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Simultaneous recordings of blood flow in the right and left aorta and carotid and coeliac artery were made in the crocodile, Crocodylus porosus at rest and during various stimuli. In resting animals the right aorta and carotid artery flow profiles resembled the recordings obtained in the caiman (Axelsson et al. 1989a), with an anterograde blood flow throughout the cardiac cycle. As in the caiman, the left aorta flow profile was complex with both anterograde and retrograde blood flow during the cardiac cycle, and a net left aorta blood flow near zero at rest. The coeliac artery blood flow profile did not show the complex pattern seen in the upper aorta, immediately suggesting that most of the coeliac artery blood originates elsewhere. We believe that coeliac artery blood flow in the resting animal derives from the right aorta via the abdominal anastomosis between the two aortas. Feeding induced an increase in the coeliac artery and left aorta blood flow, probably due to a decrease in visceral vascular resistance, and hence coeliac arterial and the left aorta blood pressure, which facilitates blood flow (from right to left aorta) through the foramen of Panizza. During short 'fright dives', heart rate fell and there was a decrease in the recorded blood flows: carotid artery blood flow did not decrease to the same extent as the RAo and coeliac artery flow, indicating some capacity for redistribution of blood to the cephalic circuits during diving. Similarly, a near-unimpaired carotid artery blood flow was maintained after adrenaline injection. Substance P increased the coeliac artery blood flow and produced a right-to-left cardiac shunt, probably by construction of the pulmonary vasculature.

Falck-Hillarp fluorescent histochemistry, analyses of catecholamine content in different organs and experiments with drug effects on isolated strip preparations were used to study the adrenergic system of the Florida spotted gar, Lepisosteus platyrhincus. The study reveals a well developed system of adrenergic nerve terminals, especially in blood vessels and in the wall of the sinus venosus and atrium. The lung (swimbladder) receives an excitatory cholinergic nerve supply which controls both smooth and striated muscle in the trabeculae of the lung. No conclusive evidence for an adrenergic inhibitory innervation of the lung was obtained. Chromaffin cells line the walls of the posterior cardinal veins, similar to the situation in the teleosts, and the dominating catecholamine in the chromaffin tissue and in the blood plasma is adrenaline. The concentration of adrenaline in the blood plasma is high enough to affect the contraction force of the heart, and circulating adrenaline may thus contribute to the adrenergic control of the heart in this species.

The effect of vasoactive intestinal polypeptide (VIP) on regional blood flows was studied with labeled microspheres in albino rabbits. Intravenous injection of 500 ng VIP/kg b.w. during 100 s did not change the arterial blood pressure significantly, but caused a rise in intraocular pressure (IOP) and an increase in the choroidal blood flow by 35%, while the blood flow through the anterior uvea was unaffected. The most pronounced vasodilation was observed in the pancreas, the thyroid gland and the parotid gland. In these tissues local blood flow increased by more than 100%. Other tissues, in which this dose of VIP produced vasodilation, were the submandibular gland, the eyelids, the nictitating membrane, the choroid plexus and the heart muscle. Ganglionic or muscarinic blockade had little or no effect on the VIP-induced vasodilation in most of the tissues. Intracameral injection of VIP (1 microgram) produced vasodilation in the iris and the ciliary body, but did not affect IOP. VIP had no apparent effect on the pupil size or the blood-aqueous barrier. In experiments with direct blood flow determination from an opened vortex vein intravenous infusion of VIP, 100 ng X kg-1 X min-1 b.w., during five minutes reduced the uveal vascular resistance by about 50%. This study shows that VIP is a potent vasodilator in many tissues at doses hardly affecting the arterial blood pressure and supports the suggestion, that VIP is responsible for the non-cholinergic vasodilation in the eye caused by facial nerve stimulation.

