Home LiteratureArticle Details
PMID: 10444069 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Two compartments for insulin-stimulated exocytosis in 3T3-L1 adipocytes defined by endogenous ACRP30 and GLUT4.

The Journal of cell biology ·Vol. 146 ·No. 3 ·1999-08-09 ·Pages 609-20

Bogan JS, Lodish HF

Abstract

Insulin stimulates adipose cells both to secrete proteins and to translocate the GLUT4 glucose transporter from an intracellular compartment to the plasma membrane. We demonstrate that whereas insulin stimulation of 3T3-L1 adipocytes has no effect on secretion of the alpha3 chain of type VI collagen, secretion of the protein hormone adipocyte complement related protein of 30 kD (ACRP30) is markedly enhanced. Like GLUT4, regulated exocytosis of ACRP30 appears to require phosphatidylinositol-3-kinase activity, since insulin-stimulated ACRP30 secretion is blocked by pharmacologic inhibitors of this enzyme. Thus, 3T3-L1 adipocytes possess a regulated secretory compartment containing ACRP30. Whether GLUT4 recycles to such a compartment has been controversial. We present deconvolution immunofluorescence microscopy data demonstrating that the subcellular distributions of ACRP30 and GLUT4 are distinct and nonoverlapping; in contrast, those of GLUT4 and the transferrin receptor overlap. Together with supporting evidence that GLUT4 does not recycle to a secretory compartment via the trans-Golgi network, we conclude that there are at least two compartments that undergo insulin-stimulated exocytosis in 3T3-L1 adipocytes: one for ACRP30 secretion and one for GLUT4 translocation.

MeSH Terms
3T3 Cells Adaptor Protein Complex gamma Subunits Adipocytes/cytology,drug effects,metabolism Adiponectin Animals Blood Proteins/analysis,metabolism Coatomer Protein Collagen/metabolism Endoplasmic Reticulum/chemistry,metabolism Exocytosis/drug effects Fluorescent Antibody Technique Glucose Transporter Type 4 Golgi Apparatus/chemistry Insulin/pharmacology Intercellular Signaling Peptides and Proteins Membrane Glycoproteins/analysis Membrane Proteins/analysis Mice Microtubule-Associated Proteins/analysis Monosaccharide Transport Proteins/analysis,metabolism Muscle Proteins Phosphatidylinositol 3-Kinases/metabolism Phosphoinositide-3 Kinase Inhibitors Proteins Receptors, Transferrin/analysis
Chemicals
Adaptor Protein Complex gamma Subunits Adiponectin Blood Proteins Coatomer Protein Glucose Transporter Type 4 Insulin Intercellular Signaling Peptides and Proteins Membrane Glycoproteins Membrane Proteins Microtubule-Associated Proteins Monosaccharide Transport Proteins Muscle Proteins Phosphoinositide-3 Kinase Inhibitors Proteins Receptors, Transferrin Slc2a4 protein, mouse endoplasmin Collagen
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bogan J S
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142-1479, USA.
Lodish H F
References (60)
60 references, click to expand
  1. Wortmannin inhibits mitogen-activated protein kinase activation by platelet-activating factor through a mechanism independent of p85/p110-type phosphatidylinositol 3-kinase.
    J Biol Chem. 1996 May 17;271(20):11684-8 PMID: 8662643
  2. GLUT4 in cultured skeletal myotubes is segregated from the transferrin receptor and stored in vesicles associated with TGN.
    J Cell Sci. 1996 Dec;109 ( Pt 13):2967-78 PMID: 9004032
  3. Immunocytochemical evidence that GLUT4 resides in a specialized translocation post-endosomal VAMP2-positive compartment in rat adipose cells in the absence of insulin.
