Apparent total tract energy and macronutrient digestibility of one- to three-day-old, adult ground, extruded, and canned chicken-based diets in domestic cats (Felis silvestris catus).

There has been a recent increase in the popularity of feeding unconventional diets, including whole prey diets, to domestic cats (Felis silvestris catus). Data are needed that allow animal caretakers to choose and formulate diets that meet the nutritional requirements of their cats. Our objective was to evaluate the effects of feeding 1- to 3-d-old whole chicks (WHO), ground adult chicken product (GRO), a chicken-based canned diet (CAN), and a chicken-based extruded diet (EXT) on apparent total tract energy and macronutrient digestibility, N balance, and blood metabolites of domestic cats (n = 11). Macronutrient, energy, and moisture concentrations of diets varied greatly (e.g., CP: 35 to 72% DM); however, cats fed all diets maintained BW and N balance. In general, cats fed WHO had lower nutrient digestibility than those fed CAN and EXT. Cats fed GRO had greater nutrient digestibility than cats fed commercial diets. For example, apparent OM and GE digestibility coefficients were greater (P ≤ 0.05) for cats fed CAN (86 and 88%, respectively), EXT (88 and 88%), and GRO (94 and 95%) compared with those fed WHO (83 and 83%) and greater (P ≤ 0.05) for cats fed GRO compared with those fed CAN and EXT. Many blood metabolites were modified by diet, but most remained within reference ranges for domestic cats. Serum cholesterol was elevated above the reference range for all treatments and greater (P ≤ 0.05) for cats fed WHO compared with those fed CAN, EXT, and GRO. Serum creatinine concentrations were above the reference range for all treatments and greater (P ≤ 0.05) for cats fed GRO compared with those fed CAN or WHO. These data indicate that the whole prey tested herein maintained short-term health and are adequately digestible for use in companion animal diets. Research is needed to determine the global and long-term health implications of feeding whole or ground diets to domestic cats, which may be different in terms of macronutrient, energy, and moisture profiles and nutrient digestibility.

Influence of dietary fiber type and amount on energy and nutrient digestibility, fecal characteristics, and fecal fermentative end-product concentrations in captive exotic felids fed a raw beef-based diet.

Little nutritional or metabolic information has been collected from captive exotic cats fed raw diets. In particular, fiber types and concentrations for use in raw meat-based diets for captive exotic felids have not been well studied. Our objective was to evaluate the effects of fiber type and concentration on apparent total tract energy and macronutrient digestibility, fecal characteristics, and fecal fermentative end-products in captive exotic felids. Four animals of each captive exotic species (jaguar (Panthera onca), cheetah (Acinonyz jubatus), Malayan tiger (Panthera tigris corbetti), and Siberian tiger (Panthera tigris altaica) were randomized in four 4 × 4 Latin square designs (1 Latin square per species) to 1 of the 4 raw beef-based dietary treatments (94.7 to 96.7% beef trimmings): 2 or 4% cellulose or 2 or 4% beet pulp. Felid species, fiber type, and fiber concentration all impacted digestibility and fecal fermentative end-products. Inclusion of beet pulp increased (P ≤ 0.05) fecal short-chain fatty acids and fecal output in all cats. Inclusion of 2 and 4% cellulose, and 4% beet pulp increased (P ≤ 0.05) fecal bulk and diluted fecal branched-chain fatty acid concentrations compared with 2% beet pulp. Apparent total tract DM, OM, fat, and GE digestibility coefficients decreased (P ≤ 0.05) linearly with BW of cats. Additionally, fecal moisture, fecal score, and concentrations of fermentative end-products increased (P ≤ 0.05) with BW. Although the response of many outcomes was dependent on cat size, in general, beet pulp increased wet fecal weight, fecal scores, and fecal metabolites, and reduced fecal pH. Cellulose generally reduced DM and OM digestibility, but increased dry fecal weight and fecal percent DM. Although beet pulp and cellulose fibers were tested individually in this study, these data indicate that the optimum fiber type and concentration for inclusion in captive exotic felid diets is likely a combination of fermentable and nonfermentable fibers, with the optimal fiber blend being dependent on species. Smaller cats, such as cheetahs and jaguars, tolerated fermentable fibers, whereas larger cats, such as Malayan and Siberian tigers, appeared to require more insoluble fibers that limit fermentation and provide fecal bulk. Further research is required to test whether these trends hold true when fed in combination.

Evaluation of four raw meat diets using domestic cats, captive exotic felids, and cecectomized roosters.

