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  • 1
    ISSN: 1432-2013
    Keywords: Colon ; Rectum ; Epithelial resistance ; AC impedance ; Stripping ; Subepithelial resistance ; Aldosterone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Epithelial and subepithelial electrical resistances of rat large intestine were measured by means of a 4-electrode AC impedance technique in three segments, colon ascendens, colon descendens and rectum. (i) Epithelial resistance of colon ascendens and colon descendens was about 35 Ω · cm2 and not different between these two segments. It was, however, about 3 times higher in rectum (99 Ω · cm2). This finding is in accord with our previous observation of about 3-fold higher net fluxes of ions and water in colon ascendens and colon descendens than in rectum. It confirms the concept of a main functional difference between the terminal part of the large intestine (rectum) and the more proximal segments (colon). (ii) The acutely (within hours) varied level of aldosterone by keeping the rats for 7 h in anaesthesia caused in the rectum a more than 10-fold increase in short circuit current (I SC) and transepithelial voltage but no significant decrease in resistance. Similarly, the decline inI SC, as regularly observed in the early phase of in vitro measurements on partially stripped large intestine, was paralleled by voltage changes but not by changes in resistance. We conclude that the wide range of resistance values published so far was caused to a great extent by including various portions of colon or rectum. (iii) By comparing intact (not stripped) and partially stripped preparations (muscularis propria removed) of the rectum it was shown that partial stripping did not alter the epithelial resistance but reduced the subepithelial resistance in this segment from 26 to 8 Ω · cm2, or by 68%. Subepithelial resistances of stripped rectum, colon ascendens and colon descendens were 8, 12 and 13 Ω · cm2, respectively. Based on these figures,I SC of conventional voltage clamp measurements is underestimated due to subepithelial tissue layers in intact rectum by 23% and in the partially stripped preparations of rectum, colon ascendens and colon descendens by 9%, 34%, and 37%, respectively.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Mammalian small intestine ; in vitro technique ; AC impedance ; epithelial resistance ; subepithelial resistance ; leaky epithelia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Epithelial and subepithelial resistance of rat jejunum was measured in vitro by two independent methods. (i) Transepithelial AC impedance data were interpreted in terms of a simple parallel RPCP element (representing the epithelial cell layer) in series with an ohmic resistor RS (representing the subepithelial layers). (ii) In separate experiments, the tip of a microelectrode was positioned between epithelium and subepithelial layers and the respective resistances were obtained from DC-pulse voltage divider ratios between both structures. The total tissue resistance as measured in conventional Ussing-chamber experiments (49±4 Ohm·cm2, mean of both methods) was formed to 81±6% (40±3 Ohm·cm2) by subepithelial layers and to only 19±3% (9±1 Ohm·cm2) by the epithelial cell line. We conclude that rat jejunum is more conductive than assumed so far. In in vitro flux studies on intact jejunal sheets a pronounced back-diffusion of absorbed substances will lead to an underestimation of the true net transport capacity of this structure. This error averages about fivefold and will be found likewise in conventional short-circuit measurements.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2013
    Keywords: Amiloride ; Bumetanide ; Cl-secretion ; Electrical field stimulation ; Large intestine ; Na-absorption ; Rat ; Stripping ; Submucosal plexus ; Tetrodotoxin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The initial phase of in vitro experiments in Ussing-type chambers on large intestine is characterized by short-circuit currents (ISC) declining from high starting values to a lower plateau within 0.5 h. The origin of this “initial ISC-transient” was investigated by ISC measurements on partially stripped segments of rat rectal colon. Transport was pre-stimulated in vivo by keeping animals in barbiturateanesthesia for 5 h prior to tissue preparation. This procedure caused by endogenous aldosterone-liberation amiloride-sensitive Na-absorption to become the predominant electrogenic transport. The initial ISC-transient was abolished by tetrodotoxin (TTX, 1 μM), indicating a neuronal mediation of this phenomenon. In order to identify the transport which was subject to neuronal control, the amiloride-sensitive Na-absorption was measured during electrical field stimulation (bipolar rectangular pulses: 5 Hz, 1 ms, ±6 mA). There was no difference to unstimulated controls. In contrast, the initial ISC-transient was dependent on Cl in the bath following Michaelis-Menten-kinetics (K M=20 mM) and could be prevented by 10 μM serosal bumetanide. Then, initial filling of the Ussing-chamber was imitated during the course of the experiment by removal and immediate readdition of the bathing fluid. This procedure caused ISC-changes of similar appearance as the initial ISC-transient. To verify that indeed mechanical stretch is the sensory stimulus triggering the initial ISC-transient, the effect of small pressure oscillations was studied. This also produced an ISC-transient which was TTX-sensitive and was abolished after removal of the submucosal plexus Meissner by total stripping. It is concluded that amiloride-sensitive Na-absorption does not contribute to the initial transient and is not affected by the enteric nervous system. Initial ISC-transients asobserved during the first half hour of Ussing experiments are due to electrogenic Cl-secretion which is stimulated by mechanical stretch during tissue preparation and filling of the chamber via a submucosal neuronal reflex pathway. The possible biological meaning of this stretch-induced secretory process could be facilitation of transit during imminent stasis of the gut contents.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2013
    Keywords: Adrenalectomy ; Aldosterone ; K absorption ; K secretion ; Na transport ; Osmolyte transport ; Proximal colon ; Rectal colon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The short-term action of aldosterone in physiological concentration on net fluxes of Na, K, Cl, HCO3, osmolytes, and water was examined in the proximal colon and rectal colon of adrenalectomized (ADX) rats in vivo. The measuring time was 12 h, divided in eight periods of 90 min. (a) Aldosterone alone (6 nmol h−1kg−1) did not stimulate transport in ADX rats. In these experiments plasma [K] increased to fatal values. A basal glucocorticoid substitution of 24 nmol h−1kg−1 corticosterone caused plasma K to stay constant throughout the experiment, so that epithelial transport was not handicapped by non-specific effects of ADX, but this also did not restore the decreased transport of ADX rats to control values. Under these conditions (absence of aldosterone) in the rectal colon Na and H2O transport was zero, whereas in the proximal colon flux rates were depressed by between 30% and 50%. In contrast, basal glucocorticoid substitution of 18 nmol h−1kg−1 corticoster-one plus infusion of 6 nmol h−1kg−1 aldosterone caused transport stimulation to values not significantly different from those of non-ADX controls. We conclude that after ADX, aldosterone at physiological concentrations increases transport if, as a prerequisite, a basal glucocorticoid substitution is provided. Transport of Na, K, and H2O is under the total control of aldosterone in the rectal colon but is only moderately altered in the proximal colon. (b) In the proximal colon aldosterone is effective on electroneutral Na transport consistent with the Na/H, Cl/HCO3 double exchanger, while in the rectal colon aldosterone controls the amiloride-sensitive Na channel, which is the sole apical Na transporter in that segment. In this respect the rectal colon is functionally distinct from the distal colon of other studies in the rat, where under unstimulated conditions the double exchanger is present. Regarding Na transport mechanisms, the rat large intestine thus consists of three distinct segments, the proximal, distal, and rectal colon. (c) In the rectal, but not the proximal colon, active net K absorption against the electrochemical gradient took place in the absence of aldosterone, but was suppressed in its presence. (d) The flux ratio of osmolytes over H2O was constantly hypertonic as compared to the plasma (387±1 mosmol l−1), independent of absolute flux size, the colonic segment examined, or the steroid concentration used. This effect may be due to a hypertonic unstirred layer within the lamina propria.
    Type of Medium: Electronic Resource
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