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Biomedical subjects

J Hardcastle

Publications and source records attributed to J Hardcastle.

At least 37 records · Page 2Linked to original sources
PubMed1995

Do 5-HT4 receptors mediate the intestinal secretory response to 5-HT in rat in-vivo?

The involvement of the recently characterized 5-HT4 receptor in the actions of 5-hydroxytryptamine (5-HT) on jejunal, ileal and colonic electrogenic ion secretion was investigated in the rat in-vivo. 5-HT and the 5-HT1-, 5-HT2- and 5-HT4-receptor agonist 5-methoxytryptamine (5-MeOT), induced a rise in transintestinal PD in all regions of the gut. However, the 5-HT4-receptor agonists renzapride and cisapride had no effect. Furthermore, the 5-HT4-receptor antagonists SDZ 205-557 (2-diethylaminoethyl-[2-methoxy-4-amino-5-chloro] benzoate), tropisetron and SB 204070 ([1-butyl-4-piperidinylmethyl]-8-amino-7-chloro-1,4- benzodioxan-5-carboxylate hydrochloride) did not affect the secretory response to either 5-HT or 5-MeOT in the jejunum, but did cause a small inhibition in the ileum and colon. It is concluded that 5-HT4 receptors do not make a contribution to the electrically monitored 5-HT intestinal secretory response in the rat jejunum in-vivo, but may play a small role in the ileum and colon.

4-Aminobenzoic Acid↗
PubMed1995

Evidence that the secretory response of rat intestine to 5-hydroxytryptamine in-vivo involves more than one 5-hydroxytryptamine-receptor subtype.

The transintestinal potential difference (PD) across rat mid-small intestine and proximal colon was measured in-vivo. The 5-hydroxytryptamine (5-HT)-induced increase in PD, which reflects a stimulation of electrogenic C1 secretion, was mimicked by both 2-methyl-5-hydroxytryptamine (2-CH3-5-HT), an agonist at 5-HT3 receptors, and 5-methoxytryptamine (5-MT), an agonist that lacks affinity for 5-HT3 receptors. The 5-HT3 antagonist granisetron caused a marked inhibition of the response to 2-CH3-5-HT in both regions, but only produced a small inhibition of the small intestinal response to 5-HT, with a more pronounced effect in the colon. The failure of granisetron to produce a marked antagonism of the 5-HT-induced rise in the transintestinal PD, coupled with the ability of 5-MT to induce a secretory response, indicates that 5-HT3 receptors are not the only ones involved in the stimulation of C1 secretion. The 5-HT2 antagonist ketanserin failed to influence the response to 5-HT in either the small intestine or the colon, but it did inhibit the action of 5-MT, having a much greater effect in the small intestine. In the presence of granisetron however, ketanserin also inhibited the small intestinal response to 5-HT, having only a minimal effect in the colon. This suggests that 5-HT2 receptors can also play a role in the activation of C1 secretion. These observations suggest that both 5-HT2 and 5-HT3 receptors contribute to the stimulation of electrogenic C1 secretion by 5-HT, with 5-HT2 receptors playing a more prominent role in the small intestine and 5-HT3 receptors being more important in the colon.

5-Methoxytryptamine↗
PubMed1994

Is desensitization of intestinal 5-hydroxytryptamine receptors an in-vitro phenomenon?

The responses of proximal jejunum and distal ileum to successive applications of 5-hydroxytryptamine (5-HT) were examined in-vitro and in-vivo by measuring the electrical changes that reflect the stimulation of Cl- secretion. In stripped intestinal sheets the second application of a maximal concentration of 5-HT failed to elicit any response, indicating that complete desensitization had occurred. If submaximal concentrations were used, a second response was observed, although it was smaller than the first, indicating partial desensitization. Replacing the bathing solutions following application of a maximal 5-HT concentration also reduced, but did not abolish, the degree of desensitization observed with a second application of 5-HT. In an in-vivo preparation, however, there was no diminution of the responses to four successive maximal doses of 5-HT. This lack of desensitization was also evident in the cardiovascular responses to 5-HT. It is concluded that desensitization to 5-HT is a phenomenon that is readily observed only in-vitro and which is probably related to the inability of a small amount of isolated tissue to eliminate 5-HT.

Animals↗
PubMed1994

Involvement of nitric oxide in the response to 5-hydroxytryptamine in the rat in-vivo.

The involvement of nitric oxide (NO) in the effects of 5-HT on intestinal secretion and cardiovascular function in anaesthetized rats was investigated using NG-nitro-L-arginine methyl ester (L-NAME), a specific NO-synthase antagonist, and its optical isomer D-NAME. L-NAME significantly reduced the prolonged hypotensive response to 5-HT. It also caused a small rightward shift in the colonic 5-HT dose-response curve. This suggests that NO plays a significant role in the prolonged hypotensive response to 5-HT, and may make a small contribution to the secretory response of the colon, but not that of the jejunum, in the rat in-vivo.

