Cloning and functional expression of a second new aquaporin abundantly expressed in testis.
A new member of water channels has been identified from rat testis. This gene, termed aquaporin 8 (AQP8), encoded a 263-amino-acid protein that contained the conserved NPA motifs of MIP family proteins. AQP8 has amino acid sequence identity with other aquaporins (approximately 35%) and highest with a plant water channel, AQP-gamma TIP (39%), suggesting that AQP8 is a unique member in mammalian aquaporins. The expression of AQP8 in Xenopus oocytes stimulated the osmotic water permeability (Pr) 8.5 folds. The increase of Pr was inhibited with 0.3 mM mercury chloride by 55%, which was reversed with mercaptoethanol. The Arrhenius activation energy for the stimulated water permeability was low (5.1 kcal/mol). AQP8 did not facilitate glycerol transport. Northern blot analysis revealed a 1.5-kb transcript of AQP8 abundantly in testis and slightly in liver. In situ hybridization of testis revealed the expression of AQP8 mRNA in all stages of spermatogenesis from primary spermatocytes to spermatids in seminiferous tubules. Together with previously cloned AQP7, AQP8 may also play an important role in spermatogenesis. The unexpected complexity of the presence of two aquaporins in testis may call for the further analysis of the role of aquaporins in the reproduction biology.
Ishibashi
K
K
Second Department of Internal Medicine, Tokyo Medical and Dental University, Japan. kishibashi.med2@med.tmd.ac.jp
Kuwahara
M
M
Kageyama
Y
Y
Tohsaka
A
A
Marumo
F
F
Sasaki
S
S
D000595:Amino Acid Sequence; D000818:Animals; D020346:Aquaporins; D001483:Base Sequence; D002463:Cell Membrane Permeability; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D015723:Gene Library; D017403:In Situ Hybridization; D007473:Ion Channels; D008297:Male; D008969:Molecular Sequence Data; D009865:Oocytes / Q000502:physiology; D016133:Polymerase Chain Reaction; D018272:Porins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013737:Testis / Q000166:cytology / Q000378:metabolism*; D014982:Xenopus laevis
D020346:Aquaporins; D018076:DNA, Complementary; D007473:Ion Channels; D018272:Porins; D011994:Recombinant Proteins; C108330:aquaporin 8