson, M. R. Rosen, P. R. Brink, and I. S. Cohen Human mesenchymal stem cells make cardiac connexins and form functional gap junctions J. Physiol., March 15, 2004; 555(3): 617 - 626. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Bao, G. A. Altenberg, and L. Reuss Mechanism of regulation of the gap junction protein connexin 43 by protein kinase C-mediated phosphorylation Am J Physiol Cell Physiol, March 1, 2004; 286(3): C647 - C654. [Abstract] [Full Text]
|
|
|
|
|
|
D. Garcia-Dorado, A. Rodriguez-Sinovas, and M. Ruiz-Meana Gap junction-mediated spread of cell injury and death during myocardial ischemia-reperfusion Cardiovasc Res, February 15, 2004; 61(3): 386 - 401. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. Sandow, R. Looft-Wilson, B. Doran, T.H. Grayson, S. S. Segal, and C. E. Hill Expression of homocellular and heterocellular gap junctions in hamster arterioles and feed arteries Cardiovasc Res, December 1, 2003; 60(3): 643 - 653. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Zaniboni, A. Rossini, P. Swietach, N. Banger, K. W. Spitzer, and R. D. Vaughan-Jones Proton Permeation Through the Myocardial Gap Junction Circ. Res., October 17, 2003; 93(8): 726 - 735. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Kronengold, E.B. Trexler, F.F. Bukauskas, T.A. Bargiello, and V.K. Verselis Single-channel SCAM Identifies Pore-lining Residues in the First Extracellular Loop and First Transmembrane Domains of Cx46 Hemichannels J. Gen. Physiol., September 29, 2003; 122(4): 389 - 405. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Yao, M. Suwa, B. Li, K. Kawamura, T. Morioka, and T. Oite ATP-Dependent Mechanism for Coordination of Intercellular Ca2+ Signaling and Renin Secretion in Rat Juxtaglomerular Cells Circ. Res., August 22, 2003; 93(4): 338 - 345. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. J. Martinez-Wittinghan, C. Sellitto, L. Li, X. Gong, P. R. Brink, R. T. Mathias, and T. W. White Dominant cataracts result from incongruous mixing of wild-type lens connexins J. Cell Biol., June 9, 2003; 161(5): 969 - 978. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. W. White Nonredundant Gap Junction Functions Physiology, June 1, 2003; 18(3): 95 - 99. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Delmar Connexin Diversity: Discriminating the Message Circ. Res., July 26, 2002; 91(2): 85 - 86. [Full Text] [PDF]
|
|