Summary
Pure cholinergic synaptosomes isolated from the electric organ ofTorpedo marmorata were stimulated by calcium ionophore A-23187. The effect of time course of stimulation on the changes in intramembrane particles (IMPs) on presynaptic membranes was studied by quickfreezing and aldehyde-fixation freeze-fracture. We showed that the decrease of small-particle density at the P-face and the increase of large-particle density at the E-face was maximum after 30 sec of A-23187 stimulation. Later, the density of synaptic vesicles decreased. We suggest that the redistribution of IMPs on the presynaptic membrane and acetylcholine (ACh) release from pure cholinergic synaptosomes have a similar time course when triggered by A-23187
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Branton D, Bullivant S, Gilula NB, Karnovsky MJ, Muhlethaler K, Northcote DH, Packer L, Satir B, Satir P, Speth V, Staehlin LA, Steere RL, Weinstein RS (1975) Freeze-etching nomenclature. Science 190:54–56
Bearer EL, Duzzunes N, Friend DS, Papahadjopoulos D (1982) Fusion of phospholipid vesicles arrested by quick-freezing. The question of lipidic particles as intermediates in membrane fusion. BBA 693:93–98
Ceccarelli B, Grohovaz F, Hurlbut WP (1979) Freeze fracture of frog neuromuscular junctions during intense release of neurotransmitter. II. Effects of electrical stimulation and high potassium. J Cell Biol 81:178–192
Dunant Y, Muller D, Garcia-Segura LM (1983) Freeze fracture of presynaptic membrane during transmission of a single giant impulse inTorpedo electric organ. Experientia 39:680
Fesce R, Grohovaz WP, Hurlbut WP, Ceccarelli B (1980) Freeze fracture studies of frog neuromuscular junctions during intense release of neurotransmitter. III. A morphometric analysis of the number and diameter of intramembrane particles. J Cell Biol 85:337–345
Fujita H, Kurihara H, Miyama J (1983) Ultrastructural aspects of the effect of calcium ionophore A-23187 on incubated anterior pituitary cells of rats. Cell Tissue Res 229:129–136
García-Segura LM, Dunant Y, Jones GJ, Muller D and Parduzc A (1985) Rapíd change of presynaptic intramembrane particles during transmission of a single nerve impulse. Experientia 41:828
Gratzl M, Dahl G, Russell JR, Thorn NA (1977) Fusion of neurohypophysial membranes in vitro. BBA 470:45–57
Gulik-Krzywicki T, Costello MJ (1978) The use of low temperature X-ray diffraction to evaluate freezing methods used in freeze fracture electron microscopy. J Microsc (Lond) 112:103–113
Hay FE, Hasty DL (1979) Extrusion of particle-free membrane blisters during glutaraldehyde fixation. In: Rash JE, Hudson CS (eds) Freeze fracture: methods, artifacts and interpretations. Raven Press New York, pp 59–66
Hays RM, Chevalier J, Gobur R, Bourguet J (1985) Fusion images and intramembrane particles aggregates during the action of antidiuretic hormone. A rapid freeze study. Cell Tissue Res 240:433–439
Heuser JE, Reese TS (1981) Structural changes after transmitter release at the frog neuromuscular junction. J Cell Biol 88:564–580
Heuser JE, Reese TS, Landis DMD (1974) Functional changes in frog neuromuscular junctions studied with freeze-fracture. J Neurocytol 5:109–131
Heuser JE, Reese TS, Dennis MJ, Jan L, Jan Y, Evans L (1979) Synaptic vesicles exocytosis captured by quick-freezing and correlated with quantal transmitter release. J Cell Biol 81:275–300
Israel M, Lesbats B (1981a) Chemiluminescent determination of ACh and continuous detection of its release fromTorpedo electric organ synapses and synaptosomes. Neurochem Int 3:81–90
Israel M, Lesbats B (1981b) Continuous determination by a chemiluminescent method of ACh release and compartmentation inTorpedo electric synaptosomes. J Neurochem 37:1475–1483
