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Diabetologia

, Volume 56, Issue 9, pp 1999–2009 | Cite as

Deficiency of APPL1 in mice impairs glucose-stimulated insulin secretion through inhibition of pancreatic beta cell mitochondrial function

  • Chen Wang
  • Xiaowen Li
  • Kaida Mu
  • Ling Li
  • Shihong Wang
  • Yunxia Zhu
  • Mingliang Zhang
  • Jiyoon Ryu
  • Zhifang Xie
  • Dongyun Shi
  • Weiping J. Zhang
  • Lily Q. DongEmail author
  • Weiping JiaEmail author
Article

Abstract

Aims/hypothesis

Adaptor protein, phosphotyrosine interaction, pleckstrin homology domain and leucine zipper containing 1 (APPL1) is an adapter protein that positively mediates adiponectin signalling. Deficiency of APPL1 in the target tissues of insulin induces insulin resistance. We therefore aimed, in the present study, to determine its role in regulating pancreatic beta cell function.

Methods

A hyperglycaemic clamp test was performed to determine insulin secretion in APPL1 knockout (KO) mice. Glucose- and adiponectin-induced insulin release was measured in islets from APPL1 KO mice or INS-1(832/13) cells with either APPL1 knockdown or overproduction. RT-PCR and western blotting were conducted to analyse gene expression and protein abundance. Oxygen consumption rate (OCR), ATP production and mitochondrial membrane potential were assayed to evaluate mitochondrial function.

Results

APPL1 is highly expressed in pancreatic islets, but its levels are decreased in mice fed a high-fat diet and db/db mice compared with controls. Deletion of the Appl1 gene leads to impairment of both the first and second phases of insulin secretion during hyperglycaemic clamp tests. In addition, glucose-stimulated insulin secretion (GSIS) is significantly decreased in islets from APPL1 KO mice. Conversely, overproduction of APPL1 leads to an increase in GSIS in beta cells. In addition, expression levels of several genes involved in insulin production, mitochondrial biogenesis and mitochondrial OCR, ATP production and mitochondrial membrane potential are reduced significantly in APPL1-knockdown beta cells. Moreover, suppression or overexproduction of APPL1 inhibits or stimulates adiponectin-potentiated GSIS in beta cells, respectively.

Conclusions/interpretation

Our study demonstrates the roles of APPL1 in regulating GSIS and mitochondrial function in pancreatic beta cells, which implicates APPL1 as a therapeutic target in the treatment of type 2 diabetes.

Keywords

Adiponectin APPL1 Beta cells Insulin Mitochondrial function 

Abbreviations

Ad

Adenovirus

AMPK

AMP activated protein kinase

APPL1

Adaptor protein, phosphotyrosine interaction, pleckstrin homology domain and leucine zipper containing 1

EGFP

Enhanced green fluorescent protein

FCCP

Carbonylcyanide p-trifluoromethoxyphenylhydrazone

GIR

Glucose infusion rate

GSIS

Glucose-stimulated insulin secretion

GTT

Glucose tolerance test

HFD

High-fat diet

α-KIC

α-Ketoisocaproate

KO

Knockout

MMP

Mitochondrial membrane potential

MOI

Multiplicity of infection

OCR

Oxygen consumption rate

PGC-1α

Peroxisome proliferator-activated receptor-γ coactivator-1α

PH

Pleckstrin homology

ROS

Reactive oxygen species

SAME

Succinic acid methyl ester

sh

Short hairpin

TFAM

Mitochondrial transcription factor A

WT

Wild-type

Notes

Acknowledgements

We thank Pengying Li and Kelei Dong (Department of Biochemistry and Molecular Biology, Shanghai Medical College of Fudan University, China) for technical help.

Funding

This work was supported by grants from the National Natural Science Foundation (30971121) (to CW), Shanghai Committee of Science and Technology (11140900900) (to CW), and National Institute of Health grant R01 DK080344 (to LQD).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Contribution statement

CW and LQD were responsible for designing the experiments. XL, KM, LL, SW, YZ, MZ, JR, ZX, DS, WJZ were responsible for acquisition of data, and analysis and interpretation of data. CW, LQD and WJ analysed and interpreted data. CW drafted the manuscript. All authors critically revised and approved the final version of the manuscript.

Supplementary material

125_2013_2971_MOESM1_ESM.pdf (16 kb)
ESM Materials and methods (PDF 16.0 kb)
125_2013_2971_MOESM2_ESM.pdf (88 kb)
ESM Table 1 (PDF 88.0 kb)
125_2013_2971_MOESM3_ESM.pdf (18 kb)
ESM Fig. 1 (PDF 18.4 kb)
125_2013_2971_MOESM4_ESM.pdf (10 kb)
ESM Fig. 2 (PDF 9.67 kb)
125_2013_2971_MOESM5_ESM.pdf (11 kb)
ESM Fig. 3 (PDF 10.7 kb)
125_2013_2971_MOESM6_ESM.pdf (14 kb)
ESM Fig. 4 (PDF 13.7 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Chen Wang
    • 1
    • 2
    • 3
  • Xiaowen Li
    • 2
    • 3
  • Kaida Mu
    • 1
    • 2
    • 3
  • Ling Li
    • 1
    • 2
    • 3
  • Shihong Wang
    • 1
    • 2
    • 3
  • Yunxia Zhu
    • 1
    • 2
    • 3
  • Mingliang Zhang
    • 1
    • 2
    • 3
  • Jiyoon Ryu
    • 4
  • Zhifang Xie
    • 5
  • Dongyun Shi
    • 6
  • Weiping J. Zhang
    • 5
  • Lily Q. Dong
    • 4
    Email author
  • Weiping Jia
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of Endocrinology and MetabolismShanghai Jiao Tong University Affiliated Sixth People’s HospitalShanghaiPeople’s Republic of China
  2. 2.Diabetes InstituteShanghai Jiao Tong UniversityShanghaiPeople’s Republic of China
  3. 3.Shanghai Key Laboratory of Diabetes MellitusShanghaiPeople’s Republic of China
  4. 4.Department of Cellular and Structural BiologyUniversity of Texas Health Science Center at San AntonioSan AntonioUSA
  5. 5.Department of PathophysiologySecond Military Medical UniversityShanghaiPeople’s Republic of China
  6. 6.Department of Biochemistry and Molecular BiologyShanghai Medical College of Fudan UniversityShanghaiPeople’s Republic of China

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