Vestibular malformation in mice lacking Na-K-2Cl cotransporter 1 and expression of Na-K-2Cl cotransporter 1 in human vestibular end organs.
CONCLUSIONS
The Na-K-2Cl cotransporter-1 (NKCCl) may be essential for the maintenance and functioning of the vestibular morphology in mice and it is strongly expressed in human vestibular end organs.


OBJECTIVE
NKCCl is a member of the cation-coupled chloride transporter which participates in salt transport and cell volume regulation in diverse tissues. NKCCl-deficient mice exhibit deafness, and show structural alterations in the cochlea. In addition to hearing loss, NKCCl-deficient mice show a shaker-waltzer behavior, which suggests a vestibular system defect. In this study we investigated the morphology of the vestibular system of NKCCl-deficient mice and also evaluated whether NKCCl mRNA and its protein are expressed in human vestibular end organs.


METHODS
NKCCl-deficient and wild-type mice aged 4-5 weeks were sacrificed. Their heads were cut in the midsagittal plane, fixed and decalcified. For light microscopy, 5-microm sections were cut and stained with hematoxylin-eosin. Human vestibular end organs were harvested during acoustic tumor surgery via a translabyrinthine approach. Some of these end organs were used for total mRNA extraction and the remainder for immunostaining. Reverse transcriptase polymerase chain reaction and immunostaining were performed for NKCCl.


RESULTS
The scala media of the cochleae of the NKCCl-deficient mice had collapsed but the bony labyrinth appeared unaffected. However, the semicircular canals (SCCs) were much smaller than those in the wild-type mice. Furthermore, the SCCs were completely missing in some NKCCl-deficient mice. NKCCl mRNA was expressed in both the human macula and crista ampullaris, and its protein was expressed mainly in the transitional and dark cell areas of the human crista ampullaris.
Young Choi
Jae
J
Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, South Korea.
Ho Jung
Sang
S
Namkung
Wan
W
Lee
Ju-Hyoung
JH
Jin Son
Eun
E
Wook Shin
Joong
J
Park
Hun Yi
HY
Sang Lee
Won
W
Kim
Hee Nam
HN
D000818:Animals; D001707:Biopsy, Needle; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D017136:Ion Transport; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D023421:Models, Animal; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D028021:Sodium-Potassium-Chloride Symporters / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity; D014722:Vestibule, Labyrinth / Q000002:abnormalities* / Q000473:pathology*
D026901:Membrane Transport Proteins; D012333:RNA, Messenger; D028021:Sodium-Potassium-Chloride Symporters