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  • 1
    ISSN: 1432-2013
    Keywords: Large intestine ; Aldosterone ; Anaesthesia ; Amiloride ; Ion transport ; Osmolyte transport ; Water transport ; Luminal steady state concentrations ; Transepithelial voltage ; Segmental heterogeneity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Thiobutabarbital anaesthetized and abdominally operated control rats develop high endogenous plasma levels of both aldosterone and corticosterone during the course of a 12 h experiment. This effect was used as a model for examining ‘acute’ steroid action (i) on net ion and water fluxes and (ii) on zero flux luminal limiting concentrations in rat upper colon (proximal 50% of large intestine) and rectum (distal 40%). Experiments of both kinds consisted of 8 independent 90 min measuring periods. (i) In rectum net fluxes of Na, K, osmolytes (sum of all solutes) and water started at low levels around zero, began to rise about 2 h after plasma levels of aldosterone had increased, and reached plateau values around the 6th hour of anaesthesia. In upper colon, fluxes of Na, K, Cl, and osmolytes were high from the beginning and did not vary significantly with time. (ii) At zero flux conditions limiting concentrations of Na in the hormonally unstimulated phase of the experiment were 20±3 mM in upper colon and 22±3 mM in rectum. After maximal endogenous aldosterone liberation zero flux concentrations were 5.2 mM in upper colon and 2.2 mM in rectum, corresponding to luminal fluid to plasma ratios (LF/P) of 0.040 and 0.016, respectively. Amiloride reduced the maximal Na gradient in rectum to aLF/P of 0.3 but was not effective in upper colon and did not prevent the stimulating effect of aldosterone in this segment. Under all experimental conditions zero flow concentrations of K were higher than consistent with a solely passive distribution, indicating simultaneous passive and active secretion in both segments. In contrast to the findings of others, the luminal fluid remained isoosmolar with plasma in all zero flux experiments. Conclusions: Under acute elevation of steroid levels not higher than reached in a living animal the rectum varies net fluxes of Na, K, osmolytes, and water between zero and high rates. At this range of steroid levels, the upper colon practically does not respond with variations in net fluxes. Under zero flux conditions aldosterone acutely increases maximal Na gradients in rectum and, to a lesser degree, also in upper colon. Thus, acutely elevated aldosterone levels affect Na transport in upper colon and rectum by different mechanisms and intensities: in rectum, which is the most sensitive site of transport regulation, via an amiloridesensitive Na-pathway, and in upper colon, via an amiloride-insensitive pathway.
    Type of Medium: Electronic Resource
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  • 2
    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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  • 3
    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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