Ablation of gap junctional communication in hepatocytes of transgenic mice does not lead to disrupted cellular homeostasis or increased spontaneous tumourigenesis

Gap junctions between murine hepatocytes are composed of two subunit proteins, connexin26 (Cx26) and connexin32 (Cx32). Previously, we found increased formation of chemically induced liver tumours but no increase in spontaneous development of preneoplastic hepatic foci in mice that lacked Cx32 and e...

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Published inEuropean journal of cell biology Vol. 85; no. 8; pp. 717 - 728
Main Authors Ott, Thomas, Jokwitz, Melanie, Lenhard, Diana, Romualdi, Alessandro, Dombrowski, Frank, Ittrich, Carina, Schwarz, Michael, Willecke, Klaus
Format Journal Article
LanguageEnglish
Published Germany Elsevier GmbH 01.08.2006
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Abstract Gap junctions between murine hepatocytes are composed of two subunit proteins, connexin26 (Cx26) and connexin32 (Cx32). Previously, we found increased formation of chemically induced liver tumours but no increase in spontaneous development of preneoplastic hepatic foci in mice that lacked Cx32 and expressed decreased amounts of Cx26. In order to clarify this tumour-suppressive effect and to overcome embryonic lethality of constitutive Cx26-deficient mice, cell type-specific targeting of the Cx26 gene was performed. Mice with loxP-flanked Cx26 coding DNA were crossed with mice expressing the Cre recombinase exclusively in hepatocytes. Progeny mice lacking Cx26 in the liver were viable and fertile with no obvious signs of phenotypic alterations. To generate mice that totally lack gap junctional intercellular coupling, these mice were crossed with constitutive Cx32-deficient mice. We found no increase in spontaneously induced liver tumour formation in Cx26 and double deficient Cx26/Cx32 mice. Occasionally, double deficient livers exhibited morphological alterations, like amyloidosis, and a slightly increased basal proliferation rate of hepatocytes. Although the absence of gap junction channels led to altered expression of adhesion-related proteins like E-cadherin and actin, microarray analyses of total liver transcripts yielded only few differences between Cx26-deficient and double deficient livers compared to control samples. Our results suggest that total lack of gap junctional communication due to hepatocytic ablation of Cx26 and Cx32 does not drastically alter basal hepatocytic function and does not lead to increased spontaneous liver tumour formation.
AbstractList Gap junctions between murine hepatocytes are composed of two subunit proteins, connexin26 (Cx26) and connexin32 (Cx32). Previously, we found increased formation of chemically induced liver tumours but no increase in spontaneous development of preneoplastic hepatic foci in mice that lacked Cx32 and expressed decreased amounts of Cx26. In order to clarify this tumour-suppressive effect and to overcome embryonic lethality of constitutive Cx26-deficient mice, cell type-specific targeting of the Cx26 gene was performed. Mice with loxP-flanked Cx26 coding DNA were crossed with mice expressing the Cre recombinase exclusively in hepatocytes. Progeny mice lacking Cx26 in the liver were viable and fertile with no obvious signs of phenotypic alterations. To generate mice that totally lack gap junctional intercellular coupling, these mice were crossed with constitutive Cx32-deficient mice. We found no increase in spontaneously induced liver tumour formation in Cx26 and double deficient Cx26/Cx32 mice. Occasionally, double deficient livers exhibited morphological alterations, like amyloidosis, and a slightly increased basal proliferation rate of hepatocytes. Although the absence of gap junction channels led to altered expression of adhesion-related proteins like E-cadherin and actin, microarray analyses of total liver transcripts yielded only few differences between Cx26-deficient and double deficient livers compared to control samples. Our results suggest that total lack of gap junctional communication due to hepatocytic ablation of Cx26 and Cx32 does not drastically alter basal hepatocytic function and does not lead to increased spontaneous liver tumour formation.
Author Willecke, Klaus
Dombrowski, Frank
Romualdi, Alessandro
Ittrich, Carina
Ott, Thomas
Jokwitz, Melanie
Lenhard, Diana
Schwarz, Michael
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Issue 8
Keywords Connexin26
GJIC, Gap junctional intercellular communication
PH, Partial hepatectomy
Gene targeting
Gap junctions
Liver
Cx26, Connexin26
db-cAMP, Dibutyryl 3,5-cyclic adenosine monophosphate
Connexin32
Proliferation
Cx32, Connexin32
Language English
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Snippet Gap junctions between murine hepatocytes are composed of two subunit proteins, connexin26 (Cx26) and connexin32 (Cx32). Previously, we found increased...
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pubmed
elsevier
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StartPage 717
SubjectTerms Animals
Apoptosis - drug effects
Carcinogens - toxicity
Cell Communication - physiology
Cell Proliferation - drug effects
Connexin 26
Connexin26
Connexin32
Connexins - deficiency
Connexins - genetics
Connexins - physiology
Cx26, Connexin26
Cx32, Connexin32
db-cAMP, Dibutyryl 3,5-cyclic adenosine monophosphate
Diethylnitrosamine - pharmacology
Female
Gap Junction beta-1 Protein
Gap junctions
Gap Junctions - genetics
Gap Junctions - physiology
Gene targeting
GJIC, Gap junctional intercellular communication
Hepatocytes - cytology
Hepatocytes - drug effects
Hepatocytes - metabolism
Homeostasis - genetics
Homeostasis - physiology
Immunoblotting
Immunohistochemistry
Liver
Liver - cytology
Liver - drug effects
Liver - metabolism
Liver Neoplasms, Experimental - chemically induced
Liver Neoplasms, Experimental - genetics
Liver Neoplasms, Experimental - pathology
Male
Mice
Mice, Transgenic
Oligonucleotide Array Sequence Analysis - methods
PH, Partial hepatectomy
Proliferation
Title Ablation of gap junctional communication in hepatocytes of transgenic mice does not lead to disrupted cellular homeostasis or increased spontaneous tumourigenesis
URI https://dx.doi.org/10.1016/j.ejcb.2006.03.004
https://www.ncbi.nlm.nih.gov/pubmed/16740338
Volume 85
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