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. 1990 Apr 1;110(4):1033–1039. doi: 10.1083/jcb.110.4.1033

Patch-clamp measurements reveal multimodal distribution of granule sizes in rat mast cells

PMCID: PMC2116067  PMID: 2182644

Abstract

Using patch-clamp techniques, we have followed the attributes of the secretory granules of peritoneal mast cells obtained from rats of different ages. The granule attributes were determined by following the step increases in the cell surface membrane area caused by the exocytosis of the granules in GTP gamma S stimulated mast cells. Our data show that the amount of granule membrane available for exocytosis depends exponentially on the weight (age) of the donor rat, reaching a maximum at approximately 300 g. The data are consistent with an exponential growth in the number of granules contained by mast cells of maturing animals. Histograms of the sizes of the step increases in surface area caused by exocytosis of the granules showed at least four equally spaced peaks of similar variance where the position of the first peak and the spacing between peaks averaged 1.3 +/- 0.4 micron2. In all cells recorded, no more than seven peaks could be found, the higher order peaks having a lower probability of occurrence. The distribution of granule sizes did not change measurably between young and adult animals. This study suggests that at least two separate steps may determine the size of a secretory granule: granule to granule fusion that may account for the subunit composition of granule sizes and traffic of microvesicles through the maturing granules that may account for the variance observed in the granule sizes. This study also demonstrates a novel way to study granulo-genesis in living cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BOYD I. A., MARTIN A. R. The end-plate potential in mammalian muscle. J Physiol. 1956 Apr 27;132(1):74–91. doi: 10.1113/jphysiol.1956.sp005503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Breckenridge L. J., Almers W. Final steps in exocytosis observed in a cell with giant secretory granules. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1945–1949. doi: 10.1073/pnas.84.7.1945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clausen J., Jahn H., Nielsen E. H. Electron microscopical study of rat mast cell maturation. Virchows Arch B Cell Pathol Incl Mol Pathol. 1983;43(2):151–158. doi: 10.1007/BF02932952. [DOI] [PubMed] [Google Scholar]
  4. Combs J. W. Maturation of rat mast cells. An electron microscope study. J Cell Biol. 1966 Dec;31(3):563–575. doi: 10.1083/jcb.31.3.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dvorak A. M., Hammond M. E., Morgan E., Orenstein N. S., Galli S. J., Dvorak H. F. Evidence for a vesicular transport mechanism in guinea pig basophilic leukocytes. Lab Invest. 1980 Feb;42(2):263–276. [PubMed] [Google Scholar]
  6. Farquhar M. G., Palade G. E. The Golgi apparatus (complex)-(1954-1981)-from artifact to center stage. J Cell Biol. 1981 Dec;91(3 Pt 2):77s–103s. doi: 10.1083/jcb.91.3.77s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fernandez J. M., Neher E., Gomperts B. D. Capacitance measurements reveal stepwise fusion events in degranulating mast cells. 1984 Nov 29-Dec 5Nature. 312(5993):453–455. doi: 10.1038/312453a0. [DOI] [PubMed] [Google Scholar]
  8. Fidler N., Fernandez J. M. Phase tracking: an improved phase detection technique for cell membrane capacitance measurements. Biophys J. 1989 Dec;56(6):1153–1162. doi: 10.1016/S0006-3495(89)82762-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Griffiths G., Simons K. The trans Golgi network: sorting at the exit site of the Golgi complex. Science. 1986 Oct 24;234(4775):438–443. doi: 10.1126/science.2945253. [DOI] [PubMed] [Google Scholar]
  10. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  11. Hammel I., Lagunoff D., Bauza M., Chi E. Periodic, multimodal distribution of granule volumes in mast cells. Cell Tissue Res. 1983;228(1):51–59. doi: 10.1007/BF00206264. [DOI] [PubMed] [Google Scholar]
  12. Hammel I., Lagunoff D., Krüger P. G. Studies on the growth of mast cells in rats. Changes in granule size between 1 and 6 months. Lab Invest. 1988 Oct;59(4):549–554. [PubMed] [Google Scholar]
  13. Helander H. F., Bloom G. D. Quantitative analysis of mast cell structure. J Microsc. 1974 Apr;100(3):315–321. doi: 10.1111/j.1365-2818.1974.tb03943.x. [DOI] [PubMed] [Google Scholar]
  14. Joshi C., Fernandez J. M. Capacitance measurements. An analysis of the phase detector technique used to study exocytosis and endocytosis. Biophys J. 1988 Jun;53(6):885–892. doi: 10.1016/S0006-3495(88)83169-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kelly R. B. Pathways of protein secretion in eukaryotes. Science. 1985 Oct 4;230(4721):25–32. doi: 10.1126/science.2994224. [DOI] [PubMed] [Google Scholar]
  16. Komuro M., Kiuchi Y., Shioda T. Membrane modification during secretory granule formation in rat somatotrophs. Eur J Cell Biol. 1987 Feb;43(1):98–103. [PubMed] [Google Scholar]
  17. Maruyama Y. Agonist-induced changes in cell membrane capacitance and conductance in dialysed pancreatic acinar cells of rats. J Physiol. 1988 Dec;406:299–313. doi: 10.1113/jphysiol.1988.sp017381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nadelhaft I. Measurement of the size distribution of zymogen granules from rat pancreas. Biophys J. 1973 Oct;13(10):1014–1029. doi: 10.1016/S0006-3495(73)86042-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Neher E., Marty A. Discrete changes of cell membrane capacitance observed under conditions of enhanced secretion in bovine adrenal chromaffin cells. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6712–6716. doi: 10.1073/pnas.79.21.6712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. PADAWER J., GORDON A. S. Peritoneal fluid mast cells: their numbers and morphology in rats of various body weights (ages). J Gerontol. 1956 Jul;11(3):268–272. doi: 10.1093/geronj/11.3.268. [DOI] [PubMed] [Google Scholar]
  21. Padawer J. Mast cells: extended lifespan and lack of granule turnover under normal in vivo conditions. Exp Mol Pathol. 1974 Apr;20(2):269–280. doi: 10.1016/0014-4800(74)90059-8. [DOI] [PubMed] [Google Scholar]
  22. Tooze J., Tooze S. A. Clathrin-coated vesicular transport of secretory proteins during the formation of ACTH-containing secretory granules in AtT20 cells. J Cell Biol. 1986 Sep;103(3):839–850. doi: 10.1083/jcb.103.3.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zimmerberg J., Curran M., Cohen F. S., Brodwick M. Simultaneous electrical and optical measurements show that membrane fusion precedes secretory granule swelling during exocytosis of beige mouse mast cells. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1585–1589. doi: 10.1073/pnas.84.6.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. von Zastrow M., Castle J. D. Protein sorting among two distinct export pathways occurs from the content of maturing exocrine storage granules. J Cell Biol. 1987 Dec;105(6 Pt 1):2675–2684. doi: 10.1083/jcb.105.6.2675. [DOI] [PMC free article] [PubMed] [Google Scholar]

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