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The Journal of Neuroscience, January 7, 2004, 24(1):96-102; doi:10.1523/JNEUROSCI.1484-03.2004

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Cellular/Molecular
Tetraspanin Protein CD9 Is a Novel Paranodal Component Regulating Paranodal Junctional Formation

Tomoko Ishibashi,¹ Lei Ding,² Kazuhiro Ikenaka,³ Yoshiro Inoue,² Kenji Miyado,⁴ Eisuke Mekada,⁴ and Hiroko Baba¹

¹Department of Molecular Neurobiology, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji 192-0392, Japan, ²Department of Molecular Neuroanatomy, Hokkaido University School of Medicine, Sapporo 060-8368, Japan, ³Laboratory of Neural Information, National Institute for Physiological Sciences, Okazaki National Research Institutes, Okazaki 444-8585, Japan, ⁴ and Department of Cell Biology, Research Institute for Microbial Disease, Osaka University, Suita 565-0871, Japan

The axoglial paranodal junction is essential for the proper localization of ion channels around the node of Ranvier. The integrity of this junction is important for nerve conduction. Although recent studies have made significant progress in understanding the molecular composition of the paranodal junction, it is not known how these membrane components are distributed to the appropriate sites and interact with each other. Here we show that CD9, a member of the tetraspanin family, is present at the paranode. CD9 is concentrated in the paranode as myelination proceeds, but CD9 clusters become diffuse, associated with disruption of the paranode, in cerebroside sulfotransferase-deficient mice. Immunohistochemical and Western blot analysis showed that CD9 is distributed predominantly in the PNS. Ablation of CD9 in mutant mice disrupts junctional attachment at the paranode and alters the paranodal components contactin-associated protein (also known as Paranodin) and neurofascin 155, although the frequency of such abnormalities varies among individuals and individual axons even in the same mouse. Electron micrographs demonstrated that compact myelin sheaths were also affected in the PNS. Therefore, CD9 is a myelin protein important for the formation of paranodal junctions. CD9 also plays a role in the formation of compact myelin in the PNS.

Key words: CD9; tetraspanin-TM4SF; myelin; paranode; axoglial interaction; node of Ranvier

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Received May 16, 2003; revised October 29, 2003; accepted October 31, 2003.

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