Chloralose-urethane Anesthesia Research Articles

Localization of serotonin receptors in the rat thalamus by electrophysiology and the action of 5-HTP-DP-hex

This study was designed to pinpoint the site at which N-hexanoyl-5-hydroxytryptophyl-5-hydroxytryptophan amide (5-HTP-DP-hex) exerts its previously reported effect on thalamic neurons in rats. The animals were prepared under chloralose-urethane anesthesia for a stereotaxic approach to either the nucleus ventralis posterolateralis (nVPL) or the centrum medianum-parafascicularis complex (CM-Pf) of the thalamus. Individual neurons in these nuclei were separately activated by single-pulse stimulation of the sciatic nerve or the thalamic fibers that form reciprocal connections between the CM-Pf and the second somatosensory (SII) region of the nVPL. Poststimulus time histograms were constructed from computer readouts of the stimulus-evoked responses of a neuron during a 500-ms period accumulated in a digital computer 100X. In addition, the number of spikes accumulated in each histogram was compared to the number of spikes accumulated under identical conditions on the same neuron after intracarotid infusion of 5-HTP-DP-hex. The effect of the drug was reversed by the infusion of 5-HTP. Statistical evaluation of the accumulated spike counts indicated that 5-HTP-DP-hex suppressed only the excitation of CM-Pf neurons from the SII of the nVPL; the input of the sciatic nerve into the CM-Pf remained unaltered. Furthermore, no effect was exerted by this dipeptide on the afferent excitation of neurons in the SII of the nVPL. Because the interaction of the CM-Pf with the neurons in the SII of the nVPL is necessary for the arousal of localized somatic pain, it is possible that interference of this interaction by 5-HTP-DP-hex might be the mechanism whereby the dipeptide produces its analgetic effect. The specificity and the exclusive localization of the action of 5-HTP-DP-hex points to the possibility that the synaptic junctions formed at the CM-Pf neurons by afferents from the nVPL may contain serotonin receptors.

Effects of chloralose-urethane anesthesia on single-axon reciprocal Ia IPSPs in the cat

Reciprocal Ia inhibitory postsynaptic potentials (IPSPs) generated by single afferents have been recorded with signal averaging in unanesthetized ischemic-decapitate cats for comparison with measurements previously obtained from preparations anesthetized with a mixture of chloralose and urethane. The results are similar to those which we obtained recently for single-axon recurrent IPSPs. Together, the studies show that chloralose-urethane anesthesia has a depressant effect on two widely studied circuits in the mammalian spinal cord.

Characteristics of tachyphylaxis to inhaled histamine in anesthetized dogs.

Three consecutive dose-response curves to aerosolized histamine were obtained in 11 anesthetized dogs. All dogs showed desensitization (i.e., tachyphylaxis) to high doses of histamine. Tachyphylaxis was highly reproducible. No tachyphylaxis occurred with inhaled acetylcholine or methacholine. Beta-Adrenergic blockade with propranolol or muscarinic blockade with atropine given intravenously had no effect on the histamine tachyphylaxis. Duration of thiamylal anesthesia did not alter the histamine responsiveness. Histamine tachyphylaxis was also seen with chloralose-urethan anesthesia. Since tachyphylaxis is not observed with acetylcholine, it cannot be attributed to a general decline in muscle contractility. We conclude that histamine tachyphylaxis in vivo is not explained by effects of cholinergic reflexes, catecholamine release, duration of anesthesia, or, probably, type of anesthetic agent.

Alterations in the microvasculature of one-kidney, one-clip hypertensive rats.

The microcirculation was studied in the cremaster muscle of one-kidney, one-clip (1K-1C) hypertensive rats and uninephrectomized controls under chloralose-urethan anesthesia 1-2, 4-5, or 8-9 wk following renal artery stenosis. With the use of television microscopy, inside and outside diameters of first (1A) through fourth-order (4A) arterioles were measured before and after vasodilation with 10(-4) M adenosine. Mean arterial blood pressure was significantly elevated in the 1K-1C rats, rising to 170 +/- 6 mmHg by 8-9 wks vs. 93 +/- 2 mmHg in controls. Enhanced vasoconstriction, resulting in closure of arterioles, appeared only in the smaller arterioles of 1K-1C and diminished throughout the development of hypertension. Structural rarefaction appeared later and increased with the development of hypertension. Vasodilated inside diameters of control, but not 1K-1C, 1A, 2A, and 3As, increased with increasing age, leaving the hypertensive arterioles with structurally reduced lumens and increased wall-to-lumen ratios, but without increases in wall cross-sectional areas. Structural lumen reduction appeared first in the 1A and advanced downstream as hypertension developed. Thus vasoconstriction of the smallest arterioles is important initially in renal hypertension, but structural alterations become more important later.