    J Histochem Cytochem. 1997 Aug;45(8):1083-96 PMID: 9267469
  4. A model of secretory protein trafficking in recombinant AtT-20 cells.
    Biotechnol Bioeng. 1991 Jul;38(3):280-95 PMID: 18600762
  5. Use of bismannose photolabel to elucidate insulin-regulated GLUT4 subcellular trafficking kinetics in rat adipose cells. Evidence that exocytosis is a critical site of hormone action.
    J Biol Chem. 1993 Aug 25;268(24):17820-9 PMID: 8349666
  6. Positional cloning of the mouse obese gene and its human homologue.
    Nature. 1994 Dec 1;372(6505):425-32 PMID: 7984236
  7. Wortmannin inhibits the action of insulin but not that of okadaic acid in skeletal muscle: comparison with fat cells.
    Endocrinology. 1995 Aug;136(8):3564-70 PMID: 7628394
  8. The endoplasmic reticulum stress protein GRP94, in addition to BiP, associates with unassembled immunoglobulin chains.
    J Biol Chem. 1992 Oct 25;267(30):21303-6 PMID: 1400441
  9. Isoform-specific subcellular targeting of glucose transporters in mouse fibroblasts.
    J Cell Biol. 1992 Feb;116(3):785-97 PMID: 1309819
  10. Intracellular targeting of GLUT4 in transfected insulinoma cells: evidence for association with constitutively recycling vesicles distinct from synaptophysin and insulin vesicles.
    J Cell Sci. 1996 Jun;109 ( Pt 6):1311-23 PMID: 8799820
  11. Beta-COP, a 110 kd protein associated with non-clathrin-coated vesicles and the Golgi complex, shows homology to beta-adaptin.
    Cell. 1991 Feb 8;64(3):649-65 PMID: 1840503
  12. Chloroquine diverts ACTH from a regulated to a constitutive secretory pathway in AtT-20 cells.
    Nature. 1983 Mar 31-Apr 6;302(5907):434-6 PMID: 6300682
  13. Inhibition of the translocation of GLUT1 and GLUT4 in 3T3-L1 cells by the phosphatidylinositol 3-kinase inhibitor, wortmannin.
    Biochem J. 1994 Jun 15;300 ( Pt 3):631-5 PMID: 8010944
  14. Lilly lecture 1995. Glucose transport: pivotal step in insulin action.
    Diabetes. 1996 Nov;45(11):1644-54 PMID: 8866574
  15. Immunoelectron microscopic demonstration of insulin-stimulated translocation of glucose transporters to the plasma membrane of isolated rat adipocytes and masking of the carboxyl-terminal epitope of intracellular GLUT4.
    Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6893-7 PMID: 1713695
  16. Phosphatidylinositol 3-kinase signals activation of p70 S6 kinase in situ through site-specific p70 phosphorylation.
    Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5744-8 PMID: 7777579
  17. Compartment ablation analysis of the insulin-responsive glucose transporter (GLUT4) in 3T3-L1 adipocytes.
    Biochem J. 1996 Apr 15;315 ( Pt 2):487-95 PMID: 8615819
  18. Insulin stimulates the acute release of adipsin from 3T3-L1 adipocytes.
    Biochim Biophys Acta. 1989 Oct 30;1014(1):83-9 PMID: 2679882
  19. Evidence for the involvement of vicinal sulfhydryl groups in insulin-activated hexose transport by 3T3-L1 adipocytes.
    J Biol Chem. 1985 Mar 10;260(5):2646-52 PMID: 3882699
  20. Insulin stimulates both leptin secretion and production by rat white adipose tissue.
    Endocrinology. 1997 Oct;138(10):4463-72 PMID: 9322964
  21. Wortmannin and its structural analogue demethoxyviridin inhibit stimulated phospholipase A2 activity in Swiss 3T3 cells. Wortmannin is not a specific inhibitor of phosphatidylinositol 3-kinase.
    J Biol Chem. 1995 Oct 27;270(43):25352-5 PMID: 7592698
  22. Cloning and expression of gamma-adaptin, a component of clathrin-coated vesicles associated with the Golgi apparatus.
