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Volume 17, Number 20, Issue of October 15, 1997 pp. 7725-7735
Copyright ©1997 Society for Neuroscience

Expression and Clustered Distribution of an Inwardly Rectifying Potassium Channel, K_AB-2/Kir4.1, on Mammalian Retinal Müller Cell Membrane: Their Regulation by Insulin and Laminin Signals

Received May 22, 1997; revised July 16, 1997; accepted July 28, 1997.

Masaru Ishii¹, Yoshiyuki Horio¹, Yoshihiko Tada¹, Hiroshi Hibino^{1, 3}, Atsushi Inanobe^{1, 4}, Minoru Ito⁴, Mitsuhiko Yamada¹, Takahiro Gotow², Yasuo Uchiyama², and Yoshihisa Kurachi^{1, 4}

Departments of ¹ Pharmacology II, ² Anatomy I, and ³ Otolaryngology, Faculty of Medicine, Osaka University, Osaka 565, Japan, and ⁴ Department of Cell Biology and Signaling, Yamagata University School of Medicine, Yamagata 990-23, Japan

Inwardly rectifying potassium (K⁺) channels (Kir) in Müller cells, the dominant glial cells in the retina, are supposed to be responsible for the spatial buffering action of K⁺ ions. The molecular properties and subcellular localization of Müller cell Kir channels in rat and rabbit retinas were examined by using electrophysiological, molecular biological, and immunostaining techniques. Only a single population of Kir channel activity, the properties of which were identical to those of K_AB-2/Kir4.1 expressed in HEK293T cells, could be recorded from endfoot to the distal portion of Müller cells. Consistently, Northern blot, in situ hybridization, and RT-PCR analyses indicated expression of Kir4.1 in Müller cells per se. The Kir4.1 immunoreactivity was distributed in clusters throughout Müller cell membrane. The Kir4.1 expression in Müller cells disappeared promptly after culturing. When the dissociated Müller cells were cultured on laminin-coated dishes in the presence of insulin, Kir4.1 immunoreactivity was detected in a clustered manner on the cell membrane. Because insulin and laminin exist in the surrounding of Müller cells in the retina, these substances possibly may be physiological regulators of expression and distribution of Kir4.1 in Müller cells in vivo.

Key words: inwardly rectifying potassium channel; retinal (glial) Müller cell; clustered distribution; insulin; laminin; cell culture

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