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Biomedical subjects

C R Green

Publications and source records attributed to C R Green.

At least 73 records · Page 4Linked to original sources
PubMed1987

Selective disruption of gap junctional communication interferes with a patterning process in hydra.

The cells that make up the body column of hydra are extensively joined by gap junctions, capable of mediating the rapid exchange of small hydrophilic molecules between the cytoplasms of neighboring cells. Both the rate of transfer of small molecules through the gap junctions and the rate of return of gap junction coupling after grafting experiments are sufficiently rapid to mediate events in the patterning of hydra tissue. Antibodies to the major rat liver gap junction protein (27,000 daltons) recognize a gap junction antigen in hydra and are effective in eliminating junctional communication between hydra cells. The antibodies perturb the head inhibition gradient in grafting operations, suggesting that cell-cell communication via gap junctions is important in this defined tissue patterning process.

Animals↗
PubMed1987

Topological analysis of the major protein in isolated intact rat liver gap junctions and gap junction-derived single membrane structures.

The topological organization of the major rat liver gap junction protein has been examined in intact gap junctions and gap junction-derived single membrane structures. Two methods, low pH and urea at alkaline pH, were used to "transform" or "split" double membrane gap junctions into single membrane structures. Low pH treatment "transforms" rat liver gap junctions into small single membrane vesicles which have an altered sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile after digestion with L-1-to-sylamido-2-phenylethylchloromethyl ketone-trypsin. Alkaline pH treatment in the presence of 8 M urea can split isolated rat liver gap junctions into single membrane sheets which have no detectable structural alteration or altered sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile after proteolytic digestion, suggesting that these single membrane sheets may be useful for topological studies of the gap junction protein. Proteolytic digestion studies have been used to localize the carboxyl terminus of the molecule on the cytoplasmic surface of the intact gap junction. However, the amino terminus does not appear to be accessible to proteases or to interaction with an antibody that is specific for the amino-terminal region of the molecule in intact or split gap junctions. Binding of antibodies, that block junctional channel conductance, can be eliminated by proteolytic digestion of intact gap junctions, suggesting that all antigenic sites for these antibodies are located on the cytoplasmic surface of the intact gap junction. In addition, calmodulin gel overlays indicate that at least two calmodulin binding sites exist on the cytoplasmic surface of the junctional protein. The information generated from these studies has been used to develop a low resolution two-dimensional model for the organization of the major rat liver gap junctional protein in the junctional membrane.

Amino Acid Sequence↗
PubMed1987

Quantitation of ischemia during total coronary occlusion with computer-assisted high resolution ST-segment monitoring: effect of collateral flow.

The effect of increasing grades of collateral flow to the distal target vessel on the clinical and electrocardiographic manifestations of ischemia was evaluated in 118 patients undergoing transluminal coronary angioplasty (TCA). A qualitative scoring system for collateralization was defined as follows: 0--no visible collaterals; 1--visualization of collateral vessels only with no filling of the distal TCA vessel; 2--partial filling of the distal TCA vessel, and; 3--complete filling of the distal TCA vessel. All patients underwent computerized ST-segment monitoring using 3-channel Holter recorders. Criteria for an ischemic ST-segment response was 1.0 mm ST-segment deviation from ST baseline measured 60 msec after the J-point. Episodes of chest pain during and after the procedure were noted. Patients with collateral filling (collateral scores 2 and 3) of the distal TCA vessel had a significantly decreased incidence of angina and diagnostic ischemic ST-segment changes (42% and 15% respectively) as compared to patients without collateral filling (collateral scores 0 and 1) (83% and 74%) (p less than .001). Fifty-eight percent of patients without collateral filling had ischemic ST-segment responses on every balloon inflation as opposed to only 9% with collateral filling (p less than .0001). After TCA, the incidence of angina and ischemic ST-segment changes was similar in both groups (20% and 17% without collateral filling vs. 9% and 9%). We conclude that: 1. increasing qualitative collateralization protects against the development of myocardial ischemia during TCA, but; 2. the similar incidence of late episodes of ischemia after successful TCA suggests that collateral flow may no longer be adequate.

Angioplasty, Balloon↗
PubMed1984

Connexon rearrangement in cardiac gap junctions: evidence for cytoskeletal control?

Using ultrarapid-freezing techniques and freeze-fracture electron microscopy, we report here a close association between cardiac gap junctions and specialized membrane domains containing regularly-spaced furrows. These specialized furrowed domains are observed only during periods of gap junction re-organisation (i.e., connexon redistribution) and may reflect the presence of underlying cytoskeletal elements controlling the position of connexons in the membrane.

