CoChat
Sign Up
Cited by 100Pancreatic function and diabetesCellular transport and secretionNeuroscience and Neuropharmacology Research

Corelease and Differential Exit via the Fusion Pore of GABA, Serotonin, and ATP from LDCV in Rat Pancreatic β Cells

Open access
Braun, Matthias, Wendt, Anna, Karanauskaite, Jovita, Galvanovskis, Juris, Clark, Anne, MacDonald, Patrick E., Rorsman, Patrik·2007·The Journal of General Physiology
DOI 10.1085/jgp.200609658PMC PMC2151613
Open this paper in CoChat — free accountOpen PDF

Abstract

The release of γ-aminobutyric acid (GABA) and ATP from rat β cells was monitored using an electrophysiological assay based on overexpression GABAA or P2X2 receptor ion channels. Exocytosis of LDCVs, detected by carbon fiber amperometry of serotonin, correlated strongly (∼80%) with ATP release. The increase in membrane capacitance per ATP release event was 3.4 fF, close to the expected capacitance of an individual LDCV with a diameter of 0.3 μm. ATP and GABA were coreleased with serotonin with the same probability. Immunogold electron microscopy revealed that ∼15% of the LDCVs contain GABA. Prespike “pedestals,” reflecting exit of granule constituents via the fusion pore, were less frequently observed for ATP than for serotonin or GABA and the relative amplitude (amplitude of foot compared to spike) was smaller: in some cases the ATP-dependent pedestal was missing entirely. An inward tonic current, not dependent on glucose and inhibited by the GABAA receptor antagonist SR95531, was observed in β cells in clusters of islet cells. Noise analysis indicated that it was due to the activity of individual channels with a conductance of 30 pS, the same as expected for individual GABAA Cl− channels with the ionic gradients used. We conclude that (a) LDCVs accumulate ATP and serotonin; (b) regulated release of GABA can be accounted for by exocytosis of a subset of insulin-containing LDCVs; (c) the fusion pore of LDCVs exhibits selectivity and compounds are differentially released depending on their chemical properties (including size); and (d) a glucose-independent nonvesicular form of GABA release exists in β cells.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1085/jgp.200609658

Cites

3 of 3 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • In both rodent and human β-cells, GABA is released both by a vesicular route (via exocytosis of insulin-containing granules) and a non-vesicular route.

    “However, it is now clear that in both rodent and human β-cells, GABA is released both by a vesicular route (via exocytosis of the insulin-containing granules) and a non-vesicular route (89, 95).”

    From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [95]

    The Discussion (Concluding Remarks; page not available) says GABA can leave β-cells through vesicular and nonvesicular pathways, with vesicular release involving insulin-containing granules. The Methods (Islet Cell Preparation; page not available) identify the experimental cells as rat cells. This source supports the rodent portion of the claim but does not establish the same routes in human β-cells. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 35

