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Biomedical subjects

M M Dewey

Publications and source records attributed to M M Dewey.

At least 37 records · Page 2Linked to original sources

The septum of the lateral axon of the earthworm: a thin section and freeze-fracture study.

Septa occur between the axonal segments in the lateral giant septate axon of the nerve cord of the earthworm. This septum is demonstrated here to be permeable to fluorescein and to exhibit a negligible time delay for impulse transmission. Periodic anastomoses between the two lateral axons of the nerve cord are revealed by fluorescein. The permeability of the septum is correlated with the demonstration that nexuses occur along the septum. In thin sections, the nexuses may appear as long septilaminar or pentalaminar membrane appositions, but most frequently appear as a series of short or punctate membrane appositions. In freeze-fracture replicas, the nexuses appear as particles 10-12 nm in diameter on the PF face and as pits on the EF face. The particles and pits are arranged in plaques, in anastomosing strands, or most frequently in small plaques with strands of particles or pits emerging from the periphery. In addition to the nexuses, a junction characterized by the presence of 31 nm diameter hemispherical densities on the cytoplasmic surfaces of the septal membranes is revealed in thin sections. The densities are paired on the adjacent septal membranes, and most frequently are shown by optical diffraction to be arranged on the membrane surfaces in hexagonal or rhomboidal lattices with a centre-to-centre spacing of 34.8 nm. In freeze-fracture replicas, an array of particles and pits with a similar lattice symmetry and spacing to the arrays of hemispherical densities is demonstrated.

Action Potentials↗

Antibody binding by native and denatured myosin and paramyosin.

By the techniques of immunodiffusion and fluorescent immunohistochemistry we show that antibodies to both the native and the SDS-denatured forms of the proteins, paramyosin and myosin, react with the native, SDS-denatured and glutaraldehyde-fixed forms of their respective antigens. Anti-denatured myosin also binds to both native and denatured forms of the proteolytic subfragments of myosin: globular subfragment-1 and alpha-helical LMM. Anti-native myosin, on the other hand, while able to bind to both native and denatured LMM or rod and to native and glutaraldehyde-fixed S-1, does not bind to SDS-denatured S-1.

Animals↗

Localization of rhodopsin in isolated, osmotically intact rod outer segment discs.

Localization of rhodopsin and its position in the membrane has been the subject of numerous studies. Most recently, immunocytochemical techniques have been employed to localize the opsin component of the molecule in in situ rod outer segments. Due to the problems inherent in localization procedures (penetration and mechanical interference) we have utilized isolated, osmotically intact rod outer segment discs in this study. Specific antibodies to chromatographically pure rhodopsin were prepared and enzymatically digested to their Fab components. The univalent Fab antibodies were conjugated to horseradish peroxidase and used to label the isolated rod outer segment discs. Discs treated with anti-opsin conjugate stained uniformly and heavily on their interdisc surfaces. Reaction product was also present on the intradisc surface in a thinner but still uniformly distributed layer. Controls treated with preimmune Fab - horseradish peroxidase conjugate showed no deposition of reaction product.

Animals↗

Nexal membrane permeability to anions.

The permeability of the septa of the earthworm in the median axon has been calculated for the anions fluorescein and its halogen derivatives. The values ranged from 5.4 X 10(-5) to 4 X 10(-6) cm/s. Previously, the septa had been shown to contain nexuses. By using freeze-fracture material, the surface area of nexus on the septal membranes was determined to be 4.5%, very similar to the percentage of nexus in the intercalated disk of mammalian myocardium. Plasma membrane permeability to these dyes was also calculated and shown to be much less than that of the septal membranes. In addition, an estimate of cytoplasmic binding for each dye was made, and most dyes showed little or no binding with the exception of aminofluorescein.

Animals↗

Glial cells in the earthworm ventral nerve cord make an A-type nexus.

Nexuses have been demonstrated between glial cell processes in the earthworm ventral nerve cord. The nexuses have been demonstrated by freeze-cleavage to be A-type neuxuses, thus demonstrating for the first time the type of nexus present in the Annelida. If the nexal cleavage pattern has physiological significance then the results suggest that physiologically the nexuses of the earthworm may be similar to those of vertebrates.

Animals↗

Nexus of frog ventricle.