    J Cell Biol. 1990 Dec;111(6 Pt 1):2319-26 PMID: 2126014
  23. A distinct class of intracellular storage vesicles, identified by expression of the glucose transporter GLUT4.
    Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12750-4 PMID: 7809115
  24. Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat.
    J Cell Biol. 1991 Apr;113(1):123-35 PMID: 2007617
  25. Protection from obesity-induced insulin resistance in mice lacking TNF-alpha function.
    Nature. 1997 Oct 9;389(6651):610-4 PMID: 9335502
  26. Phosphatidylinositol 3-kinase activation is required for insulin stimulation of pp70 S6 kinase, DNA synthesis, and glucose transporter translocation.
    Mol Cell Biol. 1994 Jul;14(7):4902-11 PMID: 8007986
  27. An endocytosed TGN38 chimeric protein is delivered to the TGN after trafficking through the endocytic recycling compartment in CHO cells.
    J Cell Biol. 1998 Aug 24;142(4):923-36 PMID: 9722606
  28. Membrane-permeant esters of phosphatidylinositol 3,4,5-trisphosphate.
    J Biol Chem. 1998 May 1;273(18):11017-24 PMID: 9556583
  29. The crystal structure of a complement-1q family protein suggests an evolutionary link to tumor necrosis factor.
    Curr Biol. 1998 Mar 12;8(6):335-8 PMID: 9512423
  30. A glucose transport protein expressed predominately in insulin-responsive tissues.
    Proc Natl Acad Sci U S A. 1989 Apr;86(8):2535-9 PMID: 2649883
  31. Structural relationships between clathrin assembly proteins from the Golgi and the plasma membrane.
    EMBO J. 1988 Apr;7(4):919-29 PMID: 3402440
  32. Comparison of GLUT4 and GLUT1 subcellular trafficking in basal and insulin-stimulated 3T3-L1 cells.
    J Biol Chem. 1993 Mar 5;268(7):4600-3 PMID: 8444835
  33. Activation of pp70/85 S6 kinases in interleukin-2-responsive lymphoid cells is mediated by phosphatidylinositol 3-kinase and inhibited by cyclic AMP.
    Mol Cell Biol. 1995 Jan;15(1):326-37 PMID: 7528328
  34. Morphological effects of wortmannin on the endosomal system and GLUT4-containing compartments in rat adipose cells.
    J Cell Sci. 1997 Nov;110 ( Pt 22):2795-806 PMID: 9427288
  35. Localization of transferrin receptors and insulin-like growth factor II receptors in vesicles from 3T3-L1 adipocytes that contain intracellular glucose transporters.
