scholarly journals Kinetics of Calcium Fluxes Across the Intestinal Mucosa of the Marine Teleost, Gadus Morhua, Measured Using an In Vitro Perfusion Method

1988 ◽  
Vol 140 (1) ◽  
pp. 171-186 ◽  
Author(s):  
KRISTINA SUNDELL ◽  
BJÖRN THRANDUR BJÖRNSSON

An in vitro technique for perfusion of the intestinal vasculature and lumen was developed and used to measure calcium (Ca2+) fluxes across the intestinal mucosa of the marine teleost, the Atlantic cod (Gadus morhua). Saturable and nonsaturable components of the calcium influx and efflux were determined. The calcium influx had one passive component and one saturable component, following Michaelis-Menten kinetics with Km = 8.41mmoll−1 and Vmax = 0.604μmol Ca2+ kg−1 h−1. At physiological Ca2+ concentrations in the vascular ([Ca2+] = l.9mmoll−1) and luminal ([Ca2+] =14.9mmoll−1) perfusion fluids, the saturable component amounted to 60% of the Ca2+ influx. The high-affinity Ca2+-ATPase inhibitor chlorpromazine (CP, 10−4moll−1) antagonized 45% of the Ca2+ influx. The Ca2+ efflux across the intestinal mucosa of the cod was a saturable process, following Michaelis-Menten kinetics with Km =6.15mmoll−1 and Vmax =3.79μmol Ca2+ kg−1h−1, but insensitive to CP (l0−5moll−1). The Ca2+ efflux was l.22μumol Ca2+ kg−1 h−1, representing about 20% of the total calcium excretion and about 50% of the extrarenal excretion of the Atlantic cod in vivo.

1988 ◽  
Vol 137 (1) ◽  
pp. 287-301 ◽  
Author(s):  
M. Axelsson

The nervous regulation of heart rate and stroke volume in the Atlantic cod Gadus morhua was investigated both in vivo, during rest and exercise, and in vitro. The cholinergic and adrenergic influences on the heart were estimated in experiments with injections of atropine and sotalol. At rest the cholinergic and adrenergic tonus on the heart were 38% and 21%, respectively (ratio 1.81:1). At the end of an exercise period, the cholinergic tonus had decreased to 15% but the adrenergic tonus had increased to 28% (ratio 0.54:1). The results suggest that variation of the cholinergic tonus on the heart is a major factor in the regulation of the heart rate. In one group of fish, cardiac output was also measured, allowing calculation of stroke volume. Cardiac output increased significantly during exercise, and this effect persisted in the presence of both atropine and sotalol, although the increase in heart rate was reduced or abolished. The persisting increase in cardiac output during exercise is due to an increase in stroke volume, reflecting a Starling relationship. In the presence of the adrenergic neurone-blocking agent bretylium, a positive inotropic effect on isolated, paced atrial and ventricular strips was observed. In the atrial preparations the effect persisted after 24 h. The effect was prevented by pretreatment with sotalol or cocaine, but potentiated by phentolamine pretreatment. This shows that bretylium exerts its neurone-blocking action after being taken up into the adrenergic nerves, and suggests that the positive inotropic effect of bretylium observed in vivo is due to release of endogenous catecholamines. The concentration-response curves for adrenaline on isolated spontaneously beating atrial preparations showed that the concentrations of catecholamines necessary to produce appreciable effects on the heart are higher than the concentrations found in cod plasma during ‘stress’ situations (handling and exhaustive swimming).


1985 ◽  
Vol 248 (1) ◽  
pp. R18-R22 ◽  
Author(s):  
B. T. Bjornsson ◽  
S. Nilsson

The calcium turnover, with particular emphasis on the routes and rates of excretion, was investigated in the Atlantic cod, Gadus morhua. A very rapid internal redistribution of injected 45Ca was demonstrated, with the decline in the plasma concentration conforming to the power function f(t) = 8.83% X t-0.585. The clearance for 45Ca and [3H]inulin were both linearly correlated to urine flow and showed a ratio (Cca/CIn) of 2.6:1. Likewise a linear correlation between the renal and extra-renal excretion of 45Ca could be demonstrated with a ratio of 1:0.47, indicating that the renal route of excretion is of major importance in this species. The renal excretion of calcium was estimated to be 4.2 mumol X kg-1 X h-1 and the total calcium excretion to be 6.2 mumol X kg-1 X h-1. Calculations are presented, suggesting that the balance between intake and excretion of calcium can be explained in terms of growth of the animal, which would allow for an increase in the amount of body calcium of approximately 5 g X yr-1. It is also concluded that because about two-thirds calcium is excreted via the renal route, the kidneys may be the most important site for the regulation of calcium excretion in the cod.


