Anis K. Mir

Cardiovascular response to 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) in the rat: site of action and pharmacological analysis.

Summary: The cardiovascular response to 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), a selective putative 5-HT1A receptor agonist, has been investigated in the rat. Comparisons were made with clonidine, a centrally acting hypotensive agent with negligible affinity for 5-HT receptors. In conscious, spontaneously hypertensive (SH) rats, 8-OH-DPAT caused dose-related and sustained falls in blood pressure and heart rate that were unaffected by depletion of brain 5-HT by p-chlorophenylalanine. 8-OH-DPAT caused hypotension and bradycardia in anesthetized normotensive rats. In pithed rats, 8-OH-DPAT neither lowered blood pressure nor affected the cardiovascular response to spinal sympathetic stimulation or to phenylephrine. The response to 8-OH-DPAT was blocked selectively by intracisternal injection of 8-methoxy-2-(N-2-chloroethyl-N-n propyl) amino tetralin (8-MeO-CIEPAT), a putative irreversible 5-HT1A receptor antagonist, and was abolished in animals whose central monoamine transmitter stores were depleted selectively by combined treatment with dl-α-monofluoromethyl-dopa and dopamine. The cardiovascular response to 8-OH-DPAT was inhibited selectively by metergoline, methiothepin, and 8-MeO-CIEPAT; it was nonselectively inhibited by (±)-pindolol, (±)-cyanopindolol, buspirone, yohimbine, idazoxan, and WY 26392; and was unaffected by prazosin and cis-flupenthixol. These results establish that the cardiovascular response to 8-OH-DPAT in the rat is centrally mediated and point to the putative 5-HT1A receptor as the key site involved. An indirect link involving a catecholaminergic mechanism is suggested by the fact that α2-adrenoceptor antagonists are also inhibitory despite 8-OH-DPAT having no direct agonist effects at α2-adrenoceptors per se.

Catecholamines in the carotid body of several mammalian species: Effects of surgical and chemical sympathectomy

The catecholamine content of the carotid body of several mammalian species has been assayed using high performance liquid chromatography coupled to electrochemical detection and radioenzymatic assays. Although there were strain differences in the content of catecholamines in the carotid body of the rat, noradrenaline was equal to or exceeded the dopamine level in this species. No apparent differences were found in carotid bodies of animals killed by cervical dislocation or those dissected from anaesthetized animals. Noradrenaline concentrations were found to be substantially higher than those of dopamine in the cat and guinea-pig carotid body, though dopamine was the predominant amine in the rabbit and ferret. Unilateral superior cervical ganglionectomy or chemical sympathectomy with 6-hydroxydopamine substantially depleted noradrenaline without influencing dopamine in the rat carotid body. A marked selective reduction in noradrenaline was also observed in the rabbit and guinea-pig following ganglionectomy, though similar procedures in the cat failed to alter the levels of either catecholamine in the carotid body. The present data highlights the marked species variation in catecholamine content and the contribution to the latter by sympathetic innervation to this organ. This information will be useful in determining the species specificity regarding the relative roles of dopamine and noradrenaline in the modulation of chemoreceptor afferent discharge.

Radioligand binding study of a series of 5-HT1A receptor agonists and definition of a steric model of this site

Abstract Agonists of the 5-HT1A recognition site have been prepared and their affinities for the [3H]8-OH-DPAT binding site have been determined in rat frontal cortex. A pharmacophore has been developed, based on the reported conformation—activity relationship. Free and forbidden zones of the receptor have been characterized with the assistance of a graphics computer, defining a receptor map which accounts for different chemical classes.

Central and peripheral effects of the dihydropyridine calcium channel activator BAY K 8644 in the rat

Following intraperitoneal (i.p.) administration BAY K 8644 (0.5-4 mg/kg) induced an increase in blood pressure associated with bradycardia, increased tail-flick latency in response to radiant heat, decreased locomotion, induced muscle contraction, postural changes and also reduced reflex activity. Only the postural changes and reduced locomotion were seen after intracerebroventricular administration (5-20 micrograms/kg), suggesting that the other effects are mediated peripherally. All the above effects were antagonised by the calcium channel blocker nifedipine. BAY K 8644 (4 mg/kg i.p.) also significantly increased homovanillic acid and 3,4-dihydroxyphenylacetic acid concentrations in the cortex and striatum, an effect which could also be reversed by nifedipine. Apart from inducing hypotension and tachycardia, nifedipine alone had no effect on any of the above parameters. The analgesic-like activity of BAY K 8644 observed in the tail-flick test appears to be related to its vasoconstrictor effects as the peripherally acting vasodilator phenylephrine had similar analgesic activity. These results show that both central and peripheral dihydropyridine-sensitive calcium channels mediate the effects of BAY K 8644. Although a physiological role for the dihydropyridine-sensitive voltage-operated calcium channel in the CNS remains to be demonstrated, activation of these channels can clearly have functional effects.

