Advertisement

Brief Guide to Patch-Clamp Current Measurements in Organelle Membranes

  • Yoshio Maruyama
  • Akihiro Hazama
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

The standard giga-seal patch-clamp techniques opened the way to monitor ion-channel activity in the native organelle membrane in situ. One has to have an organelle preparation, covered with the intact membrane carrying rightly oriented channel proteins and suitable for proper patch-clamping. Here we deal with the intact “native” organelle, the nucleus with an inner and outer nucleic membrane (nuclear envelope), and mitochondria without the outer membrane but an intact inner membrane (mitoplast). We collect them by rupturing the cell using a variety of procedures including cell homogenization, cell swelling with a hypotonic solution, cell shaking in a Na citrate solution, and their appropriate combination, usually modified by individual laboratories.

Keywords

Acinar Cell Nuclear Envelope Pancreatic Acinar Cell Hypotonic Solution Glass Pipette 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Wieckowski M, Giorgi C, Lebiedzinska M, Duszynski J, Pinton P (2009) Isolation of mitochondria-associated membranes and mitochondria from animal tissues and cells. Nature Protocol 4:1582–1590CrossRefGoogle Scholar
  2. 2.
    Borecky J, Jezek P, Siemen D (1997) 108-pS channels in brown fat mitochondria might be identical to the inner membrane anion channel. J Biol Chem 272:19282–19289PubMedGoogle Scholar
  3. 3.
    Keller B, Hedrich R (1992) Patch-clamp techniques to study ion channels from organelles. Methods Enzymol 207:673–681, 146PubMedCrossRefGoogle Scholar
  4. 4.
    Zoratti M, De Marchi U, Gulbins E, Szabo I (2009) Novel channels of the inner mitochondrial membrane. Biochim Biophys Acta 1789:351–363Google Scholar
  5. 5.
    Franco-Obregon A, Wang H, Clapham D (2000) Distinct ion channel classes are expressed on the outer nuclear envelope on T- and B-lymphocyte cell lines. Biophys J 79:202–214PubMedCrossRefGoogle Scholar
  6. 6.
    Ionescu L, Cheung K-H, Vais H, Mak D-O, White C, Foskett K (2006) Graded recruitment and inactivation of single InsP3 receptor Ca2+-release channels: implications for quantal Ca2+ release. J Physiol 573:645–662PubMedCrossRefGoogle Scholar
  7. 7.
    Maruyama Y, Shimada H, Taniguchi J (1995) Ca2+-activated K+-channels in the nuclear envelope isolated from single pancreatic acinar cells. Pflugers Arch 430:148–150PubMedCrossRefGoogle Scholar
  8. 8.
    Mak D, Foskette K (1994) Single-channel inositol 2,4,5-trisphosphate receptor currents revealed by patch clamp of isolated Xenopus oocyte nuclei. J Biol Chem 269:29375–29378PubMedGoogle Scholar
  9. 9.
    Matzke A, Weiger T, Matzke M (2010) Ion channels at the nucleus: electrophysiology meets the genome. Mol Plant 4:642–652CrossRefGoogle Scholar
  10. 10.
    Humbert J-P, Matter N, Artault J-C, Koppler P, Malviya A (1996) Inositol 1,4,5-trisphosphate receptor is located to the inner nuclear membrane vindicating regulation of nuclear calcium signaling by inositol 1,4,5-trisphosphate. J Biol Chem 271:475–478Google Scholar
  11. 11.
    Marchenko S, Yarotskyy V, Kovalenko T, Kostyuk P, Thomas R (2005) Spontaneously active and InsP3-activated ion channels in cell nuclei from rat cerebellar Purkinje and granule neurons. J Physiol 565:897–910PubMedCrossRefGoogle Scholar
  12. 12.
    Schieder M, Rotzer K, Bruggemann A, Biel M, Wahl-Schott C (2010) Characterization of two-pore channel 2 (TPCN2)-mediated Ca2+ currents in isolated lysosomes. J Biol Chem 285:21219–21222PubMedCrossRefGoogle Scholar
  13. 13.
    Lee W, Torchalski B, Roussa E, Thvenod F (2008) Evidence for KCNQ1 K+ channel expression in rat zymogen granule membranes and involvement in cholecystokinin-induced pancreatic acinar secretion. Am J Physiol 294:C879–C892CrossRefGoogle Scholar

Copyright information

© Springer 2012

Authors and Affiliations

  1. 1.Department of PhysiologyTohoku University Graduate School of MedicineSendaiJapan
  2. 2.Fukushima Medical UniversityFukushimaJapan

Personalised recommendations