Here were demonstrate in Rana pipiens ventricle a nexus with very unusual morphology. This tissue has been reported previously to lack nexuses. The nexus appears in thin sections of ventricle, fixed in aldehyde and OsO4 or permanganate as a series of punctate membrane appositions regularly alternating with regions of membrane separation. The junctional width at membrane appositions, as determined by microdensitometry and optical measurements, is 15-17 nm, and the width of the electron-translucent region between the junctional membranes is 1.8 nm. These values correspond closely to similar measurements of the more typical nexues in frog liver. Along the nexus the mean distance between punctate appositions is 74.5 nm. Freeze-cleave replicas of the nexuses between myocardial cells show particles 10.4 nm in diameter arranged in arrays of up to nine linked circles or partial circles on the PF-face and similar arrays of pits of shallow grooves on the EF-face. The mean diameter of the circles on both membrane fracture faces is 76.7 nm comparsion of the thin-sectioned and freeze-cleaved nexuses demonstrates an excellent correspondence between the spacing of membrane appositions along the junction and the diameters of the freeze-cleaved circles of particles and pits or grooves.

Animals↗

Changes in thick filament length in Limulus striated muscle.

Here we describe the change in thick filament length in striated muscle of Limulus, the horseshoe crab. Long thick filaments (4.0 microns) are isolated from living, unstimulated Limulus striated muscle while those isolated from either electrically or K+-stimulated fibers are significantly shorter (3.1 microns) (P less than 0.001). Filaments isolated from muscle glycerinated at long sarcomere lengths are long (4.4 microns) while those isolated from muscle glycerinated at short sarcomere lengths are short (2.9 microns) and the difference is significant (P less than 0.001). Thin filaments are 2.4 microns in length. The shortening of thick filaments is related to the wide range of sarcomere lengths exhibited by Limulus telson striated muscle.

Actin Cytoskeleton↗

Paramyosin in invertebrate muscles. I. Identification and localization.

By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunodiffusion, we identified paramyosin in two smooth invertebrate "catch" muscles (Mytilus anterior byssus retractor and Mercenaria opaque adductor) and five invertebrate striated muscles (Limulus telson levator, Homarus claw muscle, Balanus scutal depressor, Lethocerus air tube retractor, and Aequipecten striated adductor). We show that (a) the paramyosins in all of these muscles have the same chain weights and (b) they are immunologically similar. We stained all of these muscles with specific antibody to Limulus paramyosin using the indirect fluorescent antibody technique. Paramyosin was localized to the A bands of the glycerinated striated muscles, and diffus fluorescence was seen throughout the glycerinated fibers of the smooth catch muscles. The presence of paramyosin in Homarus claw muscle, Balanus scutal depressor, and Lethocerus air tube retractor is shown here for the first time. Of the muscles in this study, Limulus telson levator is the only one for which the antiparamyosin staining pattern has been previously reported.

Animals↗

Paramyosin in invertebrate muscles. II. Content in relation to structure and function.

By quantitative sodium dodecyl sulfate-polyacrylamide gel electrophoresis, paramyosin:myosin heavy chain molecular ratios were calculated for three molluscan muscles:Aequipecten striated adductor, Mercenaria opaque adductor, and Mytilus anterior byssus retractor; and four arthropodan muscles:Limulus telson, Homarus slow claw. Balanus scutal depressor, and Lethocerus air tube retractor. These ratios correlate positively with both thick filament dimensions and maximum active tension development in these tissues. The role of paramyosin in these muscles is discussed with respect to the following characteristics: force development, "catch," and extreme reversible changes in length.

Animals↗

Isolation and characterization of plasma membrane fractions from garfish Lepisosteus osseus olfactory nerve.

Garfish Lepisosteus osseus olfactory nerve, because of its large size and the unusually high concentration of axonal membrane, is an excellent source of axonal membrane. A procedure is described for the isolation of two types of plasma membranes from the nerve which are obtained in yields of about 20 mg (fraction I) and 1.5 mg (fraction II) per g of wet nerve. Both membrane fractions consist mostly of rounded membrane vesicles, with a unit membrane thickness of approximately 7.5 nm. The two membrane fractions are different in their lipid to protein ratios, Na-K ATPase activities, polypeptide patterns on sodium dodecyl sulfate (SDS) gel electrophoresis, and fatty acid compositions. They have similar phospholipid composition. On the basis of the relative concentration of axonal and Schwann cell plasma membranes in the nerve, the Na-K ATPase activities of the two membrane fractions and a comparison of the properties of the membrane fractions to those of squid and lobster nerve membrane preparations, fraction I seems to be the axonal membrane and fraction II the Schwann cell plasma membrane. Fraction I has a low protein to lipid ratio. Its polypeptide pattern on SDS gel appears to be much more complex as compared to that of fraction II membrane.

