Skip to main content
SpringerLink
Log in

The Functional Role of Chromogranins in Exocytosis

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Chromogranins A (CgA) and B (CgB) are the main soluble proteins of large dense-core secretory vesicles (LDCVs). Using CgA- and CgB-knockout (KO) mice, we found that the absence of chromogranins A and B induces significant changes in catecholamine (CA) accumulation and the kinetics of exocytosis. By crossing these two knockout strains, we generated a viable and fertile double CgA/B-KO mouse in which the catecholamine content in chromaffin LDCVs was halved, and the secretory response significantly reduced. Incubating cells with l-DOPA increased the vesicular CA content in wild-type (WT) but not in Cg-KO cells, which was not due to changes in amine transport, or in the synthesis or degradation of cytosolic amines. Electron microscopy revealed the presence of giant secretory vesicles exhibiting significant alterations, with little or no electrodense inner matrix. Proteomic analysis confirmed the absence of CgA and B, and revealed small changes in SgII in the LDCV-enriched fraction, as well as the overexpression of fibrinogen and other proteins. In summary, our findings indicate that the mechanisms responsible for vesicular accumulation of CA are saturated in Cgs-KO cells, in contrast to the ample capacity for further accumulation in WT cells. We conclude that Cgs contribute to a highly efficient system that directly mediates monoamine accumulation and exocytosis in LDCVs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Albillos A, Dernick G, Horstmann H, Almers W, Alvarez de Toledo G, Lindau M (1997) The exocytotic event in chromaffin cells revealed by patch amperometry. Nature 389:509–512

    Article  PubMed  CAS  Google Scholar 

  • Beuret N, Stettler H, Renold A, Rutishauser J, Spiess M (2004) Expression of regulated secretory proteins is sufficient to generate granule-like structures in constitutively secreting cells. J Biol Chem 279:20242–20249

    Article  PubMed  CAS  Google Scholar 

  • Blaschko H, Comline RS, Schneider FH, Silver M, Smith AD (1967) Secretion of a chromaffin granule protein, chromogranin, from the adrenal gland after splanchnic stimulation. Nature 215:58–59

    Article  PubMed  CAS  Google Scholar 

  • Borges R, Diaz-Vera J, Dominguez N, Arnau MR, Machado JD (2010) Chromogranins as regulators of exocytosis. J Neurochem 114:335–343

    Article  PubMed  CAS  Google Scholar 

  • Camacho M, Machado JD, Montesinos MS, Criado M, Borges R (2006) Intragranular pH rapidly modulates exocytosis in adrenal chromaffin cells. J Neurochem 96:324–334

    Article  PubMed  CAS  Google Scholar 

  • Camacho M, Machado JD, Alvarez J, Borges R (2008) Intravesicular calcium release mediates the motion and exocytosis of secretory organelles: a study with adrenal chromaffin cells. J Biol Chem 283:22383–22389

    Article  PubMed  CAS  Google Scholar 

  • Colliver TL, Pyott SJ, Achalabun M, Ewing AG (2000) VMAT-Mediated changes in quantal size and vesicular volume. J Neurosci 20:5276–5282

    PubMed  CAS  Google Scholar 

  • Day R, Gorr SU (2003) Secretory granule biogenesis and chromogranin A: master gene, on/off switch or assembly factor? Trends Endocrinol Metab 14:10–13

    Article  PubMed  CAS  Google Scholar 

  • Diaz-Vera J, Morales YG, Hernandez-Fernaud JR, Camacho M, Montesinos MS, Calegari F, Huttner WB, Borges R, Machado JD (2010) Chromogranin B gene ablation reduces the catecholamine cargo and decelerates exocytosis in chromaffin secretory vesicles. J Neurosci 30:950–957

    Article  PubMed  CAS  Google Scholar 

  • Diaz-Vera J, Camacho M, Machado J, Dominguez N, Montesinos M, Hernandez-Fernaud J, Lujan R, Borges R (2012) Chromogranins A and B are key proteins in amine accumulation but the catecholamine secretory pathway is conserved without them. FASEB J 26:430–438

