Immunofluorescence revealed the presence of NHE-1 in the nuclear membranes of rat cardiomyocytes and isolated nuclei of human, rabbit, and rat aortic and liver tissues.
Using immunofluorescence and 3-dimensional confocal microscopy techniques, the present study was designed to verify if NHE-1 is present at the level of the nuclear membrane in cells that are known to express this type of exchanger. Nuclei were isolated from aortic tissues of adult human, rabbit, and rats, as well as from liver tissues of human fetus, and adult rabbit and rat. In addition, cultured ventricular cardiomyocytes were isolated from 2-week-old rat. Our results showed the presence of NHE-1 in isolated nuclei of aortic vascular smooth muscle and liver of human, rabbit, and rat. NHE-1 seems to be distributed throughout the isolated nucleus and more particularly at the level of the nuclear membranes. The relative fluorescence density of NHE-1 was significantly higher (p < 0.05) in isolated liver nuclei of human, when compared with those of rabbit and rat. However, in isolated nuclei of aortic vascular smooth muscle, the relative fluorescence density of NHE-1 was significantly (p < 0.001) higher in the rabbit when compared with human and rat. In cultured rat ventricular cardiomyocytes, NHE-1 fluorescent labeling could be easily seen throughout the cell, including the nucleus, and more particularly at both the sarcolemma and the nuclear membranes. In rat cardiomyocytes, the relative fluorescence density of NHE-1 of the sarcolemma membrane, including the cytosol, was significantly lower than that of the whole nucleus (including the nuclear envelope membranes). In conclusion, our results showed that NHE-1 is present at the nuclear membranes and in the nucleoplasm and its distribution and density may depend on cell type and species used. These results suggest that nuclear membranes' NHE-1 may play a role in the modulation of intranuclear pH.
Bkaily
Ghassan
G
Department of Anatomy and Cell Biology, Faculty of Medicine, University of Sherbrooke, 3001-12th Avenue North, Sherbrooke, Quebec J1H 5N4, Canada. Ghassan.Bkaily@USherbrooke.ca
Nader
Moni
M
Avedanian
Levon
L
Jacques
Danielle
D
Perrault
Claudine
C
Abdel-Samad
Dima
D
D'Orléans-Juste
Pedro
P
Gobeil
Fernand
F
Hazzouri
Khaled M
KM
D000328:Adult; D000818:Animals; D001011:Aorta / Q000737:chemistry* / Q000276:immunology; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D006801:Humans; D008099:Liver / Q000737:chemistry* / Q000276:immunology; D032383:Myocytes, Cardiac / Q000737:chemistry* / Q000276:immunology; D009685:Nuclear Envelope / Q000737:chemistry* / Q000276:immunology; D011817:Rabbits; D051381:Rats; D017923:Sodium-Hydrogen Exchangers / Q000032:analysis* / Q000276:immunology; D013045:Species Specificity
D017923:Sodium-Hydrogen Exchangers; C069139:growth factor-activatable Na-H exchanger NHE-1