Skip to main content
Log in

The use of antibody and complement to gain access to the interior of presynaptic nerve terminals

  • Published:
Experimental Brain Research Aims and scope Submit manuscript

Summary

Treatment of synaptosomes with sera containing antibodies (Ab) directed against synaptosomal membranes and complement (C) alters the plasma membrane so that it becomes selectively permeable to small molecules and ions but not to proteins. When synaptosomes are incubated with Ab and C, a rapid release of intracellular K occurs. This release does not occur after treatment with antiserum alone, or with normal serum + C. Ab + C treatment releases approximately the same amount of K as does detergent treatment or hypotonic lysis, two procedures that extensively disrupt the plasma membrane.

The selectivity of the complement-induced lesion is consistent with the equivalent pore radius determined in other systems (Michaels and Mayer 1978; Sears et al. 1964). The lesions are large enough to allow the rapid permeation of small ions, but too small to permit the escape of the soluble cytoplasmic enzyme, lactate dehydrogenase. In addition, electron microscopic studies indicate that Ab+C treatment does not lead to gross morphological disruption of the synaptosomes. Ab+C treated synaptosomes are also permeable to calcium and ATP, as demonstrated by the stimulation of Ca sequestration into endoplasmic reticulum when 45Ca and ATP are added to the incubation medium.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Baker P F, Knight D E (1978) Calcium-dependent exocytosis in bovine adrenal medullary cells with leaky plasma membranes. Nature 276: 620–622

    Google Scholar 

  • Bhakdi S, Tranum-Jensen J (1978) Molecular nature of the complement lesion. Proc Natl Acad Sci USA 75: 5655–5659

    Google Scholar 

  • Blaustein M P (1974) The interrelationship between sodium and calcium fluxes across cell membranes. Rev Physiol Biochem Pharmacol 70: 33–82

    Google Scholar 

  • Blaustein M P (1975) Effects of potassium, veratridine, and scorpion venom on calcium accumulation and transmitter release by nerve terminals in vitro. J Physiol (Lond) 247: 617–655

    Google Scholar 

  • Blaustein M P, Goldring J M (1975) Membrane potentials in pinched-off presynaptic nerve terminals monitored with a fluorescent probe: evidence that synaptosomes have potassium diffusion potentials. J Physiol (Lond) 247: 589–615

    Google Scholar 

  • Blaustein M P Ratzlaff, R W, Kendrick N C, Schweitzer E S (1978a) Calcium buffering in presynaptic nerve terminals I. Evidence for involvement of a nonmitochondrial ATP-dependent sequestration mechanism. J Gen Physiol 72: 15–41

    Google Scholar 

  • Blaustein M P, Ratzlaff, R W, Schweitzer E S (1978b) Calcium buffering in presynaptic nerve terminals II. Kinetic properties of the nonmitochondrial Ca sequestration mechanism. J Gen Physiol 72: 43–66

    Google Scholar 

  • Boyle M D P, Ohanian S H, Borsos T (1976) Lysis of tumor cells by antibody and complement VII. Complement-dependent 86Rb release — a nonlethal event? J Immunol 117: 1346–1350

    Google Scholar 

  • Breckenridge W C, Combos G, Morgan L G (1972) The lipid composition of adult rat brain synaptosomal plasma membranes. Biochim Biophys Acta 266: 695–707

    Google Scholar 

  • Breckenridge W C, Morgan I G, Zanetta, J P, Vincendon G (1973) Adult rat brain synaptic vesicles II. Lipid composition. Biochim Biophys Acta 320: 681–686

    Google Scholar 

  • Burakoff S J, Martz E, Benacerrat B (1975) Is the primary complement lesion insufficient for lysis? Failure of cells damaged under osmotic protection to lyse in EDTA or at low temperature after removal of osmotic protection. Clin Immunol Immunopathol 4: 108–126

    Google Scholar 

  • Carrasco L (1978) Membrane leakiness after viral infection and a new approach to the development of antiviral agents. Nature 272: 694–699

    Google Scholar 

  • Castellot J J, Jr, Miller M R, Pardee A B (1978) Animal cells reversibly permeable to small molecules. Proc Natl Acad Sci USA 75: 351–355

    Google Scholar 

  • DeRobertis E, Lapetina E, Saavedra J P, Soto E F (1966) In vivo and in vitro action of antisera against isolated nerve endings of brain cortex. Life Sci 5: 1979–1989

    Google Scholar 

  • DiPolo R, Requena J, Brinley F J, Jr, Mullins L J, Scarpa A, Tiffert T (1976) Ionized calcium concentrations in squid axons. J Gen Physiol 67: 433–467

    Google Scholar 

  • Endo M, Kitazawa T, Yagi S, Iino M, Kakuta Y (1977) Some properties of chemically skinned smooth muscle fibers. In: Excitation — contraction coupling in smooth muscle. Casteels R, Godfraind T, Ruegg J C (eds), Elsevier/North-Holland Biomedical Press, Amsterdam, pp 199–209

    Google Scholar 

  • Fabiato A, Fabiato F (1978) Effects of pH on the myofilaments and the sarcoplasmic reticulum of skinned cells from cardiac and skeletal muscles. J Physiol (Lond) 276: 233–255

    Google Scholar 

  • Forsgren A, Sjoquist J (1967) Protein A from Staphylococcus aureus. III. Reaction with rabbit γ-globulin. J Immunol 99: 19–24

