Espace extracellulaire, perméabilité à l'inuline et accumulation de L-alanine dans l'intestin isolé de Grenouille

1973 ◽  
Vol 51 (1) ◽  
pp. 22-28
Author(s):  
Joël de la Noüe ◽  
André Gagnon
Keyword(s):  
Extracellular Space ◽  
Muscle Layer ◽  
Solute Concentration ◽  
Tissue Water ◽  
Intracellular Amino Acid ◽  
Inulin Space ◽  
Total Tissue ◽  
Frog Intestine

In order to calculate the intracellular concentration of accumulated L-alanine, the extracellular space (inulin-14C) of frog intestine was measured. To check the validity of the technique, frog liver and gastrocnemius were used too. By scraping proximal portions of intestine, the inulin space was found to be similar (around 20% of total tissue water) in both the muscle layer and the mucosa. The mucosal epithelium is an imperfect barrier to inulin while the serosa is very permeable. These results suggest that the interstitial solute concentration is best approximated by equating it to that of the serosal solution. The in vitro inulin space, compared to the in vivo one, increases with time, as does the cellular hydration. The data obtained from measurements of extracellular space and from L-alanine uptake show that the intracellular amino acid is in a free state.

Influence of pilocarpine on transport by salivary gland slices

1963 ◽  
Vol 205 (5) ◽  
pp. 1058-1062 ◽  
Author(s):  
L. H. Schneyer ◽  
C. A. Schneyer
Keyword(s):  
Salivary Gland ◽  
Submaxillary Gland ◽  
Nitrogen Atmosphere ◽  
Total Water ◽  
Early Effect ◽  
Apparent Loss ◽  
Inulin Space ◽  
Total Tissue

Effects of pilocarpine on net movements of water and electrolytes in gland cells were investigated in vitro, using slices from submaxillary gland of rat. Slices were depleted of K, and loaded with Na, Cl, and water, by incubation in Krebs-Ringer phosphate with nitrogen atmosphere. After this, the slices were transferred to Krebs-Ringer phosphate with oxygen atmosphere. During this period with O2, pilocarpine caused apparent loss of water from cells, since tissue total water decreased and inulin space remained almost unchanged. Without pilocarpine during this time, water in cells increased. Electrolyte movements were also affected by pilocarpine. Specifically, there occurred reduction in net accumulation of K in total tissue and cells. Reduction in net extrusion of Na was suggested. Since, in vivo, an early effect of stimulation involves depletion of gland K, it appears that the current observations have relevance to normal secretion to the extent, at least, that in both circumstances stimulating agents reduce the ability of the cells to maintain stores of K.

Hemorrhagic shock impairs myocardial cell volume regulation and membrane integrity in dogs

1987 ◽  
Vol 252 (6) ◽  
pp. H1203-H1210
Author(s):  
J. W. Horton
Keyword(s):  
Hemorrhagic Shock ◽  
Cell Volume ◽  
Volume Regulation ◽  
Membrane Integrity ◽  
Cell Volume Regulation ◽  
Calcium Entry ◽  
Tissue Water ◽  
Total Tissue

An in vitro myocardial slice technique was used to quantitate alterations in cell volume regulation and membrane integrity after 2 h of hemorrhagic shock. After in vitro incubation in Krebs-Ringer-phosphate medium containing trace [14C]inulin, values (ml H2O/g dry wt) for control nonshocked myocardial slices were 4.03 +/- 0.11 (SE) for total water, 2.16 +/- 0.07 for inulin impermeable space, and 1.76 +/- 0.15 for inulin diffusible space. Shocked myocardial slices showed impaired response to cold incubation (0 degrees C, 60 min). After 2 h of in vivo shock, total tissue water, inulin diffusible space, and inulin impermeable space increased significantly (+19.2 +/- 2.4, +8.1 +/- 1.9, +34.4 +/- 6.1%, respectively) for subendocardium, whereas changes in subepicardium parameters were minimal. Shock-induced cellular swelling was accompanied by an increased total tissue sodium, but no change in tissue potassium. Calcium entry blockade in vivo (lidoflazine, 20 micrograms X kg-1 X min-1 during the last 60 min of shock) significantly reduced subendocardial total tissue water as compared with shock-untreated dogs. In addition, calcium entry blockade reduced shock-induced increases in inulin impermeable space and inulin diffusible space. In vitro myocardial slice studies confirm alterations in subendocardial membrane integrity after 2 h of in vivo hemorrhagic shock. Shock-induced abnormalities in myocardial cell volume regulation are reduced by calcium entry blockade in vivo.

