从维管束植物中分离质体膜片钳技术：一种快速分离原生质体的方法，该方法具有形成千兆欧姆封接的高成功率。
Patch clamping protoplasts from vascular plants : method for the quick isolation of protoplasts having a high success rate of gigaseal formation.
机构信息
Department of Botany, University of Washington, Seattle, Washington 98195.
出版信息
Plant Physiol. 1991 Dec;97(4):1573-5. doi: 10.1104/pp.97.4.1573.
Abstract
A method is described for the isolation of protoplasts (Pisum sativum, Phaseolus vulgaris, Avena sativa, Arabidopsis thaliana) in preparation for ion flux studies using patch clamp electrophysiology. Protoplasts that have been exposed to hydrolytic, cell wall degrading, enzymes for as little as 5 minutes form gigaseals (seal resistance higher than 10 giga Ohm) with the patch pipette with success rates greater than 40%. Sealing of these protoplasts is fast, averaging less than 2 minutes. This method yields high rates of gigaseal formation in a variety of tissues from both monocots and dicots and will enhance data collection in ion flux studies of plasma membranes of vascular plants.
摘要
描述了一种分离原生质体（豌豆、菜豆、燕麦、拟南芥）的方法，以便用膜片钳电生理学进行离子通量研究。原生质体在接触到水解酶、细胞壁降解酶后，只需 5 分钟即可与膜片钳形成千兆欧姆级别的巨封（电阻高于 10 千兆欧姆），成功率大于 40%。这些原生质体的密封速度很快，平均不到 2 分钟。该方法可在单子叶植物和双子叶植物的多种组织中产生高比例的千兆欧姆封接，将提高对植物质膜离子通量研究的数据收集效率。
相似文献
1
Patch clamping protoplasts from vascular plants : method for the quick isolation of protoplasts having a high success rate of gigaseal formation.从维管束植物中分离质体膜片钳技术：一种快速分离原生质体的方法，该方法具有形成千兆欧姆封接的高成功率。
Plant Physiol. 1991 Dec;97(4):1573-5. doi: 10.1104/pp.97.4.1573.
2
Electrophysiology in the eukaryotic model cell Saccharomyces cerevisiae.真核模式细胞酿酒酵母中的电生理学
Pflugers Arch. 1998 Nov;436(6):999-1013. doi: 10.1007/s004240050735.
3
Isolation and whole-cell patch clamping of Arabidopsis guard cell protoplasts.拟南芥保卫细胞原生质体的分离与全细胞膜片钳技术
CSH Protoc. 2008 Jun 1;2008:pdb.prot5014. doi: 10.1101/pdb.prot5014.
4
Seal-promoting solutions and pipette perfusion for patch clamping plant cells.用于膜片钳记录植物细胞的促进封接溶液和移液管灌注
Plant J. 1997 Apr;11(4):891-6. doi: 10.1046/j.1365-313x.1997.11040891.x.
5
Cell marking in Arabidopsis thaliana and its application to patch-clamp studies.拟南芥中的细胞标记及其在膜片钳研究中的应用。
Plant J. 1998 Sep;15(6):843-51. doi: 10.1046/j.1365-313x.1998.00256.x.
6
Giga-ohm seals on intracellular membranes: a technique for studying intracellular ion channels in intact cells.细胞内膜上的千兆欧封接：一种研究完整细胞内离子通道的技术。
Neuron. 1997 Jul;19(1):7-13. doi: 10.1016/s0896-6273(00)80343-8.
7
Novel ion channels in the protists, Mougeotia and Saprolegnia, using sub-gigaseals.利用亚千兆封接技术研究原生生物穆氏藻和水霉中的新型离子通道。
FEBS Lett. 1992 Oct 5;310(3):219-22. doi: 10.1016/0014-5793(92)81336-k.
8
Preparation and applications of Arabidopsis thaliana guard cell protoplasts.拟南芥保卫细胞原生质体的制备及应用
New Phytol. 2002 Mar;153(3):517-526. doi: 10.1046/j.0028-646X.2001.00329.x. Epub 2002 Mar 5.
9
Reducing agents facilitate membrane patch seal integrity and longevity.还原剂促进膜片钳封接的完整性和稳定性。
Channels (Austin). 2024 Dec;18(1):2297621. doi: 10.1080/19336950.2023.2297621. Epub 2023 Dec 28.
10
A high-performance elastomeric patch clamp chip.一种高性能弹性膜片钳芯片。
Lab Chip. 2006 Oct;6(10):1338-45. doi: 10.1039/b607913j. Epub 2006 Aug 23.
引用本文的文献
1
In vivo femtosecond laser nanosurgery of the cell wall enabling patch-clamp measurements on filamentous fungi.用于丝状真菌膜片钳测量的活体细胞壁飞秒激光纳米手术
Microsyst Nanoeng. 2024 Apr 7;10:47. doi: 10.1038/s41378-024-00664-x. eCollection 2024.
2
Obtaining spheroplasts of armored dinoflagellates and first single-channel recordings of their ion channels using patch-clamping.获得甲藻的原生质球，并首次使用膜片钳技术对其离子通道进行单通道记录。
Mar Drugs. 2014 Sep 5;12(9):4743-55. doi: 10.3390/md12094743.
