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VAMP8/Endobrevin is a critical factor for the homotypic granule growth in pancreatic acinar cells

Abstract

The delivery of newly-formed secretory content to the granule inventory occurs through direct fusion of recently formed granules and mature granules. The introduction of knockout mice allowed us to investigate the characteristics of the delivery process and to determine the core protein machinery required for granule growth. The SNARE machinery mediates membrane fusion and is essential for the granule lifecycle. In the current work, we use VAMP8 knockout mice to show that the SNARE machinery plays a critical role in the process of granule homotypic fusion. Consistent with this, the mutated mouse pancreatic acinar secretory granules are significantly smaller compared to the control group, demonstrating few granule profiles that might be the result of homotypic fusion.

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Fig. 1

References

  1. Advani RJ, Bae HR, Bock JB, Chao DS, Doung YC, Prekeris R, Yoo JS, Scheller RH (1998) Seven novel mammalian SNARE proteins localize to distinct membrane compartments. J Biol Chem 273:10317–10324

  2. Antonin W, Holroyd C, Tikkanen R, Höning S, Jahn R (2000a) The R-SNARE endobrevin/VAMP-8 mediates homotypic fusion of early endosomes and late endosomes. Mol Biol Cell 11:3289–3298

  3. Antonin W, Holroyd C, Fasshauer D, Pabst S, Von Mollard GF, Jahn R (2000b) A SNARE complex mediating fusion of late endosomes defines conserved properties of SNARE structure and function. EMBO J 19:6453–6464

  4. Behrendorff N, Dolai S, Hong W, Gaisano HY, Thorn P (2011) Vesicle-associated membrane protein 8 (VAMP8) is a SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) selectively required for sequential granule-to-granule fusion. J Biol Chem 286:29627–29634

  5. Chen X, Walker AK, Strahler JR, Simon ES, Tomanicek-Volk SL, Nelson BB, Hurley MC, Ernst SA, Williams JA, Andrews PC (2006) Organellar proteomics: analysis of pancreatic zymogen granule membranes. Mol Cell Proteomics 5:306–312

  6. Collins TJ (2007) ImageJ for microscopy. Biotechniques 43(Suppl):25–30

  7. Cosen-Binker LI, Binker MG, Wang CC, Hong W, Gaisano HY (2008) VAMP8 is the v-SNARE that mediates basolateral exocytosis in a mouse model of alcoholic pancreatitis. J Clin Invest 118:2535–2551

  8. Dressel R, Elsner L, Novota P, Kanwar N, Fischer von Mollard G (2010) The exocytosis of lytic granules is impaired in Vti1b- or Vamp8-deficient CTL leading to a reduced cytotoxic activity following antigen-specific activation. J Immunol 185:1005–1014

  9. Graham GJ, Ren Q, Dilks JR, Blair P, Whiteheart SW, Flaumenhaft R (2009) Endobrevin/VAMP 8-dependent dense granule release mediates thrombus formation in vivo. Blood 114:1083–1090

  10. Grützkau A, Smorodchenko A, Lippert U, Kirchhof L, Artuc M, Henz BM (2004) LAMP-1 and LAMP-2, but not LAMP-3, are reliable markers for activation-induced secretion of human mast cells. Cytometry A 61:62–68

  11. Hammel I (1986) Progression of errors in morphometry. Estimation of particle number density. J Histocem Cytochem 34:941–944

  12. Hammel I, Alroy J (1995) The effect of lysosomal storage diseases on secretory cells: an ultrastructural study of pancreas as an example. J Submicrosc Cytol Pathol 27:143–160

  13. Hammel I, Dvorak AM, Galli SJ (1987) Defective cytoplasmic granule formation I. Abnormalities affecting tissue mast cells and pancreatic acinar cells of beige mice. Lab Invest 56:321–328

  14. Hammel I, Alroy J, Goyal V, Galli SJ (1993) Ultrastructure of human mast-cells in 29 different lysosomal storage diseases. Virchows Arch B Cell Pathol Incl Mol Pathol 64:83–89

  15. Hammel I, Dvorak AM, Fox P, Shimoni E, Galli SJ (1998) Defective cytoplasmic granule formation.II. Differences in patterns of radiolabeling of secretory granules in beige versus normal mouse pancreatic acinar cells after [3H]glycine administration in vivo. Cell Tissue Res 293:445–452

  16. Hammel I, Shor-Hazan O, Eldar T, Amihai D, Lew S (1999) Morphometric studies of secretory granule formation in mouse pancreatic acinar cells. Dissecting the early structural changes following pilocarpine injection. J Anat 194:51–60