Animals↗
PubMed1994

Comparison of a computer simulation program and a traditional laboratory practical class for teaching the principles of intestinal absorption.

Here we describe an evaluation of the effectiveness, compared with a traditional laboratory, of an interactive computer-assisted learning (CAL) program, which simulates a series of experiments performed using isolated, everted sacs of rat small intestine. The program is aimed at undergraduate students of physiology and is designed to offer an alternative student-centered learning approach to the traditional laboratory-based practical class. The evaluative study compared two groups of second-year undergraduate students studying a module on epithelial transport: one group worked independently using the CAL program and associated learning materials, and the other group followed a conventional practical class approach, working in the laboratory under supervision. Knowledge gain of each group was measured by means of a test consisting of a range of question types (e.g., short-answer factual, calculation, interpretation) given to students before and after the module. Student attitude to both approaches was assessed by questionnaire, and the resource requirements were also compared. It was found that the knowledge gain of both groups of students was the same, that students had a positive attitude toward using CAL programs of this type, and that the cost of the conventional laboratory-based approach was five times greater. The potential for integrating CAL programs into the undergraduate curriculum is discussed.

Animal Testing Alternatives↗
PubMed1994

Loperamide inhibits the enhanced intestinal glucose absorption of cystic fibrosis in vitro.

Enhanced Na(+)-linked nutrient absorption has been demonstrated in the cystic fibrosis bowel and may contribute to the dehydration of the luminal contents. The ability of loperamide to inhibit glucose transport was therefore assessed in jejunal biopsy specimens from children with cystic fibrosis by measuring the increased short-circuit current associated with active glucose absorption using a mini-Ussing chamber technique. The presence of loperamide in the luminal solution reduced the rise in short-circuit current induced by glucose over the range of concentrations tested (2.5 to 40 mmol.L-1). This finding suggests that the enhanced Na+ absorption, which is a feature of cystic fibrosis, is amenable to treatment and can be restored to normal levels. This result may have therapeutic implications for gastrointestinal function and may be applicable to other tissues where similar therapeutic approaches to reduce Na+ and thus water absorption are being pursued.

Adolescent↗
PubMed1993

Local and systemic actions of loperamide on fluid transport and transmural potential difference across rat small intestine.

The ability of loperamide to influence intestinal fluid transport was assessed using a dual loop preparation. Loperamide was applied to the lumen of the oral, but not the aboral loop, yet basal and prostaglandin-stimulated fluid transport was affected in both, indicating that the drug can act systemically in addition to any local actions. Loperamide had both pro-absorptive and anti-absorptive effects, the latter only occurring when basal fluid absorption was high. The effects of loperamide were greater in the aboral loop to which it was available only via the systemic route. This difference may reflect the absence of counteracting local influences in the aboral loop or a variation in the sensitivity of different regions of the gut to loperamide.

Animals↗
PubMed1993

Comparison of the effects of loperamide and loperamide oxide on absorptive processes in rat small intestine.

Mucosal loperamide inhibited the absorption of glycine by everted sacs of rat small intestine over 10-, 30- or 60-min incubation periods, but loperamide oxide was without effect. In stripped intestinal sheets, loperamide inhibited the rise in short-circuit current associated with Na(+)-linked glucose absorption, but this effect was not observed with loperamide oxide. It is concluded that although there is evidence that loperamide oxide is converted to loperamide in the intestinal lumen, the rate at which free loperamide appears is not sufficient to inhibit absorptive processes.

Animals↗
PubMed1992

Calcium-calmodulin-dependent activation of adenylate cyclase in prostaglandin-induced electrically-monitored intestinal secretion in the rat.

The calcium-calmodulin antagonist 5-iodo-C8-W7 inhibited the PGE2-induced stimulation of cAMP production by isolated enterocytes from rat small intestine. It also reduced the secretory response of intestinal sheets to PGE2, measured as a rise in short-circuit current. It did not however, inhibit the electrical responses to forskolin and dibutyryl cAMP, nor to acetylcholine, a secretagogue whose effect is not mediated by cAMP. It is concluded that the receptor-mediated activation of adenylate cyclase and the subsequent secretory response are dependent upon calcium-calmodulin.

Acetylcholine↗
PubMed1992

Failure of tetrodotoxin to inhibit the prostaglandin-induced secretory response of rat small intestine in-vitro.

Tetrodotoxin (TTX, 10 microM) did not inhibit prostaglandin E2 (PGE2)-stimulated increases in electrical activity in intact or stripped sheets of rat small intestine, although it reduced basal electrical activity in the intact preparation. Basal and PGE2-stimulated cAMP production by enterocytes isolated from the small intestine was unaffected by TTX. Thus its reported ability to inhibit prostaglandin-induced fluid secretion in-vivo does not appear to represent a direct interaction of the neurotoxin with the mechanism of prostaglandin action at the enterocyte.

