Comparison between pial and intraparenchymal vascular responses to cervical sympathetic stimulation in cats. Part 1. Under normal resting conditions

To investigate the role of sympathetic regulation in both resistance and capacitance vessels in cerebral circulation, the response of pial and intraparenchymal vessels to sympathetic nerve stimulation were simultaneously examined in 14 cats by means of a newly developed video camera photoelectric system. The system consisted of a video camera system for measurement of pial vascular diameters and a photoelectric apparatus for estimating regional cerebral blood volume in the intraparenchymal vessels. The ipsilateral superior cervical ganglion was electrically stimulated for 5 min. Initially, both the pial and intraparenchymal vessels constricted. The large pial arteries (173 +/- 25 micron, mean +/- SEM) remained constricted throughout the stimulation, whereas the intraparenchymal vessels began to dilate after the initial constriction and exceeded the control level at 175 +/- 25 s despite continued stimulation. In conclusion, such sympathetic nerve stimulation is considered to exert a constrictive effect on the intraparenchymal as well as the pial vessels at the early stage. The compensatory dilation of the intraparenchymal vessels was delayed 3 min after initiation of the stimulation.     

Locus coeruleus neurons in the neonatal rat: electrical activity and responses to sensory stimulation

Newborn rats, 1-3 days of age, were anesthetized with urethane and single-unit activities were recorded extracellularly in the locus coeruleus. Ca. 35% of locus coeruleus neurons recorded were antidromically activated from stimulation of the frontal cortex. Although the majority of locus coeruleus neurons were not spontaneously active, non-noxious as well as noxious sensory stimuli such as touches to the skin, air puffs and tail pinches were very effective in exciting locus coeruleus neurons.     

Comparison between pial and intraparenchymal vascular responses to sympathetic stimulation under hypercapnic conditions. With special reference to the mechanism for escape phenomenon

We have shown that secondary vasodilation ('escape' phenomenon) during sympathetic nerve stimulation occurs in the intraparenchymal vessels but not remarkable in the pial vessels. To test a possible role of CO2 accumulation in the brain tissue in this phenomenon, the responses of pial and intraparenchymal vessels to sympathetic nerve stimulation were investigated during hypercapnia in 9 cats by using a video camera photoelectric system. The ipsilateral superior cervical ganglion was electrically stimulated for 5 min during hypercapnia (PaCO2 = 50 +/- 2 mm Hg). The intraparenchymal vessels as well as pial vessels remained constricted throughout the stimulation. Secondary dilation of the intraparenchymal vessels as seen at the later stage of sympathetic stimulation during normocapnia was not observed under the hypercapnic conditions. We assume that the arterial CO2 tension was so high that the constriction of inflow vessels could not result in accumulation of CO2 in the brain parenchyma. The accumulation of chemical metabolites as represented by CO2 is therefore considered to be the most probable mechanism underlying the escape phenomenon of the intraparenchymal vessels.     

Effect of locus coeruleus stimulation on regional cerebral oxygen consumption in the cat

Regional cerebral oxygen consumption was determined during stimulation of the intra-axial noradrenergic pathway to quantitate the metabolic effects of this manipulation on cerebral oxygen extraction, cerebral blood flow (CBF) and its regional distribution. Regional arterial and venous oxygen saturation were examined microspectrophotometrically. Regional CBF was examined using radioactively tagged microspheres (15 +/- 3 microns in diameter). Oxygen consumption was calculated as the regional product of CBF and oxygen extraction. Bipolar concentric electrodes were stereotaxically implanted bilaterally in the locus coeruleus of alpha-chloralose anesthetized, artificially respired adult mongrel cats. The control group was killed after hemodynamic and CBF measurements were taken. The experimental group was sacrificed after these same measurements were taken before and during 10 min of bilateral locus coeruleus stimulation. The cats' heads were simultaneously sawed in 3 places and quickly frozen in liquid nitrogen-cooled propane. Systolic blood pressure was significantly increased during treatment. The heterogeneity of venous oxygen saturation was significantly reduced by stimulation. Average CBF and oxygen consumption were significantly decreased to 57% and 59% of control, respectively. Oxygen consumption was significantly reduced in the hypothalamus from 1.5 +/- 0.3 to 0.9 +/- 0.3 ml O2/min/100 g and from 3.5 +/- 0.9 to 1.2 +/- 0.4 ml O2/min/100 g in the cerebellum by treatment. Changes in the neuronal and/or synthetic cerebral activity produced regional decreases in cerebral oxygen consumption and secondarily altered CBF. These changes are probably due to interaction of the intraparenchymal noradrenergic pathways with other systems or processes in the brain.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of serotonin and its antagonist ketanserin on pial vessels