Postinspiratory-ramp activity of diaphragn under inspiratory resistive load

Inspiratory ramp activity, postramp acitivity (PRA) of diaphragm and respiratory flow were studied in anesthetized rabbits and conscious humans under inspiratory resisitive load. Under load with chloralose-urethane anesthesia neural inspiration lasted 124±12 msec more than under control conditions and end of mechanical inspiration lagged behind that of neural inspiration by 114±4 msec. With pentobarbital-urethane anesthesia corresponding changes under load were 67±18 msec and 44±6 msec; after SO 2 block of slowly adapting stretch receptors in bronchi corresponding changes under load were nil and 65±7 msec. In human corresponding changes under load were about 192 and 127 msec. Both in rabbits and humans time course of PRA under load was essentially unchanged. Hence, under load a considerable part of PRA (or all of it under pentobarbital-urethane anesthesia without SO 2 block) occured during inspiration and that occuring during expiration was smaller than under control conditions. Consequently, under load braking of expiratory flow was smaller and peak expiratory flow occured earlier than under control conditions.

A cardioinhibitory area in the midbrain central tegmental field of cats

A cardioinhibitory area in the central tegmental field of the midbrain (CIM) was studied in cats under chloralose-urethane anesthesia. Electrical and chemical (glutamate and DL-homocysteic acid) stimulations in this area produced marked bradycardia accompanied by mostly hypotension or minimal change in arterial pressure and by occasional hypertension particularly in the dorsal portion. CIM excitation potentiated the reflex bradycardia induced by IV phenylephrine, while bilateral electrolytic lesion of CIM neither changed the resting cardiovascular parameters nor the reflex bradycardia. The CIM bradycardia was not affected by supracollicular decerebration, but substantially reduced by unilateral vagotomy and completely eliminated by bilateral vagotomy. Destruction of the ambiguus nucleus (NA) and solitary and dorsal motor nuclei (NTS/DMV) abolished the bradycardia. Midline bisection at the midbrain-pontine level only slightly reduced the bradycardia while at the medullary level it was moderately attenuated. Electrolytic lesion of the cardioinhibitory area in gigantocellular reticular nucleus (GRN) abolished the bradycardia. These findings suggest that CIM is an independent mechanism which may send axons to GRN from which the axons may in turn synapse with the NTS/DMV complex and NA. Its final output may utilize both vagus nerves to modulate baroreceptor reflex in promoting bradycardia.

Increased arteriolar wall-to-lumen ratio in a normotensive vascular bed in coarctation hypertension.

In rats with coarctation hypertension, resting resistance and resistance at maximal vasodilation are elevated in the hindquarters, although blood pressure in this vascular bed remains normal. To assess the nature of these non-pressure-related vascular abnormalities, we observed third- to fifth-order arterioles in the cremaster microcirculation using standard techniques and chloralose-urethan anesthesia in rats with coarctation or sham coarctation of the abdominal aorta. Four weeks after clipping, carotid pressure was significantly elevated in coarcted compared with sham-coarcted rats, but femoral pressure was not. The wall-to-lumen ratio and wall thickness in cremaster arterioles was elevated by 12-33% in coarcted compared with sham-coarcted rats in the resting state. After maximal arteriolar relaxation with topical nitroprusside, differences in wall-to-lumen ratio persisted. We also studied the microcirculation of one-kidney normotensive control and one-kidney, one-clip Goldblatt hypertensive rats. Femoral pressure in Goldblatt hypertensive rats was elevated, but changes in the microcirculation were similar to those observed in coarcted rats. These non-pressure-related changes in vascular structure in hypertension may result from systemic neural or humoral influences and/or growth factors local to vascular wall tissues.