    Article  PubMed  CAS  Google Scholar 

  • Ehrhart M, Grube D, Bader MF, Aunis D, Gratzl M (1986) Chromogranin A in the pancreatic islet: cellular and subcellular distribution. J Histochem Cytochem 34:1673–1682

    Article  PubMed  CAS  Google Scholar 

  • Gasnier C, Lugardon K, Ruh O, Strub JM, Aunis D, Metz-Boutigue MH (2004) Characterization and location of post-translational modifications on chromogranin B from bovine adrenal medullary chromaffin granules. Proteomics 4:1789–801

    Google Scholar 

  • Glombik MM, Kromer A, Salm T, Huttner WB, Gerdes HH (1999) The disulfide-bonded loop of chromogranin B mediates membrane binding and directs sorting from the trans-Golgi network to secretory granules. EMBO J 18:1059–1070

    Article  PubMed  CAS  Google Scholar 

  • Gong LW, Hafez I, Alvarez de Toledo G, Lindau M (2003) Secretory vesicles membrane area is regulated in tandem with quantal size in chromaffin cells. J Neurosci 23:7917–1921

    PubMed  CAS  Google Scholar 

  • Helle KB (2004) The granin family of uniquely acidic proteins of the diffuse neuroendocrine system: comparative and functional aspects. Biol Rev Camb Philos Soc 79:769–94

    Google Scholar 

  • Helle KB, Reed RK, Pihl KE, Serck-Hanssen G (1985) Osmotic properties of the chromogranins and relation to osmotic pressure in catecholamine storage granules. Acta Physiol Scand 123:21–33

    Article  PubMed  CAS  Google Scholar 

  • Helle KB, Corti A, Metz-Boutigue MH, Tota B (2007) The endocrine role for chromogranin A: a prohormone for peptides with regulatory properties. Cell Mol Life Sci 64:2863–2886

    Article  PubMed  CAS  Google Scholar 

  • Hendy GN, Li T, Girard M, Feldstein RC, Mulay S, Desjardins R, Day R, Karaplis AC, Tremblay ML, Canaff L (2006) Targeted ablation of the chromogranin a (Chga) gene: normal neuroendocrine dense-core secretory granules and increased expression of other granins. Mol Endocrinol 20:1935–1947

    Article  PubMed  CAS  Google Scholar 

  • Huh YH, Jeon SH, Yoo SH (2003) Chromogranin B-induced secretory granule biogenesis: comparison with the similar role of chromogranin A. J Biol Chem 278:40581–40589

    Article  PubMed  CAS  Google Scholar 

  • Huttner WB, Gerdes HH, Rosa P (1991) The granin (chromogranin/secretogranin) family. Trends Biochem Sci 16:27–30

    Article  PubMed  CAS  Google Scholar 

  • Iacangelo AL, Eiden LE (1995) Chromogranin A: current status as a precursor for bioactive peptides and a granulogenic/sorting factor in the regulated secretory pathway. Regul Pept 58:65–88

    Article  PubMed  CAS  Google Scholar 

  • Kim T, Tao-Cheng JH, Eiden LE, Loh YP (2001) Chromogranin A, an "on/off" switch controlling dense-core secretory granule biogenesis. Cell 106:499–509

    Article  PubMed  CAS  Google Scholar 

  • Kopell WN, Westhead EW (1982) Osmotic pressures of solutions of ATP and catecholamines relating to storage in chromaffin granules. J Biol Chem 257:5707–5710

    PubMed  CAS  Google Scholar 

  • Koshimizu H, Kim T, Cawley NX, Loh YP (2010) Chromogranin A: a new proposal for trafficking, processing and induction of granule biogenesis. Regul Pept 160:153–159

    Article  PubMed  CAS  Google Scholar 

  • Kromer A, Glombik MM, Huttner WB, Gerdes HH (1998) Essential role of the disulfide-bonded loop of chromogranin B for sorting to secretory granules is revealed by expression of a deletion mutant in the absence of endogenous granin synthesis. J Cell Biol 140:1331–1346

