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Biomedical subjects

J Rosenbluth

Publications and source records attributed to J Rosenbluth.

At least 73 records · Page 4Linked to original sources

Synaptic membrane structure in Torpedo electric organ.

The innervated and noninnervated membranes of Torpedo electrocytes have been examined by electron microscopy of thin-sectioned and freeze-fractured specimens. The ventral innervated membrane is approximately 120 A thick and is characterized by an unusually broad outer dense lamina (approximately 60 A) in which a granular substructure can be resolved. The granules are approximately 70 A in diameter and are spaced irregularly. The same membrane specialization was noted in a previous study of amphibian myoneural junctions, and it was proposed then that the granular elements represent ACh receptor molecules. The morphologically equivalent structures presumably have the same significance in the Torpedo electric organ. However, in this case the specialized membrane covers the entire innervated surface, leading to the conclusion that high concentrations of receptors occur normally in extrajunctional as well as post-junctional regions of the innervated membrane of the electrocyte. In replicas of freeze-fractured specimens, the A face of this membrane is covered with large particles having the same distribution and approximate concentration as the granules visible in thin sections, indicating that the granules visible at the outer surface of the membrane extend at least into the hydrophobic middle layer of the membrane. The cytoplasmic surface of this membrane has an amorphous coating into which 'decorated' cytoplasmic filaments insert. Synaptic vesicle and axon terminal membranes also contain granules visible in thin sections but with a much sparser distribution. These probably correspond to the intramembranous particles seen in freeze-fractured specimens. Vesicles are occasionally attached to the axolemma by thin linear strands.

Animals↗

Substructure of amphibian motor end plate. Evidence for a granular component projecting from the outer surface of the receptive membrane.

End-plate membrane has been examined at amphibian myoneural junctions by means of transmission electron microscopy of thin tissue sections. The postjunctional membrane exhibits morphologically specialized dense, convex patches which are located superficially facing the axon terminal but do not extend into the depths of the junctional folds. In the specialized regions the plasma membrane is approximately 120 A thick and trilaminar. The outer dense lamina is thickened by the presence in it of granular elements approximately 60-120 A in diameter which are spaced semiregularly at approximately 100-150-A intervals and which border the junctional cleft directly. In these regions the concentration of the granules is of the order of approximately 10(4)/microm(2), which is in the same range as the estimated concentration of receptor sites at other vertebrate cholinergic junctions. Filamentous projections can sometimes be seen extending from the granules to the overlying basement membrane, and in oblique views a reticular pattern may appear both in these patches and in the basement membrane. The cytoplasmic surface of the specialized membrane is covered with an amorphous and filamentous dense material whose distribution coincides with that of the granules visible in the outer layer and which may be connected to them across the membrane. In unosmicated specimens stained with permanganate and uranyl acetate the specialized regions exhibit the same morphological features but stand out sharply in contrast to adjacent areas of unspecialized membrane which appear only faintly. Such preparations are particularly useful in assessing the extent of the specialized membrane. It is proposed that the granules visible at the outer surface of the end-plate membrane represent acetylcholine receptors and that in amphibians, as in annelids, the receptors at myoneural junctions are concentrated into patches which occupy less than the total postjunctional membrane surface area.

Animals↗

Membrane specialization at an insect myoneural junction.

Myoneural junctions were examined in the asynchronous basalar flight muscle of the beetle Pachnoda ephippiata. The outer surface of the postjunctional membrane exhibits an array of prominent projections spaced at approximately 200 A intervals which arise directly from the outer dense lamina of the plasma membrane and extend part way across the junctional cleft. The projections follow irregularities in the contour of the postjunctional membrane precisely and they end abruptly near the edge of the junctional region. No separation can be resolved between the projections and the underlying trilaminar plasma membrane after a variety of preparative methods, and the projections therefore appear to be a component part of the membrane. This specialization, which is distinctly different from that at desmosomes and hemidesmosomes, occurs nowhere else on the surface of the muscle and is interpreted as a mosaic of specialized membrane subunits which probably include the receptor sites for the transmitter.