Adenosine Triphosphatases↗

Structure of Limulus striated muscle. The contractile apparatus at various sarcomere lengths.

The musculature of the telson of Limulus polyphemus L. consists of three dorsal muscles: the medial and lateral telson levators and the telson abductor, and one large ventral muscle; the telson depressor, which has three major divisions: the dorsal, medioventral, and lateroventral heads. The telson muscles are composed of one type of striated muscle fiber, which has irregularly shaped myofibrils. The sarcomeres are long, with discrete A and I and discontinuous Z bands. M lines are not present. H zones can be identified easily, only in thick (1.0 microm) longitudinal sections or thin cross sections. In lengthened fibers, the Z bands are irregular and the A bands appear very long due to misalignment of constituent thick filaments. As the sarcomeres shorten, the Z lines straighten somewhat and the thick filaments become more aligned within the A band, leading to apparent decrease in A band length. Further A band shortening, seen at sarcomere lengths below 7.4 microm may be a function of conformational changes of the thick filaments, possibly brought about by alterations in the ordering of their paramyosin cores.

Animals↗

The structure of Mytilus smooth muscle and the electrical constants of the resting muscle.

The individual muscle fibers of the anterior byssus retractor muscle (ABRM) of Mytilus edulis L. are uninucleate, 1.2-1.8 mm in length, 5 microm in diameter, and organized into bundles 100-200 microm in diameter, surrounded by connective tissue. Some bundles run the length of the whole muscle. Adjacent muscle cell membranes are interconnected by nexuses at frequent intervals. Specialized attachments exist between muscle fibers and connective tissue. Electrical constants of the resting muscle membrane were measured with intracellular recording electrodes and both extracellular and intracellular current-passing electrodes. With an intracellular current-passing electrode, the time constant tau, was 4.3 +/- 1.5 ms. With current delivered via an extracellular electrode tau was 68.3 +/- 15 ms. The space constant, lambda, was 1.8 mm +/- 0.4. The membrane input resistance, R(eff), ranged from 23 to 51 MOmega. The observations that values of tau depend on the method of passing current, and that the value of lambda is large relative to fiber length and diameter are considered evidence that the individual muscle fibers are electrically interconnected within bundles in a three-dimensional network. Estimations are made of the membrane resistance, R(m), to compare the values to fast and slow striated muscle fibers and mammalian smooth muscles. The implications of this study in reinterpreting previous mechanical and electrical studies are discussed.

Animals↗

Immunohistochemical localization of contractile proteins in limulus striated muscle.

Limulus paramyosin and myosin were localized in the A bands of glycerinated Limulus striated muscle by the indirect horseradish peroxidase-labeled antibody and direct and indirect fluorescent antibody techniques. Localization of each protein in the A band varied with sarcomere length. Antiparamyosin was bound at the lateral margins of the A bands in long ( approximately 10.0 micro) and intermediate ( approximately 7.0 micro) length sarcomeres, and also in a thin line in the central A bands of sarcomeres, 7.0- approximately 6.0 micro. Antiparamyosin stained the entire A bands of short sarcomeres (<6.0). Conversely, antimyosin stained the entire A bands of long sarcomeres, showed decreased intensity of central A band staining except for a thin medial line in intermediate length sarcomeres, and was bound only in the lateral A bands of short sarcomeres. These results are consistent with a model in which paramyosin comprises the core of the thick filament and myosin forms a cortex. Differential staining observed using antiparamyosin and antimyosin at various sarcomere lengths and changes in A band lengths reflect the extent of thick-thin filament interaction and conformational change in the thick filament during sarcomeric shortening.

Actomyosin↗

Electrical transmission at the nexus between smooth muscle cells.

The hypothesis that nexuses between cells are responsible for the core conductor properties of tissues was tested using smooth muscle preparations from the taenia coli of guinea pigs. Action potentials recorded from small diameter preparations across a sucrose gap change from monophasic to diphasic when a shunt resistor is connected across the gap. This indicates that transmission between smooth muscle cells is electrical, because the resistor only allows current to flow. Nexal fusion of cell membranes occurs especially where one cell sends a large bulbous projection into a neighbor. Hypertonic solutions rupture the nexuses between smooth muscle cells. Hypertonicity also increases the resistance of a bundle across the sucrose gap and blocks propagation of action potentials. Thus the structural and functional changes in smooth muscle due to hypertonicity correlate with the hypothesis.

Action Potentials↗