    Article  PubMed  CAS  Google Scholar 

  • Lee JC, Hook V (2009) Proteolytic fragments of chromogranins A and B represent major soluble components of chromaffin granules, illustrated by two-dimensional proteomics with NH2-terminal Edman peptide sequencing and MALDI-TOF MS. Biochemistry 48:5254–5262

    Article  PubMed  CAS  Google Scholar 

  • Machado JD, Diaz-Vera J, Dominguez N, Alvarez CM, Pardo MR, Borges R (2010) Chromogranins A and B as regulators of vesicle cargo and exocytosis. Cell Mol Neurobiol 30:1181–1187

    Article  PubMed  CAS  Google Scholar 

  • Mahapatra NR, O'Connor DT, Vaingankar SM, Hikim AP, Mahata M, Ray S, Staite E, Wu H, Gu Y, Dalton N, Kennedy BP, Ziegler MG, Ross J, Mahata SK (2005) Hypertension from targeted ablation of chromogranin A can be rescued by the human ortholog. J Clin Invest 115:1942–1952

    Article  PubMed  CAS  Google Scholar 

  • Montero-Hadjadje M, Elias S, Chevalier L, Benard M, Tanguy Y, Turquier V, Galas L, Yon L, Malagon MM, Driouich A, Gasman S, Anouar Y (2009) Chromogranin A promotes peptide hormone sorting to mobile granules in constitutively and regulated secreting cells: role of conserved N- and C-terminal peptides. J Biol Chem 284:12420–12431

    Article  PubMed  CAS  Google Scholar 

  • Montesinos MS, Machado JD, Camacho M, Diaz J, Morales YG, Alvarez de la Rosa D, Carmona E, Castaneyra A, Viveros OH, O'Connor DT, Mahata SK, Borges R (2008) The crucial role of chromogranins in storage and exocytosis revealed using chromaffin cells from chromogranin A null mouse. J Neurosci 28:3350–3358

    Article  PubMed  CAS  Google Scholar 

  • Mosharov EV, Gong LW, Khanna B, Sulzer D, Lindau M (2003) Intracellular patch electrochemistry: regulation of cytosolic catecholamines in chromaffin cells. J Neurosci 23:5835–5845

    PubMed  CAS  Google Scholar 

  • Nanavati C, Fernandez JM (1993) The secretory granule matrix: a fast-acting smart polymer. Science 259:963–965

    Article  PubMed  CAS  Google Scholar 

  • Natori S, Huttner WB (1996) Chromogranin B (secretogranin I) promotes sorting to the regulated secretory pathway of processing intermediates derived from a peptide hormone precursor. Proc Natl Acad Sci U S A 93:4431–4436

    Article  PubMed  CAS  Google Scholar 

  • Obermuller S, Calegari F, King A, Lindqvist A, Lundquist I, Salehi A, Francolini M, Rosa P, Rorsman P, Huttner WB, Barg S (2010) Defective secretion of islet hormones in chromogranin-B deficient mice. PLoS One 5:e8936

    Article  PubMed  Google Scholar 

  • Park HY, So SH, Lee WB, You SH, Yoo SH (2002) Purification, pH-dependent conformational change, aggregation, and secretory granule membrane binding property of secretogranin II (chromogranin C). Biochemistry 41:1259–1266

    Article  PubMed  CAS  Google Scholar 

  • Rosa P, Gerdes HH (1994) The granin protein family: markers for neuroendocrine cells and tools for the diagnosis of neuroendocrine tumors. J Endocrinol Invest 17:207–225

    PubMed  CAS  Google Scholar 

  • Rosa P, Hille A, Lee RW, Zanini A, De Camilli P, Huttner WB (1985) Secretogranins I and II: two tyrosine-sulfated secretory proteins common to a variety of cells secreting peptides by the regulated pathway. J Cell Biol 101:1999–2011

    Article  PubMed  CAS  Google Scholar 

  • Sombers LA, Maxson MM, Ewing AG (2007) Multicore vesicles: hyperosmolarity and l-DOPA induce homotypic fusion of dense core vesicles. Cell Mol Neurobiol 27:681–685

