Abstract
Protoplasts isolated from root cap cells of maize were shown to secrete fucose-rich polysaccharides and were used in a patch-clamp study to monitor changes in whole-cell capacitance. Ca2+ was required for exocytosis, which was measured as an increase in cell capacitance during intracellular dialysis with Ca2+ buffers via the patch pipette. Exocytosis was stimulated significantly by small increases above normal resting [Ca2+]. In the absence of Ca2+, protoplasts decreased in size. In situ hybridization showed significant expression of the maize annexin p35 in root cap cells, differ-entiating vascular tissue, and elongating cells. Dialysis of protoplasts with maize annexins stimulated exocytosis at physiological [Ca2+], and this could be blocked by dialysis with antibodies specific to maize annexins. Dialysis with milli-molar concentrations of GTP strongly inhibited exocytosis, causing protoplasts to decrease in size. GTPgammaS and GDPbetaS both caused only a slight inhibition of exocytosis at physiological Ca2+. Protoplasts were shown to internalize plasma membrane actively. The results are discussed in relation to the regulation of exocytosis in what is usually considered to be a constitutively secreting system; they provide direct evidence for a role of annexins in exocytosis in plant cells.
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- Battey N. H., James N. C., Greenland A. J. cDNA isolation and gene expression of the maize annexins p33 and p35. Plant Physiol. 1996 Nov;112(3):1391–1396. doi: 10.1104/pp.112.3.1391. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blackbourn H. D., Barker P. J., Huskisson N. S., Battey N. H. Properties and partial protein sequence of plant annexins. Plant Physiol. 1992 Jul;99(3):864–871. doi: 10.1104/pp.99.3.864. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bradley D., Carpenter R., Sommer H., Hartley N., Coen E. Complementary floral homeotic phenotypes result from opposite orientations of a transposon at the plena locus of Antirrhinum. Cell. 1993 Jan 15;72(1):85–95. doi: 10.1016/0092-8674(93)90052-r. [DOI] [PubMed] [Google Scholar]
- Burgess T. L., Kelly R. B. Constitutive and regulated secretion of proteins. Annu Rev Cell Biol. 1987;3:243–293. doi: 10.1146/annurev.cb.03.110187.001331. [DOI] [PubMed] [Google Scholar]
- Burgoyne R. D., Handel S. E. Activation of exocytosis by GTP analogues in adrenal chromaffin cells revealed by patch-clamp capacitance measurement. FEBS Lett. 1994 May 16;344(2-3):139–142. doi: 10.1016/0014-5793(94)00361-0. [DOI] [PubMed] [Google Scholar]
- Burgoyne R. D., Morgan A. Regulated exocytosis. Biochem J. 1993 Jul 15;293(Pt 2):305–316. doi: 10.1042/bj2930305. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clark G. B., Roux S. J. Annexins of plant cells. Plant Physiol. 1995 Dec;109(4):1133–1139. doi: 10.1104/pp.109.4.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coorssen J. R., Schmitt H., Almers W. Ca2+ triggers massive exocytosis in Chinese hamster ovary cells. EMBO J. 1996 Aug 1;15(15):3787–3791. [PMC free article] [PubMed] [Google Scholar]
- Creutz C. E. The annexins and exocytosis. Science. 1992 Nov 6;258(5084):924–931. doi: 10.1126/science.1439804. [DOI] [PubMed] [Google Scholar]
- Donnelly S. R., Moss S. E. Annexins in the secretory pathway. Cell Mol Life Sci. 1997 Jun;53(6):533–538. doi: 10.1007/s000180050068. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Díaz-Muñoz M., Hamilton S. L., Kaetzel M. A., Hazarika P., Dedman J. R. Modulation of Ca2+ release channel activity from sarcoplasmic reticulum by annexin VI (67-kDa calcimedin). J Biol Chem. 1990 Sep 15;265(26):15894–15899. [PubMed] [Google Scholar]
- 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]
- Fiedler K., Lafont F., Parton R. G., Simons K. Annexin XIIIb: a novel epithelial specific annexin is implicated in vesicular traffic to the apical plasma membrane. J Cell Biol. 1995 Mar;128(6):1043–1053. doi: 10.1083/jcb.128.6.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fischer von Mollard G., Stahl B., Li C., Südhof T. C., Jahn R. Rab proteins in regulated exocytosis. Trends Biochem Sci. 1994 Apr;19(4):164–168. doi: 10.1016/0968-0004(94)90278-x. [DOI] [PubMed] [Google Scholar]
- Gilroy S., Bethke P. C., Jones R. L. Calcium homeostasis in plants. J Cell Sci. 1993 Oct;106(Pt 2):453–461. doi: 10.1242/jcs.106.2.453. [DOI] [PubMed] [Google Scholar]
- Goda Y., Südhof T. C. Calcium regulation of neurotransmitter release: reliably unreliable? Curr Opin Cell Biol. 1997 Aug;9(4):513–518. doi: 10.1016/s0955-0674(97)80027-0. [DOI] [PubMed] [Google Scholar]
