Use of carboxyfluorescein diacetate to study formation of permeable channels between mouse blastomeres

Goodall, Harry W.; Johnson, Martin H.

doi:10.1038/295524a0

Cited by 113 publications

(34 citation statements)

References 15 publications

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“…After excision, the fruitbearing detached shoots were immediately recut under water with or without 100 g mL Ϫ1 of CFDA and illuminated at a photon flux density of 500 mol m Ϫ2 sec Ϫ1 at 22 to 25ЊC for 24 hr, unless otherwise stated. Upon entering cells, the CFDA is cleaved by cytoplasmic esterase to produce the membrane-impermeant symplastic fluorescent probe CF (Goodall and Johnson, 1982). Because the subsequent confocal imaging of CF in seed took place ‫02ف‬ cm away from the loading end of shoots, the disturbance to the CF movement into the seed was minimized.…”

Section: Loading Of Cf and Confocal Laser Scanning And Fluorescent Mmentioning

confidence: 99%

The Control of Single-Celled Cotton Fiber Elongation by Developmentally Reversible Gating of Plasmodesmata and Coordinated Expression of Sucrose and K⁺ Transporters and Expansin

2001

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Each cotton fiber is a single cell that elongates to 2.5 to 3.0 cm from the seed coat epidermis within ‫ف‬ 16 days after anthesis (DAA). To elucidate the mechanisms controlling this rapid elongation, we studied the gating of fiber plasmodesmata and the expression of the cell wall-loosening gene expansin and plasma membrane transporters for sucrose and K ؉ , the major osmotic solutes imported into fibers. Confocal imaging of the membrane-impermeant fluorescent solute carboxyfluorescein (CF) revealed that the fiber plasmodesmata were initially permeable to CF (0 to 9 DAA), but closed at ‫ف‬ 10 DAA and re-opened at 16 DAA. A developmental switch from simple to branched plasmodesmata was also observed in fibers at 10 DAA. Coincident with the transient closure of the plasmodesmata, the sucrose and K ϩ transporter genes were expressed maximally in fibers at 10 DAA with sucrose transporter proteins predominately localized at the fiber base. Consequently, fiber osmotic and turgor potentials were elevated, driving the rapid phase of elongation. The level of expansin mRNA, however, was high at the early phase of elongation (6 to 8 DAA) and decreased rapidly afterwards. The fiber turgor was similar to the underlying seed coat cells at 6 to 10 DAA and after 16 DAA. These results suggest that fiber elongation is initially achieved largely by cell wall loosening and finally terminated by increased wall rigidity and loss of higher turgor. To our knowledge, this study provides an unprecedented demonstration that the gating of plasmodesmata in a given cell is developmentally reversible and is coordinated with the expression of solute transporters and the cell wall-loosening gene. This integration of plasmodesmatal gating and gene expression appears to control fiber cell elongation. INTRODUCTIONA unique feature of cotton seed development is that ‫ف‬ 30% of the ovule epidermal cells initiate into fibers from the outermost layer of integument at anthesis (See Figure 1A). Each cotton fiber is a single cell and elongates from 10 to 15 m up to 2.5 to 3.0 cm by ‫ف‬ 16 days after anthesis (DAA) before it switches to secondary cell wall cellulose synthesis (Basra and Malik, 1984;Tiwari and Wilkins, 1995). The rate of fiber elongation and the final length attained are well above that commonly seen for plant cells (Cosgrove, 1997) and render it perhaps the longest single cell in higher plants. Thus, the cotton fiber represents a unique system in which to study not only carbon partitioning to cellulose synthesis (Delmer and Amor, 1995;Ruan et al., 1997) but also the control of cell elongation without the complication of cell division and multicellular development. Apart from its significance in understanding basic cell biology, elucidating the cellular and molecular basis of fiber elongation could also identify potential targets for genetic manipulation of fiber length, a key determinant of fiber yield and quality.The rapid fiber elongation is believed to be driven by high turgor (Dhindsa et al., 1975;Ruan and Chourey, 1998;Smart et al., 19...

