Isolation of Synaptic Vesicles and Structural Organization of the Acetylcholine System Within Brain Nerve Endings*

Robertis, E. De; Arnaiz, Georgina Rodrı́guez de Lores; Iraldi, L. Salganicoff Amanda Pellegrino de; Zieher, Luis Marı́a

doi:10.1111/j.1471-4159.1963.tb05038.x

Cited by 419 publications

(42 citation statements)

References 2 publications

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“…ACh-esterase has been concentrated in subcellular fractions of brain containing nerve ending ghosts (5). Also, in subcellular fractions of the electric tissue of Torpedo ocellata the presence of ACh-esterase has been correlated with the presence of membrane (6).…”

Section: Introductionmentioning

confidence: 93%

THE ASSOCIATION OF ACETYLCHOLINESTERASE AND MEMBRANE IN SUBCELLULAR FRACTIONS OF THE ELECTRIC TISSUE OF ELECTROPHORUS

Karlin

1965

The Journal of Cell Biology

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Subcellular fractions of the electric tissue of the main organ of the eel Electrophorus electricus were prepared in sucrose media by differential centrifugation and differential discontinuous gradient centrifugation. The distributions of acetylcholinesterase, cytochrome oxidase, DNA, and protein were determined. The appearance of the fractions was determined by phase contrast microscopy and by electron microscopy. A fraction prepared by differectial centrifugation at 30,000 g for 20 minutes in 0.89 M sucrose contained 63 per cent of the total acetylcholinesterase activity at 4 times the specific activity of that of the tissue homogenate. A subfraction prepared by centrifugation in a discontinuous density gradient showed a peak of total and relative specific acetylcholinesterase activity of 35 per cent and 1.9, respectively. The average over-all purification was 7 times. The acetylcholinesterase peak was below the cytochrome oxidase peak and above the DNA peak in the density gradient. The presence of acetylcholinesterase in the fractions was correlated with the presence of large fragments of the cell membrane; however, the presence of other tissue components was noted. The acetylcholinesterase associated with membrane was found to be activated by incubation with sodium deoxycholate. The possible use of the peak fraction containing membranes rich in acetylcholinesterase for the investigation of other components of the acetylcholine system and of other properties of the membrane is discussed.

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Section: Introductionmentioning

confidence: 93%

THE ASSOCIATION OF ACETYLCHOLINESTERASE AND MEMBRANE IN SUBCELLULAR FRACTIONS OF THE ELECTRIC TISSUE OF ELECTROPHORUS

Karlin

1965

The Journal of Cell Biology

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“…In the CNS, ACh synthesis (Quastel et al 1936) and release (Macintosh and Oborin 1953) were demonstrated. Bound ACh, the ACh-synthesizing enzyme cholineacetyltransferase (CHAT) and the ACh-cleaving enzyme acetylcholinesterase (ACHE) were shown to be contained in CNS synaptosome terminals (DeRobertis et al 1962(DeRobertis et al , 1963Whittaker et al 1963Whittaker et al , 1964.…”

Section: Cholinergic and Cholinoceptive Structures Are Involved In A mentioning

confidence: 99%

Immunohistochemistry of cholinergic receptors

Schröder

1992

Anat Embryol

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Acetylcholine and its receptors are involved in a variety of important signal transduction processes. As shown here paradigmatically for the human neuromuscular junction and the cerebral cortex, acetylcholine receptors can be visualized immunohistochemically at the cellular and subcellular level under physiological and pathological conditions. At normal motor endplates nicotinic cholinoceptors are localized at the surface of the postsynaptic junctional folds. In myasthenic syndromes investigation of muscle biopsies enables the diagnosis of receptor deficiencies at the ultrastructural level. In normal cerebral cortex pyramidal neurons are equipped with both nicotinic and muscarinic acetylcholine receptors localized to postsynaptic densities. In neuropsychiatric diseases cholinoceptor expression can be monitored at the cellular level by quantitative assessment of immunolabeled cortical neurons.

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“…To characterize 2 of the mAbs, 6-4E and 3-l lG, obtained from a fusion in which GCPs had served as the antigen, crude synaptosomes were prepared from hamster brain tissue, in lieu of GCPs, essentially as described by Cotman (1974) and DeRobertis et al (1963) up to the discontinuous sucrose density gradient step. In brief, hamsters at postnatal ages 3, 6, 9, 12, 15, 21, and 37 d were anesthetized and sacrificed by either an overdose of Nembutal (Abbott Labs, IL) or hypothermia on ice for infant animals.…”

Section: Methodsmentioning

confidence: 99%

Expression of two developmentally regulated brain-specific proteins is correlated with late outgrowth of the pyramidal tract

Kalil

Perdew

1988

J. Neurosci.

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The regulation of axon outgrowth is not well understood. In previous studies, however, axon elongation has been well correlated with expression of a small number of growth-associated proteins (GAPs). To identify other proteins whose expression could be correlated with axon outgrowth during development of CNS pathways, monoclonal antibodies were raised against growth cone particles isolated from neonatal hamster brains. Two of these antibodies recognized a brain-specific 33 kDa protein associated with intracellular membranes of axons and growth cones. Immunoblotting demonstrated a sharp developmental decline in levels of the protein in hamster brain during the first postnatal week and a more gradual decline thereafter. Immunocytochemical studies with the antibodies revealed ubiquitous staining of the neuropil during the first several days, which by the end of the first week became restricted to a few later-maturing pathways. Staining was most intense in the pyramidal tract and was well correlated with axon outgrowth, which continues until 14 d in this pathway. These results suggest that the 33 kDa protein may, like previously identified GAPs, play a role in axon elongation. Late outgrowth of the hamster pyramidal tract is also correlated with expression of another developmentally regulated protein, the high-molecular-weight neurofilament subunit (NF-H). Immunostaining with a monoclonal antibody that recognized phosphorylated NF-H demonstrated that this subunit does not begin to appear in the late-maturing pyramidal tract fibers until several weeks after birth, in striking contrast to intense immunoreactivity of other spinal cord pathways from postnatal day 1. This finding suggests that specific pathways may have a highly idiosyncratic time course for expression of neurofilament subunits.

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Isolation of Synaptic Vesicles and Structural Organization of the Acetylcholine System Within Brain Nerve Endings*

Cited by 419 publications

References 2 publications

THE ASSOCIATION OF ACETYLCHOLINESTERASE AND MEMBRANE IN SUBCELLULAR FRACTIONS OF THE ELECTRIC TISSUE OF ELECTROPHORUS

THE ASSOCIATION OF ACETYLCHOLINESTERASE AND MEMBRANE IN SUBCELLULAR FRACTIONS OF THE ELECTRIC TISSUE OF ELECTROPHORUS

Immunohistochemistry of cholinergic receptors

Expression of two developmentally regulated brain-specific proteins is correlated with late outgrowth of the pyramidal tract

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