Expression and localization of aquaporin 7 in normal skeletal myofiber

We examined whether AQP7 molecules are expressed in the 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 normal human or mouse muscles by using oligonucleotide primers for human or m...

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Bibliographic Details
Published in:Cell and tissue research 2004-04, Vol.316 (1), p.123-129
Main Authors: Wakayama, Yoshihiro, Inoue, Masahiko, Kojima, Hiroko, Jimi, Takahiro, Shibuya, Seiji, Hara, Hajime, Oniki, Hiroaki
Format: Article
Language:English
Subjects:
Animals
Antibodies
Aquaporins - biosynthesis
Aquaporins - immunology
Gene Expression
Humans
Immunohistochemistry
Mice
Muscle Fibers, Skeletal - metabolism
RNA, Messenger - biosynthesis
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Summary:We examined whether AQP7 molecules are expressed in the 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 normal human or mouse muscles by using oligonucleotide primers for human or mouse AQP7 showed a band of 328 or 369 basepairs, which corresponded to the basepair length between two primers of AQP7. The nucleotide sequence of these RT-PCR products coincided with those of human and mouse AQP7. Immunoblot, immunohistochemical and immunoelectron-microscopic studies of the protein were done by using the rabbit antibody against the synthetic peptide of the N-terminal cytoplasmic domain of the human AQP7 molecule. Immunoblot analysis showed that the rabbit antibody against human AQP7 reacted with a protein of approximately 30 kDa molecular weight in extracts of normal human and mouse skeletal muscles, and normal mouse liver. Immunohistochemistry with our anti-AQP7 antibody showed an immunoreaction at the myofiber surface of type 1 and type 2 fibers in human muscles and of type 2 fibers in mouse muscles.
ISSN:0302-766X
1432-0878
DOI:10.1007/s00441-004-0857-y