1964 ◽  
Vol 21 (2) ◽  
pp. 347-354
Author(s):  
P. M. Townsley ◽  
M. L. Hughes

The early stages in the recovery of the dorsal fin of the Atlantic cod (Gadus morhua) to a "clean cut injury" are described. It is concluded that the observed rapid epidermal migration, wound closure and cell mitosis are essentially the same in in-vivo as in in-vitro experiments. An accumulation of carbohydrate material occurs in the outermost layer of epidermal cells. There is a change in the carbohydrate composition or structure in the dermal layers at the site of injury. The basal epidermal cells rapidly divide in the in-vitro culture whereas only those basal epidermal cells in an in-vivo injury in close proximity to the injury divide. The surrounding nutrient medium in in-vitro cultures does not appear to be involved in the initial cell migration. However, ascorbic acid does stimulate epidermal migration, mucous secretion, and basal epidermal cell mitosis.


Cells ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 66
Author(s):  
Rashmita Pradhan ◽  
Phuong A. Ngo ◽  
Luz d. C. Martínez-Sánchez ◽  
Markus F. Neurath ◽  
Rocío López-Posadas

Rho proteins operate as key regulators of the cytoskeleton, cell morphology and trafficking. Acting as molecular switches, the function of Rho GTPases is determined by guanosine triphosphate (GTP)/guanosine diphosphate (GDP) exchange and their lipidation via prenylation, allowing their binding to cellular membranes and the interaction with downstream effector proteins in close proximity to the membrane. A plethora of in vitro studies demonstrate the indispensable function of Rho proteins for cytoskeleton dynamics within different cell types. However, only in the last decades we have got access to genetically modified mouse models to decipher the intricate regulation between members of the Rho family within specific cell types in the complex in vivo situation. Translationally, alterations of the expression and/or function of Rho GTPases have been associated with several pathological conditions, such as inflammation and cancer. In the context of the GI tract, the continuous crosstalk between the host and the intestinal microbiota requires a tight regulation of the complex interaction between cellular components within the intestinal tissue. Recent studies demonstrate that Rho GTPases play important roles for the maintenance of tissue homeostasis in the gut. We will summarize the current knowledge on Rho protein function within individual cell types in the intestinal mucosa in vivo, with special focus on intestinal epithelial cells and T cells.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anderson B. Guimaraes-Costa ◽  
John P. Shannon ◽  
Ingrid Waclawiak ◽  
Jullyanna Oliveira ◽  
Claudio Meneses ◽  
...  

AbstractApart from bacterial formyl peptides or viral chemokine mimicry, a non-vertebrate or insect protein that directly attracts mammalian innate cells such as neutrophils has not been molecularly characterized. Here, we show that members of sand fly yellow salivary proteins induce in vitro chemotaxis of mouse, canine and human neutrophils in transwell migration or EZ-TAXIScan assays. We demonstrate murine neutrophil recruitment in vivo using flow cytometry and two-photon intravital microscopy in Lysozyme-M-eGFP transgenic mice. We establish that the structure of this ~ 45 kDa neutrophil chemotactic protein does not resemble that of known chemokines. This chemoattractant acts through a G-protein-coupled receptor and is dependent on calcium influx. Of significance, this chemoattractant protein enhances lesion pathology (P < 0.0001) and increases parasite burden (P < 0.001) in mice upon co-injection with Leishmania parasites, underlining the impact of the sand fly salivary yellow proteins on disease outcome. These findings show that some arthropod vector-derived factors, such as this chemotactic salivary protein, activate rather than inhibit the host innate immune response, and that pathogens take advantage of these inflammatory responses to establish in the host.