  1. Ämmälä, C., F.M. Ashcroft, and P. Rorsman. 1993. Calcium-independent potentiation of insulin release by cyclic AMP in single β cells. Nature. 363: 356–358. 7684514
  2. Aspinwall, C.A., L. Huang, J.R. Lakey, and R.T. Kennedy. 1999. Comparison of amperometric methods for detection of exocytosis from single pancreatic β cells of different species. Anal. Chem. 71: 5551–5556. 10624159
  3. Barg, S. 2003. Mechanisms of exocytosis in insulin-secreting B-cells and glucagon-secreting A-cells. Pharmacol. Toxicol. 92: 3–13. 12710591
  4. Birnir, B., M.L. Tierney, N.P. Pillai, G.B. Cox, and P.W. Gage. 1995. Rapid desensitization of α1 β1 GABAA receptors expressed in Sf9 cells under optimized conditions. J. Membr. Biol. 148: 193–202. 8606368
  5. Braun, M., A. Wendt, K. Buschard, A. Salehi, S. Sewing, J. Gromada, and P. Rorsman. 2004b. GABAB-receptor activation inhibits exocytosis in rat pancreatic β cells by G-protein-dependent activation of calcineurin. J. Physiol. 559: 397–409. 15235087 PMC1665126
  6. Bruns, D., D. Riedel, J. Klingauf, and R. Jahn. 2000. Quantal release of serotonin. Neuron. 28: 205–220. 11086995
  7. Conley, E.C. 1996. Ion Channel Factsbook. Vol. I: Extracellular Ligand-gated Channels. Academic Press, London. 860 pp.
  8. Chow, R.H., L. von Ruden, and E. Neher. 1992. Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. Nature. 356: 60–63. 1538782
  9. Eliasson, L., P. Proks, C. Ammala, F.M. Ashcroft, K. Bokvist, E. Renstrom, P. Rorsman, and P.A. Smith. 1996. Endocytosis of secretory granules in mouse pancreatic beta-cells evoked by transient elevation of cytosolic calcium. J. Physiol. 493: 755–767. 8799897 PMC1159023
  10. Finnegan, J.M., K. Pihel, P.S. Cahill, L. Huang, S.E. Zerby, A.G. Ewing, R.T. Kennedy, and R.M. Wightman. 1996. Vesicular quantal size measured by amperometry at chromaffin, mast, pheochromocytoma, and pancreatic β cells. J. Neurochem. 66: 1914–1923. 8780018
  11. Gammelsaeter, R., M. Froyland, C. Aragon, N.C. Danbolt, D. Fortin, J. Storm-Mathisen, S. Davanger, and V. Gundersen. 2004. Glycine, GABA and their transporters in pancreatic islets of Langerhans: evidence for a paracrine transmitter interplay. J. Cell Sci. 117: 3749–3758. 15252115
  12. Gandhi, S.P., and C.F. Stevens. 2003. Three modes of synaptic vesicular recycling revealed by single-vesicle imaging. Nature. 423: 607–613. 12789331
  13. Gillis, K.D. 1995 Techniques for membrane capacitance measurements. In Single-channel recording. B. Sakmann, and E. Neher, editors. Plenum Press, New York. 155–198.
  14. Hazama, A., S. Hayashi, and Y. Okada. 1998. Cell surface measurements of ATP release from single pancreatic β cells using a novel biosensor technique. Pflugers Arch. 437: 31–35. 9817782
  15. Hille, B. 2001. Ion Channels of Excitable Membranes. Third edition. Sinauer, Sunderland, MA. 814 pp.
  16. Hiriart, M., and D.R. Matteson. 1988. Na channels and two types of Ca channels in rat pancreatic B cells identified with the reverse hemolytic plaque assay. J. Gen. Physiol. 91: 617–639. 2458427 PMC2216150
  17. Hollins, B., and S.R. Ikeda. 1997. Heterologous expression of a P2x-purinoceptor in rat chromaffin cells detects vesicular ATP release. J. Neurophysiol. 78: 3069–3076. 9405526
  18. Hutton, J.C. 1989. The insulin secretory granule. Diabetologia. 32: 271–281. 2526768
  19. Kasai, H. 1999. Comparative biology of Ca2+-dependent exocytosis: implications of kinetic diversity for secretory function. Trends Neurosci. 22: 88–93. 10092049
  20. Kennedy, R.T., L. Huang, M.A. Atkinson, and P. Dush. 1993. Amperometric monitoring of chemical secretions from individual pancreatic β cells. Anal. Chem. 65: 1882–1887. 8368536
  21. MacDonald, P.E., M. Braun, J. Galvanovskis, and P. Rorsman. 2006. Release of small transmitters through kiss-and-run fusion pores in rat pancreatic β cells. Cell Metab. 4: 283–290. 17011501
  22. MacDonald, P.E., S. Obermüller, J. Vikman, J. Galvanovskis, P. Rorsman, and L. Eliasson. 2005. Regulated exocytosis and kiss-and-run of synaptic-like microvesicles in INS-1 and primary rat β cells. Diabetes. 54: 736–743. 15734850
  23. Obermüller, S., A. Lindqvist, J. Karanauskaite, J. Galvanovskis, P. Rorsman, and S. Barg. 2005. Selective nucleotide-release from dense-core granules in insulin-secreting cells. J. Cell Sci. 118: 4271–4282. 16141231
  24. Reetz, A., M. Solimena, M. Matteoli, F. Folli, K. Takei, and P. De Camilli. 1991. GABA and pancreatic β cells: colocalization of glutamic acid decarboxylase (GAD) and GABA with synaptic-like microvesicles suggests their role in GABA storage and secretion. EMBO J. 10: 1275–1284. 2022191 PMC452782
  25. Renstrom, E., L. Eliasson, and P. Rorsman. 1997. Protein kinase A-dependent and -independent stimulation of exocytosis by cAMP in mouse pancreatic B-cells. J. Physiol. 502: 105–118. 9234200 PMC1159575
  26. Rorsman, P., P.O. Berggren, K. Bokvist, H. Ericson, H. Mohler, C.G. Ostenson, and P.A. Smith. 1989. Glucose-inhibition of glucagon secretion involves activation of GABAA-receptor chloride channels. Nature. 341: 233–236. 2550826
  27. Salehi, A., S.S. Qader, E. Grapengiesser, and B. Hellman. 2005. Inhibition of purinoceptors amplifies glucose-stimulated insulin release with removal of its pulsatility. Diabetes. 54: 2126–2131. 15983214
  28. Smismans, A., F. Schuit, and D. Pipeleers. 1997. Nutrient regulation of γ-aminobutyric acid release from islet β cells. Diabetologia. 40: 1411–1415. 9447948
  29. Smith, P.A., P. Proks, and F.M. Ashcroft. 1999. Quantal analysis of 5-hydroxytryptamine release from mouse pancreatic β cells. J. Physiol. 521: 651–664. 10601496 PMC2269697
  30. Takahashi, N., T. Kadowaki, Y. Yazaki, Y. Miyashita, and H. Kasai. 1997. Multiple exocytotic pathways in pancreatic β cells. J. Cell Biol. 138: 55–64. 9214381 PMC2139953
  31. Thomas-Reetz, A., J.W. Hell, M.J. During, C. Walch-Solimena, R. Jahn, and P. De Camilli. 1993. A gamma-aminobutyric acid transporter driven by a proton pump is present in synaptic-like microvesicles of pancreatic β cells. Proc. Natl. Acad. Sci. USA. 90: 5317–5321. 8506380 PMC46707
  32. Tsuboi, T., and G.A. Rutter. 2003. Insulin secretion by ‘kiss-and-run’ exocytosis in clonal pancreatic islet beta-cells. Biochem. Soc. Trans. 31: 833–836. 12887316
  33. Wendt, A., B. Birnir, K. Buschard, J. Gromada, A. Salehi, S. Sewing, P. Rorsman, and M. Braun. 2004. Glucose inhibition of glucagon secretion from rat β cells is mediated by GABA released from neighbouring β cells. Diabetes. 53: 1038–1045. 15047619
  34. Winnock, F., Z. Ling, R. De Proft, S. Dejonghe, F. Schuit, F. Gorus, and D. Pipeleers. 2002. Correlation between GABA release from rat islet β cells and their metabolic state. Am. J. Physiol. Endocrinol. Metab. 282: E937–E942. 11882516
  35. Zhou, Z., and S. Misler. 1996. Amperometric detection of quantal secretion from patch-clamped rat pancreatic β cells. J. Biol. Chem. 271: 270–277. 8550572

