Aquaporin 9 expression and its localization in normal skeletal myofiber

• Published in: Journal of molecular histology Vol. 40; no. 3; pp. 165 - 170
• Main Authors: Inoue, Masahiko, Wakayama, Yoshihiro, Kojima, Hiroko, Shibuya, Seiji, Jimi, Takahiro, Hara, Hajime, Iijima, Shoji, Masaki, Hisatsugu, Oniki, Hiroaki, Matsuzaki, Yoko
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01.06.2009; Springer Netherlands; Springer Nature B.V
• Subjects: Animals; Antibodies - immunology; Aquaporins - genetics; Aquaporins - metabolism; Biomedical and Life Sciences; Biomedicine; Cell Biology; Developmental Biology; Gene Expression Regulation; Humans; Immunoblotting; Immunohistochemistry; Life Sciences; Mice; Mice, Inbred C57BL; Muscle Fibers, Skeletal - cytology; Muscle Fibers, Skeletal - metabolism; Muscle Fibers, Skeletal - ultrastructure; Original Paper; Protein Transport; Reverse Transcriptase Polymerase Chain Reaction; Immunoblot; Skeletal myofiber; Aquaporin 9; Immunocytochemistry; RT-PCR
• ISSN: 1567-2379; 1567-2387
• DOI: 10.1007/s10735-009-9226-1

The examination was performed whether aquaporin (AQP) 9 is expressed in normal skeletal muscle at mRNA and protein levels. Gel electrophoresis of the reverse transcription-polymerase chain reaction (RT-PCR) product of total RNA samples of human normal muscles by oligonucleotide primers for human AQP9 showed a band of 221 basepairs, which corresponded to the basepair length between two primers of AQP9. The nucleotide sequence of RT-PCR product coincided with that of human AQP9. Immunoblot analysis revealed that the rabbit and sheep antibodies against the synthetic peptide of the C-terminal cytoplasmic domain of human AQP9 molecule reacted with a protein of approximately 30 kDa molecular weight in extracts of human normal skeletal muscles. Immunohistochemistry with our anti-AQP9 antibodies showed an immunoreaction at the myofiber surface of both type 1 and type 2 fibers with almost equal staining intensity in human skeletal muscles. The implication of AQP9 expression in skeletal myofibers was discussed.