Reply from Vladimir V. Matchkov and Christian Aalkjaer : Letter

Key points Substantial information on rat mesenteric small artery physiology and pharmacology based on in vitro experiments is available. Little is known about the relevance of this for artery function in vivo . We here present an intravital model where rat mesenteric small artery diameters are studied under isolated and controlled conditions in situ with simultaneous measurement of blood flow. The responses of the isolated arteries vary with the anaesthetic used, and they are quantitatively but not qualitatively different from the responses seen in vitro . Abstract Functional characteristics of rat mesenteric small arteries (internal diameter ∼150–200 μm) have been extensively studied in vitro using isometric and isobaric myographs. In vivo , precapillary arterioles (internal diameter < 50 μm) have been studied, but only a few studies have investigated the function of mesenteric small arteries. We here present a novel approach for intravital studies of rat mesenteric small artery segments (∼5 mm long) isolated in a chamber. The agonist‐induced changes in arterial diameter and blood flow were studied using video imaging and laser speckle analysis in rats anaesthetized by isoflurane, pentobarbital, ketamine–xylazine, or by a combination of fentanyl, fluanison and midazolam (rodent mixture). The arteries had spontaneous tone. Noradrenaline added to the chamber constricted the artery in the chamber but not the downstream arteries in the intestinal wall. The constriction was smaller when rats were anaesthetized by rodent mixture in comparison with other anaesthetics, where responses were qualitatively similar to those reported in vitro . The contraction was associated with reduction of blood flow, but no flow reduction was seen in the downstream arteries in the intestinal wall. The magnitude of different endothelium‐dependent relaxation pathways was dependent on the anaesthesia. Vasomotion was present under all forms of anaesthesia with characteristics similar to in vitro . We have established an intravital method for studying the tone and flow in rat mesenteric arteries. The reactivity of the arteries was qualitatively similar to the responses previously obtained under in vitro conditions, but the choice of anaesthetic affects the magnitude of responses. Substantial information on rat mesenteric small artery physiology and pharmacology based on in vitro experiments is available. Little is known about the relevance of this for artery function in vivo . We here present an intravital model where rat mesenteric small artery diameters are studied under isolated and controlled conditions in situ with simultaneous measurement of blood flow. The responses of the isolated arteries vary with the anaesthetic used, and they are quantitatively but not qualitatively different from the responses seen in vitro .

1. Blood pressure has been measured in the aorta and at four points in the mesenteric circulation of conscious, freely moving rats under physiological, resting conditions. 2. Using small polythene catheters, blood pressure was measured simultaneously in the aorta and either distally in the superior mesenteric artery (group A), at the base of a mesenteric arterial arcade (vessel diameter ca 100 microns) (group B), at the base of a mesenteric venous arcade (group C) or distally in the superior mesenteric vein (group D). Local blood flow distribution proximal and distal to the measurement point was restored after the cannulations through appropriate ligations. 3. In conscious animals 5-17 h after surgery, systemic mean blood pressure was 121 +/- 2 mmHg. Local pressures at the four locations (as a percentage of systemic pressure) were: 95 +/- 1% in group A, 64 +/- 2% in group B, 13 +/- 1% in group C and 7 +/- 1% in group D. Thus, large arteries dissipated 5% of the total pressure drop, arcade small arteries 31%, the intramural circulation 51%, arcade veins 6% and the remaining veins plus the hepatic circulation 7%. 4. Immediately after surgery, the corresponding pressure drops were 4, 16, 66, 5 and 9%, respectively, thus emphasizing that the pressure profile can be profoundly affected by surgery and anaesthesia. 5. The data indicate that under resting conditions in conscious, freely moving rats, half the mesenteric vascular resistance resides outside the intramural circulation, primarily in the arcade small arteries.