Animals↗
PubMed1984

Gap junction connexon configuration in rapidly frozen myocardium and isolated intercalated disks.

By using two ultrarapid freezing techniques, we have captured the structure of rat and rabbit cardiac gap junctions in a condition closer to that existing in vivo than to that previously achieved. Our results, which include those from fully functional hearts frozen in situ in the living animal, show that the junctions characteristically consist of multiple small hexagonal arrays of connexons. In tissue frozen 10 min after animal death, however, unordered arrays are common. Examination of junction structure at intervals up to 40 min after death reveals a variety of configurations including dispersed and close-packed unordered arrays, and hexagonal arrays. By use of an isolated intercalated disk preparation, we show that the configuration of cardiac gap junctions in vitro cannot be altered by factors normally considered to induce functional uncoupling. These experiments demonstrate that, contrary to the conclusions of some earlier studies (Baldwin, K. M., 1979, J. Cell Biol., 82:66-75; Peracchia, C., and L. L. Peracchia, 1980, J. Cell Biol., 87:708-718), the arrangement of gap junction connexons, in cardiac tissue at least, cannot be used as a reliable guide to the functional state of the junctions.

Animals↗
PubMed1983

A simplified method for the rapid isolation of cardiac intercalated discs.

A simple and rapid method for the isolation of intercalated discs from a single rat heart is described. The outstanding features of this method are (i) a large quantity of starting material is not necessary, (ii) ultracentrifugation and separating gradient steps are not required, (iii) the disc fraction is relatively pure as verified by electron microscopy, and (iv) the starting tissue is rapidly homogenized into buffer, reducing possible damage to membrane structures after animal death. The fraction is rich in intercellular junctions, with gap junctions, fasciae adherentes and maculae adherentes all appearing well preserved. It should prove a useful starting base for further purification of these junctions and of sarcolemma from a specific portion of the cell surface membrane, namely the major area of structural interaction between adjacent cardiac cells.

Animals↗
PubMed1983

Isolation and characterization of invertebrate smooth septate junctions.

Using modifications of techniques used for the isolation of macula type intercellular junctions (gap junctions and desmosomes) the arthropod smooth septate junction has been isolated from insect midgut tissue. Midguts from cockroaches or mealworms were used and membrane fractions were obtained by sucrose gradient and ultracentrifugation techniques. Preparations with reasonable concentrations of septate junction were obtained and have been studied by thin-section, negative-stain and freeze-fracture electron microscopy. The junctions appeared to be well preserved, although there was evidence that the junction strands were able to slide within the plane of the membrane. Septa were seen to have a cross-striated appearance when viewed after negative staining but their exact structure remained difficult to determine. Polyacrylamide gel electrophoretic studies demonstrated the reproducibility of the isolation procedure and showed that septa may have a 47 000 molecular weight glycoprotein component. Gel electrophoresis also gave some indication of the intramembrane biochemistry of the smooth septate junction, with proteins of 31 000 and 32 000 molecular weight always occurring in the junction fractions. The junctions were, however, very sensitive to both mechanical and chemical treatments, the septa were destroyed by rough homogenization or by treatment with urea at a concentration as low as 1 M. Freeze-fracture of untreated, isolated junctions demonstrated no differences from junctions in intact tissue, while replicas of urea-treated material were more difficult to interpret as the component parts of the junctions became separated once the septa had been destroyed. Gap junctions were also obtained and resisted both mechanical and chemical treatment, which destroyed the septate junctions. Their major protein component appeared to have a molecular weight of 36 000. Attempts to isolate pleated septate junctions (from insects, molluscs and annelids) by the same techniques failed, implying a significant difference in the structures of the two types of septate junction.

Animals↗
PubMed1982

A new type of gap junction in the phylum Brachiopoda.

Gap junctions in the phylum Brachiopoda are shown to be of the P type as has been previously reported in invertebrates only in molluscs and possibly bryozoans. However, the mollusc gap junctions appear identical to vertebrae gap junctions in the fracture face on which particles remain, in particle spacing and in particle arrangement. The brachiopod gap junction, on the other hand, has general appearance traits which are more commonly seen in other invertebrate gap junction types. The freeze-fracture appearance of the brachiopod gap junction is described and discussed in relation to other types of gap junction which have been reported.

Animals↗
PubMed1981

A clarification of the two types of invertebrate pleated septate junction.

