Skip to main content

Advertisement

Springer Nature Link
Log in

How do patch clamp seals form? A lipid bleb model

  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

Individual ion channels are electrically isolated and studied in living cells with the tight patch voltage clamp method. Channels are identified, categorized, and sometimes named on the basis of the biophysical properties obtained with this method. Although it is usually presumed that these recordings are from native, undisturbed membrane, the physical basis of this technique is not well established. Observations that lipid blebs readily form when suction is applied to patch clamp electrodes suggest that many single channel recordings are from ion channels in these blebs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Canada)

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ascher P, Bregestovski P, Nowak L (1988) N-Methyl-D aspartate-activated channels of mouse central neurones in magnesium-free solutions. J Physiol (Lond) 399:207–226

    Google Scholar 

  • Auerbach A, Sachs F (1985) High-resolution patch-clamp techniques. In: Smith TG Jr, Lecar H, Redman SJ, Gage PW (eds) Voltage and patch clamping with microelectrodes. Waverly Press, Baltimore, pp 121–149

    Google Scholar 

  • Bekoff A, Betz WJ (1977) Physiological properties of dissociated muscle fibres obtained from innervated and denervated adult rat muscle. J Physiol (Lond) 271:25–40

    Google Scholar 

  • Benz R, Fröhlich O, Läuger P, Montai M (1975) Electrical capacity of black lipid films and of lipid bilayers made from monolayers. Biochem Biophys Acta 394:323–334

    Google Scholar 

  • Caldwell JH, Campbell DT, Beam KG (1986) Na channel distribution in vertebrate skeletal muscle. J Gen Physiol 87:907–932

    Google Scholar 

  • Camerino D, Bryant SH (1976) Effects of denervation and colchicine treatment on the chloride conductance of rat skeletal muscle fibers. J Neurobiol 7:221–228

    Google Scholar 

  • Coleman HA, Parkington HC (1987) Single channel Cl− and K+ currents from cells of uterus not treated with enzymes. Pflügers Arch 410:560–562

    Google Scholar 

  • Corey DP, Stevens CF (1983) Science and technology of patch-recording electrodes. In: Sakmann B, Neher E (eds) Single channel recording. Plenum Press, New York, pp 53–68

    Google Scholar 

  • Coronado R, Latorre R (1983) Phospholipid bilayers made from monolayers on patch-clamp pipettes. Biophys J 43:231–236

    Google Scholar 

  • Dudel J, Franke C, Hatt H (1989) Artefacts in recording glutamate-activated single channel currents from crayfish muscle. Neurosci Lett 97:350–353

    Google Scholar 

  • Dunlap K, Brehm P (1988) Control of a light emitting photoprotein by calcium channels in a hydrozoan coelenterate. In: Grinnell AD, Armstrong D, Jackson MB (eds) Calcium and ion channel modulation. Plenum Press, New York, pp 89–101

    Google Scholar 

  • Edwards FA, Konnerth A, Sakmann B, Takahashi T (1989) A thin slice preparation for patch clamp recordings from neurones of the mammalian central nervous system. Pflügers Arch 414:600–612

    Google Scholar 

  • Elgsaeter A, Shotton DM, Branton D (1976) Intramembrane particle aggregation in erythrocyte ghosts. II. The influence of spectrin aggregation. Biochim Biophys Acta 426:101–122

    Google Scholar 

  • Fahlke Ch, Ruppersberg JP, Rüdel R (1989) Simultaneous measurements of the sodium currents in the cell-attached and the whole-cell clamp modes lead to different results. Pflügers Arch 412:R17

    Google Scholar 

  • Farnbach GC, Barchi RL (1977) Determination of muscle cable parameters from a single membrane voltage response. J Membr Biol 32:133–149

    Google Scholar 

  • Fenwick EM, Marty A, Neher E (1982) A patch-clamp study of bovine chromaffin cells and of their sensitivity to acetylcholine. J Physiol (Lond) 331:577–597

    Google Scholar 

  • Fernandez JM, Fox AP, Krasne S (1984) Membrane patches and whole-cell membranes: a comparison of electrical properties in rat clonal pituitary (GH3) cells. J Physiol (Lond) 356: 565–585

    Google Scholar 

  • Fischmeister R, Ayer RK Jr, DeHaan RL (1986) Some limitations of the cell-attached patch clamp technique: a two-electrode analysis. Pflügers Arch 406:73–82

    Google Scholar 

  • Franke Ch, Dudel J (1987) Single glutamate-gated synaptic channels at the crayfish neuromuscular junction. I. The effect of enzyme treatment. Pflügers Arch 408:300–306

    Google Scholar 

  • Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100

    Google Scholar 

  • Hanai T, Haydon DA, Taylor J (1965) The influence of lipid composition and of some adsorbed proteins on the capacitance of black hydrocarbon membranes. J Theor Biol 9:422–432