Animals↗

Myoneural junctions of two ultrastructurally distinct types in earthworm body wall muscle.

THE LONGITUDINAL MUSCLE OF THE EARTHWORM BODY WALL IS INNERVATED BY NERVE BUNDLES CONTAINING AXONS OF TWO TYPES WHICH FORM TWO CORRESPONDING TYPES OF MYONEURAL JUNCTION WITH THE MUSCLE FIBERS TYPE I JUNCTIONS RESEMBLE CHOLINERGIC NEUROMUSCULAR JUNCTIONS OF VERTEBRATE SKELETAL MUSCLE AND ARE CHARACTERIZED BY THREE FEATURES: (a) The nerve terminals contain large numbers of spherical, clear, approximately 500 A vesicles plus a small number of larger dense-cored vesicles (b) The junctional gap is relatively wide ( approximately 900 A), and it contains a basement membrane-like material, (c) The postjunctional membrane, although not folded, displays prominent specializations on both its external and internal surfaces The cytoplasmic surface is covered by a dense matrix approximately 200 A thick which appears to be the site of insertion of fine obliquely oriented cytoplasmic filaments The external surface exhibits rows of projections approximately 200 A long whose bases consist of hexagonally arrayed granules seated in the outer dense layer of the plasma membrane The concentration of these hexagonally disposed elements corresponds to the estimated concentration of both receptor sites and acetylcholinesterase sites at cholinergic junctions elsewhere. Type II junctions resemble the adrenergic junctions in vertebrate smooth muscle and exhibit the following structural characteristics: (a) The nerve fibers contain predominantly dense-cored vesicles approximately 1000 A in diameter (b) The junctional gap is relatively narrow ( approximately 150 A) and contains no basement membrane-like material, (c) Postjunctional membrane specialization is minimal. It is proposed that the structural differences between the two types of myoneural junction reflect differences in the respective transmitters and corresponding differences in the mechanisms of transmitter action and/or inactivation.

Animals↗

Myosin-like aggregates in trypsin-treated smooth muscle cells.

Segments of the lower small intestine of the toad Bufo marinus were excised and soaked for approximately 2 hr in Ringer's solution (pH 7.4 or 7.8) containing crystalline trypsin and then fixed for electron microscopy at approximately the same pH. Thin sections of the tunica muscularis of these specimens show smooth muscle cells ranging in appearance from severely damaged at one extreme to apparently unaffected at the other. Among these are cells at intermediate stages, including some which exhibit large and conspicuous populations of thick filaments closely resembling artificially prepared aggregates of smooth muscle myosin. The thick filaments have the form of tactoids approximately 250-300 A in diameter in their middle regions and are approximately 0.5-1.0 micro in length. In some preparations they also display an axial periodicity approximating 143 A. They are usually randomly oriented and segregated from the thin filaments, which tend to form closely packed, virtually crystalline bundles at the periphery of these cells. "Dense bodies" are absent from cells showing these changes. The simplest interpretation of these data is that smooth muscle myosin normally exists among the actin filaments in a relatively disaggregated state and that trypsin induces aggregation by altering the conformation of the myosin molecule. Alternatively, trypsin may act indirectly through an effect on some other smooth muscle protein which normally forms a stable complex with relatively disaggregated myosin.

Acid-Base Equilibrium↗

Characterization of an unusual catecholamine-containing cell type in the toad hypothalamus. A correlated ultrastructural and fluorescence histochemical study.