Effects of halothane on the ventilatory response to hypoxia and hypercapnia in cats.

The influence of halothane 0.8-1.2% inspired on the peripheral hypoxic chemoreflex was investigated in 13 cats subjected to artificial brain stem perfusion (ABP). This technique allows for an independent control of blood gas tensions and halothane concentration between blood perfusing the brain stem (central) and the systemic circulation (peripheral). In six cats the ventilatory response to isocapnic hypoxia was assessed during overall halothane anesthesia (HO) before and during ABP. Before ABP, systemic and brain stem circulations both were rendered hypoxic. During ABP, hypoxia was induced systemically while the brain stem was maintained hyperoxic. The ventilatory response in non-ABP cats (mean 698 ml . min-1 at PaO2 6.6 kPa; 50 mmHg) was about half the response in ABP cats (mean 1,194 ml . min-1 at PaO2 6.5 kPa; 49 mmHg), indicating that in the presence of halothane, central hypoxia depressed ventilation appreciably. Compared with chloralose-urethane anesthesia (CU), halothane reduced the ventilatory response to hypoxia in both perfusion conditions but never abolished it. To assess the influence of halothane on peripheral and central mediation of the CO2 response during hypoxia, each was assessed during CU anesthesia, during HO, and with halothane applied exclusively peripherally against a background of CU (CUHP). In all drug states, the periphery was kept hypoxic and brain stem hyperoxic. Compared with CU anesthesia, HO and CUHP anesthesia reduced both peripheral (Sp) and central (Sc) CO2 sensitivity but not the Sp/Sc ratio. Similarly, the extrapolated PaCO2 at zero ventilation was not detectably different among these three states.(ABSTRACT TRUNCATED AT 250 WORDS)

Response to hypercapnia and exercise hyperpnea in graded anesthesia.

We examined the relationship between response to hypercapnia and ventilatory response to exercise under graded anesthesia in eight dogs. The response to hypercapnia was measured by the CO2 rebreathing method under three grades of chloralose-urethan anesthesia. The degrees of response to hypercapnia (delta VE/delta PETCO2, 1 X min-1 X Torr-1) in light (L), moderate (M), and deep (D) anesthesia were 0.40 +/- 0.05 (mean +/- SE), 0.24 +/- 0.03, and 0.10 +/- 0.02, respectively, and were significantly different from each other. Under each grade of anesthesia, exercise was performed by electrically stimulating the bilateral femoral and sciatic nerves for 4 min. The time to reach 63% of full response of the increase in ventilation (tauVE) after beginning of exercise was 28.3 +/- 1.5, 38.1 +/- 5.2, and 56.0 +/- 6.1 s in L, M, and D, respectively. During steady-state exercise, minute ventilation (VE) in L, M, and D significantly increased to 6.17 +/- 0.39, 5.14 +/- 0.30, and 3.41 +/- 0.16 1 X min-1, from resting values of 3.93 +/- 0.34, 2.97 +/- 0.17, and 1.69 +/- 0.14 1 X min-1, respectively, while end-tidal CO2 tension (PETCO2) in L decreased significantly to 34.8 +/- 0.9 from 35.7 +/- 0.9, did not change in M (38.9 +/- 1.1 from 38.9 +/- 0.8), and increased significantly in D to 47.3 +/- 1.9 from 45.1 +/- 1.7 Torr.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of histamine inhalation and airway vibration on phrenic nerve activity in rabbits

Experiments were performed on 8 rabbits under urethane-chloralose anesthesia. Histamine aerosol (5%) administered into the trachea by a cannula produced tonic activity of the phrenic nerve, with discharges also being evident during the expiratory phase. Airway vibration (100 Hz) inhibited this tonic activity and caused silence during the expiratory phase.

Impaired central mediation of the arterial baroreflex in chronic renal hypertension.

