Histochemical and Biochemical Demonstration of Sialic Acid and Sulphate in Vesicles and Membranes Isolated from Nerve Endings of Rat Brain

1973 ◽  
Vol 13 (1) ◽  
pp. 237-255
Author(s):  
RUTH MARX ◽  
ELKE GRAF ◽  
W. WESEMANN
Keyword(s):  
Sialic Acid ◽  
Rat Brain ◽  
Sulphuric Acid ◽  
Nerve Ending ◽  
Synaptic Vesicles ◽  
Iron Hydroxide ◽  
Acid Methyl Ester ◽  
Nerve Endings ◽  
Uniform Size ◽  
Colloidal Iron

The reaction of colloidal iron hydroxide (CIH) with acidic groups was applied for an ultra-structural study of the distribution of sulphuric acid monoesters and sialic acid in synaptic vesicles and external nerve ending membranes isolated from rat brain. At pH 1.7 CIH was precipitated as electron-dense granules with a uniform size of 6-7 nm specifically labelling the carboxyl group of sialic acid and the sulphate group of monoesters of sulphuric acid. The differentiation of these 2 groups was achieved by treatment with neuraminidase and methylation followed by saponification. After preincubation with neuraminidase, which released 90-100% of the sialic acid from the membranes of the synaptic vesicles and the nerve endings, the electron-dense deposits marked the reaction sites of sulphate with CIH. The sulphate groups which were present at a concentration of 2.3 and 2.2 µmol/mg protein for the synaptic vesicle and nerve ending membrane preparations, respectively, were rendered soluble as methyl monosulphate by trans-esterification with acid/methanol and quantitatively removed from the structures. By this treatment membrane-bound sialic acid was blocked as sialic acid methyl ester and partly split off by acid hydrolysis. About 55% of the sialic acid found in the nerve ending membranes remained attached to the structure as compared with about 35% of the sialic acid of the vesicles. The acid-resistant proportion of the sialic acid could be localized with CIH after saponification of the esterified preparations. The method described allows the electron-microscopical demonstration of acid-resistant, neuraminidase-sensitive sialic acid in synaptic structures and the discrimination from sulphated mucopolysaccharides.

scholarly journals Binding of 1,1,1-trichloro-2,2-di-p-chlorophenylethane (DDT) with subcellular fractions of rat brain

1969 ◽  
Vol 114 (1) ◽  
pp. 135-139 ◽  
Author(s):  
Hans Brunnert ◽  
Fumio Matsumura
Keyword(s):  
Rat Brain ◽  
Nerve Ending ◽  
Soluble Proteins ◽  
Subcellular Fractions ◽  
Nerve Endings ◽  
Sephadex Column ◽  
Column Method ◽  
Synaptic Complexes ◽  
Soluble Components

DDT (1,1,1-trichloro-2,2-di-p-chlorophenylethane) and DDE (1,1-dichloro-2,2-di-p-chlorophenylethylene, a non-insecticidal analogue of DDT) were found to bind with various nerve components of rat brain. The amount of DDT binding exceeded that of DDE only in the fraction containing mainly the nerve endings. Among various components in the nerve-ending fraction, a subfraction containing mainly the pre- and post-synaptic complexes had the highest affinity for DDT in comparison with that for DDE. By using an ‘acetone-powdering’ technique on brain preparations, the Sephadex-column method was shown to provide reliable results for studies on the binding of DDT or DDE with soluble proteins in the nerve tissues. From these results it was concluded that DDE had a higher affinity for soluble components of the rat brain than did DDT.

Studies On Plasma Membranes

1967 ◽  
Vol 2 (4) ◽  
pp. 499-512
Author(s):  
E. L. BENEDETTI ◽  
P. EMMELOT
Keyword(s):  
Sialic Acid ◽  
Tight Junctions ◽  
Plasma Membranes ◽  
Iron Hydroxide ◽  
Chelating Agent ◽  
Colloidal Iron ◽  
Outer Leaflet ◽  
Liver Membranes ◽  
Junctional Complexes ◽  
Pre Treatment

Plasma membranes were isolated from rat liver and a transplanted rat hepatoma of the hepatocellular type. After glutaraldehyde fixation the membranes were treated with colloidal iron hydroxide (CIH) at pH 1.7, which was found to react specifically with the neuraminidase-sensitive sialic acid of the liver membranes. The CIH-reactive, neuraminidase-sensitive sialic acid, comprising 70% of the membrane-bound sialic acid, was exclusively located in the outer leaflet of the liver membranes as shown by the rather regular distribution of electron-dense CIH granules. This granular, asymmetric type of staining was also observed in the hepatoma membranes, which contained some 50% more sialic acid than did the liver membranes. In addition, the hepatoma membranes showed an intense and uniform staining by CIH of short segments of both membrane leaflets; the latter type of staining was but little impaired by neuraminidase pre-treatment. None of the junctional complexes of the liver membranes was stained by CIH. Tight junctions were very rarely observed in the hepatoma membrane preparations, and the desmosomes and intermediate junctions of these membranes not infrequently exhibited a loosened appearance exposing CIH-reactive neuraminidase-sensitive sialic acid at their opposite plates. This aspect could be induced in the desmosomes and intermediate junctions, but not in the tight junctions, by pre-treatment of the liver membranes with the chelating agent ethylenediaminetetra-acetate.

scholarly journals GTPase activities of synaptic vesicles in nerve ending of rat brain tissues.