    J Cell Biol. 1989 Apr;108(4):1537-45 PMID: 2538483
  36. Protein folding in the cell.
    Nature. 1992 Jan 2;355(6355):33-45 PMID: 1731198
  37. Characterization of the intracellular GLUT-4 compartment.
    Mol Membr Biol. 1995 Jul-Sep;12(3):263-9 PMID: 8520627
  38. Sequential interaction of the chaperones BiP and GRP94 with immunoglobulin chains in the endoplasmic reticulum.
    Nature. 1994 Aug 4;370(6488):373-5 PMID: 7913987
  39. GLUT4 heterozygous knockout mice develop muscle insulin resistance and diabetes.
    Nat Med. 1997 Oct;3(10):1096-101 PMID: 9334720
  40. A novel serum protein similar to C1q, produced exclusively in adipocytes.
    J Biol Chem. 1995 Nov 10;270(45):26746-9 PMID: 7592907
  41. Endoplasmic reticulum contains a common, abundant calcium-binding glycoprotein, endoplasmin.
    J Cell Sci. 1986 Dec;86:217-32 PMID: 3308928
  42. Regulation of ob gene expression and leptin secretion by insulin and dexamethasone in rat adipocytes.
    Diabetes. 1999 Feb;48(2):272-8 PMID: 10334301
  43. Biochemical dissection of AP-1 recruitment onto Golgi membranes.
    J Cell Biol. 1993 Nov;123(3):561-73 PMID: 8227126
  44. Cloning of cell-specific secreted and surface proteins by subtractive antibody screening.
    Nat Biotechnol. 1998 Jun;16(6):581-6 PMID: 9624692
  45. AdipoQ is a novel adipose-specific gene dysregulated in obesity.
    J Biol Chem. 1996 May 3;271(18):10697-703 PMID: 8631877
  46. Glucose transporter (GLUT-4) is targeted to secretory granules in rat atrial cardiomyocytes.
    J Cell Biol. 1997 Jun 16;137(6):1243-54 PMID: 9182659
  47. The binding of AP-1 clathrin adaptor particles to Golgi membranes requires ADP-ribosylation factor, a small GTP-binding protein.
    Cell. 1993 Jun 4;73(5):999-1005 PMID: 8500185
  48. Targeting of the "insulin-responsive" glucose transporter (GLUT4) to the regulated secretory pathway in PC12 cells.
    J Cell Biol. 1993 Aug;122(3):579-88 PMID: 8335686
  49. Immunocytochemical localization of beta-COP to the ER-Golgi boundary and the TGN.
    J Cell Sci. 1995 Aug;108 ( Pt 8):2839-56 PMID: 7593324
  50. Dissociation of pp70 ribosomal protein S6 kinase from insulin-stimulated glucose transport in 3T3-L1 adipocytes.
    J Biol Chem. 1993 Feb 5;268(4):3005-8 PMID: 7679106
  51. Identification and characterization of two distinct intracellular GLUT4 pools in rat skeletal muscle: evidence for an endosomal and an insulin-sensitive GLUT4 compartment.
    Biochem J. 1997 Aug 1;325 ( Pt 3):727-32 PMID: 9271094
  52. Analysis of the co-localization of the insulin-responsive glucose transporter (GLUT4) and the trans Golgi network marker TGN38 within 3T3-L1 adipocytes.
    Biochem J. 1994 Jun 15;300 ( Pt 3):743-9 PMID: 8010955
  53. Beta-COP localizes mainly to the cis-Golgi side in exocrine pancreas.
    J Cell Biol. 1993 Apr;121(1):49-59 PMID: 8458872
  54. Differential sorting of two glucose transporters expressed in insulin-sensitive cells.
    Am J Physiol. 1991 Mar;260(3 Pt 1):C570-80 PMID: 2003579
  55. Moving GLUT4: the biogenesis and trafficking of GLUT4 storage vesicles.
    Diabetes. 1997 Nov;46(11):1667-77 PMID: 9356011
  56. Isolation and molecular cloning of wortmannin-sensitive bovine type III phosphatidylinositol 4-kinases.
    J Biol Chem. 1997 Jul 18;272(29):18358-66 PMID: 9218477
  57. Biogenesis of storage granules and vesicles.
    Curr Opin Cell Biol. 1992 Aug;4(4):616-22 PMID: 1419043
  58. Identification of two types of beta-COP vesicles in the Golgi complex of rat spermatids.
    Eur J Cell Biol. 1996 Oct;71(2):137-43 PMID: 8905290
  59. Cytosolic and membrane-associated proteins involved in the recruitment of AP-1 adaptors onto the trans-Golgi network.
    J Biol Chem. 1996 Oct 11;271(41):25446-51 PMID: 8810314
  60. Differentiation of 3T3-L1 fibroblasts to adipocytes induced by transfection of ras oncogenes.
    Science. 1991 Aug 2;253(5019):565-8 PMID: 1857988
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.146.3.609
Published
1999-08-09
Pages
609-20
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2150549
Subset
IM
Grants
NIDDK NIH HHS · DK02371 · United States
NIDDK NIH HHS · DK47618 · United States
External Links
PubMed source DOI: 10.1083/jcb.146.3.609 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2