After 6 wk of renal hypertension in rabbits, the arterial baroreflex control of heart rate (HR) is impaired but the baroreflex control of lumbar sympathetic nerve activity ( LSNA ) is preserved. This selective impairment may reflect a predominant abnormality in the baroreceptors. In this study, we tested the hypothesis that renal hypertension of longer duration may impair baroreflex control of LSNA through a defect in the central nervous system mediation of the reflex. Four months after induction of renal hypertension, baroreflex responses were determined during increases in arterial pressure with intravenous phenylephrine or decreases in pressure with vena caval occlusion under chloralose-urethan anesthesia. Reflex control of LSNA and HR was impaired markedly in hypertensive rabbits. Reflex inhibition of LSNA and HR in response to afferent electrical stimulation of the left aortic depressor nerve (all arterial baroreceptor afferents cut) was attenuated in hypertensive in contrast to normotensive rabbits. This attenuation was noted when the medullated fibers only were stimulated or when both medullated and nonmedullated fibers were stimulated. We conclude that baroreflex control of LSNA that is preserved after 6 wk of hypertension is impaired after 4 mo of hypertension. The impairment reflects an abnormality in the central nervous system mediation of the reflex.

Microvascular alterations in the one-kidney, one-clip renal hypertensive rat.

The microcirculation was studied in one-kidney, one-clip renal hypertensive rats ( 1KG ) and uninephrectomized controls at 4-6 and 8-10 wk postoperation. Under chloralose-urethan anesthesia the gracilis muscle was transilluminated in situ with a light pipe. Measurements of arteriolar and capillary density and arteriolar diameter were made in three consecutive states: innervated, denervated, and vasodilated with nitroprusside. Arteriolar wall-to-lumen ratio was measured after vasodilation. At 4-6 wk vasoconstriction was significantly greater in 1KG and vasodilated arteriolar diameter was significantly smaller, partially because of an elevated wall-to-lumen ratio. Capillary density in 1KG was reduced in the innervated and denervated states compared with controls. At 8-10 wk in the 1KG , arteriolar vasoconstriction was no longer significantly elevated. However, the vasodilated arteriolar diameter was smaller, wall-to-lumen ratio had increased further, and rarefaction of arterioles and capillaries was present. Thus, with time, structural mechanisms for increasing vascular resistance were seen to displace the active mechanism of vasoconstriction.

Reticular formation of the lower brainstem. A common system for cardiorespiratory and somatomotor functions: discharge patterns of neighboring neurons influenced by cardiovascular and respiratory afferents

Experiments were done in dogs with chloralose-urethane anesthesia. Long-lasting extracellular recordings were made from the medial parts of the reticular formation of the lower brainstem for up to 250 min. The study is based on reactions of 103 neurons. The activities of 2 or 3 neighbouring neurons recorded under identical conditions with one electrode or of neurons recorded with two electrodes at the same time could be changed regularly and synchronously by experimental changes of hemodynamic or ventilatory parameters. Action potentials were separated by amplitude discrimination. Rhythmic pulsatile modulations were proved to be present in 78% of all neurons by post-event-time histograms triggered by the R-wave of the ECG. In the 96 neurons tested 86% changed their activity when arterial pressure was raised by inflating a balloon in the abdominal aorta (79% decreased and 7% increased their activity). In post-event-time histograms triggered by the start of inspiration, 83% of the neurons showed modulations of their activity with respiratory rhythm. Experimental lung inflation decreased the activity in 75% of the tested neurons, while experimental lung deflation activated 47% of the tested neurons. Stimulation of arterial chemoreceptors activated 77% of the tested neurons. It was thus demonstrated that receptors in the cardiovascular and respiratory systems exert an influence on nearly all neurons from which recordings were made in that part of the reticular formation. Arterial baroreceptors and lung stretch receptors revealed a generalized depressing effect on the neuronal activity while chemoreceptors exert a generalized augmenting effect. At different times of recording these neurons did not always react to the same extent to comparable stimulations of afferents.