Our objective was to evaluate raw meat diets for captive exotic and domestic carnivores containing traditional and alternative raw meat sources, specifically, beef trimmings, bison trimmings, elk muscle meat, and horse trimmings. We aimed to examine diet composition and protein quality; apparent total tract energy and macronutrient digestibility in domestic cats, African wildcats, jaguars, and Malayan tigers; and ME and fecal fermentative end-products in domestic cats. Because of variation in the meat sources, dietary proximate, AA, and long-chain fatty acid composition were variable. Our analyses indicated that all diets had essential fatty acid deficiencies, and the elk diet (i.e., trimmed muscle meat) was deficient in total fat. Standardized AA digestibilities measured using the cecectomized rooster assay were high (>87%). Using the NRC minimum requirements for the growth of kittens, the first limiting AA of all diets was the combined requirement of Met and Cys (AA score: 81 to 95; protein digestibility corrected AA score: 75 to 90). All diets were highly digestible (88 to 89% OM digestibility). There was no effect of diet or felid species on DM (85 to 87%), OM, and GE (90 to 91%) digestibilities. Apparent CP digestibility was greater (P≤0.05) in cats fed elk (97%) compared with those fed bison (96%), and greater (P≤0.05) in wildcats (97%) and domestic cats (97%) compared with tigers (95%). The diet and species interaction (P≤0.05) was observed for apparent fat digestibility. In domestic cats, the fresh fecal pH and proportions of acetate and butyrate were altered (P≤0.05) due to diet. Diet also affected (P≤0.05) fresh fecal concentrations of total branched-chain fatty acids, valerate, and Lactobacillus genus. In conclusion, although the raw meat diets were highly digestible, because of variation in raw meat sources the nutrient composition of the diets was variable. Thus, compositional analysis of raw meat sources is necessary for proper diet formulation. The types of meat commonly used in raw meat diets may be deficient in total fat (trimmed muscle meat) and essential fatty acids (trimmings and muscle meats). Additionally, differences in raw meat source nutrient composition and digestibility affect the beneficial and putrefactive fermentative end-products found in feces.

Effects of chronic sympatho-inhibition on reflex control of renal blood flow and plasma renin activity in renovascular hypertension.

BACKGROUND AND PURPOSE: We determined if chronic sympatho-inhibition with rilmenidine has functional significance for the kidney by altering responses of renal blood flow (RBF) and plasma renin activity (PRA) to stress and acute hypotension in rabbits with renovascular hypertension. EXPERIMENTAL APPROACH: RBF to each kidney and renal sympathetic nerve activity (RSNA) to the left kidney were measured in rabbits in which a renal artery clip induced hypertension (2K1C) and in sham-operated rabbits. After 2 weeks, a subcutaneous minipump was implanted to deliver rilmenidine (2.5 mg.kg(-1).day(-1)) to 2K1C rabbits for 3 weeks. KEY RESULTS: After 5 weeks of renal artery stenosis, mean arterial pressure (MAP) was 23% higher and PRA 3-fold greater than in sham-operated rabbits. Blood flow and renal vascular conductance in the stenosed kidney were lower (-75% and -80%) compared with sham, and higher in the non-clipped kidney (68% and 39%). Responses of RBF and PRA to hypotension were similar in 2K1C and sham rabbits. Airjet stress evoked a greater increase in MAP in 2K1C rabbits than sham controls. Chronic rilmenidine normalized MAP, reduced RSNA and PRA, and did not reduce RBF in the stenosed kidney. Responses of RBF (clipped and non-clipped kidney), RSNA and PRA to hypotension and airjet were little affected by rilmenidine. CONCLUSIONS AND IMPLICATIONS: Our observations suggest that chronic sympatho-inhibition is an effective antihypertensive therapy in renovascular hypertension. It normalizes MAP and reduces basal PRA without compromising blood flow in the stenosed kidney or altering responses of MAP, haemodynamics and PRA to acute hypotension and stress.

Imidazoline receptors associated with noradrenergic terminals in the rostral ventrolateral medulla mediate the hypotensive responses of moxonidine but not clonidine.