3
Extracellular Ca2+ ameliorates NaCl-induced K+ loss from Arabidopsis root and leaf cells by controlling plasma membrane K+ -permeable channels.细胞外钙离子通过控制质膜钾离子通透通道，改善氯化钠诱导的拟南芥根和叶细胞钾离子流失。
Plant Physiol. 2006 Aug;141(4):1653-65. doi: 10.1104/pp.106.082388. Epub 2006 Jun 23.
4
Localized Patch Clamping of Plasma Membrane of a Polarized Plant Cell : Laser Microsurgery of the Fucus spiralis Rhizoid Cell Wall.植物极化细胞质膜的局域片钳：石莼根状细胞细胞壁的激光微手术。
Plant Physiol. 1992 Aug;99(4):1686-8. doi: 10.1104/pp.99.4.1686.
5
Gadolinium effects on gigaseal formation and the adhesive properties of a fungal amoeboid cell, the slime mutant of Neurospora crassa.钆对巨大封接形成及粗糙脉孢菌黏液突变体这种真菌变形细胞黏附特性的影响。
J Membr Biol. 2004 Mar 15;198(2):77-87. doi: 10.1007/s00232-004-0661-3.
6
Differences in whole-cell and single-channel ion currents across the plasma membrane of mesophyll cells from two closely related Thlaspi species.两种近缘遏蓝菜属植物叶肉细胞质膜上全细胞离子电流和单通道离子电流的差异。
Plant Physiol. 2003 Feb;131(2):583-94. doi: 10.1104/pp.011932.
7
ATP-Dependent Regulation of an Anion Channel at the Plasma Membrane of Protoplasts from Epidermal Cells of Arabidopsis Hypocotyls.拟南芥下胚轴表皮细胞原生质体质膜上阴离子通道的ATP依赖性调控
Plant Cell. 1995 Dec;7(12):2091-2100. doi: 10.1105/tpc.7.12.2091.
8
Inward-Rectifying K+ Channels in Root Hairs of Wheat (A Mechanism for Aluminum-Sensitive Low-Affinity K+ Uptake and Membrane Potential Control).小麦根毛中的内向整流钾离子通道（铝敏感型低亲和力钾离子吸收及膜电位调控机制）
Plant Physiol. 1994 Aug;105(4):1399-1408. doi: 10.1104/pp.105.4.1399.
9
Characterization of a Light-Controlled Anion Channel in the Plasma Membrane of Mesophyll Cells of Pea.豌豆叶肉细胞质膜中光控阴离子通道的特性分析
Plant Physiol. 1997 Apr;113(4):1419-1426. doi: 10.1104/pp.113.4.1419.
10
Laser microsurgery of higher plant cell walls permits patch-clamp access.高等植物细胞壁的激光显微手术可实现膜片钳接入。
Plant Physiol. 1996;110(4):1063-8. doi: 10.1104/pp.110.4.1063.
本文引用的文献
1
Substrate regulation of single potassium and chloride ion channels in Arabidopsis plasma membrane.拟南芥质膜中单个钾离子和氯离子通道的底物调节
Plant Physiol. 1991 Feb;95(2):642-7. doi: 10.1104/pp.95.2.642.
2
Properties of Single K and Cl Channels in Asclepias tuberosa Protoplasts.肉质叶烟草质体中单 K 和 Cl 通道的特性。
Plant Physiol. 1987 Oct;85(2):413-8. doi: 10.1104/pp.85.2.413.
3
Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection.利用卡那霉素筛选进行拟南芥根外植体的根癌农杆菌介导转化。
Proc Natl Acad Sci U S A. 1988 Aug;85(15):5536-40. doi: 10.1073/pnas.85.15.5536.
4
Light-stimulated cell expansion in bean (Phaseolus vulgaris L.) leaves. I. Growth can occur without photosynthesis.菜豆（Phaseolus vulgaris L.）叶片中光刺激引起的细胞扩张。I. 生长可在无光合作用的情况下发生。
Planta. 1990 Aug;182(1):72-6. doi: 10.1007/BF00239986.
5
Ion channels in plasmalemma of wheat protoplasts.小麦原生质体质膜中的离子通道。
Science. 1984 Nov 16;226(4676):835-8. doi: 10.1126/science.6093255.
6
Evaluation of pollen viability by enzymatically induced fluorescence; intracellular hydrolysis of fluorescein diacetate.通过酶促诱导荧光评估花粉活力；荧光素二乙酸酯的细胞内水解
Stain Technol. 1970 May;45(3):115-20. doi: 10.3109/10520297009085351.
7
K+ transport properties of K+ channels in the plasma membrane of Vicia faba guard cells.蚕豆保卫细胞质膜中钾离子通道的钾离子转运特性
J Gen Physiol. 1988 Nov;92(5):667-83. doi: 10.1085/jgp.92.5.667.
8
Effects of abscisic acid on K+ channels in Vicia faba guard cell protoplasts.脱落酸对蚕豆保卫细胞原生质体中钾离子通道的影响。
Biochem Biophys Res Commun. 1987 May 29;145(1):284-90. doi: 10.1016/0006-291x(87)91318-0.
9
How do patch clamp seals form? A lipid bleb model.膜片钳封接是如何形成的？脂质泡模型。
Pflugers Arch. 1990 Aug;416(6):758-62. doi: 10.1007/BF00370626.
Zotero 插件