  17. Hammel I, Lagunoff D, Galli SJ (2010a) Regulation of secretory granule size by the precise generation and fusion of unit granules. J Cell Mol Med 14:1904–1916

  18. Hammel I, Shoichetman T, Amihai D, Galli SJ, Skutelsky E (2010b) Localization of anionic constituents in mast cell granules of brachymorphic (bm/bm) mice by using avidin-conjugated colloidal gold. Cell Tissue Res 339:561–570

  19. Imai A, Nashida T, Yoshie S, Shimomura H (2003) Intracellular localisation of SNARE proteins in rat parotid acinar cells: SNARE complexes on the apical plasma membrane. Arch Oral Biol 48:597–604

  20. James DE, Piper RC, Luzio JP, Simpson RJ, Connolly LM, Bryant NJ, Wade N (2001) Syntaxin 7 complexes with mouse Vps10p tail interactor 1b, syntaxin 6, vesicle-associated membrane protein (VAMP)8, and VAMP7 in b16 melanoma cells. J Biol Chem 276:19820–19827

  21. Jena BP (2011) Role of SNAREs in membrane fusion. Adv Exp Med Biol 713:13–32

  22. Jones LC, Moussa L, Fulcher LM, Zhu Y, Hudson EJ, O'Neal WK, Randell SH, Lazarowski ER, Boucher RC, Kreda SM (2012) VAMP8 is a vesicle SNARE that regulates mucin secretion in airway goblet cells. J Physiol 590:545–562

  23. Kalesnikoff J, Galli SJ (2008) New developments in mast cell biology. Nat Immunol 9:1215–1223

  24. Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolarity for use in electron microscopy. J Cell Biol 27:137a

  25. Kondkar AA, Bray MS, Leal SM, Nagalla S, Liu DJ, Jin Y, Dong JF, Ren Q, Whiteheart SW, Shaw C, Bray PF (2010) VAMP8/endobrevin is overexpressed in hyperreactive human platelets: suggested role for platelet microRNA. J Thromb Haemost 8:369–378

  26. Lagunoff D (1968) The properties of mast cell proteases. Biochem Pharmacol Suppl 1968:221–227

  27. Lagunoff D, Pritzl P (1976) Characterization of rat mast cell granule proteins. Arch Biochem Biophys 173:554–563

  28. Lew S, Hammel I, Galli SJ (1994) Cytoplasmic granule formation in mouse pancreatic acinar cells. Evidence for formation of immature granules (condensing vacuoles) by aggregation and fusion of progranules of unit size, and for reductions in membrane surface area and immature granule volume during granule maturation. Cell Tissue Res 278:327–336

  29. Lippert U, Ferrari DM, Jahn R (2007) Endobrevin/VAMP8 mediates exocytotic release of hexosaminidase from rat basophilic leukaemia cells. FEBS Lett 581:3479–3484

  30. Loo LS, Hwang LA, Ong YM, Tay HS, Wang CC, Hong W (2009) A role for endobrevin/VAMP8 in CTL lytic granule exocytosis. Eur J Immunol 39:3520–3528

  31. Nagamatsu S, Nakamichi Y, Watanabe T, Matsushima S, Yamaguchi S, Ni J, Itagaki E, Ishida H (2001) Localization of cellubrevin-related peptide, endobrevin, in the early endosome in pancreatic beta cells and its physiological function in exo-endocytosis of secretory granules. J Cell Sci 114:219–227

  32. Nitzany E, Hammel I, Meilijson I (2010) Quantal basis of vesicle growth and information content, a unified approach. J Theor Biol 266:202–209

  33. Paumet F, Le Mao J, Martin S, Galli T, David B, Blank U, Roa M (2000) Soluble NSF attachment protein receptors (SNAREs) in RBL-2H3 mast cells: functional role of syntaxin 4 in exocytosis and identification of a vesicle-associated membrane protein 8-containing secretory compartment. J Immunol 164:5850–5857

  34. Payne CM (1989) Phylogenetic considerations of neurosecretory granule contents: role of nucleotides and basic hormone/transmitter packaging mechanisms. Arch Histol Cytol 52(Suppl):277–292

  35. Pickett JA, Campos-Toimil M, Thomas P, Edwardson JM (2007) Identification of SNAREs that mediate zymogen granule exocytosis. Biochem Biophys Res Commun 359:599–603

  36. Polgár J, Chung SH, Reed GL (2002) Vesicle-associated membrane protein 3 (VAMP-3) and VAMP-8 are present in human platelets and are required for granule secretion. Blood 100:1081–1083

  37. Puri N, Roche PA (2008) Mast cells possess distinct secretory granule subsets whose exocytosis is regulated by different SNARE isoforms. Proc Natl Acad Sci USA 105:2580–2585