We determined whether the cardiovascular actions of central anti-hypertensive agents clonidine and moxonidine are dependent on noradrenergic or serotonergic innervation of the rostral ventrolateral medulla (RVLM) in conscious rabbits. 6-Hydroxydopamine (6-OHDA) or 5,6-dihydroxytriptamine (5,6-DHT) was injected into the RVLM to deplete noradrenergic and serotonergic terminals respectively. One, 2 and 4 weeks later, responses to fourth ventricular (4V) clonidine (0.65 microg/kg) and moxonidine (0.44 microg/kg) were examined. Destruction of noradrenergic pathways in the RVLM by 6-OHDA reduced the hypotensive response to 4V moxonidine to 62%, 47% and 60% of that observed in vehicle treated rabbits at weeks 1, 2 and 4 respectively. The moxonidine induced bradycardia was similarly attenuated (to 46% of vehicle). Conversely, 6-OHDA had no effect on the hypotensive or bradycardic effects of 4V clonidine. Efaroxan (I(1)-imidazoline receptor/alpha(2)-adrenoceptor antagonist; 3.5, 11, 35 microg/kg) and 2-methoxyidazoxan (alpha(2)-adrenoceptor antagonist; 0.3, 0.9, 3 microg/kg) equally reversed the hypotension to 4V clonidine, suggesting a mainly alpha(2)-adrenoceptor mechanism. Efaroxan preferentially reversed responses to moxonidine in both vehicle and 5,6-DHT groups and in the 1st week after 6-OHDA, suggesting a mechanism involving mainly I(1)-imidazoline receptors. This selectivity was subsequently lost in the 2nd and 4th weeks when the remaining hypotension was mainly mediated by alpha(2)-adrenoceptors. Depletion of serotonergic terminals did not alter the responses to either agonist nor did it change the relative effectiveness of the antagonists. Western blots of RVLM tissues probed with imidazoline and alpha(2)-adrenoceptor antisera showed a pattern of bands close to that reported in other species. The main effect of 6-OHDA was an 18% lower level of the 42 kDa imidazoline protein (P<0.05). We conclude that the hypotensive and bradycardic actions of moxonidine but not clonidine are mediated through imidazoline receptors and are dependent on intact noradrenergic pathways within the RVLM. Furthermore, the noradrenergic innervation may be associated with a 42 kDa imidazoline receptor protein.

Involvement of imidazoline receptors in the baroreflex effects of rilmenidine in conscious rabbits.

OBJECTIVE: It has been suggested that imidazoline receptors rather than alpha2-adrenoceptors are involved in the sympathoinhibitory action of centrally acting antihypertensive drugs such as rilmenidine. In the present study, we examined the relative importance of alpha2-adrenoceptors and imidazoline receptors in modulating the renal sympathetic and heart rate (HR) baroreflex in response to central administration of rilmenidine in conscious normotensive rabbits. METHODS: In seven conscious rabbits, chronically instrumented with a fourth ventricular (4V) catheter, aortic and vena caval cuff occluders and a renal nerve electrode, we continuously recorded renal sympathetic nerve activity (RSNA), mean arterial pressure (MAP) and HR and assessed baroreflex MAP-RSNA and MAP-HR relationships with balloon-induced ramp rises and falls in MAP. Rabbits were treated with 4V rilmenidine (22 microg/kg) followed by 4V idazoxan (30 microg/kg; a mixed alpha2-adrenoceptor and imidazoline receptor antagonist) or 4V 2-methoxy-idazoxan (1 microg/kg; an alpha2-adrenoceptor antagonist with little affinity for imidazoline receptors). RESULTS: Rilmenidine lowered blood pressure by 24% and reduced both upper and lower plateaus of the renal sympathetic baroreflex curve, such that the RSNA range (difference between plateaus) was reduced by 40% (-32 +/- 10 normalized units). Curves were shifted to the left with the fall in MAP. Idazoxan restored MAP, maximum RSNA and the RSNA baroreflex range. By contrast the alpha2-adrenoceptor antagonist 2-methoxy-idazoxan caused only a partial recovery of MAP and RSNA baroreflex upper plateau and range (-9 +/- 2 mmHg, 29 and 33% lower than control). Both antagonists partially restored the HR baroreflex. CONCLUSION: These findings suggest that in conscious rabbits, both imidazoline receptors and alpha2-adrenoceptors are involved in the central antihypertensive and baroreflex actions of rilmenidine, but that activation of imidazoline receptors is more important for its renal sympathoinhibitory action.

Relative importance of rostral ventrolateral medulla in sympathoinhibitory action of rilmenidine in conscious and anesthetized rabbits.

The pressor region of the rostral ventrolateral medulla (RVLM) is a critical site in the sympathoinhibitory action of imidazoline receptor agonists as shown by studies in anesthetized animals. The aim of this study was to compare the importance of the RVLM in mediating the inhibitory action of rilmenidine on renal sympathetic nerve activity (RSNA) and arterial pressure in urethane-anesthetized rabbits (n = 11) and in conscious, chronically instrumented rabbits (n = 6). Bilateral microinjection of rilmenidine (4 nmol in 100 nl) into the RVLM caused a greater decrease in resting arterial pressure in anesthetized animals (-19 mm Hg) than in conscious animals (-8 mm Hg). By contrast, the decrease in resting RSNA evoked by rilmenidine was similar in conscious (-27%) and anesthetized (-36%) rabbits. Furthermore, rilmenidine microinjection into the RVLM was equally effective in inhibiting the RSNA baroreflex in both groups of animals. The upper plateau of the RSNA baroreflex decreased by 37% and 42%, and gain decreased by 41% and 44% after rilmenidine treatments in conscious and anesthetized rabbits, respectively. We conclude that the RVLM plays an equally important role in the inhibitory action of rilmenidine on RSNA in conscious and anesthetized rabbits either at rest or during baroreflex responses. A relatively moderate effect of rilmenidine on arterial pressure in conscious, chronically instrumented rabbits may relate to a lower level of sympathetic drive compared with anesthetized animals.

