Organelle Identification and Characterization in Plant Cells: Using a Combinational Approach of Confocal Immunofluorescence and Electron Microscope

다국어 초록 (Multilingual Abstract)

The plant secretory and endocytic pathways

consist of several functionally distinct membrane-bounded

compartments. The ultra structures of the endoplasmic

reticulum, the Golgi apparatus, and central vacuoles have

been well characterized via traditional structural electron

microscope (EM). However, the identification of plant

prevacuolar compartments (PVCs) and early endosomes

(EEs) had not been achieved until more recently because of

the lack of specific markers for these organelles. Recent

development of fluorescent reporters for PVCs and EEs

expressing in transgenic tobacco BY-2 cells and Arabidopsis

plants has allowed their dynamic characterization in

living cells via confocal microscopy and drug treatment,

which led to their subsequent morphological identification

via structural and immunogold EM. Thus, in this review,

we will use our studies on PVCs and EEs as examples to

present an efficient approach for organelle identification in

plant cells via primary characterization of fluorescentmarked

organelles in living cells and their dynamic

response to drug treatments, which then serves as the

basis for subsequent immunogold and structural EM

studies for organelle identification. Such strategy thus

represents a powerful approach in future research for the

identification of novel organelles and transport vesicles in

plant cells.

참고문헌 (Reference)

1 Tse YC, "ldentification of multivesicular bodies as prevacuolar compartments in Nicotiana tabacum BY-2 cells" 16 (16): 672-693, 2004

2 Thelen M, "Wortmannin binds specifically to l-phosphatidylinositol 3-kinase while inhibiting guanine nucleotide-binding protein-coupled receptor signaling in neutrophil leukocytes" 91 (91): 4960-4964, 1994

3 Dettmer J, "Vaeuolar -APTase activity is required for endocytic and secretory trafficking in Arabidopsis" 18 (18): 715-730, 2006

4 Bethke PC, "Vacuoles and prevacuolar compartments" 3 (3): 469-475, 2000

5 Oparka KJ, "Using GFP to study virus invasion and spread in plant tissues" 388 (388): 401-402, 1997

6 Emans N, "Uptake of a fluorescent marker in plant cells is sensitive to brefeldin A and wortmannin" 14 (14): 71-86, 2002

7 Hara-Nishimura H, "Transport of storage proteins to protein storage vacuoles is mediated by large precursor-accumulating vesicles" 10 (10): 825-836, 1998

8 Miao Y, "Transient expression of fluorescent fusion proteins in protoplasts of suspension cultured cells" 2 (2): 2348-2353, 2007

9 Ortiz-Zapater E, "Trafficking of the human transferrin receptor in plant cells: effects of tyrphostin A23 and brefeldin A" 48 (48): 757-770, 2006

10 Lam SK, "Tracking down the elusive early endosome" 12 (12): 497-505, 2007

11 Miao Y, "The vacuolar transport of aleurain-GFP and 2S albumin-GFP fusions is mediated by the same pre-vacuolar compartments in tobacco BY-2 and Arabidopsis suspension cultured cells" 56 : 824-839, 2008

12 daSilva Conceicao A, "The syntaxin homolog AtPEP12P resides on a late post-Golgi compartment in plants" 9 (9): 571-582, 1997

13 Marcusson EG, "The sorting receptor for yeast vacuolar carboxy-peptidase Y is encoded by the VPS10 gene" 77 (77): 579-586, 1994

14 Bright NA, "The relationship between lumenal and limiting membranes in swollen late endocytic compartments formed after wortmannin treatment or sucrose accumulation" 2 (2): 631-642, 2001

15 Robinson DG, "The endosomal system of plants: charting new and familiar territories" 147 (147): 1482-1492, 2008

16 Sanderfoot AA, "The Arabidopsis genome. An abundance of soluble N-ethylmaleimide-sensitive factor adaptor protein receptors" 124 (124): 1558-1569, 2000

17 daSilva LL, "Targeting of the plant vacuolar sorting receptor BP80 is dependent on multiple sorting signals in the cytosolic tail" 18 (18): 1477-1497, 2006

18 Komfeld S, "Structure and function of the mannose 6-phosphate/insulinlike growth factor II receptors" 61 : 307-330, 1992

19 Nebenfuhr A, "Stop-and-go movements of plant Golgi stacks are mediated by the acto-myosin system" 121 (121): 1127-1142, 1999

20 Lam SK, "SCAMPs highlight the developing cell plate during cytokinesis in tobacco BY-2 cells" 147 (147): 1637-1645, 2008

21 Lam SK, "Rice SCAMP1 defines clathrin-coated, trans-Golgilocated tubular-vesicular st겨ctures as an early endosome in tobacco BY-2 cells" 19 (19): 296-319, 2007

22 Ritzenthaler C, "Reevaluation of the effects of brefeldin A on plant cells using tobacco Bright Yellow 2 cells expressing Golgi-targeted green fluorescent protein and COPI antisera" 14 (14): 237-261, 2002

23 daSilva LL, "Receptor salvage from the prevacuolar compartment is essential for efficient vacuolar protcin targeting" 17 (17): 132-148, 2005

24 Chow CM, "Rab-A2 and Rab-A3 GTPases define a trans-Golgi endosomal membrane domain in Arabidopsis that contributes substantially to the cell plate" 20 (20): 101-123, 2008