Drug Research ◽  
2018 ◽  
Vol 69 (03) ◽  
pp. 173-180 ◽  
Author(s):  
Bilal Houshaymi ◽  
Nadine Nasreddine ◽  
Mamdouh Kedees ◽  
Zeina Soayfane

AbstractThe bioavailability of ivermectin is modulated by lipid-based formulations and membrane efflux transporters such as Breast Cancer Resistance Protein and P-glycoprotein (BCRP and P-gp). We have investigated the effect of oleic acid on the uptake of ivermectin in vitro using Caco-2 cells and in vivo in the intestines of wild-type mice. Complex micelles (M) with oleic acid induced a significant increase (e. g. for M3 was 7-fold, p≤0.001) in the uptake of the drug in a time-dependent manner with no involvement of cholesterol in the mechanism. In vivo results showed a significant increase in the concentration of plasma and intestinal mucosa ivermectin (p≤0.01) in mice receiving oleic acid-based drug formulation. We also examined the expression of the drug efflux transporter, BCRP and P-gp in Caco-2 cells and found a significant decrease (p≤0.001) in their level in the presence of 5 mM oleic acid. Treatment of mice with oleic acid-based formulation showed a significant decrease in the activity of P-gp in the intestinal mucosa (p≤0.01). This study highlighted the effect of oleic acid in decreasing the expression and the activity of P-gp-mediated ivermectin efflux and in limiting the drug absorption by increasing its uptake and bioavailability in Caco-2 cells and intestine, respectively.


2013 ◽  
Vol 305 (8) ◽  
pp. R864-R876 ◽  
Author(s):  
Douglas A. Syme ◽  
A. Kurt Gamperl ◽  
Gordon W. Nash ◽  
Kenneth J. Rodnick

We employed the work loop method to study the ability of ventricular and atrial trabeculae from Atlantic cod to sustain power production during repeated contractions at acclimation temperatures (10°C) and when acutely warmed (20°C). Oxygen tension (Po2) was lowered from 450 to 34% air saturation to augment the thermal stress. Preparations worked under conditions simulating either a large stroke volume (35 contractions/min rate, 8–12% muscle strain) or a high heart rate (70 contractions/min, 2–4% strain), with power initially equal under both conditions. The effect of declining Po2 on power was similar under both conditions but was temperature and tissue dependent. In ventricular trabeculae at 10°C (and atria at 20°C), shortening power declined across the full range of Po2 studied, whereas the power required to lengthen the muscle was unaffected. Conversely, in ventricular trabeculae at 20°C, there was no decline in shortening power but an increase in lengthening power when Po2 fell below 100% air saturation. Finally, when ventricular trabeculae were paced at rates of up to 115 contractions/min at 20°C (vs. the maximum of 70 contractions/min in vivo), they showed marked increases in both shortening and lengthening power. Our results suggest that although elevated heart rates may not impair ventricular power as they commonly do isometric force, limited atrial power and the increased work required to expand the ventricle during diastole may compromise ventricular filling and hence, stroke volume in Atlantic cod at warm temperatures. Neither large strains nor high contraction rates convey an apparent advantage in circumventing this.


1999 ◽  
Vol 82 (6) ◽  
pp. 2936-2946 ◽  
Author(s):  
Mario B. Lips ◽  
Bernhard U. Keller

A quantitative analysis of activity-related calcium dynamics was performed in motoneurons of the nucleus hypoglossus in the brain stem slice preparation from mouse by simultaneous patch-clamp and microfluorometric calcium measurements. Motoneurons were analyzed under in vitro conditions that kept them in a functionally intact state represented by rhythmic, inspiratory-related bursts of excitatory postsynaptic currents and associated action potential discharges. Bursts of electrical activity were paralleled by somatic calcium transients resulting from calcium influx through voltage-activated calcium channels, where each action potential accounted for a calcium-mediated charge influx around 2 pC into the somatic compartment. Under in vivo conditions, rhythmic-respiratory activity in young mice occurred at frequencies up to 5 Hz, demonstrating the necessity for rapid calcium elevation and recovery in respiratory-related neurons. The quantitative analysis of hypoglossal calcium homeostasis identified an average extrusion rate, but an exceptionally low endogenous calcium binding capacity as cellular parameters accounting for rapid calcium signaling. Our results suggest that dynamics of somatic calcium transients 1) define an upper limit for the maximum frequency of respiratory-related burst discharges and 2) represent a potentially dangerous determinant of intracellular calcium profiles during pathophysiological and/or excitotoxic conditions.


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