Oct 12, 2000

Immunolocalization of AQP9 in liver, epididymis, testis, spleen, and brain

Biochemical and Biophysical Research Communications
M ElkjaerS Nielsen

Abstract

The aims of this study were to determine the cellular and subcellular localization of aquaporin-9 (AQP9) in different rat organs by immunoblotting, immunohistochemistry and immunoelectron microscopy. To analyze this, we used rabbit antibodies to rat AQP9 raised against three different AQP9 peptides (amino acids 267-287, 274-295, and 278-295). In Cos7 cells transfected with rat AQP9, the affinity-purified antibodies exhibited marked labeling, whereas nontransfected cells and cells transfected with aquaporin-8 (AQP8) exhibited no labeling, indicating the specificity of the AQP9 antibodies. Immunoblotting revealed a predominant band of 28 kDa in membranes of total rat liver, epididymis, testes, spleen, and brain. Preabsorption with the immunizing peptides eliminated the labeling. Immunohistochemistry showed strong anti-AQP9 labeling in liver hepatocytes. The labeling was strongest at the sinusoidal surface, and there was little intracellular labeling. Immunoelectron microscopy revealed that the labeling was associated with the plasma membrane of the hepatocytes. In testes Leydig cells exhibited anti-AQP9 labeling, and in epididymis, the stereocilia of the ciliated cells (principal cells) exhibited significant labeling, whereas the...Continue Reading

  • References2
  • Citations160

References

Mentioned in this Paper

Establishment and Maintenance of Localization
Aqp9
Splenic Red Pulp
Benign Neoplasm of Testis
AQP7 gene
Transfection
Tissue Membrane
Immunohistochemistry
Benign Neoplasm of Epididymis
Epididymis Disorders

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