Capillary pressure gradients in cremaster muscle of normotensive and spontaneously hypertensive rats

The purpose of this investigation is to test the hypothesis that capillary pressure gradients are elevated in spontaneously hypertensive rats (SHR) and to determine the mechanism for the elevation. The cremaster muscle was prepared for microscopic examination under chloralose-urethane anesthesia in seven SHR and eight Wistar-Kyoto (WKY) rats 4–6 weeks of age. Capillary hematocrit, diameter, and red cell velocity were measured. Capillary flow induced by a time-varying pressure gradient was treated mathematically. A finite Hankel transformation was applied to the Navier-Stokes equation for capillary vessels. The solution was expressed as a Fourier-Bessel series, and the fluctuation of capillary flow induced by a time-varying pressure gradient was studied. It was shown that if the velocity fluctuation depended only on the pressure gradient, then the velocity fluctuation would be diminished almost instantly after the capillary started to flow. Capillary pressure gradient and shear stress were evaluated according to two different flow models, Newtonian and Casson. The capillary viscosity was obtained from the capillary hematocrit based on the empirical correlation of viscosity vs hematocrit. Calculations based on both flow models indicate that the capillary pressure gradient and shear stress of SHR is higher than in WKY, especially in vessels near 6 μm in diameter. The elevated pressure gradient is due to a combination of reduced capillary density, causing a higher red cell velocity and a tendency toward smaller capillary diameters in the SHR. Capillary hematocrit and viscosity were not elevated in the SHR.

Sites of action of halothane on respiratory pattern and ventilatory response to CO2 in cats.

To assess the major sites of action of halothane on the control of breathing, the ventilatory response to CO2 was studied in 11 cats and partitioned into tidal volume and frequency response. In these cats artificial perfusion of the ponto-medullary region was applied. In essence, this technique allows one to deliver to the brainstem blood-gas tensions and anesthetic concentrations at predetermined levels which are independent from those in the systemic circulation; thus the central and peripheral effects of halothane and CO2 can be determined separately. In cats exposed both centrally and peripherally to halothane (1.0-1.6%) tachypnea was observed which disappeared when the blood perfusing the brainstem was purged of halothane. From these results is follows that the tachypnea is exclusively due to an action of halothane on structures in the brainstem. In these cats the extrapolated Paco2 at zero ventilation was significantly lower during general halothane anesthesia than during light chloralose-urethane anesthesia (P less than 0.05). In cats lightly anesthetized with chloralose-urethane, halothane (0.5-1.5%) was either administered centrally or peripherally. In these experiments the "overall" ventilatory CO2 sensitivity of both the peripheral and central chemorereflex pathways decreased significantly (P less than 0.01). However, the ratio between these two sensitivities remained the same (P less than 0.5). The extrapolated Paco2 at zero ventilation was not affected by halothane provided its concentration was below 1% (P less than 0.7). From these results we conclude that the depressant effect of halothane on ventilation originates centrally as well as peripherally. Furthermore, from the findings that the ratio of the CO2 sensitivities and the extrapolated Paco2 at zero ventilation remained constant, the authors argue that halothane acts on the processing part of the neural respiratory drive (integrating centers) rather than on the neural activity of the peripheral and central chemoreceptors per se. The peripheral effect is mainly on the neuromechanical link between integrating centers and respiratory movements.

Development of microvascular rarefaction in the spontaneously hypertensive rat.

Using stereological methods in vivo, we have investigated the rarefaction of arterioles and capillaries in male spontaneously hypertensive rats (SHR) and the Wistar-Kyoto controls (WKY) at 6-8, 12-14, and 16-18 wk of age. Under chloralose-urethan anesthesia, the gracilis muscle was isolated for microscopic observation. Vessel length and surface area per unit volume of tissue (density) were determined during three consecutive states: innervation, denervation, and vasodilation with nitroprusside. Arteriolar wall-to-lumen ratio was measured after vasodilation. At 6-8 wk capillary density was reduced in the SHR. At 12-14 wk there was a reduction of arteriole and capillary density under innervated and denervated conditions but not after vasodilation (a state of functional rarefaction). At 16-18 wk there was a reduction of arteriolar and capillary density under all three conditions (a state of anatomical rarefaction). At 12-14 and 16-18 wk there was an elevated level of arteriolar vasoconstriction in the SHR that was masked in any one state by the closure of the smaller arterioles. Arteriolar wall-to-lumen ratio was not elevated in the SHR at any time. Arteriolar closure was not reversed by acute denervation.