Animals↗
PubMed1991

The influence of the gamma-amino butyric acid (GABA) antagonist bicuculline on transport processes in rat small intestine.

GABAergic involvement in the regulation of intestinal ion transport is suggested by the reduction in the short-circuit current generated by sheets of rat small intestine by the GABA antagonist bicuculline, although GABA itself caused no change. Bicuculline action was abolished by the inhibitor of Cl- secretion frusemide, suggesting that it involved a change in endogenous Cl- secretory tone. The effect of bicuculline appeared to be specific as it did not affect the electrical responses to glucose or exogenous secretagogues. The action of bicuculline was not observed in stripped intestinal sheets where the myenteric plexus is absent, and it was reduced by tetrodotoxin and atropine. It is suggested that endogenous GABA could be involved in the maintenance of a secretory tone in rat small intestine by acting on cholinergic mechanisms within the myenteric plexus.

Animals↗
PubMed1991

Measurement of intracellular mediators in enterocytes isolated from jejunal biopsy specimens of control and cystic fibrosis patients.

A method that maximises the yield of viable enterocytes has been developed for the isolation of enterocytes from human jejunal biopsy specimens. These enterocytes have been used to study the values of intracellular free calcium and the rises in adenosine 3'5'-cyclic monophosphate (cAMP) induced by secretagogues in normal and cystic fibrosis cells. Basal intracellular free calcium of cystic fibrosis enterocytes, measured fluorimetrically with fura-2, was within the range of the basal intracellular free calcium of non-cystic fibrosis enterocytes (cystic fibrosis 263 nmol/l; non-cystic fibrosis 287 nmol/l). Changes in intracellular free calcium were observed after exposure to ionomycin: a 100 nmol/l solution induced a 2.5 fold increase in intracellular free calcium in the cystic fibrosis enterocytes and a 2.2 fold increase in the intracellular free calcium concentration of the non-cystic fibrosis enterocytes. Basal cAMP values were not significantly different between cystic fibrosis and non-cystic fibrosis enterocytes (cystic fibrosis 575 fmol/100,000 cells; non-cystic fibrosis 716 fmol/100,000 cells, p greater than 0.05) and the enterocyte cAMP value increased in response to stimulation with prostaglandin E2 (7 mumol/l) (cystic fibrosis 2.2 fold increase over basal, p less than 0.05; non-cystic fibrosis 1.9 fold stimulation over basal, p less than 0.05) and vasoactive intestinal polypeptide (100 nmol/l) (cystic fibrosis 7.1 fold increase over basal, p less than 0.05; non-cystic fibrosis 5.8 fold increase over basal, p less than 0.05). There was no significant difference in the magnitude of the response between cystic fibrosis and non-cystic fibrosis enterocytes (p greater than 0.05). These results indicate that the cystic fibrosis defect in the small intestine, as in other affected epithelia, seems to be distal to the production of second messengers. The small intestine is therefore an appropriate model in which to study the biochemical defect in cystic fibrosis.

Animals↗
PubMed1991

Failure of cholinergic stimulation to induce a secretory response from the rectal mucosa in cystic fibrosis.

The secretory response to cholinergic stimulation was investigated in rectal biopsy specimens from children with cystic fibrosis and a control group using a modified Ussing chamber technique. Acetylcholine (10(-3) mol/l) increased the short circuit current in 12 control specimens by mean (SEM) 83.0 (16.4) microA/cm2, but samples from five children with cystic fibrosis failed to exhibit such a response (-1.4 (3.2) microA/cm2). Amiloride (10(-4) mol/l), which will inhibit electrogenic sodium absorption in viable tissues, caused similar reductions in the short circuit current of both control and cystic fibrosis tissues (control = -37.7 (7.7) microA/cm2; cystic fibrosis = -44.0 (9.3) microA/cm2). Thus, the failure of chloride secretion observed in the small intestine also exists in the rectal mucosa. This observation could be used both to aid diagnosis and to study the basic defect.

Acetylcholine↗
PubMed1990

Stimulatory effects of 5-hydroxytryptamine on fluid secretion and transmural potential difference in rat small intestine are mediated by different receptor subtypes.

The rise in transmural potential difference (PD) and the fluid secretion induced by 5-hydroxytryptamine (5-HT) were measured in rat small intestine in-vivo. Both cisapride and ketanserin abolished the 5-HT-induced rise in systolic blood pressure mediated by 5-HT2 receptors. Cisapride inhibited the 5-HT-induced increases in the transintestinal PD, but over the same dose range it had no effect on the fluid secretion induced by 5-HT. In contrast, ketanserin caused a dose-dependent reduction in 5-HT-induced fluid secretion at doses that failed to influence the rise in PD. It is concluded that different receptors are responsible for the effects of 5-HT on fluid secretion and electrical activity in the rat small intestine.

Animals↗