The effects of drugs and nerve stimulation on the spleen of 2 species of dogfish have been examined by experiments with perfused spleens and isolated spleen and artery strips. Adrenaline, noradrenaline and phenylephrine, acting via alpha adrenoceptors, constrict the perfused spleen of both species, thereby releasing erythrocytes, and contract the isolated spleen and artery strips. Phenotolamine competitively antagonizes the excitatory effects of adrenergic agonists. The responses of the spleen to acetylcholine are very irregular, and a rapid desensitization makes evaluation of the mode of action of this drug difficult. In the artery strips acetylcholine produces a dose-dependent contraction. Fluorescent histochemistry reveals well developed adrenergic innervation of the arteries, and a few adrenergic terminals in the spleen. Stimulation of splenic nerves produces normally splenoconstriction in Squalus, which can be blocked by phentolamine but not by atropine. The nervous control of the Scyliorhinus spleen seems to be poor or lacking. It is concluded that the dogfish spleen, and maybe also the arteries, are to a large extent controlled by circulating catecholamines and (in Squalus) also by sympathetic adrenergic fibres.

The activity of phenylethanolamine-N-methyl transferase (PNMT) has been measured in the chromaffin tissue of the head kidney and in the sympathetic neurons of the coeliac ganglion/splanchnic nerve in the cod. The content of adrenaline and noradrenaline in these tissues and in other adrenergically innervated tissues and blood plasma was also determined. Adrenaline dominates over noradrenaline in most sympathetically innervated tissues, in the chromaffin tissue and in blood plasma, but not in the muscularis mucosae of the swimbladder. High PNMT activity was found in the chromaffin tissue in the walls of the posterior cardinal veins, and also in sympathetic neurons. It is concluded that the adrenaline found in the sympathetic nerves may originate from intraneuronal adrenaline synthesis, but also from circulating adrenaline which is taken up and stored.

The activities of dopamine-beta-hydroxylase (DBH, E.C. 1.14.17.1) and phenylethanol-N-methyl transferase (PNMT, E.C.1.1.1.10) were determined in tissue homogenates from the chromaffin tissue containing atrium and intercostal arteries from the African lungfish. Clearly measurable activities of DBH were found in both kinds of tissues, while PNMT activity was ascertained only in the intercostal arteries. The activity of DBH in the atrium indicates a possibility for local synthesis of the large amounts of catecholamines, especially noradrenaline, found in this tissue. Catecholamines synthetized in and released from the chromaffin tissues of Protopterus, may be of great importance for the adrenergic control in this animal in the absence of a well developed sympathetic nervous system.

The activities of dopamine-beta-hydroxylase (DBH; E.C. 1.14.17.1) and phenylethanolamine-N-methyl transferase (PNMT; E.C. 2.1.1.10) were estimated in homogenates from the heart, the lung and the posterior cardinal veins from Lepisosteus platyrhincus. Both enzymes could be detected in all three tissues, the activity being highest in the chromaffin tissue of the posterior cardinal veins. The activity of DBH estimated in the present in vitro experiments was about 100-fold higher than the PNMT activity. Preliminary experiments with substrate specificity for the PNMT shows a low (heart) or undetectable (lung and cardinal veins) methylation of phenylethylamine, and the specific PNMT inhibitor SK&F 64139 (1 micro M) lowered the PNMT activity in all three tissues by 55-75%. The presence of PNMT in the adrenergically innervated tissues puts Lepisosteus with the teleosts and the amphibians among vertebrates with capacity for truly adrenergic, as opposed to noradrenergic, transmission.

The neuronal and extraneuronal uptake of adrenaline and noradrenaline in the isolated perfused spleen of the cod has been studied using radiolabelled amines. The neuronal uptake of adrenaline shows an optimum at 0.5 microgram/ml, but is less efficient than for noradrenaline in the concentration range 0.01-2.5 micrograms/ml. The neuronal uptake of noradrenaline accounts for 60-70% of the total (neuronal plus extraneuronal) uptake in the concentration range 0.01-1.0 microgram/ml. At higher concentrations (2.5-5.0 micrograms/ml) the importance of the extraneuronal uptake increases for both amines, to reach more than 90% of the total uptake at 5.0 micrograms/ml. The extraneuronal uptake shows equal efficiency for the two amines throughout the concentration range studied. It is concluded that the cod spleen possesses a neuronal uptake mechanism which shows a higher uptake efficiency for noradrenaline than for adrenaline, while the extraneuronal uptake mechanism shows no preference.

