Genetic and pharmacological inhibition of rho-associated kinase II enhances adipogenesis

Michio Noguchi, Kiminori Hosoda, Junji Fujikura, Muneya Fujimoto, Hiroshi Iwakura, Tsutomu Tomita, Takako Ishii, Naoki Arai, Masakazu Hirata, Ken Ebihara, Hiroaki Masuzaki, Hiroshi Itoh, Shuh Narumiya, Kazuwa Nakao

Research output: Contribution to journalArticle

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Abstract

Rho-associated kinase (ROCK) regulates reorganization of actin cytoskeleton. During adipogenesis, the structure of filamentous actin is converted from long stress fibers to cortical actin, suggesting that the ROCK is involved in adipogenesis. Two ROCK isoforms have been identified: ROCK-I and ROCK-II. However, pharmacological inhibitors of ROCK cannot distinguish two ROCK isoforms. In the present study, we examined the role of ROCK in adipogenesis and actin cytoskeleton using genetic and pharmacological approaches. Y-27632, which inhibits the activity of both ROCK isoforms, enhanced adipogenesis through the up-regulation of adipogenic transcription factors in 3T3-L1 cells. Furthermore, Y-27632 restored inhibition of adipogenesis by lysophosphatidic acid, which activates Rho. Regarding actin cytoskeleton, Y-27632 disrupted stress fibers in 3T3-L1 preadipocytes. Next, we analyzed adipogenesis of mouse embryonic fibroblasts (MEFs) derived from ROCK-I and ROCK-II knock-out mice, respectively. Adipogenesis of ROCK-II (-/-) MEFs was markedly enhanced compared with wild-type MEFs while that of ROCK-I (-/-) MEFs was not. In contrast to pharmacological approaches, no obvious alteration was found in actin cytoskeleton of ROCK-II (-/-) MEFs compared with wild-type MEFs. In 3T3-L1 cells, knockdown of ROCK-II by RNA interference enhanced the expression of adipogenic transcription factors while that of ROCK-I did not. Moreover, Y-27632 inhibited IRS-1 serine phosphorylation and enhanced Akt phosphorylation in 3T3-L1 preadipocytes. Similarly, Akt phosphorylation in ROCK-II (-/-) MEFs was augmented compared with wild-type MEFs. In conclusion, inhibition of ROCK-II, not ROCK-I, enhances adipogenesis accompanied by the up-regulation of adipogenic transcription factors. Augmentation of insulin signaling may contribute to the enhancement of adipogenesis.

Original languageEnglish
Pages (from-to)29574-29583
Number of pages10
JournalJournal of Biological Chemistry
Volume282
Issue number40
DOIs
Publication statusPublished - 2007 Oct 5
Externally publishedYes

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Adipogenesis
rho-Associated Kinases
Pharmacology
Fibroblasts
Actins
Actin Cytoskeleton
Phosphorylation
3T3-L1 Cells
Stress Fibers
Protein Isoforms
Transcription Factors
Up-Regulation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Noguchi, M., Hosoda, K., Fujikura, J., Fujimoto, M., Iwakura, H., Tomita, T., ... Nakao, K. (2007). Genetic and pharmacological inhibition of rho-associated kinase II enhances adipogenesis. Journal of Biological Chemistry, 282(40), 29574-29583. https://doi.org/10.1074/jbc.M705972200

Genetic and pharmacological inhibition of rho-associated kinase II enhances adipogenesis. / Noguchi, Michio; Hosoda, Kiminori; Fujikura, Junji; Fujimoto, Muneya; Iwakura, Hiroshi; Tomita, Tsutomu; Ishii, Takako; Arai, Naoki; Hirata, Masakazu; Ebihara, Ken; Masuzaki, Hiroaki; Itoh, Hiroshi; Narumiya, Shuh; Nakao, Kazuwa.

In: Journal of Biological Chemistry, Vol. 282, No. 40, 05.10.2007, p. 29574-29583.

Research output: Contribution to journalArticle

Noguchi, M, Hosoda, K, Fujikura, J, Fujimoto, M, Iwakura, H, Tomita, T, Ishii, T, Arai, N, Hirata, M, Ebihara, K, Masuzaki, H, Itoh, H, Narumiya, S & Nakao, K 2007, 'Genetic and pharmacological inhibition of rho-associated kinase II enhances adipogenesis', Journal of Biological Chemistry, vol. 282, no. 40, pp. 29574-29583. https://doi.org/10.1074/jbc.M705972200
Noguchi M, Hosoda K, Fujikura J, Fujimoto M, Iwakura H, Tomita T et al. Genetic and pharmacological inhibition of rho-associated kinase II enhances adipogenesis. Journal of Biological Chemistry. 2007 Oct 5;282(40):29574-29583. https://doi.org/10.1074/jbc.M705972200
Noguchi, Michio ; Hosoda, Kiminori ; Fujikura, Junji ; Fujimoto, Muneya ; Iwakura, Hiroshi ; Tomita, Tsutomu ; Ishii, Takako ; Arai, Naoki ; Hirata, Masakazu ; Ebihara, Ken ; Masuzaki, Hiroaki ; Itoh, Hiroshi ; Narumiya, Shuh ; Nakao, Kazuwa. / Genetic and pharmacological inhibition of rho-associated kinase II enhances adipogenesis. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 40. pp. 29574-29583.
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AU - Tomita, Tsutomu

AU - Ishii, Takako

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