Cited by

Sorted by citations

Authors
Braun, M., Wendt, A., Karanauskaite, J., Galvanovskis, J., Clark, A., MacDonald, P. E., & Rorsman, P.
Affiliations
  • Churchill Hospital (GB)
  • Lund University (SE)
Corresponding author
Braun, Matthias
ORCID
Venue
The Journal of General Physiology
Publisher
Rockefeller University Press
Published
2007-02-12
Volume
129
Issue
3
Pages
221-231
ISSN
  • 0022-1295 (ISSN-L)
  • 1540-7748
DOI
10.1085/jgp.200609658
PMC
PMC2151613
OpenAlex
W2103829969
Cited by
100
References
3
Citations by year
2026: 2 · 2025: 2 · 2024: 1 · 2023: 3 · 2022: 3 · 2021: 4 · 2020: 6 · 2019: 6 · 2018: 1 · 2017: 5 · 2016: 9 · 2015: 5 · 2014: 7 · 2013: 10 · 2012: 7
FWCI
8.55
Type
journal article
Language
en
Open access
Locations
  • The Journal of General Physiology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • Lithuanian University of Health Sciences (submittedVersion)
  • Lund University Publications (Lund University) (submittedVersion)
Funders
  • Wellcome Trust (WT072289)
  • European Commission (LSHM-CT-2006-518153, LSHB-CT-2004-005137, 005137)
Topics
  • Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
GABA release, serotonin release, ATP release, large dense core vesicles, pancreatic beta cells, corelease, exocytosis, GABAA receptor, P2X2 receptor
MeSH terms
Adenosine Triphosphate, Animals, Exocytosis, gamma-Aminobutyric Acid, Kinetics, Membrane Fusion, Models, Biological, Molecular Weight, Pyridazines, Serotonin, Microscopy, Immunoelectron, Secretory Vesicles, Insulin-Secreting Cells, Rats, Receptors, Purinergic P2X2, GABA-A Receptor Antagonists, Purinergic P2 Receptor Antagonists

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review