The responses of I β cells to increases in the rate of lung inflation

The activity of inspiratory cells in the region of the nucleus of the tractus solitarius (NTS) was recorded extracellularly in paralyzed, artifically ventilated cats either during chloralose-urethane anesthesia or following midcollicular decerebration. Twenty-three of the 68 inspiratory cells recorded in the region of the NTS were classified as I β cells on the basis of their response to withholding lung flation. The dynamic sensitivity of I β cells was determined by studying their response to increases in the rate of lung inflation at contant peak volume. The I β cells in this study showed 3 distinct patterns of response to increases in the rate of inflation. Five cells showed no change in firing pattern (fixed firing pattern). Ten cells showed an increase in the rate of rise of cell activity but no change in peak frequency (low dynamic sensitivity). Eight cells showed increases in both the rate of rise of cell activity and peak frequency (high dynamic sensitivity). It was concluded that I β cells are not a functionally homogeneous population, at least in terms of their dynamic sensitivity. Cells showing fixed firing patterns have the characteristics of off-switch neurons. Cells with low levels of dynamic sensitivity may receive afferents from pulmonary stretch receptors. Cells showing a high degree of dynamic sensitivity may received afferents from rapidly adapting receptors. The fact that I β cells are not a functionally homogenous population may explain the many divergent observations reported from studies of these cells.

Analysis of temperature dependence of muscle cell membrane repolarization in the diaphragm of rats of different ages

In experiments on cats with dissected vagus and aortal nerves under chloralose-urethane anesthesia, ventricular disorders of the cardiac rhythm were induced by ligation of the common carotid arteries. Appearance of arrhythmias was preceded by an increase in the sympathetic activity (recorded from the inferior cardiac or renal nerve) accompanied by a rise of the arterial blood pressure and of the heart rate. Intravenous injection of lithium chloride or hydroxybutyrate resulted in lowering of the sympathetic activity, arterial blood pressure, and heart rate, and led to the recovery of the sinus rhythm.

The respiratory role of the ventral surface of the medulla studied in the anaesthetized rat.

1. The respiratory role of the ventral surface of the medulla was studied in rats anaesthetized with a urethane-chloralose mixture. 2. In fifty-eight studies on twelve animals, direct superfusion of the medullary surface with artificial c.s.f. made acid by the reduction of bicarbonate content or by the increase of PCO2 produced no significant stimulation of respiration provided that the temperature of the brain surface was unaltered. 3. Superperfusion of the medullary surface with c.s.f. of low bicarbonate content produced an inhibition of respiration in fourteen of thirty-eight experiments. 4. Electrical stimulation on the surface revealed a localized area lateral to the pyramids and rostral to the XIIth nerve where stimulation at low intensity produced an increase in the frequency and depth of respiration. 5. The application of carbachol to a similar region increased both the frequency and amplitude of ventilation at lower concentrations than were required to obtain effects from surrounding areas. 6. Sudden switching between perfusates at different temperatures produced changes of ventilation within 1-2 sec of a change of surface temperature. The Q10 for the ventilation/temperature relationship was approximately 6. 7. The experiments confirm that the ventral surface of the medulla contains neural elements which, at least during urethane-chloralose anaesthesia, have a significant effect on respiration. The stimulus for these effects in the rat does not appear to be a change in H+ concentration. It appears more probable that the primary role of the area lies in the link between thermal and respiratory regulation.

Baroreceptor influence on a spinal cardiovascular reflex.

A stretch of the walls of the thoracic aorta, performed in vagotomized cats without obstructing aortic flow, induces increases in heart rate, myocardial contractility, and arterial pressure. These reflex responses are still present after high spinal section. Cats under chloralose-urethane anesthesia were vagotomized and one carotid sinus was isolated and perfused with arterial blood at constant flow. The contralateral carotid sinus nerve and both aortic nerves were sectioned. A stretch of the walls of the thoracic aorta between the 7th and 10th intercostal arteries induced a reflex increase in mean arterial pressure 29 +/- 2 mmHg (mean +/- SE). Stepwise increases of carotid sinus pressure (CSP) or electrical stimulation of the carotid sinus nerve induced stepwise decreases of this reflex response. At maximal baroreceptor stimulation (CSP 212 +/- 9 mmHg) the reflex response to aortic stretch was reduced by 42%. These experiments show that this spinal cardiovascular reflex is at least partially under the inhibitory control of the baroreceptor input.