Renal and cardiac sympathetic baroreflexes in hypertensive rabbits.

1. The purpose of the present study was to assess the changes to renal sympathetic nerve activity (RSNA) baroreflexes during the development of hypertension after renal clipping in conscious rabbits. 2. Rabbits were fitted with a clip on the right renal artery or underwent a sham operation under halothane anaesthesia. A recording electrode was implanted on the left renal nerve 1 week before the experiment, 3 or 6 weeks after the initial operation. During the experiment, drug-induced ramp rises and falls in mean arterial pressure (MAP) were used to produce RSNA and heart rate (HR) baroreflex curves. The RSNA for each experiment was calibrated against maximum RSNA evoked by stimulation of baroreceptor-independent trigeminal afferents. 3. Mean arterial pressure was 20 and 36% higher 3 and 6 weeks after clip implantation, respectively. Renal sympathetic nerve activity baroreflex curves were reset rightwards accordingly, but the shape of the RSNA curves was differentially affected. 4. At both hypertensive periods, MAP-HR baroreflex gain was markedly reduced due to a reduction in curvature. The HR baroreflex range was increased. The RSNA baroreflex gain was reduced at 3 weeks, which was due to a 35% lower RSNA baroreflex range, but was similar to sham animals at 6 weeks. 5. The results show that, in established two kidney, one clip hypertension in rabbits, the sympathetic baroreflex is relatively well preserved but sensitivity of cardiac baroreflexes is attenuated. Therefore, the short-term inhibition of RSNA baroreflexes is not related to the level of blood pressure or the development of secondary changes, such as cardiac or vascular hypertrophy, but may be related to circulating angiotensin, which is known to increase at this time.

Comparison of renal sympathetic baroreflex effects of rilmenidine and alpha-methylnoradrenaline in the ventrolateral medulla of the rabbit.

OBJECTIVE: To determine the influence of imidazoline receptors and alpha2-adrenoceptors in the rostral ventrolateral medulla (RVLM) on the renal sympathetic baroreflex. METHODS: The effects of rilmenidine (4 nmol) and alpha-methylnoradrenaline (alpha-MNA, 80 nmol) micro-injected into the RVLM of urethane-anaesthetized rabbits previously implanted with renal nerve recording electrodes were examined before and after micro-injection of the imidazoline receptor/alpha2-adrenoceptor antagonist idazoxan and the alpha2-adrenoceptor antagonist 2-methoxyidazoxan (2-MI). RESULTS: Rilmenidine and alpha-MNA both lowered mean arterial pressure (MAP) by 28% and renal sympathetic nerve activity (RSNA) by 35%, and reduced RSNA upper plateaus and ranges by 30-70%. Rilmenidine decreased both sympathetic burst frequency and amplitude while alpha-MNA reduced amplitude only. Rilmenidine shifted the RSNA baroreflex curve to the left while alpha-MNA shifted the curve to the right Idazoxan (13 nmol) reversed the hypotension and all RSNA effects of rilmenidine, while 2-MI (4 nmol) increased MAP 18% above the control and also reversed all RSNA parameters. By contrast, 2-MI reversed the alpha-MNA-induced hypotension and partially restored RSNA and the upper plateau of the RSNA baroreflex curve. Idazoxan treatment only partially reversed the hypotension after alpha-MNA and had no effect on any of the baroreflex curves. CONCLUSION: Both alpha-MNA and rilmenidine injected into the RVLM of rabbits produce renal sympathetic inhibition, but differences in the location of the baroreflex curve and the pattern of effects on burst amplitude and frequency suggest different mechanisms of action. The effects of idazoxan suggest that rilmenidine acts via imidazoline receptors. Since 2-MI reversed the actions of alpha-MNA and also rilmenidine, this suggests that alpha2-adrenoceptor hypotension can be produced in the rabbit RVLM and that rilmenidine may activate alpha2-adrenoceptors, possibly as a result of activating imidazoline receptors.

I1 imidazoline receptors in cardiovascular regulation: the place of rilmenidine.

