Reduced cell-cell communication in experimentally induced autoimmune thyroid disease.
We have recently described a spontaneous murine model of autoimmune thyroid disease. The disorder was in part characterized by reduced thyroid epithelial cell-cell communication that was associated with abnormalities in three major connexins. To compare whether this finding was a common secondary occurrence in autoimmune thyroid disease, or unique to the spontaneous development in the MRL mice, we induced thyroiditis in Lewis rats. Immunization with thyroid extract and thyroglobulin resulted in extensive lymphocytic infiltration and increased expression of major histocompatibility gene complex (MHC) class II surface antigen in the diseased thyroid. Both experimental and control rat thyroid tissues produced gap junction proteins connexin 43, connexin 32, and connexin 26. The connexins in nondiseased tissue was located in the plasma membrane at points of cell-cell contact and labeled as discrete arrays of punctate fluorescence. The quantity of all three connexins were reduced in the diseased thyroid tissue. More importantly, the connexin proteins were not distributed as gap junctions at contacting cell interfaces. Both nondiseased and diseased thyroid tissue expressed messenger RNA (mRNA) for the three connexins, but the diseased tissue had reduced levels of mRNA for connexin 43 (45%), and to a lesser extent, connexin 26 (25%) and connexin 32 (20%). The reduced connexin mRNA, protein, and lack of assembled gap junctions measured in the diseased tissue were obtained under conditions where the infiltrating cells and their potent cytokine products were continuously present. To determine if this difference persisted when these inflammatory components were absent, primary cultures of thyroid cells from control and experimental rats were established and connexin localization experiments repeated. The diseased thyroid cells, like the diseased tissue, lacked plasma membrane associated connexin protein. The lack of gap junction assembly in the thyrocytes cultured from the diseased tissue was accompanied by a loss of functional coupling. Collectively, the data document that autoimmune diseased thyroid tissue from both the spontaneous mouse and induced rat models have reduced plasma membrane assembled gap junctions and deficient intercellular communication as determined by the inability to transfer lucifer yellow dye to contiguous cells. Nondiseased cultured thyrocyte monolayers and follicles transferred dye to second and third order neighboring cells in 80 and 95% of trials, respectively. In contrast, only 5-10% of the diseased thyrocytes transferred microinjected dye, and in these cases the transit was limited to primary contacting cells. Culturing removed inequities introduced by the infiltrating cells and their products. However, the established cultures of diseased thyroid cells retained their communication deficiency. This suggests that the loss of communication may be a common abnormality in autoimmune disease, and furthermore, this uncoupling could contribute to the loss of coordinated hormonal regulation (hypothyroidism) in the diseased thyroid gland in the absence of thyroid cell destruction.
Green
L M
LM
Research Service, J.L. Pettis Memorial Veterans Medical Center, Loma Linda, California 92357.
LaBue
M
M
Lazarus
J P
JP
Jennings
J C
JC
D000818:Animals; D002450:Cell Communication; D002478:Cells, Cultured; D000072259:Connexin 26; D018031:Connexin 43 / Q000096:biosynthesis; D017630:Connexins / Q000096:biosynthesis*; D004594:Electrophysiology; D005260:Female; D017629:Gap Junctions / Q000502:physiology; D015870:Gene Expression; D005802:Genes, MHC Class II; D000949:Histocompatibility Antigens Class II / Q000096:biosynthesis; D007150:Immunohistochemistry; D051379:Mice; D008817:Mice, Mutant Strains; D051381:Rats; D011917:Rats, Inbred Lew; D012016:Reference Values; D013961:Thyroid Gland / Q000276:immunology* / Q000473:pathology / Q000503:physiopathology; D013967:Thyroiditis, Autoimmune / Q000276:immunology / Q000473:pathology / Q000503:physiopathology*; D014020:Tissue Extracts / Q000276:immunology; D000097002:Gap Junction beta-1 Protein
D000072259:Connexin 26; D018031:Connexin 43; D017630:Connexins; D000949:Histocompatibility Antigens Class II; D014020:Tissue Extracts; D000097002:Gap Junction beta-1 Protein; C000607043:Gjb2 protein, mouse; C000607044:Gjb2 protein, rat