Localization of ATP-sensitive K+ channel subunits in rat liver

Ming Zhou; Kiwamu Yoshikawa; Hideo Akashi; Mitsutaka Miura; Ryoji Suzuki; Tao-Sheng Li; Hiroshi Abe; Yoshio Bando

doi:10.5493/wjem.v9.i2.14

Localization of ATP-sensitive K⁺ channel subunits in rat liver

World J Exp Med. 2019 Dec 19;9(2):14-31. doi: 10.5493/wjem.v9.i2.14.

Authors

Ming Zhou¹, Kiwamu Yoshikawa², Hideo Akashi³, Mitsutaka Miura², Ryoji Suzuki³, Tao-Sheng Li⁴, Hiroshi Abe⁵, Yoshio Bando³

Affiliations

¹ Department of Anatomy, Akita University Graduate School of Medicine, Akita 010-8543, Japan. mzhou@med.akita-u.ac.jp.
² Department of Cell Biology and Morphology, Akita University Graduate School of Medicine, Akita 010-8543, Japan.
³ Department of Anatomy, Akita University Graduate School of Medicine, Akita 010-8543, Japan.
⁴ Department of Stem Cell Biology, Atomic Bomb Disease Institute, Nagasaki University, Nagasaki 852-8523, Japan.
⁵ TRUST, A Long-Term Care Health Facility, Sendai 980-0011, Japan.

Abstract

Background: ATP-sensitive K⁺ (K_ATP) channels were originally found in cardiac myocytes by Noma in 1983. K_ATP channels were formed by potassium ion-passing pore-forming subunits (Kir6.1, Kir6.2) and regulatory subunits SUR1, SU2A and SUR2B. A number of cells and tissues have been revealed to contain these channels including hepatocytes, but detailed localization of these subunits in different types of liver cells was still uncertain.

Aim: To investigate the expression of K_ATP channel subunits in rat liver and their localization in different cells of the liver.

Methods: Rabbit anti-rat SUR1 peptide antibody was raised and purified by antigen immunoaffinity column chromatography. Four of Sprague-Dawley rats were used for liver protein extraction for immunoblot analysis, seven of them were used for immunohistochemistry both for the ABC method and immunofluorescence staining. Four of Wistar rats were used for the isolation of hepatic stellate cells (HSCs) and Kupffer cells for both primary culture and immunocytochemistry.

Results: Immunoblot analysis showed that the five kinds of K_ATP channel subunits, i.e. Kir6.1, Kir6.2, SUR1, SUR2A, and SUR2B, were detected in liver. Immunohistochemical staining showed that Kir6.1 and Kir6.2 were weakly to moderately expressed in parenchymal cells and sinusoidal lining cells, while SUR1, SUR2A, and SUR2B were mainly localized to sinusoidal lining cells, such as HSCs, Kupffer cells, and sinusoidal endothelial cells. Immunoreactivity for SUR2A and SUR2B was expressed in the hepatocyte membrane. Double immunofluorescence staining further showed that the pore-forming subunits Kir6.1 and/or Kir6.2 colocalized with GFAP in rat liver sections and primary cultured HSCs. These K_ATP channel subunits also colocalized with CD68 in liver sections and primary cultured Kupffer cells. The SUR subunits colocalized with GFAP in liver sections and colocalized with CD68 both in liver sections and primary cultured Kupffer cells. In addition, five K_ATP channel subunits colocalized with SE-1 in sinusoidal endothelial cells.

Conclusion: Observations from the present study indicated that K_ATP channel subunits expressed in rat liver and the diversity of K_ATP channel subunit composition might form different types of K_ATP channels. This is applicable to hepatocytes, HSCs, various types of Kupffer cells and sinusoidal endothelial cells.

Keywords: ATP-sensitive K+ channel; Hepatic stellate cells; Kupffer cells; Liver; Rat; Sinusoidal endothelial cells.