From the very earliest suggestion of a distinction between imidazoline receptors and alpha2-adrenoceptors, there has been much debate as to their contribution to the antihypertensive actions of clonidine-like agents. However, with the development of drugs such as rilmenidine that are more selective for I1 imidazoline receptors, their role and also their close relationship with alpha2-adrenoceptors has become clearer. We have examined this question using a range of imidazoline and alpha2-adrenoceptor antagonists given centrally and peripherally to conscious rabbits. We found that second-generation agents such as rilmenidine preferentially act via imidazoline receptors but that alpha2-adrenoceptors are important for the hypotension produced by the first-generation agents clonidine and alpha-methyldopa. In addition to the hypotension, rilmenidine facilitates cardiac vagal baroreflexes and inhibits cardiac sympathetic baroreflexes and diminishes the increase in renal sympathetic activity produced by environmental stress. In other studies using anesthetized rabbits and direct measures of sympathetic nerve activity, we confirmed that the major site of sympathoinhibitory actions and sympathetic baroreflex effects of rilmenidine is the rostral ventrolateral medulla. Our results also suggest that alpha2-adrenoceptors are activated as a consequence of imidazoline receptor activation by rilmenidine. Thus, though imidazoline receptors appear to be the primary target of rilmenidine, "downstream" alpha2-adrenoceptors within the brainstem are also involved and need to be considered in developing pharmacologic strategies for antihypertensive treatment involving imidazoline agents.

Relationship between imidazoline and alpha2-adrenoceptors involved in the sympatho-inhibitory actions of centrally acting antihypertensive agents.

Since the first suggestion of the existence of imidazoline receptors, there has been a continuing and yet unresolved debate as to their contribution to the antihypertensive actions of clonidine-like agents. In this review we bring together a number of studies from our laboratory which have examined the importance and interdependence of imidazoline receptors and alpha2-adrenoceptors in the mechanism of action of centrally acting antihypertensive drugs. Using conscious rabbits and a range of imidazoline and specific alpha2-adrenoceptor antagonists we have consistently found that second generation agents rilmenidine and moxonidine preferentially act via imidazoline receptors but that alpha2-adrenoceptors are important for the hypotension produced by clonidine and alpha-methyldopa. Despite this difference in receptor mechanism, the hypotension produced by all these drugs is dependent on central noradrenergic pathways. In other studies using anaesthetised rabbits and direct measures of sympathetic nerve activity we confirmed the generally held view that the major site of sympatho-inhibitory actions and sympathetic baroreflex effects of centrally acting antihypertensive agents is the rostral ventrolateral medulla (RVLM). We also found, using microinjection of specific antagonists, that alpha2-adrenoceptors in this nucleus appear to be activated as a consequence of imidazoline receptor activation. Thus, there appears to be a close relationship between imidazoline receptors and alpha2-adrenoceptors located in the RVLM in mediating the hypotension and inhibition of renal sympathetic nerve activity. Furthermore in recent studies using a noradrenergic neurotoxin microinjected into the RVLM we found that this treatment selectively blocked the actions of moxonidine but not clonidine, suggesting that I1-imidazoline receptors may be located on adrenergic terminals in situ. By contrast, clonidine acts predominantly via alpha2-adrenoceptors, perhaps located on cell bodies in the nucleus. We conclude that there is indeed a close nexus between 'presynaptic' imidazoline receptors on noradrenergic terminals and 'downstream' alpha2-adrenoceptors within the RVLM. Our hypothesis brings together opposing points of view that the mechanism for hypotension must involve either the imidazoline receptor or the alpha2-adrenoceptor. Clearly both are important.

Interaction of the dopamine D2 receptor agonist quinpirole with sympathetic vasomotor tone and the central action of rilmenidine in conscious rabbits.

Previous studies in conscious rats have shown that systemic administration of the dopamine D2 receptor agonist quinpirole causes a centrally-mediated increase in blood pressure which is associated with increased plasma levels of noradrenaline and adrenaline. In addition, treatment with quinpirole caused a marked inhibition of the antihypertensive effect of centrally-acting sympatho-inhibitory drugs such as clonidine, rilmenidine and alpha-methyldopa, suggesting an interaction at the level of sympathetic vasomotor tone. The main aim of the present study was investigate in conscious rabbits the effect of quinpirole on renal sympathetic nerve activity. In addition, we studied the effect of pretreatment with quinpirole on responses to additional quinpirole injections or rilmenidine treatment. Quinpirole treatment caused a prolonged dose-dependent increase in blood pressure and heart rate. Additional injection of quinpirole, 30 min after the first treatment, caused a significantly smaller pressor response (7+/-2 vs. 17+/-2 mm Hg). Injection of rilmenidine caused a larger decrease in blood pressure in rabbits which had been pretreated with quinpirole than in controls (-28+/-3 vs. -14+/-3 mm Hg). Total renal sympathetic nerve activity was markedly increased by quinpirole treatment (3.5-fold), an effect which could be attributed to both increased amplitude and increased frequency of the renal nerve signal. After a second injection of quinpirole, 30 min after the first treatment, only total renal sympathetic nerve activity and amplitude were increased and the effects were reduced. These results show marked actions of quinpirole on renal sympathetic nerve activity in conscious rabbits. However, the previously described apparent desensitisation to the antihypertensive effect of rilmenidine could not be observed in rabbits, suggesting marked species differences in the mechanism and site of action of rilmenidine.