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Section: Loading Of Cf and Confocal Laser Scanning And Fluorescent Mmentioning

confidence: 99%

The Control of Single-Celled Cotton Fiber Elongation by Developmentally Reversible Gating of Plasmodesmata and Coordinated Expression of Sucrose and K⁺ Transporters and Expansin

2001

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“…Root tips that shrivelled and became brown were considered dead. The visual criteria were confirmed by staining with 6-carboxyfluorescein diacetate (40 µg ml −" in HEPES buffer, pH 7 ; Goodall & Johnson, 1982) or with acridine orange (0n001 % (w\w) in water ; Henry & Deacon, 1981 ;Wenzel & McCully, 1991) for 2-5 min before rinsing and viewing with a BH-22 microscope (Olympus, Lake Success, NY) equipped for epifluorescence ; live roots had bright fluorescence of nuclei and cytoplasm compared to the weak fluorescence of brown tips. The number of lateral root primordia was determined in three root regions : distal, medial, and proximal.…”

Section: Root Growthmentioning

confidence: 99%

Root growth, developmental changes in the apex, and hydraulic conductivity for Opuntia ficus‐indica during drought

Dubrovsky¹,

North

Nobel

1998

New Phytologist

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Developmental changes in the root apex and accompanying changes in lateral root growth and root hydraulic conductivity were examined for Opuntia ficus-indica (L.) Miller during rapid drying, as occurs for roots near the soil surface, and more gradual drying, as occurs in deeper soil layers. During 7 d of rapid drying (in containers with a 3-cm depth of vermiculite), the rate of root growth decreased sharply and most root apices died ; such a determinate pattern of root growth was not due to meristem exhaustion but rather to meristem mortality after 3 d of drying. The length of the meristem, the duration of the cell division cycle, and the length of the elongation zone were unchanged during rapid drying. During 14 d of gradual drying (in containers with a 6-cm depth of vermiculite), root mortality was relatively low ; the length of the elongation zone decreased by 70 %, the number of meristematic cells decreased 30 %, and the duration of the cell cycle increased by 36 %. Root hydraulic conductivity (L P ) decreased to one half during both drying treatments ; L P was restored by 2 d of rewetting owing to the emergence of lateral roots following rapid drying and to renewed apical elongation following gradual drying. Thus, in response to drought, the apical meristems of roots of O. ficus-indica near the surface die, whereas deeper in the substrate cell division and elongation in root apices continue. Water uptake in response to rainfall in the field can be enhanced by lateral root proliferation near the soil surface and additionally by resumption of apical growth for deeper roots.

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Section: Embryo Collection and Culture In Vitromentioning

confidence: 99%

“…During this process, blastomeres flatten upon each other to maximize cell contact [1], and a rearrangement of many components of the cell surface [2][3][4][5][6][7], cytoplasm and cytoskeleton [8][9][10][11][12] occurs. In this stage, gap junctional communication is established between blastomeres [13][14][15][16][17].Transcription necessary for compaction is completed by the mid 4-cell stage [18][19][20]. Membrane proteins necessary for compaction, such as E-cadherin and connexin 43, are also synthesized from the late 4-cell stage onward [21][22][23][24] and are then transported to the plasma membrane.…”

mentioning

confidence: 99%

Effect of Brefeldin-A on Compaction of Preimplantation Mouse Embryos.