The effect of serotonin (5-HT) and its antagonist ketanserin on the cerebral circulation was investigated in two series totaling 24 cats using the cranial window technique. 5-HT elicited a marked dilatation of small pial arteries, whereas large arteries tended to constrict. Intravenous administration of ketanserin reversed the constriction of large arteries, causing dilatation, and reduced the extent of small arteries' dilatation. In a randomized study, i.v. administration of ketanserin in its solvent versus the solvent alone revealed a strong dilatatory effect of the solvent on pial arteries (17 +/- 1.8%), which partly jeopardized a possible constrictory effect of ketanserin, as ketanserin plus solvent induced less dilatation of small pial arteries than the solvent alone. The present data support the view that serotonin exerts a dual effect on cerebral arteries, namely, dilatation of small and constriction of large vessels. The antagonist ketanserin reverses this effect, but the strong dilatatory effect of the solvent alone masks the antiserotoninergic effect.     

Concentrations of putative neurovascular transmitters in major cerebral arteries and small pial vessels of various species

The levels of noradrenaline, neuropeptide Y, 5-hydroxytryptamine, and substance P were measured and compared between the large arteries of the circle of Willis and the small cerebral vessels of the pia mater in the rat, rabbit, cat, and monkey. In all species, noradrenaline and neuropeptide Y concentrations were greater in the larger arteries than in small pial vessels. Noradrenaline concentrations were dramatically reduced following cervical sympathectomy, with the extent of diminution differing greatly in the various species; the effects of cervical ganglionectomy on neuropeptide Y concentrations were less pronounced. 5-Hydroxytryptamine concentrations in rats, cats, and rabbits were significantly greater in the small pial vessels, although measurable concentrations existed in the circle of Willis. In cats and monkeys, substance P was found in major arteries, but was not detectable at the level of the small pial vessels. The differences in the regional distribution of the various neurotransmitter candidates in the cerebrovascular bed may reflect their physiological significance.     

The effect of lesions in the locus coeruleus on the physiological responses of the cerebral blood vessels in cats

The effects on cerebral blood flow (CBF) of lesions placed stereotactically in or near the locus coeruleus were studied in 15 lightly anesthetized cats; 5 control cats in which the electrode was placed but no lesion created, and 10 experimental cats in which a lesion was created. The response of CBF to changes in Pa_co2 and in mean arterial blood pressure was determined by¹³³Xe-washout studies 10 days after the stereotactic procedures. The sites of the lesions were studied histologically, and their effects on catecholamine concentrations in the paraventricular hypothalamic nucleus, anterior ventral nucleus of the thalamus, and parietal cortex were determined by radiochemical assay.Control animals and those with lesions near, but not in, the locus coeruleus had normal Pa_co2-CBF response curves and normal catecholamine concentrations in the areas of biopsy. Bilateral destruction of the locus coeruleus was confirmed in 3 animals on histological examination and in these animals there were decreased levels of catecholamines in the areas of assay, higher resting CBFs at normocapnia, and significantly abnormal CBF-Pa_co2 response curves. The autoregulatory response to changes in perfusion pressure was preserved. Thus, noradrenergic neurons originating in the locus coeruleus may contribute to the control of intraparenchymal cerebral vessels and disturbance of this control may be important in the pathology of cerebral ischemia.     

Serotonin-norepinephrine interactions: a voltammetric study on the effect of serotonin receptor stimulation followed in the N. raphe dorsalis and the Locus coeruleus of the rat

In vivo voltammetry with carbon fibre electrodes was used to study the effect of the serotoninergic (5-HT) neuronal system on the noradrenergic (NE) system in the Locus coeruleus of the rat. The voltammetric DOPAC signal in the Locus coeruleus, used as a measure of NE neuronal activity, was increased after systemic application of the 5-HT1B agonist CGS-12066B, the 5-HT2 antagonist ritanserin, and, to a lesser extent, by ipsapirone, a 5-HT1A agonist. The findings suggest that the NE neuronal system of the Locus coeruleus is stimulated by 5-HT1A and 5-HT1B receptor activation and inhibited by 5-HT2 receptors. Likewise the 5-HT releaser and uptake inhibitor fenfluramine increased the DOPAC level in the Locus coeruleus. In contrast to the 5-HT1 agonists, which reduced 5-hydroxyindoleacetic acid (5-HIAA) in the Nucleus raphe dorsalis, ritanserin increased the 5-HIAA signal in this nucleus. This finding could help to explain the action of ritanserin as sleep-modulating substance.     