scholarly journals Water Compartmentalization and Spread of Ischemic Injury in Thick-Slice Ischemia Model

2002 ◽  
Vol 22 (1) ◽  
pp. 80-88 ◽  
Author(s):  
Sabina Hrabětová ◽  
Kevin C. Chen ◽  
Daniel Masri ◽  
Charles Nicholson
Keyword(s):  
Water Content ◽  
Extracellular Space ◽  
Cell Swelling ◽  
Extracellular Potassium ◽  
Tissue Water ◽  
Diffusion Analysis ◽  
Artificial Cerebrospinal Fluid ◽  
Water Redistribution ◽  
Total Tissue

Water compartmentalization was studied in a thick-slice (1000 μm) model of ischemia by combining water-content measurements with extracellular diffusion analysis. Thick slices bathed in artificial cerebrospinal fluid continually gained water. Total tissue water content was increased by 67% after 6 hours of the incubation. Diffusion measurements using the tetramethylammonium method showed that the extracellular space, typically occupying 20% of brain tissue in vivo, was decreased to 10% at 30 minutes and 15% at 6 hours in both deep and superficial layers of thick slices. Quantification of water compartmentalization revealed that water moved initially from the extracellular space into the cells. Later, however, both compartments gained water. The initial cell swelling was accompanied by dramatic shifts in potassium. An initial rise of extracellular potassium to about 50 mmol/L was measured with a potassium-selective microelectrode positioned in the center of the thick slice; the concentration decreased slowly afterwards. Potassium content analysis revealed a 63% loss of tissue potassium within two hours of the incubation. In thick slices, ionic shifts, water redistribution, and a loss of synaptic transmission occur in both deep and superficial layers, indicating the spread of ischemic conditions even to areas with an unrestricted supply of nutrients.

THE LEVATOR ANI MUSCLE OF THE RAT AS AN INTACT PREPARATION SUITABLE FOR IN VITRO INVESTIGATIONS

1966 ◽  
Vol 52 (2) ◽  
pp. 325-336 ◽  
Author(s):  
A. Arvill ◽  
K. Ahrén
Keyword(s):  
Levator Ani ◽  
Male Rats ◽  
Levator Ani Muscle ◽  
List Type ◽  
Tissue Water Content ◽  
Tissue Water ◽  
Incubation Periods ◽  
Total Tissue

ABSTRACT A method is described of dissecting out and incubating the levator ani muscle of immature male rats, keeping its normal connections to the rest of the perineal complex. In order to find out whether this isolated preparation remained intact with undamaged cell membranes, comparisons were made between this preparation of the levator ani muscle and a cut preparation of the same muscle, and also between the levator ani muscle and the intact and cut preparations of the diaphragm. The following determinations were made: Distribution of sucrose-14C in vivo and in vitro Distribution of inulin-14C in vitro Distribution of D-xylose-14C in vitro Total tissue water content Potassium concentration in the medium after different incubation periods. It is concluded that it is possible to dissect out the levator ani muscle with undamaged cells and that this preparation can be both suitable and useful for in vitro investigations.