25 Robinson DG, "Protein sorting to the storage vacuoles of plants: a critical appraisal" 6 (6): 615-625, 2005

26 Pesacreta TC, "Presence of a partially-coated reticulum in angiosperms" 125 : 173-184, 1985

27 Reggiori F, "Polar transmembrane domains target proteins to the interior of the yeast vacuole" 11 (11): 3737-3749, 2000

28 Mo B, "Plant prevacuolar/endosomal compartments" 253 : 95-129, 2006

29 Foresti O, "Overexpression of the Arabidopsis syntaxin PEP12/SYP21 inhibits transport from the prevacuolar compartment to the lytîc vacuole in vivo" 18 (18): 2275-2293, 2006

30 Henandez LE, "Novel expression patterns of phosphatidylinositol 3-hydroxy kinase in nodulated Medicago spp. plants" 55 (55): 957-959, 2004

31 Paris N, "Molecular cloning and further characterization of a probable plant vacuolar sorting receptor" 115 (115): 29-39, 1997

32 Brandizzi F, "Membrane protein transport between the endoplasmic reticulum and the Golgi in tobacco leaves is energy dependent but cytoskeleton independent: evidence from selective photobleaching" 14 (14): 1293-1309, 2002

33 Miao Y, "Localization of green fluorescent protein fusions with the seven Arabidopsis vacuolar sorting receptors to prevacuolar compartments in tobacco BY-2 cells" 142 (142): 945-962, 2006

34 Jiang L, "Integral membrane protein sorting to vacuoles in plant cells‘evidence for two pathways" 143 (143): 1183-1199, 1998

35 Sciaky N, "Golgi tubule traffic and the effects of brefeldin A visualized in living cells" 139 (139): 1137-1155, 1997

36 Brandizzi F, "GFP is the way to glow: bioimaging of the plant endomembrane system" 214 (214): 138-158, 2004

37 Bolte S, "FM-dyes as experimental probes for dissecting vesicle trafficking in living plant cells" 214 (214): 159-173, 2004

38 Battey NH, "Exocytosis and endocytosis" 11 (11): 643-660, 1999

39 Jaillais Y, "Evidence for a sorting endosome in Arabidopsis root cells" 53 (53): 237-247, 2008

40 Tanchak MA, "Endocytosis of cationized ferritin by coated vesic1es of soybean protoplasts" 162 : 481-486, 1984

41 Hubner R, "Endocytosis in maize root cap cells: evidence obtained using heavy metal salt solutions" 129 : 214-222, 1985

42 Tse YC, "Dynamic response of prevacuolar compartments to brefeldin a in plant cells" 142 (142): 1442-1459, 2006

43 Matsuoka K, "Different sensitivity to wortmannin of two vacuolar sorting signals indicates the presence of distinct sorting machineries in tobacco cells" 130 (130): 1307-1318, 1995

44 Das S, "Cloning of Brassica napus phospholipase C2 (BnPLC2), phosphatidylinositol 3-kinase (BnVPS34) and phosphatidylinositol synthasel (BnPtdIns SI)-comparative analysis of the effect of abiotic stresses on the expression of phosphatidylinositol signal transduction-related genes in B. napus" 220 (220): 777-784, 2005

45 Ahmed SU, "Cloning and subcellular location of an Arabidopsis receptor -like protein that shares common features with protein-sorting receptors of eukaryotic cells" 114 (114): 325-336, 1997

46 Dhonukshe P, "Clathrin-mediated consitutive endocytosis of PIN auxin efflux carriers in Arabidopsis" 17 (17): 520-527, 2007

47 Segui-Simarro JM, "Cell cycle-dependent changes in Golgi stacks, vacuoles, clathrin-coated vesicles and multivesicular bodies in meristematic cells of Arabidopsis thaliana: a quantitative and spatial analysis" 223 (223): 223-236, 2006

48 Nebenfuhr A, "Brefeldin A: deciphering an enigmatic inhibitor of secretion" 130 (130): 1102-1108, 2002

49 Satiat-Jeunemaitre B, "Brefeldin A effects in plant and fungal cells: something new about vesicle trafficking?" 181 (181): 162-177, 1996

50 Horn MA, "Biotin-mediated delivery of exogenous macromolecules into soybean cells" 93 (93): 1492-1496, 1990

51 Paris N, "BP-80 as a vacuolar sorting receptor" 50 (50): 903-914, 2002

52 Li YB, "BP-80 and homologs are concentrated on post-Golgi, probable lytic prevacuolar compartments" 43 (43): 726-742, 2002

53 Welters P, "AtVPS34, a phosphatidylinositol 3-kinase of Arabidopsis thaliana, is an essential protein with homology to a calcium-dependent lipid binding domain" 91 (91): 11398-11402, 1994

54 Jaillais Y, "AtSNXI defines an endosome for auxin-carrier trafficking in Arabidopsis" 443 (443): 106-109, 2006

55 Ueda T, "Ara6, a plant-unique novel type Rab GTPase, functions in the endocytic pathway of Arabidopsis thaliana" 20 (20): 4730-4741, 2001

56 Hong Z, "A phosphatidylinositol 3-killase is induced during soybcan nodule organogenesis and is associatcd with membrane proliferation" 91 (91): 9617-9621, 1994

57 Vida TA, "A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast" 128 (128): 779-792, 1995