    Article  PubMed  CAS  Google Scholar 

  • Stettler H, Beuret N, Prescianotto-Baschong C, Fayard B, Taupenot L, Spiess M (2009) Determinants for chromogranin A sorting into the regulated secretory pathway are also sufficient to generate granule-like structures in non-endocrine cells. Biochem J 418:81–91

    Article  PubMed  CAS  Google Scholar 

  • Strub JM, Sorokine O, Van Dorsselaer A, Aunis D, Metz-Boutigue MH (1997) Phosphorylation and O-glycosylation sites of bovine chromogranin A from adrenal medullary chromaffin granules and their relationship with biological activities. J Biol Chem 272:11928–11936

    Article  PubMed  CAS  Google Scholar 

  • Taupenot L, Harper KL, O'Connor DT (2003) The chromogranin–secretogranin family. N Engl J Med 348:1134–1149

    Article  PubMed  CAS  Google Scholar 

  • Taupenot L, Harper KL, O'Connor DT (2005) Role of H + −ATPase-mediated acidification in sorting and release of the regulated secretory protein chromogranin A: evidence for a vesiculogenic function. J Biol Chem 280:3885–3897

    Article  PubMed  CAS  Google Scholar 

  • Videen JS, Mezger MS, Chang YM, O'Connor DT (1992) Calcium and catecholamine interactions with adrenal chromogranins. Comparison of driving forces in binding and aggregation. J Biol Chem 267:3066–3073

    PubMed  CAS  Google Scholar 

  • Winkler H, Fischer-Colbrie R (1992) The chromogranins A and B: the first 25 years and future perspectives. Neuroscience 49:497–528

    Article  PubMed  CAS  Google Scholar 

  • Yoo SH (1996) pH- and Ca(2+)-dependent aggregation property of secretory vesicle matrix proteins and the potential role of chromogranins A and B in secretory vesicle biogenesis. J Biol Chem 271:1558–1565

    PubMed  CAS  Google Scholar 

  • Yoo SH (2010) Secretory granules in inositol 1, 4, 5-trisphosphate-dependent Ca2+ signaling in the cytoplasm of neuroendocrine cells. FASEB J 24:653–64

    Google Scholar 

  • Yoo SH, Albanesi JP (1990) Ca2(+)-induced conformational change and aggregation of chromogranin A. J Biol Chem 265:14414–21

    PubMed  CAS  Google Scholar 

  • Yoo SH, Albanesi JP (1991) High capacity, low affinity Ca2+ binding of chromogranin A. Relationship between the pH-induced conformational change and Ca2+ binding property. J Biol Chem 266:7740–7745

    PubMed  CAS  Google Scholar 

  • Zhao E, Zhang D, Basak A, Trudeau VL (2009) New insights into granin-derived peptides: evolution and endocrine roles. Gen Comp Endocrinol 164:161–174

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

ND is the recipient of an FPU fellowship from the Spanish Ministry of Education. JE-H and DP are recipients of an FPI (Spanish Ministry of Science and Innovation (MICINN). JDM is funded by a Ramon y Cajal contract (R&C-2010-06256, MICINN/ERDF). This work was supported by a MICINN Grant BFU2010-15822, CONSOLIDER (CSD2008-00005), the Canary Islands’ Agency for Research, Innovation and Society of Information (ACIISI/ERDF) and C2008/01000239 (RB).

Author information

Authors and Affiliations

  1. Unidad de Farmacología, Facultad de Medicina, Universidad de La Laguna, 38071, La Laguna, Tenerife, Spain

    Natalia Domínguez, Judith Estévez-Herrera, Marta R. Pardo, Daniel Pereda, José David Machado & Ricardo Borges

Authors
  1. Natalia Domínguez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Judith Estévez-Herrera
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marta R. Pardo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Daniel Pereda
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. José David Machado
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ricardo Borges
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ricardo Borges.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Domínguez, N., Estévez-Herrera, J., Pardo, M.R. et al. The Functional Role of Chromogranins in Exocytosis. J Mol Neurosci 48, 317–322 (2012). https://doi.org/10.1007/s12031-012-9736-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12031-012-9736-2

Keywords

Search

Navigation