It is confirmed that there are two distinct variations septate junction. The first of these, the 'lower invertebrate pleated septate junction', is described fully using conventional thin section, lanthanum tracer and freeze-fracture techniques. The second type, the well-known pleated septate junction characteristic of the molluscs and arthropods, is renamed the 'mollusc-arthropod pleated septate junction', and is described briefly to allow easier comparison between the two variations. As both types have not been studied in a range of invertebrate phyla the results can be used as a basis for discussing their respective phylogenetic positions. The lower invertebrate pleated septate junction occurs in several groups in the minor phyla immediately above the Coelenterata and in the lower phyla of both the deuterostome and proterostome lineages. The mollusc-arthropod pleated septate junction is restricted to the Mollusca and Arthropoda as its name implies.

Animals↗
PubMed1981

A variation of the smooth septate junction in sea spiders (pycnogonida).

A new type of septate junction considered to be a variation of the arthropod smooth septate junction is described in pycnogonid (sea spider) endothermal tissue based on the use of conventional thin-section, lanthanum tracer and freeze-fracture techniques. This new type of septate junction is apparently unique to the Pycnogonida but closely resembles septate junctions previously described in the Merostomata and Collembola. This in conjunction with previous work suggests that the septa of smooth septate junctions may not be as 'smooth' as generally thought and probably have a complex substructure.

Animals↗
PubMed1980

Two new septate junctions in the phylum Coelenterata.

Freeze-fracture of fixed and unfixed tissue, lanthanum tracer and conventional thin-section studies have revealed 2 new types of septate junction in the class Anthozoa, phylum Coelenterata. These new junctions have the 15-18-nm intercellular spacing of all other described septate junctions and are found around the apical circumference of cells lining a lumen or outside edge. However, in freeze-fracture replicas and tangential views of lanthanum-impregnated tissue, they are seen to be quite different from other known septate junction types. One of the new junctions is found in endothelial tissue such as that lining the gut or the inside of the tentacles. In tangential view it is seen to consist of relatively short, straight, double septa, again with lateral projections. In feeeze-fracture of unfixed tissue, the junction consists of double rows of particles on the P face, the particles of one row being rounded, those of the other being elongated at right angles to the line of the septum. This dichotomy in particle size is unexpected, as the 2 halves of the septa as seen in tangential view are symmetrical. In freeze-fracture of fixed material the particle arrays remain on the P face and appear similar to those of unfixed material, but never as clear. In fixed tissue, some distortion had occurred and in extreme cases septa appear as a single broad jumbled row of particles. In this double septa junction, the rows of particles seen in freeze-fracture are occasionally seen to anastomose with a septum dividing into 2 and a third row of particles aligning with the 2 new septa to form their double particle rows. In both fixed and unfixed tissues, the E face of the junction consists of wide, shallow grooves. The second of the new junctions occurs in epithelial tissue, such as around the outer edge of sea-anemone tentacles, and consists of long wavy septa with lateral projections. In views where these projections appear longest, they arise predominantly from one side of the septa. In freeze-fracture of both fixed and unfixed tissue, this junction appears as rows of closely spaced particles on the P face. Occasionally rows of particles are seen on the E face, but usually this face is characterized by shallow grooves. In some aspects these 2 new junctions have features in common with the Hydra type junction also found in the Coelenterata. In all 3 types septa are relatively straight, rather than pleated, and there are lateral projections on the septa.

Animals↗
PubMed1977

The morphology of cilia in sponge larvae.

Sponge larvae posses cillia with unusual terminal expansions which are curled or biconcave in shape externally, as seen by scanning electron microscopy. Thin sections, passing through the point where the ciliary shaft enters the expanded area reveal the ciliary axoneme to be surrounded by many membranous folds, some of which are vesicular. The 'club footed' cilia occur in disparate groups of Demospongiae and most frequently, all larval cilia are of this type. There is no immediately obvious correlation between the type of movement displayed by the larvae and the occurrence of terminally expanded cilia.

Animals↗
PubMed1977

An experimental model of analgesic-induced renal damage--some effects of p-aminophenol on rat kidney mitochondria.

1. p-Aminophenol, a known nephrotoxin, has been studied as a model for phenacetin-induced renal damage. 2. Respiration, oxidative phosphorylation and ATPase activity were inhibited in mitochondria isolated from the kidneys of treated rats; this could not be reversed by the addition of exogenous loosely bound cofactors and bovine serum albumin to the assay medium. 3. After treatment the mitochondrial levels of sodium and calcium were increased, potassium decreased and magnesium unaltered. 4. Mitochondria isolated from treated rats showed ultrastructural damage. 5. The results are interpreted to indicate that renal tubular cell mitochondrial injury is important in triggering cortical analgesic renal damage.

Adenosine Triphosphatases↗