1996 ◽  
Vol 71 ◽  
pp. 280
Author(s):  
Kiyofumi Ito ◽  
Eri Takai ◽  
Tatsuo Suzuki ◽  
Kazuo Nagai
Keyword(s):  
Rat Brain ◽  
Nerve Ending ◽  
Synaptic Vesicles ◽  
Brain Tissues

Isolation of Synaptic Vesicles from Nerve Endings of the Rat Brain

Nature ◽  
1962 ◽  
Vol 194 (4830) ◽  
pp. 794-795 ◽  
Author(s):  
E. DE ROBERTIS ◽  
GEORGINA RODRÍGUEZ DE LORES ARNAIZ ◽  
AMANDA PELLEGRINO DE IRALDI
Keyword(s):  
Rat Brain ◽  
Synaptic Vesicles ◽  
Nerve Endings

The Fine Structure of Glycocalyx in Human Spermatozoa. A High Resolution Cytochemical Study

1973 ◽  
Vol 31 ◽  
pp. 640-641
Author(s):  
P. Hernández-Jáuregui ◽  
A. Sosa ◽  
A. González Angulo
Keyword(s):  
Negative Charge ◽  
Iron Hydroxide ◽  
Human Spermatozoa ◽  
Surface Coat ◽  
Cell Surfaces ◽  
Colloidal Iron ◽  
Cytochemical Study ◽  
Specific Permeability ◽  
Extracellular Glycoprotein ◽  
Mammalian Spermatozoa

Glycocalyx is the name given by Bennett to the extracellular glycoprotein coat present in some cell surfaces. It appears to play an important role in cell properties such as antigenicity, cell adhesivity, specific permeability, and ATP ase activity. In the sperm this coat can be directly related to such important phenomena as capacitation and fertilization. The presence of glycocalyx in invertebrate spermatozoa has already been demonstrated. Recently Yanagimachi et al. has determined the negative charges on sperm surfaces of mammalian spermatozoa including man, using colloidal iron hydroxide. No mention was made however of the outer surface coat as composed of substances other than those confering a negative charge. The purpose of this work was therefore to determine the presence of a glycocalyx in human spermatozoa using alcian blue and lanthanum staining.

scholarly journals Characterization of a H+-ATPase in rat brain synaptic vesicles

1989 ◽  
Vol 264 (14) ◽  
pp. 8281-8288 ◽  
Author(s):  
S Cidon ◽  
T S Sihra
Keyword(s):  
Rat Brain ◽  
Synaptic Vesicles

Uptake of L-[3H] glutamic acid by crude and purified synaptic vesicles from rat brain

1982 ◽  
Vol 108 (3) ◽  
pp. 1221-1227 ◽  
Author(s):  
Jeffrey K. Disbrow ◽  
Mitchell J. Gershten ◽  
James A. Ruth
Keyword(s):  
Glutamic Acid ◽  
Rat Brain ◽  
Synaptic Vesicles

scholarly journals THE ORGANIZATION AND INNERVATION OF THE LUMINESCENT ORGAN IN A FIREFLY, PHOTURIS PENNSYLVANICA (COLEOPTERA)

1963 ◽  
Vol 16 (2) ◽  
pp. 323-359 ◽  
Author(s):  
David S. Smith
Keyword(s):  
Electron Microscope ◽  
Synaptic Vesicles ◽  
Light Emission ◽  
Nerve Endings ◽  
Tracheal Epithelium ◽  
Nerve Terminals ◽  
Tracheal System ◽  
Physiological Evidence ◽  
Effector System

The organization of the luminescent organ of an adult firefly has been studied with the electron microscope, and particular attention has been given to the disposition of nerve terminals within the organ. The cytological structure of the cells of the tracheal system, the peripheral and terminal axons, the photocytes and the cells of the dorsal ("reflecting") layer is described. Previous observations on the peripheral course of nerve branches alongside the tracheal trunks at the level of the dorsal layer and photocyte epithelium have been confirmed, and specialised nerve endings containing axoplasmic components structurally identical with "synaptic vesicles" and "neurosecretory droplets" have been identified, not in association with the surface of the photocytes, but lying between the apposed surfaces of two components of the tracheal epithelium: the tracheal end-cell and the tracheolar cell. These cytological findings are discussed in terms of available biochemical and physiological evidence concerning the mechanism of light emission in the firefly, especially with respect to the possible role of chemical "transmitter" action in triggering a response in a luminescent effector system.

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