Israel M, Manaranche R, Morel N, Dedieu JC, Gulik-Krzywicki T, Lesbats B (1981) Redistribution of intramembrane particles related to acetylcholine release by cholinergic synaptosomes. J Ultrastruct Res 75:162–178
Israel M, Lesbats B, Manaranche R, Morel N, Gulik-Krzywicki T, Dedieu JC (1982) Rearrangement of intramembrane particles as a possible mechanism for the release of acetylcholine. J Physiol (Paris) 78:348–356
Jost P, Brooks UJ, Griffith OH (1973) Fluidity of phospholipid bilayer and membranes after exposure to osmium tetroxide and glutaraldehyde. J Mol Biol 76:313–318
Llados FT, Ross-Canada J, Pappas GD (1984) Ultrastructural and physiological effects of the ionophore A-23187 at identified frog neuromuscular junctions. Neuroscience 13:237–247
Marsal J, Esquerda JE, Fiol C, Solsona C, Tomas J (1980) Calcium fluxes in isolated pure cholinergic nerve endings from the electric organ ofTorpedo marmorata. J Physiol (Paris) 76:443–457
Magul G (1979) Temperature-dependent changes in intramembrane particle distribution. In: Rash JE, Hudson CS (eds) Freeze fracture: methods, artifacts and interpretations. Raven Press New York, pp 37–42
Michaelson DM, Bilen J, Vobsky D (1978)Torpedo synaptosomes: evidence for synaptic vesicle fusion accompanying Ca2+-induced ionophore (A-23187)-mediated ACh release. Brain Res 154:409–414
Miller TM, Heuser JE (1984) Endocytosis of synaptic vesicle membrane at the neuromuscular junction. J Cell Biol 98:685–698
Morel N, Israel M, Manaranche R, Mastour-Frachon P (1977) Isolation of pure cholinergic nerve endings fromTorpedo electric organ. Evaluation of their metabolic properties. J Cell Biol 75:43–55
Morel N, Manaranche R, Gulik-Krzywicki T, Israel M (1980) Ultrastructural changes and transmitter release induced by depolarization of cholinergic synaptosomes. A freeze fracture study of a synaptosomal fraction fromTorpedo electric organ. J Ultrastruc Res 70:342–362
Orci L (1981) Membrane cycling in secretion: a morphological approach. In: Estrabrook RW, Steere P (eds) Currents Topics in Cellular Regulation, Vol 18. Academic Press, New York
Orci L, Perrelet A, Friend DS (1977) Freeze-fraction of membrane fusion during exocytosis in pancreatic B-cells. J Cell Biol 75:23–31
Pinto da Silva P, Kachar P (1980) Quick-freezing versus chemical fixation: capture and identification of membrane fusion intermediates. Cell Biol Int Rep 4:625–640
Pumplin DW, Reese TS (1977) Action of brown widow spider venom and botulinum toxin on the frog neuromuscular junction examined with freeze fracture technique. J Physiol (Lond) 273:443–457
Quastel DMJ (1974) Excitation-secretion coupling release at the mammalian neuromuscular junction. In: Bennett MVL (ed) Synaptic transmission and Neural Interaction. Raven Press, New York
Smith JE, Reese TS (1980) Use of aldehyde fixatives to determine the rate of synaptic transmitter release. J Exp Biol 89:19–29
Takano Y, Kamiya H (1979) Effect of ionic environment on densities of freeze-fractured synaptosomes. Experientia 35:1076–1078
Torri-Tarelli F, Grohovaz H, Fesce R, Ceccarelli B (1985) Temporal coincidence between synaptic vesicle fusion and quantal secretion of acetylcholine. J Cell Biol 101:1386–1399
Triller A, Korn H (1985) Activity-dependent deformations of presynaptic grids at central synapses. J Neurocytol 14:177–192
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Egea, G., Esquerda, J.E., Calvet, R. et al. Structural changes at pure cholinergic synaptosomes during the transmitter release induced by A-23187 inTorpedo marmorata . Cell Tissue Res. 248, 207–214 (1987). https://doi.org/10.1007/BF01239982
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01239982