Mats Jodal

Evaluation of laser Doppler flowmetry in the assessment of intestinal blood flow in cat.

Laser Doppler flowmetry with a differential detector system has been used in the assessment of blood flow in the feline small intestine. Simultaneous mucosal and serosal laser Doppler flowmeter recordings were compared with total blood flow of a bowel segment measured by an optical drop-recorder unit in 6 cats. Blood flow through the muscularis layer was estimated using the 85Kr washout technique. A correlation coefficient of r = 0.96 (mucosal recordings = 90, serosal recordings = 80, p less than 0.001) was obtained between laser Doppler flowmeter output signals and total blood flow at different levels of vascular tone, regardless of whether the flowmeter recordings were made from the mucosal or the serosal side of the bowel wall. At intense vasodilation, the flowmeters showed a tendency to underestimate blood flow. The flowmeter signals were at variance with the muscularis blood flow but were clearly correlated to the calculated mucosal-submucosal blood flow. The uneven blood flow distribution in the intestinal wall did not affect the ability of the flowmeters to reflect total blood flow from either side of the bowel wall. A calibration curve could be constructed for approximate interpretation of the laser Doppler signals in absolute flow units. However, further experiments in humans and further development of the technique must be performed to elucidate clinical applications of the method.

5-Hydroxytryptamine and Cholera Secretion: Physiological and Pharmacological Studies in Cats and Rats

The intestinal secretion evoked by close intra-arterial infusion of 5-hydroxytryptamine (5-HT) in cats was inhibited by tetrodotoxin, a drug abolishing action potentials. Furthermore, the intestinal secretion produced by placing a 2-mM 5-HT solution in the intestinal lumen of rats was inhibited by hexamethonium, a ganglionic receptor-blocking agent. These observations strongly indicate that 5-HT-induced secretion is, at least in part, neurally mediated. It was also shown that 5-HT receptors are involved in the pathophysiology of choleraic secretion, since the secretion was inhibited by making the experimental animal tachyphylactic against 5-HT. No effects of 5-HT tachyphylaxis were noted on fluid transport in normal intestines. The results are discussed in relation to a new hypothesis for the pathophysiology of cholera secretion.

On the Role of Intramural Nerves in the Pathogenesis of Cholera Toxin-induced Intestinal Secretion

Intestinal secretion was produced in anesthetized cats and rats by exposing isolated intestinal segments to cholera enterotoxin. Giving, for example, tetrodotoxin, a nerve-conduction-blocking agent, or adding lidocaine, a local anesthetic agent, to the solution in the intestinal segments markedly inhibited the rate of choleraic secretion, and in most experiments a net absorption of fluid was observed. The results suggest that intramural nervous mechanisms are involved in the pathogenesis of choleraic secretion.

Involvement of the myenteric plexus in the cholera toxin-induced net fluid secretion in the rat small intestine.

BACKGROUND The enteric nervous system is responsible in vivo for most of the change in fluid transport induced by cholera toxin. The aim of the present study was to investigate the importance of the myenteric plexus in the Intramural reflex responsible for this secretion. METHODS Long-term ablation of the myenteric plexus was achieved by serosal application of benzalkonium chloride on jejunal segments in rats. RESULTS The treated segments without functioning myenteric plexus showed a normal net fluid absorption. Cholera toxin in this segment only induced a reduction of fluid absorption, whereas in a nontreated ileal segment it concomitantly induced a conspicuous net fluid secretion. Intravenous hexamethonium did not change the cholera toxin response in the treated jejunal segments, whereas vasoactive intestinal polypeptide elicited a marked secretion. CONCLUSIONS Benzalkonium chloride treatment eliminated the ability of cholera toxin to induce intestinal secretion. Thus, all afferent fibers in the intramural secretory reflex activated by cholera toxin are probably conveyed via the myenteric plexus, which functions as the integrating center in the enteric nervous system. The Ussing chamber technique using stripped intestinal preparations cannot be used when studying effects of luminal secretagogues.

Involvement of serotonin and calcium channels in the intestinal fluid secretion evoked by bile salt and cholera toxin

The enteric nervous system (ENS) is activated when exposing the intestinal mucosa to cholera toxin or certain bile salts. Cholera toxin stimulates ENS, at least in part, by the release of 5‐hydroxytryptamine (5‐HT) from the enterochromaffin cells. Calcium channel blockers of the L‐type markedly attenuate the fluid secretion and the luminal release of 5‐HT caused by cholera toxin. The objective of the present study was to elucidate if sodium deoxycholate activated ENS in a similar manner as cholera toxin. Furthermore, the effect of several calcium channel blockers was tested on the fluid secretion caused by cholera toxin or bile salt. Sodium deoxycholate (4 mM) caused a release of 5‐HT into the intestinal lumen, which was inhibited by calcium channel blockade. Granisetron, a 5‐HT3 receptor blocker, partly inhibited the fluid secretion caused by bile salt. The effects of nifedipine, felodipine, R‐felodipine, H186/86 (t‐butyl analogue of felodipine) on the fluid secretion caused by cholera toxin or sodium deoxycholate were studied. Both secretory states were markedly attenuated in a dose dependent manner by all calcium channel blockers tested regardless of their effects on arterial pressure. It is concluded that both cholera toxin and bile salt activate ENS, at least in part, via a release of 5‐HT from the enterochromaffin cells. The antisecretory effect calcium channel blockers is partly explained by an inhibition of this release of 5‐HT.