    Google Scholar 

  • Frank M M, Rapp H J, Borsos T (1964) Studies on the terminal steps of immune hemolysis. I. Inhibition by trisodium ethylenediaminetetraacetic acid (EDTA). J Immunol 93: 409–413

    Google Scholar 

  • Fried R C, Blaustein M P (1978) Retrieval and recycling of synaptic vesicle membrane in pinched-off nerve terminals (synaptosomes). J Cell Biol 78: 685–700

    Google Scholar 

  • Gray E G, Whittaker V P (1962) The isolation of nerve terminals from brain: an electron-microscopic study of cell fragments derived by homogenization and centrifugation. J Anat 96: 79–88

    Google Scholar 

  • Gordon A R (1978) Contraction of detergent-treated smooth muscle. Proc Natl Acad Sci USA 75: 3527–3530

    Google Scholar 

  • Green H, Barrow P, Goldberg B (1959) Effect of antibody and complement on permeability control in ascites tumor cells and erythocytes. J Exp Med 110: 699–713

    Google Scholar 

  • Hajos F (1975) An improved method for preparation of synaptosomal fractions in high purity. Brain Res 93: 485–489

    Google Scholar 

  • Henry R J, Chaimori N, Golub, O J, Berkman S (1960) Revised spectrophotometric methods for the determination of glutamic-oxalacetic transaminase, glutamic-pyruvic transaminase, and lactic acid dehydrogenase. Am J Clin Pathol 4: 381–398

    Google Scholar 

  • Kabat E A, Mayer M M (1967) Experimental immunochemistry. Complement and complement fixation, Thomas, Springfield, pp 133–240

    Google Scholar 

  • Karnovsky M J (1965) A formaldehyde-gluteraldehyde fixative of high osmolarity for use in electron microscopy. J Cell Biol 27: 137a-138a

    Google Scholar 

  • Karnovsky M J (1971) Use of ferocyanide-reduced osmium tetroxide in electron microscopy. Abstracts of papers. Am Soc Cell Biol, llth Annual Meeting p 146

  • Krueger B K, Ratzlaff, R W, Strichartz, G R, Blaustein M P (1979) Saxitoxin binding to synaptosomes, membranes, and solubilized binding sites from rat brain. J Membr Biol 50: 287–310

    Google Scholar 

  • Levine L (1973) Micro-complement fixation. In: Handbook of experimental Immunology. Wier, D M (ed), Blackwell, Oxford, pp 22.1–22.8

    Google Scholar 

  • Lowry O H, Rosebrough N J, Far A L, Randall R J (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265–275

    CAS  PubMed  Google Scholar 

  • Mayer M M (1972) Mechanism of cytolysis by complement. Proc Natl Acad Sci USA 69: 2954–2958

    Google Scholar 

  • McGraw C F, Somlyo A V, Blaustein M P (1978) Ultrastructural localization of calcium by electron probe analysis in presynaptic nerve terminals. Neurosci Abst 4: 332

    Google Scholar 

  • Michaels D, Mayer M (1978) Nature of the complement lesion: formation of membrane channels. Biophys J 21: 125a

  • Miledi R (1973) Transmitter release induced by injection of calcium ions into nerve terminals. Proc Roy Soc Lond B 183: 421–425

    Google Scholar 

  • Miller D J (1978) Are cardiac muscle cells “skinned” by EGTA or EDTA? Nature 277: 142–143

    Google Scholar 

  • Müller-Eberhard H J (1975) Complement. Ann Rev Biochem 44: 697–724

    Google Scholar 

  • Nadarajah A, Leese B, Joplin G F (1969) Triton X-100 scintillant for counting calcium-45 in biological fluids. Int J Appl Rad Isotopes 20: 733–735

    Google Scholar 

  • Naitoh Y, Kaneko H (1972) Reactivated Triton-extracted models of paramecium: modification of ciliary movement by calcium ions. Science 176: 523–524

    Google Scholar 

  • Raiteri M, Bertollini A, LaBella R (1972) Synaptosome antisera affect permeability of synaptosomal membranes in vitro. Nature New Biol 238: 242–243

    Google Scholar 

  • Raiteri M, Levi G (1973) Antisynaptosome antibodies affect synaptosomal permeability to neurotransmitters. Nature New Biol 245: 90–92

    Google Scholar 

  • Schweitzer E S, Blaustein M P (1978) The use of antibody and complement to gain access to the interior of presynaptic terminals. Neurosci Abst 4: 249

    Google Scholar 

  • Sears D A, Weed R I, Swisher S N (1964) Differences in the mechanism of in vitro immune hemolysis related to antibody specificity. J Clin Invest 43: 975–985

    Google Scholar 

  • Winegrad S (1971) Studies of cardiac muscle with a high permeability to calcium produced by treatment with ethylenediaminetetra-acetic acid. J Gen Physiol 58: 71–93

    Google Scholar 

  • Wobschall D, McKeon C (1975) Step conductance increases in bilayer membranes induced by antibody-antigen-complement action. Biochim Biophys Acta 413: 317–321

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schweitzer, E.S., Blaustein, M.P. The use of antibody and complement to gain access to the interior of presynaptic nerve terminals. Exp Brain Res 38, 443–453 (1980). https://doi.org/10.1007/BF00237525

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00237525

Key words

Navigation