Effect of rilmenidine on the cadiovascular responses to stress in the conscious rabbit.

Environmental stress can cause an increase in sympathetic nerve activity both in humans and animals. While centrally acting antihypertensive drugs such as rilmenidine are known to reduce sympathetic tone, it is not clear whether they also influence the cardiovascular responses to acute stress. In the present study we examined the effects of systemic treatment with rilmenidine on the sympathetic and haemodynamic responses to air jet or noise stress. Twelve conscious rabbits previously implanted with a renal nerve recording electrode were subjected to an 8 l/min stream of air directed at their face for 10 min or exposure to 10 min of white noise (approximately 85 dB). Both air jet and noise stress elicited increases in renal sympathetic nerve activity (RSNA) which were greatest in the first minute (+55+/-9% and +40+/-6%, respectively), but which quickly reached a stable level over the subsequent 9 min (+24+/-6% and +9+/-5%, respectively). This was accompanied by a small increase in heart rate (HR) and mean arterial pressure (MAP). Intravenous rilmenidine (273 microg/kg) reduced MAP from 85+/-3 mm Hg to 68+/-2 mm Hg and HR from 203+/-10 b/min to 188+/-10 b/min and lowered basal RSNA by 54%. Rilmenidine reduced the increase in RSNA seen during the first minute of air jet stress by 35% and reduced the average increase over the next 9 min by 68%. However, rilmenidine had little effect on either the initial or stable RSNA responses to noise stress. Saline treatment did not alter the RSNA responses to either air jet or noise stress. The results show that centrally-acting antihypertensive agents not only lower basal RSNA, but can differentially influence environmentally induced sympathetic responses. In addition, the differential effect of rilmenidine on noise and air jet stress suggests that they may involve quite different central processing.

Relative importance of medullary brain nuclei for the sympatho-inhibitory actions of rilmenidine in the anaesthetized rabbit.

OBJECTIVE: To determine the contribution of the rostral ventrolateral medulla and the nucleus of the solitary tract in mediating the attenuation of the renal sympathetic baroreflex produced by administration of rilmenidine to anaesthetized rabbits and to examine the relative contribution of alpha2-adrenoceptors and imidazoline receptors at these sites to the cardiovascular effects of rilmenidine. METHODS AND RESULTS: Rilmenidine micro-injected into the rostral ventrolateral medulla produced hypotension and inhibition of renal sympathetic nerve activity with doses an order of magnitude lower than those required in the nucleus tractus solitarius. Alpha-methylnoradrenaline, however, was similarly potent at producing hypotension when it was injected into the rostral ventrolateral medulla or nucleus tractus solitarius but, unlike rilmenidine, did not lower renal sympathetic nerve activity when it was injected into the nucleus tractus solitarius. The alpha2-adrenoceptor antagonist 2-methoxyidazoxan partially reversed the hypotension and renal sympathetic nerve activity inhibition due to alpha-methylnoradrenaline when it was administered into the rostral ventrolateral medulla, whereas the mixed alpha2-adrenoceptor/imidazoline receptor antagonists, idazoxan and efaroxan, did not. 2-Methoxyidazoxan, but not idazoxan, also reversed the hypotension when alpha-methylnoradrenaline was administered into the nucleus tractus solitarius. The hypotension induced by rilmenidine in the rostral ventrolateral medulla was completely reversed both by 2-methoxyidazoxan and by idazoxan, as was the sympathetic inhibition. To assess any interaction between the nucleus tractus solitarius and the rostral ventrolateral medulla in mediating the baroreflex effects of rilmenidine, we injected rilmenidine into the rostral ventrolateral medulla, the nucleus tractus solitarius or both nuclei and determined renal baroreflex responses of sympathetic nerve activity using drug-induced changes in blood pressure. Injection of 0.5 nmol rilmenidine into the rostral ventrolateral medulla reduced mean arterial pressure and basal renal sympathetic nerve activity as well as renal sympathetic baroreflex range (by 27%) and gain (by 35%). In contrast, injection of rilmenidine into the nucleus tractus solitarius had no effect on basal renal sympathetic nerve activity and renal sympathetic baroreflex parameters. The effect of combined injection was similar to that of administration into the rostral ventrolateral medulla alone. CONCLUSION: Our results show that the rostral ventrolateral medulla, rather than the nucleus tractus solitarius, is the major site involved in the hypotension and inhibition of the renal sympathetic baroreflex by rilmenidine. Comparison of the actions of alpha2-adrenoceptor and imidazoline receptor antagonists on the effects of rilmenidine and alpha-methylnoradrenaline suggests that these agents are acting at different receptors, presumably imidazoline and alpha2-adrenoceptors receptors, respectively, and that both are important in lowering sympathetic tone and blood pressure in the rostral ventrolateral medulla.