Ogawa

Mori

Shimizu

1997

Journal of Reproduction and Development

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Abstract. The effect of the protein transport inhibitor brefeldin A (BFA) on compaction of early mouse embryos was investigated morphologically and immunocytochemically. After placing 4-and 8-cell mouse embryos in a medium with BFA for 3 or 6 h, the features of compaction were investigated. It was found that the effects of BFA on compaction were dependent upon the cell stage when this drug was applied. Compaction was delayed when embryos were placed in BFA during the mid or late 8-cell stage (3-6 and 6-9 h post division to the 8-cell stage). In embryos treated with BFA, Roh-ConA binding was weakened and detected at the cell contact regions. The distribution of microvilli seemed to be altered. It was concluded that protein transport for compaction was initiated at the early 8-cell stage just before the development of the compacted state. Key words: Mouse embryo, Protein transport, Compaction, Cell polarity.(J. Reprod. Dev. 43: [303][304][305][306][307][308][309][310] 1997) ompaction of the 8-cell mouse embryo is an important process leading to blastocyst formaBrefeldin A (BFA) is an inhibitor of vesicular transport from the endoplasmic reticulum to the Golgi apparatus [25]. BFA inhibited compaction of mouse embryos [24]. In the present study, the effects of BFA treatment of 4-and 8-cell stage embryos on compaction were examined. The main objective of the present study is to determine the time when intracellular protein transport necessary for compaction is initiated in mouse embryos cultured in vitro. Materials and Methods Embryo collection and culture in vitroF1 (C57BL/6 × C3H/He) female mice (6-10 weeks old) were superovulated by intraperitoneal injection of 8 IU pregnant mare's serum gonadotrophin (PMSG, Sankyo, Japan) followed 48 h later by 8 IU human chorionic gonadotrophin (hCG, Teikoku Hormone Mfg., Japan). The female mice C tion. During this process, blastomeres flatten upon each other to maximize cell contact [1], and a rearrangement of many components of the cell surface [2][3][4][5][6][7], cytoplasm and cytoskeleton [8][9][10][11][12] occurs. In this stage, gap junctional communication is established between blastomeres [13][14][15][16][17].Transcription necessary for compaction is completed by the mid 4-cell stage [18][19][20]. Membrane proteins necessary for compaction, such as E-cadherin and connexin 43, are also synthesized from the late 4-cell stage onward [21][22][23][24] and are then transported to the plasma membrane. These proteins are phosphorylated and assembled into a restricted array. However, these temporally regulated steps of post-translational events are not well known.

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Use of carboxyfluorescein diacetate to study formation of permeable channels between mouse blastomeres

Cited by 113 publications

References 15 publications

The Control of Single-Celled Cotton Fiber Elongation by Developmentally Reversible Gating of Plasmodesmata and Coordinated Expression of Sucrose and K⁺ Transporters and Expansin

The Control of Single-Celled Cotton Fiber Elongation by Developmentally Reversible Gating of Plasmodesmata and Coordinated Expression of Sucrose and K⁺ Transporters and Expansin

Root growth, developmental changes in the apex, and hydraulic conductivity for Opuntia ficus‐indica during drought

Effect of Brefeldin-A on Compaction of Preimplantation Mouse Embryos.

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Use of carboxyfluorescein diacetate to study formation of permeable channels between mouse blastomeres

Cited by 113 publications

References 15 publications

The Control of Single-Celled Cotton Fiber Elongation by Developmentally Reversible Gating of Plasmodesmata and Coordinated Expression of Sucrose and K+ Transporters and Expansin

The Control of Single-Celled Cotton Fiber Elongation by Developmentally Reversible Gating of Plasmodesmata and Coordinated Expression of Sucrose and K+ Transporters and Expansin

Root growth, developmental changes in the apex, and hydraulic conductivity for Opuntia ficus‐indica during drought

Effect of Brefeldin-A on Compaction of Preimplantation Mouse Embryos.

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The Control of Single-Celled Cotton Fiber Elongation by Developmentally Reversible Gating of Plasmodesmata and Coordinated Expression of Sucrose and K⁺ Transporters and Expansin

The Control of Single-Celled Cotton Fiber Elongation by Developmentally Reversible Gating of Plasmodesmata and Coordinated Expression of Sucrose and K⁺ Transporters and Expansin