Chronic MAO A and MAO B inhibition decreases the 5-HT1A receptor-mediated inhibition of forskolin-stimulated adenylate cyclase

The effect of chronic administration of various monoamine oxidase (MAO) inhibitors on the ability of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) to inhibit forskolin-stimulated adenylate cyclase activity was studied. Groups of 12 rats were given either saline, (E)-beta-fluoromethylene-m-tyrosine (MDL 72394 0.25 mg/kg p.o.), clorgyline (1 mg/kg p.o.), selegiline (1 mg/kg p.o.) or tranylcypromine (5 mg/kg p.o.) once a day for 21 days. Biochemical determinations were made 72 h after the final dose. MDL 72394 and tranylcypromine produced a nonselective inhibition of MAO but clorgyline and selegiline selectively inhibited MAO A and MAO B respectively. All treatments that inhibited MAO A also increased tissue levels of 5-HT. Chronic treatment with MDL 72394, clorgyline or tranylcypromine reduced the ability of 8-OH-DPAT to inhibit forskolin-stimulated adenylate cyclase activity. These data suggest that chronic nonselective and chronic MAO A inhibition causes a down-regulation of the 5-HT1A-mediated inhibition of forskolin-stimulated adenylate cyclase activity.

Monoamine receptor sensitivity changes following chronic administration of MDL 72394, a site-directed inhibitor of monoamine oxidase.

(E)-beta-Fluoromethylene-m-tyrosine (MDL 72394) is not per se an inhibitor of monoamine oxidase (MAO) but is a substrate of aromatic L-amino acid decarboxylase (AADC) which liberates the potent MAO inhibitor (E)-beta-fluoromethylene-m-tyramine (MDL 72392). When co-administered to animals with the peripherally selective AADC inhibitor, carbidopa, MDL 72394 inhibited MAO selectively in the brain. Chronic (14 days plus 3 days withdrawal) administration of 0.5 mg/kg per day p.o. MDL 72394, 0.1 mg/kg per day p.o. MDL 72394 combined with 10 mg/kg per day p.o. carbidopa or 50 mg/kg per day p.o. pargyline produced equivalent inhibition of rat brain MAO and decreased the binding of [3H]clonidine and [3H]RX 781094 to the alpha 2-adrenoceptor and of [3H]dihydroalprenolol to the beta-adrenoceptor without changing binding of [3H]prazosin to the alpha 1-adrenoceptor. The locomotor depressant effect of clonidine was attenuated without attenuation of the hypotensive effect in rats treated chronically with the MAO inhibitors. Neither the sensitivity of the alpha 2-autoreceptor nor of the alpha 2-heteroreceptor was decreased in brain slices. However, the sensitivity of adenylate cyclase to activation by both noradrenaline and isoprenaline was significantly reduced. The number of 5-HT2 and 5-HT1A binding sites was decreased: the 5-HT1B binding sites remained unchanged. The effect of chronic MAO inhibitor treatment on 5-HT1A receptors was associated with a decrease in the behavioural response to 8-hydroxy-2-(di-n-propylamino)tetralin and the decrease in 5-HT2 binding was related to a small reduction in the sensitivity of the inositol phosphate system to stimulation by 5-HT. The lack of effect of chronic MAO treatment on the 5-HT autoreceptor measured in cortical slices corresponded to a lack of effect on the 5-HT1B binding site except that chronic administration of pargyline produced a small but significant decrease in 5-HT autoreceptor sensitivity. Overall, the data show that chronic administration of MDL 72394 has a profile of effects on central monoamine receptor binding and function similar to that seen following chronic administration of a number of clinically effective antidepressants.

Biogenic amine-stimulated cyclic adenosine-3',5'-monophosphate formation in the rat carotid body.

Abstract: The subcutaneous injection of isoprenaline, salbutamol, histamine, and adrenaline to rats, which were subsequently killed by microwave irradiation, resulted in a rapid increase in the cyclic AMP content of the carotid body. On the other hand, noradrenaline, dopamine, adenosine, and 5‐hydroxytryptamine, at doses at least 100 times greater than that of isoprenaline, did not significantly alter the cyclic nucleotide content in vivo. The response to isoprenaline was dose related, with an ED50 of 15 μg · kg−1, and reached a peak level 1–1.5 min after injection. Incubation of intact carotid bodies with isoprenaline (10‐−5M)in vitro also resulted in a 10‐fold increase in cyclic AMP content. The in vivo response to isoprenaline could be blocked stereo‐selectively by propranolol, and ICI 118.551, a β2‐selective antagonist, blocks the isoprenaline‐elicited increase in cyclic AMP completely at a dose of 30 μg · kg−1; whereas betaxolol, a β1‐selective antagonist, was ineffective, even at a dose of 300 μg · kg−1. Hypoxia (5% oxygen in 95% N2) did not result in a significant increase in the cyclic AMP content, nor did it significantly alter the isoprenaline‐stimulated increase in the cyclic AMP content of the rat carotid body. These results suggest that some catecholamines may stimulate cyclic AMP formation by interacting with a β2‐adrenoceptor in the rat carotid body.

Increased sensitivity of rabbit carotid body chemoreceptors to dopamine after chronic treatment with domperidone

An increase in specific dopamine D2 receptor binding sites was observed in membranes prepared from the carotid bodies of rabbits treated for 8 weeks and then withdrawn for 4-9 days from the D2 antagonist domperidone (2-5 mg/kg per day). Recordings of chemoreceptor afferent discharge from the carotid body also revealed that this change in receptor density was accompanied by an increased sensitivity to the chemodepressant effects of exogenous dopamine. The chemoreceptor responsiveness of the carotid body to hypoxia is blunted in rabbits treated chronically with domperidone, but this can be restored to normal by an acute dose of the D2 antagonist. These experiments provide evidence that is compatible with a chemo-inhibitory role for endogenous dopamine in the rabbits carotid body. Furthermore, these results suggest that the carotid body provides a useful model for the functional studies of dopamine D2 receptors.