The autonomic nerve control of the swimbladder of the goldsinny wrasse has been studied by experiments with isolated strips from the muscularis mucosae, drug effects on gas secretion in vivo, fluorescent histochemistry and quantitative analysis of catecholamine content. Vagotomy, atropine and mecamylamine significantly inhibited induced gas secretion, suggesting a cholinergic vagal secretory innervation of the gas gland. The anterior part of the mucosa (secretory part) is contracted by acetylcholine and alpha-adrenoceptor agonists. These responses are competitively antagonized by atropine and phentolamine or yohimbine respectively. The posterior (resorbent) part of the mucosa is contracted by acetylcholine, but beta-adrenoceptor agonists relax the acetylcholine-precontracted preparations. These responses are competitively blocked by atropine and propranolol respectively. The pA2-values from all experiments with antagonists are well comparable with values obtained for the same drugs in other teleost or mammalian tissues. Tyramine, 5-hydroxytryptamine or phenylephrine had mixed effects on the posterior part. This effect is at least in part due to release of nervously stored catecholamines. Fluorescent histochemistry revealed a strong innervation by both smooth and varicose adrenergic fibres in all parts of the swimbladder, noradrenaline being the dominant catecholamine. Yellow fluorescent cells, which may contain 5-hydroxytryptamine, were also seen in the swimbladder mucosa. No conclusive evidence for a cholinergic innervation of the muscularis mucosae was obtained.

The axonal transport of dopamine-beta-hydroxylase (DBH; E.C. 1.14.17.1) was studied in the splanchnic nerve of the cod in vivo, and the subcellular localization of the same enzyme was studied in the chromaffin tissue from the cod head kidney. The mean rate of axonal transport for cod DBH was 18.6 mm/24 h at 10 degrees C. The mobile fraction was estimated to 22%, giving an absolute rate of transport of 85 mm/24 h at 10 degrees C. Evidence for a retrograde transport of DBH was also obtained, with an accumulation distal to a ligature of 12% of the accumulation proximal to the ligature at 3 days. DBH from the chromaffin tissue appeared to be strongly bound to the adrenergic granules, with only a small amount (ca 4%) recovered in the soluble phase.

The distribution of chromaffin tissue was studied in the African lungfish using Falck-Hillarp fluorescent histochemistry together with quantitative analysis of catecholamines in plasma and tissue extracts. Intensely fluorescent cells form chromaffin tissue in the wall of the atrium, the wall of the most anterior part of the left cardinal vein and the walls of the segmentally arranged intercostal arteries. The arrangement thus appears to be a combination of the situations in cyclostomes, elasmobranchs and teleosts. Adrenaline is present in larger quantities than noradrenaline in the intercostal arteries and the cardinal vein, while noradrenaline dominates in the atrium. During "stress" induced by physical disturbance of the animals, a strong increase in especially the noradrenaline concentration of the plasma was detected.

Effects of adrenergic and cholinergic drugs were studied on isolated preparations from the heart, the lung and the spleen of the African lungfish. In addition, a nerve-lung preparation was employed for the study of the autonomic nervous control of the lung. Falck-Hillarp fluorescent histochemistry was used in the search for adrenergic neurons in the sympathetic chain and tissues. The Protopterus lung received a cholinergic excitatory innervation via the pulmonary branches of the vagi. The presence of parasympathetic ganglia in the lung tissue is indicated. Adrenergic drugs, when producing a response, either contracted or relaxed lung strips. Spleen strips were contracted by cholinergic drugs acting on muscarinic receptors and, to a lesser extent, by adrenergic drugs. A strong negative inotropic effect was produced by carcachol on paced atrial but not ventricular strips, while adrenaline increased the contraction force in ventricular but not atrial strips. The sympathetic ganglion cells did not contain enough catecholamines to give off any visible fluorescence, nor was it possible to show any adrenergic nerve terminals in the tissues studied. It is concluded that the adrenergic control in the lungfish is mainly due to circulating catecholamines, rather than adrenergic nerve fibres, since concentrations of catecholamines in protopterus plasma are high enough to affect the spleen, lung and ventricle.