Site and receptors involved in the sympathoinhibitory actions of rilmenidine.

INTRODUCTION: It is well accepted that centrally acting antihypertensive agents such as rilmenidine reduce blood pressure through inhibition of the sympathetic nervous system. The central receptor involved, be it a central imidazoline receptor or an alpha2-adrenoceptor and its location, is less certain. METHODS AND RESULTS: The present paper reviews studies from our laboratory examining these questions by using antagonists with differing affinity for imidazoline receptors and alpha2-adrenoceptors. In addition, we have used various routes of administration in conscious and anaesthetized normotensive rabbits. We found that rilmenidine was more potent in its hypotensive action when administered into the fourth ventricle than when given intravenously and considerably more potent when injected into the rostroventrolateral medulla (RVLM) compared to the nucleus of the solitary tract (NTS). By contrast, alpha-methylnoradrenaline, which acts solely through alpha2-adrenoceptors to produce hypotension, was similarly potent in both the NTS and RVLM. Injections of rilmenidine into the RVLM reduced renal sympathetic tone and produced a marked inhibition of renal sympathetic baroreflex responses. The pattern of renal sympathetic baroreflex effects of rilmenidine administered into the RVLM was similar to the effects of the fourth ventricular or intravenous administration. These studies together support the view that the RVLM is the major site of action. We have determined the relative importance of imidazoline receptors and alpha2-adrenoceptors in the inhibition of renal sympathetic nerve activity produced by rilmenidine administered into the RVLM, the fourth ventricle or intravenously. Initial studies in conscious rabbits showed that intravenous or fourth ventricular administration of rilmenidine induced renal sympatho-inhibition which was preferentially reversed by idazoxan or efaroxan (imidazoline/alpha2-adrenoceptor antagonist) compared to 2-methoxyidazoxan (alpha2-adrenoceptor antagonist). In anaesthetized rabbits, administration of idazoxan into the RVLM reversed the inhibition of the renal sympathetic activity induced by RVLM or intravenous administration of rilmenidine. In contrast, idazoxan had no effect on the responses produced by the alpha2-adrenoceptor agonist alpha-methylnoradrenaline. CONCLUSION: Our studies suggest that rilmenidine given systemically acts primarily on imidazoline receptors in the RVLM to reduce sympathetic tone and to modulate sympathetic baroreflexes.

Frequency-dependent modulation of renal blood flow by renal nerve activity in conscious rabbits.

To examine the influence of the various frequency components of renal sympathetic nerve activity (RSNA) on renal blood flow (RBF) dynamics, a Doppler flow probe and renal nerve electrode were implanted on the left renal artery of 10 rabbits. Experiments were performed 4-9 days after surgery. Physiological changes in RSNA were induced by subjecting the rabbits to periods of breathing hypoxic gas mixtures. Signals were sampled at 1 kHz and analyzed by spectral analysis. During moderate hypoxia (arterial PO2 = 44 +/- 1 mmHg), arterial pressure and heart rate did not change, averaged RSNA increased by 90 +/- 7%, and RBF fell by 18 +/- 3%. In a separate group of renal-denervated rabbits (n = 6), no changes in RBF occurred during hypoxia. In intact rabbits, 53 +/- 4% of spectral density power of RSNA was found at the cardiac frequency and the remainder was predominantly coupled to respiration (approximately 0.9 Hz). During moderate hypoxia the amplitude of the RSNA oscillations increased 17 +/- 6 times at the cardiac frequency and 10 +/- 3 times at the respiration-related frequency. Modulation of RBF variability by the fluctuations of RSNA at the cardiac- and respiration-related frequency was, however, small. The normalized transfer gain between RSNA and RBF was approximately 0.1 at > 0.5 Hz. This means that, at > 0.5 Hz, maximally 10% of the amplitude of the RSNA oscillations is transmitted to corresponding RBF fluctuations. These transfer properties did not change during hypoxia. At < 0.5 Hz the transfer gain between RSNA and RBF increased. During moderate hypoxia, 0.3-Hz coherent oscillations of RSNA and RBF were found. In renal-denervated rabbits, 0.3-Hz oscillations in RBF were absent. Thus the renal vasculature was able to follow relatively low-frequency (< 0.5-Hz) fluctuations of RSNA and responded with corresponding oscillations in RBF. In contrast, the renal vasculature responded with increased constriction at the high-frequency (> 0.5-Hz) fluctuations of RSNA. These findings suggest that, in conscious rabbits, high-frequency oscillations of RSNA contribute to the vasoconstrictor tone, whereas the lower frequencies of RSNA contribute to the variability of RBF.