Effects of bradykinin on permeability and diameter of pial vessels in vivo

The effect of bradykinin on the permeability and vasomotor response of pial vessels has been studied to enhance our understanding of the pathophysiological role of the kallikrein-kinin system in cerebral tissue. Intravital fluorescence microscopy of the pia arachnoidea was conducted using Na+-fluorescein, FITC-dextran, and FITC-albumin as low and high molecular weight blood-brain barrier indicators. Massive arterial dilatation evolved immediately upon administration of bradykinin by superfusion of the exposed cerebral surface. An increase of the arterial diameter by 40% was the maximal response found at bradykinin concentrations of 4 x 10(-5) M. Arterial dilatation became attenuated with continuous superfusion of the preparation with bradykinin. In pial veins, a moderate reduction of the vessel diameter was observed, however, only after prolonged superfusion of the preparation. Bradykinin led to selective opening of the blood-brain barrier for Na+-fluorescein at superfusate concentrations of greater than or equal to 4 x 10(-7) M, but not for FITC-dextran or FITC-albumin. Topical administration of l-isoproterenol (10(-4) M) was found to prevent extravasation of Na+-fluorescein in the presence of bradykinin concentrations of 4 x 10(-6) M. Protection of the blood-brain barrier by isoproterenol was not observed when higher concentrations of bradykinin were employed. Intracarotid infusion of bradykinin were employed. Intracarotid infusion of bradykinin led also to a selective opening of the blood-brain barrier for Na+-fluorescein, but not for FITC-dextran or FITC-albumin. In contrast to superfusion, this route of administration did not induce changes of the vasomotor behavior of the arteries or veins. Additional experiments with B1-agonists and -antagonists suggest that bradykinin causes the openings of the blood-brain barrier th rough an interaction with B2-receptors on endothelial cells, and arterial dilatation via interaction with B2-receptors on vascular smooth muscle cells. Our findings support the concept that the release of kinins in the brain during an acute cerebral lesion mediates secondary damaging processes by the enhancement of blood-brain barrier dysfunction.     

Locus coeruleus effects on baroreceptor responsiveness and activity of neurosecretory vasopressin cells

The locus coeruleus (LC) has previously been implicated in the regulation of vasopressin secretion. To further investigate this issue experiments were done in which extracellular recordings were obtained from functionally identified neurosecretory vasopressin (VP) cells of the rat supraoptic nucleus. Electrolytic lesions of the ipsilateral LC reduced the proportion of VP cells inhibited by carotid baroreceptor activation from 93% to 35%; the inhibitory effect of aortic depressor nerve stimulation was unchanged. Electrical stimulation of the LC altered the discharge probability of 20% of VP cells tested, the predominant effect being excitation. In contrast to the effects of electrolytic lesions and electrical stimulation, neither chemical inhibition nor stimulation of the LC, by local injection of neuroactive amino acids, altered VP cell baroreceptor responsiveness or spontaneous discharge. These data indicate that while fibres of passage in the LC region can influence VP cell excitability, particularly responses to carotid baroreceptor activation, LC cells do not regulate VP cell function or, by implication, the secretion of this vasoactive and antidiuretic hormone.     

The effects of dipyridamole and theophylline on rat pial vessels during hypocarbia

Hypocarbia results in an increase in brain adenosine concentrations, presumably because of brain hypoxia associated with hypocarbic vasoconstriction. It was hypothesized that adenosine limits the degree of hypocarbic vasoconstriction. To test this hypothesis, the effects of dipyridamole and theophylline on CO2 reactivity during hypocarbia were investigated in anesthetized rats. Dipyridamole should reduce the vasoconstriction by potentiating adenosine action, whereas theophylline should increase the vasoconstriction by blocking adenosine receptors. Cortical pial arterioles of mechanically ventilated and anesthetized rats were displayed on a video monitor system through a closed cranial window. Arterial blood pressure and oxygen tension were stable. CO2 reactivity, formulated as 100 X [delta diameter (micron)/resting diameter (micron)]/delta PaCO2 (mmHg), in the hypocarbic phase was calculated before and after topical superfusion of dipyridamole (10(-6) M; n = 7) and theophylline (5 X 10(-5) M; n = 6). CO2 reactivity was significantly decreased after superfusion of dipyridamole (0.57 +/- 0.08; mean +/- SEM) as compared with mock cerebrospinal fluid (CSF) (0.97 +/- 0.17, p less than 0.05, n = 7). On the other hand, CO2 reactivity after superfusion of theophylline was increased (1.63 +/- 0.28) as compared with mock CSF (1.00 +/- 0.20, p less than 0.05, n = 6), indicating that adenosine is involved in hypocarbic vasoconstriction.     