    Google Scholar 

  • Haydon DA, Urban BW (1985) The admittance of the squid giant axon at radio frequencies and its relation to membrane structure. J Physiol (Lond) 360:275–291

    Google Scholar 

  • Horn R, Patlak J (1980) Single channel currents from excised patches of muscle membrane. Proc Natl Acad Sci USA 77:6930–6934

    Google Scholar 

  • Martinac B, Buechner M, Delcour AH, Adler J, Kung C (1987) Pressure-sensitive ion channel in Escherichia coli. Proc Natl Acad Sci USA 84:2297–2301

    Google Scholar 

  • Methfessel C, Witzemann V, Takahashi T, Mishina M, Numa S, Sakmann B (1986) Patch clamp measurements on Xenopus laevis oocytes: currents through endogenous channels and implanted acetylcholine receptor and sodium channels. Pflügers Arch 407:577–588

    Google Scholar 

  • Milton RL, Caldwell JH (1986) Vesicles extracted from skeletal muscle fibers via suction: possible implications for loose and tight patch voltage clamping. Biophys J 49:172a

    Google Scholar 

  • Milton RL, Caldwell JH (1990) Na current in membrane blebs: implications for channel mobility and patch clamp recording. J Neurosci 10:885–893

    Google Scholar 

  • Miranda AF, Godman GC, Tanenbaum SW (1974) Action of cytochalasin D on cells of established lines. II. Cortex and microfilaments. J Cell Biol 62:406–423

    Google Scholar 

  • Rae JL, Levis RA (1984) Patch voltage clamp of lens epithelial cells: theory and practice. Mol Physiol 6:115–162

    Google Scholar 

  • Roberts WM (1987) Sodium channels near end-plates and nuclei of snake skeletal muscle. J Physiol (Lond) 388:213–232

    Google Scholar 

  • Roberts WM, Hudspeth AJ (1987) Co-localization of Ca channels with Ca-activated K channels in hair cells of the bullfrog's sacculus. Soc Neurosci Abstr 13:177

    Google Scholar 

  • Sachs F, Sokabe M (1990) Stress and strain in patch clamped membranes. Quantitative video microscopy, single channel recording and capacitance. Biophys J 57:265a

    Google Scholar 

  • Sakmann B, Neher E (1983) Geometric parameters of pipettes and membrane patches. In: Sakmann B, Neher E (eds) Single channel recording. Plenum Press, New York, pp 37–51

    Google Scholar 

  • Scott RE (1976) Plasma membrane vesiculation: a new technique for isolation of plasma membranes. Science 194:743–745

    Google Scholar 

  • Stockbridge LL, French AS (1988) Stretch-activated cation channels in human fibroblasts. Biophys J 54:187–190

    Google Scholar 

  • Strickholm A (1961) Impedance of a small electrically isolated area of the muscle cell surface. J Gen Physiol 44:1073–1088

    Google Scholar 

  • Stühmer W, Roberts WM, Almers W (1983) The loose patch clamp. In: Sakmann B, Neher E (eds) Single-channel recording. Plenum Press, New York, pp 123–132

    Google Scholar 

  • Tank DW, Wu ES, Webb WW (1982a) Enhanced molecular diffusibility in muscle membrane blebs: Release of lateral constraints. J Cell Biol 92:207–212

    Google Scholar 

  • Tank DW, Miller C, Webb WW (1982b) Isolated-patch recording from liposomes containing functionally reconstituted chloride channels from Torpedo electroplax. Proc Natl Acad Sci USA 79:7749–7753

    Google Scholar 

  • Vicini S, Schuetze SM (1985) Gating properties of acetylcholine receptors at developing rat endplates. J Neurosci 5:2212–2224

    Google Scholar 

  • Zollinger HU (1948) Cytologic studies with the phase microscope. I. The formation of “blisters” on cells in suspension (potocytosis), with observations on the nature of the cellular membrane. Am J Pathol 24:545–567

    Google Scholar 

Download references

Author information

Author notes

  1. R. L. Milton

    Present address: Indiana University School of Medicine, Muncie Center for Medical Education, Ball State University, 47306, Muncie, IN, USA

Authors and Affiliations

  1. Departments of Cellular and Structural Biology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA

    J. H. Caldwell

  2. Department of Physiology, University of Colorado Health Sciences Center, 80262, Denver, CO, USA

    R. L. Milton & J. H. Caldwell

Authors
  1. R. L. Milton
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. H. Caldwell
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Milton, R.L., Caldwell, J.H. How do patch clamp seals form? A lipid bleb model. Pflügers Arch 416, 758–765 (1990). https://doi.org/10.1007/BF00370626

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00370626

Key words

Search

Navigation