- Griffing L. R. Comparisons of Golgi structure and dynamics in plant and animal cells. J Electron Microsc Tech. 1991 Feb;17(2):179–199. doi: 10.1002/jemt.1060170206. [DOI] [PubMed] [Google Scholar]
- Harder T., Simons K. Caveolae, DIGs, and the dynamics of sphingolipid-cholesterol microdomains. Curr Opin Cell Biol. 1997 Aug;9(4):534–542. doi: 10.1016/s0955-0674(97)80030-0. [DOI] [PubMed] [Google Scholar]
- Hess S. D., Doroshenko P. A., Augustine G. J. A functional role for GTP-binding proteins in synaptic vesicle cycling. Science. 1993 Feb 19;259(5098):1169–1172. doi: 10.1126/science.8438167. [DOI] [PubMed] [Google Scholar]
- Homann U., Tester M. Ca2+-independent and Ca2+/GTP-binding protein-controlled exocytosis in a plant cell. Proc Natl Acad Sci U S A. 1997 Jun 10;94(12):6565–6570. doi: 10.1073/pnas.94.12.6565. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoshino T., Mizutani A., Chida M., Hidaka H., Mizutani J. Plant annexin form homodimer during Ca(2+)-dependent liposome aggregation. Biochem Mol Biol Int. 1995 Apr;35(4):749–755. [PubMed] [Google Scholar]
- Ikonen E., Tagaya M., Ullrich O., Montecucco C., Simons K. Different requirements for NSF, SNAP, and Rab proteins in apical and basolateral transport in MDCK cells. Cell. 1995 May 19;81(4):571–580. doi: 10.1016/0092-8674(95)90078-0. [DOI] [PubMed] [Google Scholar]
- 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]
- McClung A. D., Carroll A. D., Battey N. H. Identification and characterization of ATPase activity associated with maize (Zea mays) annexins. Biochem J. 1994 Nov 1;303(Pt 3):709–712. doi: 10.1042/bj3030709. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moore P. J., Swords K. M., Lynch M. A., Staehelin L. A. Spatial organization of the assembly pathways of glycoproteins and complex polysaccharides in the Golgi apparatus of plants. J Cell Biol. 1991 Feb;112(4):589–602. doi: 10.1083/jcb.112.4.589. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nuoffer C., Balch W. E. GTPases: multifunctional molecular switches regulating vesicular traffic. Annu Rev Biochem. 1994;63:949–990. doi: 10.1146/annurev.bi.63.070194.004505. [DOI] [PubMed] [Google Scholar]
- Pollard H. B., Rojas E., Pastor R. W., Rojas E. M., Guy H. R., Burns A. L. Synexin: molecular mechanism of calcium-dependent membrane fusion and voltage-dependent calcium-channel activity. Evidence in support of the "hydrophobic bridge hypothesis" for exocytotic membrane fusion. Ann N Y Acad Sci. 1991;635:328–351. doi: 10.1111/j.1749-6632.1991.tb36503.x. [DOI] [PubMed] [Google Scholar]
- Pusch M., Neher E. Rates of diffusional exchange between small cells and a measuring patch pipette. Pflugers Arch. 1988 Feb;411(2):204–211. doi: 10.1007/BF00582316. [DOI] [PubMed] [Google Scholar]
- Raynal P., Pollard H. B. Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium- and phospholipid-binding proteins. Biochim Biophys Acta. 1994 Apr 5;1197(1):63–93. doi: 10.1016/0304-4157(94)90019-1. [DOI] [PubMed] [Google Scholar]
- Scheres B., McKhann H. I., Van Den Berg C. Roots Redefined: Anatomical and Genetic Analysis of Root Development. Plant Physiol. 1996 Aug;111(4):959–964. doi: 10.1104/pp.111.4.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith C. B., Betz W. J. Simultaneous independent measurement of endocytosis and exocytosis. Nature. 1996 Apr 11;380(6574):531–534. doi: 10.1038/380531a0. [DOI] [PubMed] [Google Scholar]
- Taylor A. R., Brownlee C. Localized Patch Clamping of Plasma Membrane of a Polarized Plant Cell : Laser Microsurgery of the Fucus spiralis Rhizoid Cell Wall. Plant Physiol. 1992 Aug;99(4):1686–1688. doi: 10.1104/pp.99.4.1686. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor A. R., Manison NFH., Fernandez C., Wood J., Brownlee C. Spatial Organization of Calcium Signaling Involved in Cell Volume Control in the Fucus Rhizoid. Plant Cell. 1996 Nov;8(11):2015–2031. doi: 10.1105/tpc.8.11.2015. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tester M., Zorec R. Cytoplasmic calcium stimulates exocytosis in a plant secretory cell. Biophys J. 1992 Sep;63(3):864–867. doi: 10.1016/S0006-3495(92)81662-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Verma DPS., Cheon C., 3rd, Hong Z. Small GTP-Binding Proteins and Membrane Biogenesis in Plants. Plant Physiol. 1994 Sep;106(1):1–6. doi: 10.1104/pp.106.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vitale N., Mukai H., Rouot B., Thiersé D., Aunis D., Bader M. F. Exocytosis in chromaffin cells. Possible involvement of the heterotrimeric GTP-binding protein G(o). J Biol Chem. 1993 Jul 15;268(20):14715–14723. [PubMed] [Google Scholar]
- Wilson K. L. NSF-independent fusion mechanisms. Cell. 1995 May 19;81(4):475–477. doi: 10.1016/0092-8674(95)90067-5. [DOI] [PubMed] [Google Scholar]