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Inducible Coexpression of Connexin37 or Connexin40 with Connexin43 Selectively Affects Intercellular Molecular Transfer

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Abstract

Many tissues express multiple gap junction proteins, or connexins (Cx); for example, Cx43, Cx40, and Cx37 are coexpressed in vascular cells. This study was undertaken to elucidate the consequences of coexpression of Cx40 or Cx37 with Cx43 at different ratios. EcR-293 cells (which endogenously produce Cx43) were transfected with ecdysone-inducible plasmids encoding Cx37 or Cx40. Immmunoblotting showed a ponasterone dose-dependent induction of Cx37 or Cx40 while constant levels of Cx43 were maintained. The coexpressed connexins colocalized at appositional membranes. Double whole-cell patch clamp recordings showed no significant change in total junctional conductances in cells treated with 0, 0.5, or 4 μM ponasterone; however, they did show a diversity of unitary channel sizes consistent with the induced connexin expression. In cells with induced expression of either Cx40 or Cx37, intercellular transfer of microinjected Lucifer yellow was reduced, but transfer of NBD-TMA (2-(4-nitro-2,1,3-benzoxadiol-7-yl)[aminoethyl]trimethylammonium) was not affected. In cocultures containing uninduced EcR cells together with cells induced to coexpress Cx37 or Cx40, Lucifer yellow transfer was observed only between the cells expressing Cx43 alone. These data show that induced expression of either Cx37 or Cx40 in Cx43-expressing cells can selectively alter the intercellular exchange of some molecules without affecting the transfer of others.

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Acknowledgments

This work was supported by National Institutes of Health grants HL59199 (ECB), 5R01HL058732 (JMB), and 5R01HL077675 (JMB).

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Authors and Affiliations

  1. Department of Pediatrics, University of Chicago, Chicago, IL, USA

    Joanna Gemel & Eric C. Beyer

  2. Department of Physiology, University of Arizona, Tucson, AZ, USA

    Tasha K. Nelson & Janis M. Burt

  3. Section of Pediatric Hematology/Oncology, University of Chicago, 900 E 57th St., KCBD 5152, Chicago, IL, 60637, USA

    Eric C. Beyer

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  1. Joanna Gemel
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  2. Tasha K. Nelson
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  3. Janis M. Burt
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  4. Eric C. Beyer
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Correspondence to Eric C. Beyer.

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Gemel, J., Nelson, T.K., Burt, J.M. et al. Inducible Coexpression of Connexin37 or Connexin40 with Connexin43 Selectively Affects Intercellular Molecular Transfer. J Membrane Biol 245, 231–241 (2012). https://doi.org/10.1007/s00232-012-9444-4

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