Characterization of connexin 30.3 and 43 in thymocytes

Paula Candida Fonseca, Oscar Kenji Nihei, Márcia Urban-Maldonado, Simone Abreu, Antonio Carlos Campos De Carvalho, David C. Spray, Wilson Savino, Luiz Anastacio Alves

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

During maturation, thymocytes interact directly and indirectly with different cell types of the thymic microenvironment. Such a cellular communication has been basically ascribed to soluble factors and surface receptors. However, little attention has been given to cellular communication mediated by gap junctions. The existence of these intercellular channels in the immune system remained a controversial issue since the 1970s until recently, when a growing body of evidence has indicated their presence and physiological roles in the immune system. In this work, we investigated whether thymocytes express gap junction-forming proteins (connexins, Cx) and are capable of forming functional intercellular channels. Using RT-PCR, we demonstrated that thymocytes express the mRNA for two Cx isoforms: Cx30.3 and Cx43, but not for Cx26, Cx30, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx45, Cx46, and Cx50. In addition, the presence of Cx30.3 and Cx43 was confirmed using different techniques (RNase protection assay, western blot and immunofluorescence). However, despite the expression of these two Cxs, we did not detect functional homocellular coupling between thymocytes or between EL-4 cells (a Cx43 expressing thymic lymphoma-derived cell line) or heterocellular coupling between thymocytes and thymic epithelial cells (TEC) or between EL-4 and TEC in unstimulated conditions. Concluding, in this study, we described for the first time the expression of connexins in thymocytes, which may constitute a new molecule having a functional role in thymocytes maturation.

Original languageEnglish (US)
Pages (from-to)65-75
Number of pages11
JournalImmunology Letters
Volume94
Issue number1-2
DOIs
StatePublished - Jun 15 2004

Fingerprint

Connexin 43
Thymocytes
Connexins
Immune System
Epithelial Cells
Cellular Microenvironment
Gap Junctions
Ribonucleases
connexin 30.3
Fluorescent Antibody Technique
Lymphoma
Protein Isoforms
Western Blotting
Cell Line
Polymerase Chain Reaction
Messenger RNA

Keywords

  • Connexins
  • Gap junctions
  • Thymocytes

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Fonseca, P. C., Nihei, O. K., Urban-Maldonado, M., Abreu, S., De Carvalho, A. C. C., Spray, D. C., ... Alves, L. A. (2004). Characterization of connexin 30.3 and 43 in thymocytes. Immunology Letters, 94(1-2), 65-75. https://doi.org/10.1016/j.imlet.2004.03.019

Characterization of connexin 30.3 and 43 in thymocytes. / Fonseca, Paula Candida; Nihei, Oscar Kenji; Urban-Maldonado, Márcia; Abreu, Simone; De Carvalho, Antonio Carlos Campos; Spray, David C.; Savino, Wilson; Alves, Luiz Anastacio.

In: Immunology Letters, Vol. 94, No. 1-2, 15.06.2004, p. 65-75.

Research output: Contribution to journalArticle

Fonseca, PC, Nihei, OK, Urban-Maldonado, M, Abreu, S, De Carvalho, ACC, Spray, DC, Savino, W & Alves, LA 2004, 'Characterization of connexin 30.3 and 43 in thymocytes', Immunology Letters, vol. 94, no. 1-2, pp. 65-75. https://doi.org/10.1016/j.imlet.2004.03.019
Fonseca PC, Nihei OK, Urban-Maldonado M, Abreu S, De Carvalho ACC, Spray DC et al. Characterization of connexin 30.3 and 43 in thymocytes. Immunology Letters. 2004 Jun 15;94(1-2):65-75. https://doi.org/10.1016/j.imlet.2004.03.019
Fonseca, Paula Candida ; Nihei, Oscar Kenji ; Urban-Maldonado, Márcia ; Abreu, Simone ; De Carvalho, Antonio Carlos Campos ; Spray, David C. ; Savino, Wilson ; Alves, Luiz Anastacio. / Characterization of connexin 30.3 and 43 in thymocytes. In: Immunology Letters. 2004 ; Vol. 94, No. 1-2. pp. 65-75.
@article{bb7ea8867d6c4d4cb037dc3698f5a1b0,
title = "Characterization of connexin 30.3 and 43 in thymocytes",
abstract = "During maturation, thymocytes interact directly and indirectly with different cell types of the thymic microenvironment. Such a cellular communication has been basically ascribed to soluble factors and surface receptors. However, little attention has been given to cellular communication mediated by gap junctions. The existence of these intercellular channels in the immune system remained a controversial issue since the 1970s until recently, when a growing body of evidence has indicated their presence and physiological roles in the immune system. In this work, we investigated whether thymocytes express gap junction-forming proteins (connexins, Cx) and are capable of forming functional intercellular channels. Using RT-PCR, we demonstrated that thymocytes express the mRNA for two Cx isoforms: Cx30.3 and Cx43, but not for Cx26, Cx30, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx45, Cx46, and Cx50. In addition, the presence of Cx30.3 and Cx43 was confirmed using different techniques (RNase protection assay, western blot and immunofluorescence). However, despite the expression of these two Cxs, we did not detect functional homocellular coupling between thymocytes or between EL-4 cells (a Cx43 expressing thymic lymphoma-derived cell line) or heterocellular coupling between thymocytes and thymic epithelial cells (TEC) or between EL-4 and TEC in unstimulated conditions. Concluding, in this study, we described for the first time the expression of connexins in thymocytes, which may constitute a new molecule having a functional role in thymocytes maturation.",
keywords = "Connexins, Gap junctions, Thymocytes",
author = "Fonseca, {Paula Candida} and Nihei, {Oscar Kenji} and M{\'a}rcia Urban-Maldonado and Simone Abreu and {De Carvalho}, {Antonio Carlos Campos} and Spray, {David C.} and Wilson Savino and Alves, {Luiz Anastacio}",
year = "2004",
month = "6",
day = "15",
doi = "10.1016/j.imlet.2004.03.019",
language = "English (US)",
volume = "94",
pages = "65--75",
journal = "Immunology Letters",
issn = "0165-2478",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Characterization of connexin 30.3 and 43 in thymocytes