A nucleus of catecholamine-containing cells bordering the preoptic recess of the toad hypothalamus has been studied by both fluorescence histochemical and electron microscopic methods. The perikarya of these cells form one to three rows immediately subjacent to the ependyma. They send brightly fluorescent apical processes between the ependymal cells to the ventricular surface, and also give rise to long basal processes, the proximal portions of which are also fluorescent. These cells contain two distinctive constitutents: juxtanuclear bundles of tightly packed filaments, the members of which are separated from one another by only approximately 100 A, and large numbers of dense-cored vesicles (400-2200 A in diameter), which appear to arise from an agranular tubular reticulum distinct from the Golgi apparatus. Axons containing either clear vesicles alone or clear and dense-cored vesicles form synapses on the subependymal cells, but no evidence has been found that the subependymal cells themselves form presynaptic contacts, or that axons originate from them. The cytological characteristics of these catecholamine-containing cells, plus the fact that they border directly on the cerebrospinal fluid, suggest that they may be more closely related to peripheral chromaffin cells than to the other cell types intrinsic to the central nervous system, and the name "encephalo-chromaffin cells" is therefore proposed for them. The possible functions of such cells in the central nervous system are discussed.

Animals↗

Sarcoplasmic reticulum of an unusually fast-acting crustacean muscle.

The fast-acting, synchronous "remotor" muscle of the lobster second antenna was examined by light and electron microscopy and was found to have a more profuse sarcoplasmic reticulum (SR) than any other muscle known. Myofibrils are widely separated from one another and occupy only about one-fourth of the volume of the muscle; most of the remaining volume is taken up by the SR, which resembles the smooth-surfaced reticulum of steroid-secreting cells. Dense granules (0.03-0.1 micro in diameter) are scattered through the reticulum. T-tubules penetrate into the fibers and form dyads along the A bands of myofibrils; however, ferritin-labeling experiments show that the volume of the T-system is very small compared with that of the SR. Myofibrils are approximately 0.5 micro x 1.0 micro in cross section and consist of thick filaments, which appear tubular except at the M region, and thin filaments, which are situated midway between neighboring thick filaments. The ratio of thin to thick filaments is 3:1. The extreme development of the SR in this muscle is discussed in relation to the exceedingly short duration of the contraction-relaxation cycle.

Animals↗

Ultrastructure of dyads in muscle fibers of Ascaris lumbricoides.

The dyads of Ascaris body muscle cells consist of flattened intracellular cisternae applied to the sarcolemma at the cell surface and along the length of T-tubules. In specimens prepared by conventional methods (glutaraldehyde fixation, osmium tetroxide postfixation, double staining of sections with uranyl acetate and lead hydroxide), both the sarcolemma and the limiting membrane of the cisterna exhibit unit membrane structure and the space between them is occupied by a layer of peg-shaped densities which is referred to as the subsarcolemmal lamina. The lumen of the cisterna contains a serrated layer of dense material referred to as the intracisternal lamina. In specimens fixed in glutaraldehyde, dehydrated, and then postfixed in phosphotungstic acid, with no exposure to osmium tetroxide or heavy metal stains, the membranous components of the dyads appear only as negative images, but the subsarcolemmal and intracisternal laminae still appear dense. Except for the lack of density in membranes and in glycogen deposits, the picture produced by the latter method is very much like that of tissue prepared by conventional methods.

Animals↗

Obliquely striated muscle. 3. Contraction mechanism of Ascaris body muscle.

Segments of the obliquely striated body muscle of Ascaris were fixed at minimum body length after treatment with acetylcholine and at maximum body length after treatment with piperazine citrate and then studied by light and electron microscopy. Evidence was found for two mechanisms of length change: sliding of thin filaments with respect to thick filaments such as occurs in cross-striated muscle, and shearing of thick filaments with respect to each other such that the degree of their stagger increases with extension and decreases with shortening. The shearing mechanism could account for great extensibility in this muscle and in nonstriated muscles in general and could underlie other manifestations of "plasticity" as well. In addition, it is suggested that the contractile apparatus is attached to the endomysium in such a way that the sarcomeres can act either in series, as in cross-striated muscle, or individually. Since the sarcomeres are virtually longitudinal in orientation and are almost coextensive with the muscle fiber, it would, therefore, be possible for a single sarcomere contracting independently to develop tension effectively between widely separated points on the fiber surface, thus permitting very efficient maintenance of isometric tension.

Acetylcholine↗