Central imidazoline receptors and centrally acting anti-hypertensive agents.

We have examined the location and contribution of imidazoline receptors (IR) in mediating the hypotensive and sympatholytic actions of first and second generation anti-hypertensive agents in rabbits. We found that the hypotension produced by rilmenidine and moxonidine given intravenously (i.v.) or into the fourth ventricle (4V) was preferentially reversed by the IR antagonists idazoxan and efaroxan (compared to a selective alpha(2)-adrenoceptor antagonist 2-methoxy-idazoxan), suggesting that IR are important in the sympatho-inhibition produced by these agents. Clonidine was not preferentially reversed by the IR antagonists suggesting an action via alpha(2)-adrenoceptors. In anaesthetised rabbits, the rostral ventrolateral medulla (RVLM) was the most potent site for rilmenidine to produce the sympatho-inhibition and modulation of sympathetic baroreflexes. alpha-Methylnoradrenaline was also sympatholytic suggesting alpha(2)-adrenoceptors are also present in this site. Microinjection of the IR and alpha(2)-adrenoceptor antagonists showed that rilmenidine activates IR in the RVLM but that alpha(2)-adrenoceptors are also activated as a consequence. These studies suggest that rilmenidine acts primarily via IR in the RVLM to reduce sympathetic tone but also imply an important association of alpha(2)-adrenoceptors and IR in the region.

Multiple negative and positive cis-acting elements control the expression of the murine CD4 gene.

The cis-acting elements located within 15 kb 5' of the murine CD4 gene transcriptional start site and the first intron of the CD4 gene have been investigated using deletion constructs. Our transient transfection data indicate that the expression of the murine CD4 gene is controlled by multiple positive and negative regulatory cis-acting elements. There are at least two cis-acting elements that have a positive effect on the expression of the CD4 gene and at least four regions of DNA that have a negative effect. The positive control elements are located about 13.5 kb 5' of the promoter and within the flanking sequences of the first intron. The DNA between the 5' enhancer and the promoter contains at least two regions that exert a negative effect on CD4 expression. In addition to the positive effect that the first intron has on CD4 expression, there are two regions within the first intron that have a negative effect. These two negative regulatory elements correspond to two T-cell-specific DNase I hypersensitive sites found in the first intron.

Selective autonomic nervous control of thyroid hormone and calcitonin secretion during metabolic and cardiorespiratory activation by intracisternal thyrotropin-releasing hormone (TRH).

Injections of 10 micrograms/kg thyrotropin-releasing hormone (TRH) 150 microliter intracisternally (i.c.) in conscious rabbits evoked behavioral excitation and compulsive scratching, tachypnoea, an increase of heart rate and blood pressure, oxygen consumption and hyperthermia. TRH i.c. significantly increased free thyroid hormone and calcitonin secretion during depressed thyrotropin (TSH) secretion. The rise of calcitonin correlated with a fall of serum calcium. The ergotropic function of TRH i.c. was further demonstrated by rapid increases of glucagon, serum glucose, free fatty acid and free glycerol, with a delayed rise of insulin depending on glucose levels. The increases of free thyroid hormones, calcitonin, cortisol and lipolysis following TRH i.c. were augmented after spinal transection, while glucagon secretion increased at a slower rate, however, not accompanied by rises of glucose and insulin. Behavioral excitation and lipolysis were augmented by TRH i.c. after total thyroidal denervation, which completely prevented the rise in thyroid hormone and calcitonin secretion, although the thyroid follicles and C cells responded properly to TSH. Section of all thyroidal nerves except the recurrent laryngeal nerve reduced mainly calcitonin secretion following TRH i.c., while the behavioral, autonomic and other endocrine responses were augmented. Additional abdominal vagotomy in these rabbits diminished glucagon secretion by about 50% without significantly changing the other effector responses. Taking 125I-labelled TRH concentration in the cerebrospinal fluid at the site of i.c. injection as 100%, then 58% of TRH penetrated into outer parts of the dorsal and ventral medulla oblongata and pons, and 8% into the neuropil of the aqueductal region. Radioactivity in other brain areas including the hypothalamus was below 1%, while the hypophysis was practically devoid of radiolabelled TRH. It is suggested that the observed behavioral, autonomic and endocrine activity pattern elicited by injection of TRH into the cisterna magna was caused by excitation of neurons confined to that compartment and was mediated by pathways of the reticular formation of the lower brainstem, with the concept that TRH-containing neurons are intrinsic excitatory constituents of the 'activating reticular system'.