Low frequency stimulation of the locus coeruleus reduces regional cerebral blood flow in the spinalized cat

Regional cerebral blood flow (RCBF) was studied during low frequency (15/s) and high frequency (50/s) electrical stimulation of the locus coeruleus (LC) in the alpha-chloralose-anesthetized cat using the freely diffusible tracer [14C]iodoantipyrine and regional brain dissection. The responses were determined in animals spinalized at the C1/C2 level to eliminate systemic effects of pontine stimulation such as alterations in blood pressure and heart rate. The spinalization, itself, did not alter resting RCBF or reactivity to hypercapnia. Low frequency stimulation reduced regional cerebral blood flow in the cortex, basal ganglia and white matter of the corpus callosum. The reductions in RCBF were maximal (35%) in the occipital cortex whereas no changes were seen in the colliculi. No changes were seen in any brain areas with high frequency stimulation. The relevance of this brainstem effect on cerebral blood flow to pathological states such as stroke and migraine is discussed.     

Membrane excitability changes in hindlimb motoneurons induced by stimulation of the locus coeruleus in cats

The present analysis describes the cellular mechanisms underlying the heightened membrane excitability of hindlimb flexor and extensor motoneurons upon stimulation of the locus coeruleus (LC) in unanesthetized, decerebrate cats. In a total of 73 cells, brief train stimuli to the LC at 50-300 microA intensity evoked one of 4 patterns of motoneuronal responses: a simple excitatory postsynaptic potential (EPSP) with weak trailing depolarization, a double-peak EPSP, an EPSP succeeded by a weak hyperpolarization, or a slow rising EPSP. As the initial dominant EPSP was a consistent finding among all cells and the ensuing potentials were variable in polarity, quantitative characterization was focused on the initial EPSP only. In all cells tested (n = 11), the LC-EPSP was accompanied by a decrease in input resistance. The excitatory LC action was further demonstrated by the consistent (n = 25 cells) motoneuron rheobase decrease when the latter was measured coincident with the summit of an LC-EPSP. Furthermore, the time course of the single-spike afterhyperpolarization became shortened during the LC conditioning stimuli (n = 16 motoneurons). Our data show that the descending LC action on motoneurons is typified by an EPSP accompanied by a net decrease in input resistance as well as a concurrent increase in motoneuron electrical excitability.     

Telencephalic projections from midbrain and isthmal cell groups in the pigeon. I. Locus coeruleus and subcoeruleus

Horseradish peroxidase (HRP) and amino acid autoradiography were used in pigeon to determine the trajectories and projection patterns of neurons within the locus coeruleus and subcoeruleus nuclei upon the cerebral hemispheres. The specific cell groups investigated include the locus coeruleus (LoC), nucleus subcoeruleus dorsalis, and nucleus subcoeruleus ventralis. Efferents from each of these nuclei ascend to the telencephalon via the medial and lateral forebrain bundles, ansa lenticularis, and the quintofrontal and occipitomesencephalic tracts. A separate dorsally situated bundle derived from LoC neurons reaches many dorsal thalamic nuclei. The telencephalic projections of the LoC and subcoeruleus nuclei are bilateral and symmetrical, although projections to the contralateral hemisphere are sparse. Crossing fibers project to contralateral targets primarily via the dorsal supraoptic decussation and along the dorsal and ventral margins of the anterior commissure. Within the telencephalon, the following neural structures receive input from neurons in the LoC and subcoeruleus cell groups: the paleostriatal complex including the paleostriatum augmentatum and lobus parolfactorius, septal nuclei, nucleus accumbens, olfactory tubercle, hippocampus and parahippocampal area, nucleus taeniae, dorsal archistriatum, lateral neostriatum, hyperstriatum dorsale, hyperstriatum ventrale, and preoptic area. Large portions of the cerebral hemispheres including the hyperstriatum accessorium, much of the neostriatum and hyperstriatum ventrale, and all but the dorsal portion of the archistriatum receive little or no input from either the locus coeruleus or subcoeruleus cell groups. This is apparently different from the condition in mammals in which virtually all cortical fields receive input from neurons within the LoC. Moreover, the pattern of projections of the subcoeruleus nuclei upon telencephalic fields described here as well as recent histochemical data suggest that these cell groups are comparable to the lateral tegmental (A8) cell group of mammals rather than to the mammalian subcoeruleus nuclei.     

Determinants of response of pial arteries to norepinephrine and sympathetic nerve stimulation.

Feline pial arteries larger than 100 mu in diameter constricted in response to cervical sympathetic nerve stimulation suggests or in response to topical application of norepinephrine. Smaller pial arteries were unresponsive to norepinephrine. This unresponsiveness persisted when norepinephrine was dissolved in CSF with high calcium ion concentration, or in CSF with both high calcium ion and zero magnesium ion concentration, or when it was dissolved in the acid fluid used by Wahl et al. and applied by constant infusion or by intermittent application. Comparison of the responses of the larger pial vessels to norepinephrine and to sympathetic nerve stimulation that maximal activation of sympathetic nerves achieves a concentration of released norepinephrine equal to 5.9 x 10(-6) M. The constriction of the larger pial vessels in response to sympathetic nerve stimulation could account for modest reductions in cerebral blood flow.