[Extracellular space and blood volume of DS-carcinosarcoma on the chorioallantoic membrane of the chick embryo].
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A characteristic event during reperfusion after cerebral ischemia in vivo, and reoxygenation after anoxia in vitro, is hyperoxidation of the electron carriers of the mitochondrial respiratory chain. Current studies have tested the hypothesis that there is a relation among calcium molecules derived from extracellular sources, mitochondrial hyperoxidation, and electrical recovery after anoxia in hippocampal slices. Rat hippocampal slices were superfused with artificial cerebrospinal fluids (ACSF) containing calcium chloride (CaCl2) in concentrations of: 0.5, 1, 2, and 4 mmol/L. Slices were made anoxic and then allowed to recover for 60 minutes. Reduction-oxidation shifts of NADH were measured by rapid-scanning spectrofluorometry. Synaptic activity was indicated by population spike amplitudes in the CA1 pyramidal cell subfield of the hippocampus in response to stimulation of the Schaffer collaterals. Low calcium ACSF concentrations ameliorated NADH hyperoxidation and improved synaptic transmission recovery after anoxia. High calcium ACSF concentrations had opposite effects. These data suggest a link between mitochondrial hyperoxidation and electrical recovery after postanoxia reoxygenation and support the hypothesis that cytosolic calcium overload promotes mitochondrial hyperoxidation and limits electrical recovery.
1. The spaces occupied by isotopically labelled inulin, polyethylene glycol, mol. wt. 4000 (PEG 4000), polyethylene glycol, mol. wt. 1000 (PEG 1000) and sucrose in metabolizing mammalian kidney and liver slices and in toad bladder epithelial cell preparations incubated in vitro have been examined.2. In slices of mammalian tissue, and in homogenized liver, it proved impossible to extract inulin completely from tissue which had been dried. However, inulin was recovered as completely from both dried and undried toad bladder epithelial cells scraped from hemibladders incubated in vitro.3. PEG 4000 occupied a space in all preparations similar to that from which inulin was extracted in dried tissue.4. PEG 1000 and sucrose entered cellular water in mammalian slices, but PEG 1000 occupied a similar space to inulin in toad bladder epithelial cell preparations.6. It is concluded that inulin enters cellular water in mammalian slices from which after drying of the slices it cannot be extracted. It thus rather fortuitously provides a measure of extracellular water under these conditions. In preparations of toad bladder epithelial cells inulin seems to be a satisfactory extracellular marker. PEG 4000, which did not appear to enter cellular water also allows a reasonable estimate of extracellular water. PEG 1000 is a suitable extracellular marker for toad bladder epithelial cell preparations but not for mammalian slices. Sucrose entered cellular water in both slices and toad bladder epithelial cells and is not a satisfactory extracellular marker in these tissues.
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By means of in vivo microdialysis combined with HPLC/EC analysis it was shown that presentation to a rat of an inedible object (a piece of rubber) or an aversive object (food of bitter taste) instead of expected food caused a marked increase in extracellular glutamate level in n. accumbens. In rats not expecting food reinforcement, extracellular glutamate remained unchanged during presentation of these objects. Our findings suggest that dissociation between the expected biological value of a presented object and its real significance may be an important determinant for glutamate release control in n. accumbens during food behavior.
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1. Uptake of a number of isotopically labelled extracellular tracers by rat liver in vivo was studied.2. A residual intravascular space of 3.0-3.5% following exsanguination was measured by use of [(51)Cr]red cells. This space was significantly greater than that of skeletal muscle and was also greater in nephrectomized as compared to intact rats.3. [(131)I]albumin and polyvinylpyrolidine (PVP) were found to distribute initially in a space of 6-7% and this was followed by a delayed rise that was greater for PVP. The extravascular portion of this space is termed the large-molecule accessible space and is believed to correspond to the histological space of Disse.4. [(14)C]sucrose and inulin initially distribute in a space of 10-12% followed by a continued late increase. This initial space is termed the small molecule accessible space and is thought to include most of interstitial space.5. (36)Cl(-) and stable Na(+) and Cl(-) spaces were constant at 19-21% and can be used to put an upper limit on extracellular space.6. Operationally, the 1 hr inulin space in the nephrectomized rat was felt to be the best indicator of extracellular space.7. Total electrolytes were measured and intracellular values of 22, 165 and 22 m-mole/l. cell water for Na(+), K(+) and Cl(-), respectively, were calculated.8. By use of [(14)C]DMO, an intracellular pH of 7.25 and an intracellular HCO(3) (-) of 13 m-mole/l. cell water were calculated.
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Physical forces elicit biochemical signalling in a diverse array of cells, tissues and organisms, helping to govern fundamental biological processes. Several hypotheses have been advanced that link physical forces to intracellular signalling pathways, but in many cases the molecular mechanisms of mechanotransduction remain elusive. Here we find that compressive stress shrinks the lateral intercellular space surrounding epithelial cells, and triggers cellular signalling via autocrine binding of epidermal growth factor family ligands to the epidermal growth factor receptor. Mathematical analysis predicts that constant rate shedding of autocrine ligands into a collapsing lateral intercellular space leads to increased local ligand concentrations that are sufficient to account for the observed receptor signalling; direct experimental comparison of signalling stimulated by compressive stress versus exogenous soluble ligand supports this prediction. These findings establish a mechanism by which mechanotransduction arises from an autocrine ligand-receptor circuit operating in a dynamically regulated extracellular volume, not requiring induction of force-dependent biochemical processes within the cell or cell membrane.
The ECS of guinea-pig atria and frog sartorii can be determined using hemoglobin. For guinea-pig atria an ECS of (32.2 +/- 2.6)% wet wt for frog sartorii an ECS of (12.4 +/- 1.0)% wet wt can be measured.
Calreticulin was first isolated 26 years ago. Since its discovery as a minor Ca(2+)-binding protein of the sarcoplasmic reticulum, the appreciation of its importance has grown, and it is now recognized to be a multifunctional protein, most abundant in the endoplasmic reticulum (ER). The protein has well-recognized physiological roles in the ER as a molecular chaperone and Ca(2+)-signalling molecule. However, it has also been found in other membrane-bound organelles, at the cell surface and in the extracellular environment, where it has recently been shown to exert a number of physiological and pathological effects. Here, we will focus on these less-well-characterized functions of calreticulin.