Effect ofSchistosoma mansoni-induced granulomatous inflammation on murine gastrointestinal motility

2001 ◽  
Vol 280 (5) ◽  
pp. G1030-G1042 ◽  
Author(s):  
Tom G. Moreels ◽  
Joris G. De Man ◽  
Johannes J. Bogers ◽  
Benedicte Y. De Winter ◽  
Gunther Vrolix ◽  
...  
Keyword(s):  
Structural Changes ◽  
Gastrointestinal Transit ◽  
Granulomatous Inflammation ◽  
Muscle Layer ◽  
Gastric Fundus ◽  
Smooth Muscle Contractility ◽  
Functional Changes ◽  
Mucosal Thickness

In Schistosoma mansoni-infected mice, gastrointestinal transit was measured in vivo and the neuromuscular function of longitudinal muscle strips of inflamed ileum and noninflamed gastric fundus was assessed in vitro. Eight weeks after infection, the ileal wall was acutely inflamed, as shown by a mucosal inflammatory infiltrate, leading to an increase in mucosal thickness, in myeloperoxidase (MPO) activity, and in interleukin (IL)-1β production. At that time, both gastrointestinal transit and in vitro ileal contractility were normal. Twelve weeks after infection, chronic granulomatous inflammation led to proliferation of the muscle layer and to a further increase in MPO activity, whereas IL-1β production normalized. Gastrointestinal transit was decreased, whereas in vitro ileal contractility was increased irrespective of the contractile stimulus. In vitro incubation with IL-1β (10 ng/ml for 60 min) significantly increased ileal contractility only at 8 wk after infection. Indomethacin, tetrodotoxin, and atropine had no differential effect on ileal contractility in controls and infected mice. In vitro contractility of noninflamed gastric fundus was normal both 8 and 12 wk after infection. We conclude that intestinal schistosomiasis 8 wk after infection is associated only with structural changes of the ileum, whereas 12 wk after infection, both structural and functional changes are present. These changes are characterized by increased ileal wall thickness, decreased gastrointestinal transit, and increased smooth muscle contractility restricted to the inflamed gut segment.

Tissue spaces in rat heart, liver, and skeletal muscle in vivo

1998 ◽  
Vol 275 (5) ◽  
pp. R1530-R1536 ◽  
Author(s):  
Julie Cieslar ◽  
Ming-Ta Huang ◽  
Geoffrey P. Dobson
Keyword(s):  
Skeletal Muscle ◽  
Rat Heart ◽  
Gastrocnemius Muscle ◽  
Interstitial Space ◽  
Glyceric Acid ◽  
Sprague Dawley ◽  
Tissue Water ◽  
Intracellular Space ◽  
Total Tissue

Tissue spaces were determined in rat heart, liver, and skeletal muscle in vivo using isotopically labeled [14C]inulin. Tracer was injected into the jugular vein of pentobarbital-anesthetized male Sprague-Dawley rats. After a 30-min equilibration period, a blood sample was taken, and heart, liver, and gastrocnemius muscle were excised and immediately freeze clamped at liquid nitrogen temperatures. The extracellular inulin space was 0.209 ± 0.006 ( n = 13), 0.203 ± 0.080 ( n = 7), and 0.124 ± 0.006 (SE) ml/g wet wt tissue ( n = 8) for heart, liver, and skeletal muscle, respectively. Total tissue water was 0.791 ± 0.005 ( n = 9), 0.732 ± 0.002 ( n = 9), and 0.755 ± 0.005 ml/g wet wt tissue ( n = 10) for heart, liver, and skeletal muscle, respectively. Expressed as a percentage of total tissue water, the intracellular space was 73.6, 72.2, and 83.7% for heart, liver, and skeletal muscle, respectively. With use of 2,3-diphospho-d-glyceric acid as a vascular marker, the interstitial space was calculated by subtracting the counts in tissue due to whole blood from total tissue counts and dividing by plasma counts. The interstitial space was 18.8, 22.4, and 14.5% of total tissue water, with accompanying plasma spaces of 7.7, 5.3, and 1.8% for heart, liver, and gastrocnemius muscle, respectively. The tracer method used in this study provides a quantitative assessment of water distribution in tissues of nonnephrectomized rats that has applications for calculation of tissue ion and metabolite concentrations, gradients, and fluxes under normal and pathophysiological conditions.