  38. Ren Q, Barber HK, Crawford GL, Karim ZA, Zhao C, Choi W, Wang CC, Hong W, Whiteheart SW (2007) Endobrevin/VAMP-8 is the primary v-SNARE for the platelet release reaction. Mol Biol Cell 18:24–33

  39. Sander LE, Frank SP, Bolat S, Blank U, Galli T, Bigalke H, Bischoff SC, Lorentz A (2008) Vesicle associated membrane protein (VAMP)-7 and VAMP-8, but not VAMP-2 or VAMP-3, are required for activation-induced degranulation of mature human mast cells. Eur J Immunol 38:855–863

  40. Schwartz LB, Austen KF (1980) Enzymes of the mast cell granule. J Invest Dermatol 74:349–353

  41. Schwenk F, Baron U, Rajewsky K (1995) A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells. Nucleic Acids Res 23:5080–5081

  42. Sokal RR, Rohlf FJ (1995) Biometry: the principles and practice of statistics in biological research, 3rd edn. Freeman, New York

  43. Tiwari N, Wang CC, Brochetta C, Ke G, Vita F, Qi Z, Rivera J, Soranzo MR, Zabucchi G, Hong W, Blank U (2008) VAMP-8 segregates mast cell-preformed mediator exocytosis from cytokine trafficking pathways. Blood 111:3665–3674

  44. Tiwari N, Wang CC, Brochetta C, Scandiuzzi L, Hong W, Blank U (2009) Increased formation of VAMP-3-containing SNARE complexes in mast cells from VAMP-8 deficient cells. appetite. Inflamm Res 58(Suppl 1):13–14

  45. Wang CC, Ng CP, Lu L, Atlashkin V, Zhang W, Seet LF, Hong W (2004) A role of VAMP8/endobrevin in regulated exocytosis of pancreatic acinar cells. Dev Cell 7:359–371

  46. Wang CC, Shi H, Guo K, Ng CP, Li J, Gan BQ, Chien Liew H, Leinonen J, Rajaniemi H, Zhou ZH, Zeng Q, Hong W (2007) VAMP8/endobrevin as a general vesicular SNARE for regulated exocytosis of the exocrine system. Mol Biol Cell 18:1056–1063

  47. Wang CC, Ng CP, Shi H, Liew HC, Guo K, Zeng Q, Hong W (2010) A role for VAMP8/endobrevin in surface deployment of the water channel aquaporin 2. Mol Cell Biol 30:333–343

  48. Weintraub H, Abramovici A, Amichai D, Eldar T, Ben-Dor L, Pentchev PG, Hammel I (1992) Morphometric studies of pancreatic acinar granule formation in NCTR-Balb/c mice. J Cell Sci 102:141–147

  49. Wendler F, Page L, Urbé S, Tooze SA (2001) Homotypic fusion of immature secretory granules during maturation requires syntaxin 6. Mol Biol Cell 12:1699–1709

  50. Williams D, Pessin JE (2008) Mapping of R-SNARE function at distinct intracellular GLUT4 trafficking steps in adipocytes. J Cell Biol 180:375–387

  51. Wong SH, Zhang T, Xu Y, Subramaniam VN, Griffiths G, Hong W (1998) Endobrevin, a novel synaptobrevin/VAMP-like protein preferentially associated with the early endosome. Mol Biol Cell 9:1549–1563

  52. Zhao P, Yang L, Lopez JA, Fan J, Burchfield JG, Bai L, Hong W, Xu T, James DE (2009) Variations in the requirement for v-SNAREs in GLUT4 trafficking in adipocytes. J Cell Sci 122:3472–3480

  53. Zong H, Wang CC, Vaitheesvaran B, Kurland IJ, Hong W, Pessin JE (2011) Enhanced energy expenditure, glucose utilization, and insulin sensitivity in VAMP8 null mice. Diabetes 60:30–38

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Acknowledgements

We thank the staff of the Biological Resource Center (BRC, Singapore) for maintaining the VAMP8 knockout mice. This work is funded by The Singapore Agency for Science, Technology and Research (H.W.J. and W.C.C.) and by Tel Aviv University Intramural Research Program Funds (I.H.).

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Correspondence to Ilan Hammel.

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Hammel, I., Wang, C., Hong, W. et al. VAMP8/Endobrevin is a critical factor for the homotypic granule growth in pancreatic acinar cells. Cell Tissue Res 348, 485–490 (2012). https://doi.org/10.1007/s00441-012-1400-1

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Keywords

  • Unit granule
  • Homotypic fusion
  • SNARE