Blood Flow in the Small Intestine of Cat and Man as Analyzed by an Inert Gas Washout Technique

Using a recently developed 85Kr-elimination technique, blood flow and flow distribution of the human small intestine have been investigated in patients during abdominal surgery. Total intestinal blood flow was estimated to be 38 +/- 4 ml per min and 100 g of intestinal tissue (mean +/- SE: n = 19), jejunal blood flow being slightly higher than ileal. In 14 of these patients muscularis blood flow was determined to be 21 +/- 2 ml per min and 100 g of muscularis tissue and mucosal-submucosal blood flow was calculated to be 51 +/- 6 ml per min and 100 g of mucosal-submucosal tissue. Seventy-five +/- 3% of total blood flow was distributed to the mucosa-submucosa and the remaining 25 +/- 3% to the muscularis. It was demonstrated that an increasing fraction of flow was diverted to the mucosa-submucosa with enhanced total intestinal blood flow. The human gut exhibited great qualitative and quantitative circulatory similarities with the feline small intestine.

The Importance of the Enteric Nervous System for the Bile-Salt-Induced Secretion in the Small Intestine of the Rat

We have investigated the possible involvement of the enteric nervous system in the intestinal secretion induced by sodium deoxycholate. Hexamethonium, lidocaine, and tetrodotoxin significantly inhibited the fluid secretion in extrinsically innervated and denervated rat jejunal segments. Atropine had no effect. We conclude that the sodium-deoxycholate-induced intestinal secretion is partly caused by the activation of local nervous reflexes.

THE ENTERIC NERVOUS SYSTEM AND CHOLERA TOXIN-INDUCED SECRETION

This article reviews briefly some general aspects of the enteric nervous system (ENS). Furthermore, the ENS control of epithelial transport is exemplified by a description of the enteric nervous reflexes activated by cholera toxin.

COLONIC BLOOD FLOW IN CAT AND MAN AS ANALYZED BY AN INERT GAS WASHOUT TECHNIQUE

An inert gas elimination technique for studying blood flow and flow distribution in the colon was developed on the cat and applied to patients during abdominal surgery. The method involves recording of the elimination of intra-arterially injected 85Kr from a colonic segment whereby the gamma- and beta-radiation of the tracer is registered simultaneously by a scintillation detector and Geiger-Muller tube, respectively. Total blood flow was determined from the recording of gamma-radioactivity using a modification of Zierlers formula, and muscularis blood flow as calculated according to Kety from the mono-exponential elimination recorded by the Geiger-Muller tube. The relative weights of the muscularis and mucosa-submucosa were determined from dissections or histological sections. With these weights and total and muscularis blood flows, flow in the mucosa-submucosa was calculated. Total colonic blood flow was 18 +/- 2 ml per min and 100 g of colonic tissue (mean +/- SE; n = 21). In the muscularis layer blood flow amounted to 11 +/- 1 ml per min and 100 g of muscularis tissue (n = 12), and flow in the mucosa-submucosa was calculated to be 28 +/- 5 ml per min and 100 g of mucosal-submucosal tissue. A major fraction (66 +/- 6%) of total blood flow was distributed to the mucosa-submucosa. As total colonic blood flow was enhanced the increase in flow was diverted only to the mucosa-submucosa. The feline and human large bowel exhibited great qualitative and quantitative circulatory similarities.

Intestinal Epithelial Stem/Progenitor Cells Are Controlled by Mucosal Afferent Nerves

Background The maintenance of the intestinal epithelium is of great importance for the survival of the organism. A possible nervous control of epithelial cell renewal was studied in rats and mice. Methods Mucosal afferent nerves were stimulated by exposing the intestinal mucosa to capsaicin (1.6 mM), which stimulates intestinal external axons. Epithelial cell renewal was investigated in the jejunum by measuring intestinal thymidine kinase (TK) activity, intestinal 3H-thymidine incorporation into DNA, and the number of crypt cells labeled with BrdU. The influence of the external gut innervation was minimized by severing the periarterial nerves. Principal Findings Luminal capsaicin increased all the studied variables, an effect nervously mediated to judge from inhibitory effects on TK activity or 3H-thymidine incorporation into DNA by exposing the mucosa to lidocaine (a local anesthetic) or by giving four different neurotransmitter receptor antagonists i.v. (muscarinic, nicotinic, neurokinin1 (NK1) or calcitonin gene related peptide (CGRP) receptors). After degeneration of the intestinal external nerves capsaicin did not increase TK activity, suggesting the involvement of an axon reflex. Intra-arterial infusion of Substance P (SP) or CGRP increased intestinal TK activity, a response abolished by muscarinic receptor blockade. Immunohistochemistry suggested presence of M3 and M5 muscarinic receptors on the intestinal stem/progenitor cells. We propose that the stem/progenitor cells are controlled by cholinergic nerves, which, in turn, are influenced by mucosal afferent neuron(s) releasing acetylcholine and/or SP and/or CGRP. In mice lacking the capsaicin receptor, thymidine incorporation into DNA and number of crypt cells labeled with BrdU was lower than in wild type animals suggesting that nerves are important also in the absence of luminal capsaicin, a conclusion also supported by the observation that atropine lowered thymidine incorporation into DNA by 60% in control rat segments. Conclusion Enteric nerves are of importance in maintaining the intestinal epithelial barrier.