Release of endo-lysosomal cathepsins B, D, and L from IEC6 cells in a cell culture model mimicking intestinal manipulation

2009 ◽  
Vol 390 (5/6) ◽  
Author(s):  
Kristina Mayer ◽  
Anna Vreemann ◽  
Hong Qu ◽  
Klaudia Brix
Keyword(s):  
Epithelial Cells ◽  
Extracellular Space ◽  
Inflammatory Responses ◽  
Cell Damage ◽  
Mechanical Damage ◽  
Model System ◽  
Surgical Trauma ◽  
Mechanical Manipulation

AbstractIEC6 cells were used as anin vitromodel system to study the effects of cell damage caused by mechanical manipulation of intestine epithelial cells. We constructed an apparatus that allowed analyzing the consequences of mechanical compression in a standardized and reproducible manner. Manipulation of IEC6 cells induced necrosis rather than apoptosis, and resulted in release of HMGB1, which is known to function as a trigger of inflammatory responsesin vivo. Mechanical damage by traumatic injury of the intestine is accompanied by altered protease activities in the extracellular space, but only little is known about the possible contribution of endo-lysosomal cathepsins. Therefore, we tested the supernatants of manipulated cells in ourin vitromodel system for proteolytic activity and determined release rates by fluorimetric assays. Endo-lysosomal proteases, such as cathepsins B, D, and L, were released from damaged cells within the first 3 h after manipulation. While cathepsin L re-associated with the surfaces of neighboring cells, cathepsins B and D were present in the extracellular space as soluble enzymes. We conclude that our apparatus for mechanical manipulation can be used to approach surgical trauma, thereby focusing on epithelial cells of the intestine mucosa.

scholarly journals Monosaccharide transport in the mammary gland of the intact lactating rat

1984 ◽  
Vol 218 (1) ◽  
pp. 213-219 ◽  
Author(s):  
L C Threadgold ◽  
N J Kuhn
Keyword(s):  
Mammary Gland ◽  
Glucose Transport ◽  
Intracellular Water ◽  
Limiting Factor ◽  
Transport Capacity ◽  
Tissue Water ◽  
Monosaccharide Transport ◽  
Rate Limiting ◽  
Total Tissue

The Michaelis-Menten equation for the utilization of competing substrates was applied to the uptake of 2-deoxy[3H]glucose into the mammary gland of anaesthetized lactating rats. Intracellular water was calculated from total tissue water and sucrose space. Fed rats had a mean transport capacity of 2.2 mumol/min per g of tissue, giving an actual glucose transport in vivo of 1.1 mumol/min per g. Transport decreased by 90% on overnight starvation and returned to normal by 2 h of re-feeding. Similar changes were observed in the 1 min or 5 min transport of circulating 3-O-methylglucose. Transport of 3-O-methylglucose in starved rats was restored towards normal by insulin. In fed rats it increased between parturition and day 12 of lactation. The findings support the proposal that transport is a rate-limiting factor in the mammary utilization of carbohydrate.

scholarly journals Indirect regulation of HMGB1 release by gasdermin D

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Allen Volchuk ◽  
Anna Ye ◽  
Leon Chi ◽  
Benjamin E. Steinberg ◽  
Neil M. Goldenberg
Keyword(s):  
Bone Marrow ◽  
Knockout Mice ◽  
Extracellular Space ◽  
High Mobility ◽  
High Mobility Group ◽  
Inflammasome Activation ◽  
Inflammatory Stimuli ◽  
Lps Treatment

Abstract The protein high-mobility group box 1 (HMGB1) is released into the extracellular space in response to many inflammatory stimuli, where it is a potent signaling molecule. Although research has focused on downstream HMGB1 signaling, the means by which HMGB1 exits the cell is controversial. Here we demonstrate that HMGB1 is not released from bone marrow-derived macrophages (BMDM) after lipopolysaccharide (LPS) treatment. We also explore whether HMGB1 is released via the pore-forming protein gasdermin D after inflammasome activation, as is the case for IL-1β. HMGB1 is only released under conditions that cause cell lysis (pyroptosis). When pyroptosis is prevented, HMGB1 is not released, despite inflammasome activation and IL-1β secretion. During endotoxemia, gasdermin D knockout mice secrete HMGB1 normally, yet secretion of IL-1β is completely blocked. Together, these data demonstrate that in vitro HMGB1 release after inflammasome activation occurs after cellular rupture, which is probably inflammasome-independent in vivo.

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