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Impaired contractile responses and altered expression and phosphorylation of Ca2+ sensitization proteins in gastric antrum smooth muscles from ob/ob mice

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Abstract

Diabetic gastroparesis is a common complication of diabetes, adversely affecting quality of life with symptoms of abdominal discomfort, nausea, and vomiting. The pathogenesis of this complex disorder is not well understood, involving abnormalities in the extrinsic and enteric nervous systems, interstitial cells of Cajal (ICCs), smooth muscles and immune cells. The ob/ob mouse model of obesity and diabetes develops delayed gastric emptying, providing an animal model for investigating how gastric smooth muscle dysfunction contributes to the pathophysiology of diabetic gastroparesis. Although ROCK2, MYPT1, and CPI-17 activities are reduced in intestinal motility disorders, their functioning has not been investigated in diabetic gastroparesis. We hypothesized that reduced expression and phosphorylation of the myosin light chain phosphatase (MLCP) inhibitory proteins MYPT1 and CPI-17 in ob/ob gastric antrum smooth muscles could contribute to the impaired antrum smooth muscle function of diabetic gastroparesis. Spontaneous and carbachol- and high K+-evoked contractions of gastric antrum smooth muscles from 7 to 12 week old male ob/ob mice were reduced compared to age- and strain-matched controls. There were no differences in spontaneous and agonist-evoked intracellular Ca2+ transients and myosin light chain kinase expression. The F-actin:G-actin ratios were similar. Rho kinase 2 (ROCK2) expression was decreased at both ages. Basal and agonist-evoked MYPT1 and myosin light chain 20 phosphorylation, but not CPI-17 phosphorylation, was reduced compared to age-matched controls. These findings suggest that reduced MLCP inhibition due to decreased ROCK2 phosphorylation of MYPT1 in gastric antrum smooth muscles contributes to the antral dysmotility of diabetic gastroparesis.

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Acknowledgments

This work was supported by NIH grant GM103513 (B.A.P.). The authors thank Sean M. Ward, and Kathy D. Keef for helpful comments and suggestions, and gratefully acknowledge Steven A. Fisher (U. of Maryland) for the gift of the MYPT1 LZ∓ antibodies.

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Authors and Affiliations

  1. Department of Physiology & Cell Biology, Center of Biomedical Research Excellence, University of Nevada School of Medicine, CMM 203E—MS 0575, 1664 N Virginia St, Reno, NV, 89557, USA

    Bhupal P. Bhetwal, Changlong An, Salah A. Baker, Kristin L. Lyon & Brian A. Perrino

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  1. Bhupal P. Bhetwal
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  2. Changlong An
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  3. Salah A. Baker
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Correspondence to Brian A. Perrino.

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Bhetwal, B.P., An, C., Baker, S.A. et al. Impaired contractile responses and altered expression and phosphorylation of Ca2+ sensitization proteins in gastric antrum smooth muscles from ob/ob mice. J Muscle Res Cell Motil 34, 137–149 (2013). https://doi.org/10.1007/s10974-013-9341-1

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