AU - Fonseca, Paula Candida

AU - Nihei, Oscar Kenji

AU - Urban-Maldonado, Márcia

AU - Abreu, Simone

AU - De Carvalho, Antonio Carlos Campos

AU - Spray, David C.

AU - Savino, Wilson

AU - Alves, Luiz Anastacio

PY - 2004/6/15

Y1 - 2004/6/15

N2 - During maturation, thymocytes interact directly and indirectly with different cell types of the thymic microenvironment. Such a cellular communication has been basically ascribed to soluble factors and surface receptors. However, little attention has been given to cellular communication mediated by gap junctions. The existence of these intercellular channels in the immune system remained a controversial issue since the 1970s until recently, when a growing body of evidence has indicated their presence and physiological roles in the immune system. In this work, we investigated whether thymocytes express gap junction-forming proteins (connexins, Cx) and are capable of forming functional intercellular channels. Using RT-PCR, we demonstrated that thymocytes express the mRNA for two Cx isoforms: Cx30.3 and Cx43, but not for Cx26, Cx30, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx45, Cx46, and Cx50. In addition, the presence of Cx30.3 and Cx43 was confirmed using different techniques (RNase protection assay, western blot and immunofluorescence). However, despite the expression of these two Cxs, we did not detect functional homocellular coupling between thymocytes or between EL-4 cells (a Cx43 expressing thymic lymphoma-derived cell line) or heterocellular coupling between thymocytes and thymic epithelial cells (TEC) or between EL-4 and TEC in unstimulated conditions. Concluding, in this study, we described for the first time the expression of connexins in thymocytes, which may constitute a new molecule having a functional role in thymocytes maturation.

AB - During maturation, thymocytes interact directly and indirectly with different cell types of the thymic microenvironment. Such a cellular communication has been basically ascribed to soluble factors and surface receptors. However, little attention has been given to cellular communication mediated by gap junctions. The existence of these intercellular channels in the immune system remained a controversial issue since the 1970s until recently, when a growing body of evidence has indicated their presence and physiological roles in the immune system. In this work, we investigated whether thymocytes express gap junction-forming proteins (connexins, Cx) and are capable of forming functional intercellular channels. Using RT-PCR, we demonstrated that thymocytes express the mRNA for two Cx isoforms: Cx30.3 and Cx43, but not for Cx26, Cx30, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx45, Cx46, and Cx50. In addition, the presence of Cx30.3 and Cx43 was confirmed using different techniques (RNase protection assay, western blot and immunofluorescence). However, despite the expression of these two Cxs, we did not detect functional homocellular coupling between thymocytes or between EL-4 cells (a Cx43 expressing thymic lymphoma-derived cell line) or heterocellular coupling between thymocytes and thymic epithelial cells (TEC) or between EL-4 and TEC in unstimulated conditions. Concluding, in this study, we described for the first time the expression of connexins in thymocytes, which may constitute a new molecule having a functional role in thymocytes maturation.

KW - Connexins

KW - Gap junctions

KW - Thymocytes

UR - http://www.scopus.com/inward/record.url?scp=3242686827&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3242686827&partnerID=8YFLogxK

U2 - 10.1016/j.imlet.2004.03.019

DO - 10.1016/j.imlet.2004.03.019

M3 - Article

C2 - 15234537

AN - SCOPUS:3242686827

VL - 94

SP - 65

EP - 75

JO - Immunology Letters

JF - Immunology Letters

SN - 0165-2478

IS - 1-2

ER -