Reactive changes in adrenergic innervation of the cerebral arteries after activation of the cholinergic mechanisms]

Magistral arteries of the brain and pia mater have been studied in cats 24-72 h after administration of the cholinesterase inhibitor (phosphacol, 600 mcg/kg). Cholinesterase activity in blood has been checked by means of the potentiometric titration method, and acetylcholinesterase (AChE) content in varicosities of the perivascular nervous fibers--cytophotometrically in preparations treated after Karnovsky--Roots histochemical method. Cholinesterase activity of blood homogenates in test animals is 42 +/- 10%, and acetylcholinesterase content in varicosities of the perivascular nervous fibers--23.5 +/- 2.3% in comparison to the norm. Catecholamines in adrenergic nervous elements are revealed treating them with glyoxylic acid. Distribution density (DD) of histochemically active nervous elements is determined, as well as their specific content of the mediator according to luminescent intensity (LI) of varicosities of nervous fibers. The data obtained in intact animals serve as the control. In the experiment DD and LI reach 127 +/- 9% and 154 +/- 15%, respectively, as compared to the control. Signs of the adrenergic nervous apparatus activation in the experiment reflect a compensatory reaction in response to increase of dilatatory cholinergic influences to vessels under conditions of AChE activity.     

Functional recuperation of the serotoninergic innervation in the rat locus coeruleus

1. 5,6-dihydroxytryptamine (5,6-DHT) or a lesion of the raphe centralis superior (RCS) cause significant decreases in the serotonin (5-HT) content and significant increases in the tyrosine hydroxylase activity in the locus coeruleus (LC) of the rat. This suggests that noradrenaline (NA) synthesis is controlled by serotonin-containing neurons in the raphe system via their terminals in the LC. 2. Radioautography after intraventricular infusion of tritiated serotonin (3H-5-HT) and biochemical determinations of endogenous 5-HT content showed an almost complete disappearance of serotoninergic axonal varicosities and content in the LC region 10-15 days after intraventricular administration of 75 micrograms of 5,6-DHT. Two to 4 months after neurotoxin administration, 5-HT fibers had regrown in the LC but, contrary to the normal innervation pattern, the majority of them invaded the medial most portion of the nucleus and the adjacent subependymal region. The LC region regained almost all of its endogenous 5-HT content in the same time period. 3. Functional recuperation of these 5-HT fibers was demonstrated by the fact that the RCS had, after regeneration, the same functional control on NA synthesis as in the normal animal.     

烫伤大鼠脑底动脉壁内含内皮素-1能神经纤维的形态学变化

目的探讨烫(烧)伤损伤时大鼠脑血管内皮素-1能神经纤维分布与脑血管神经源性调节的关系,以及烫(烧)伤对脑血管损伤的影响。方法应用免疫组织化学技术观察烫(烧)伤大鼠脑底动脉(包括大脑前动脉、大脑中动脉、大脑后动脉和基底动脉)内皮素-1能神经纤维的分布。结果烫(烧)伤大鼠和正常大鼠脑底动脉均可见棕褐色的内皮素-1能免疫反应阳性神经纤维,似细线状,攀附于血管壁上,烫(烧)伤大鼠脑底动脉各主要分支内皮素-1能免疫反应阳性纤维密度较正常大鼠明显增加,纤维走行大多呈网状。结论烫(烧)伤可引起大鼠脑底动脉内皮素-1能免疫反应阳性神经纤维增加,增加的内皮素-1能神经纤维可能诱发脑血管痉挛和脑血液循环紊乱。提示内皮素-1能神经纤维在烫(烧)伤后在脑血管的神经源性调节中可能起重要的作用。     

Low nNOS protein in the locus coeruleus in major depression

Disruptions of glutamatergic and noradrenergic signaling have been postulated to occur in depressive disorders. Glutamate provides excitatory input to the noradrenergic locus coeruleus (LC). In this study, the location of immunoreactivity against neuronal nitric oxide synthase (nNOS), an intracellular mediator of glutamate receptor activation, was examined in the normal human LC, and potential changes in nNOS immunoreactivity that might occur in major depression were evaluated. Tissue containing LC, and a non-limbic, LC projection area (cerebellum) was obtained from 11 to 12 matched pairs of subjects with major depression and control subjects lacking major psychiatric diagnoses. In the LC region, nNOS immunoreactivity was found in large neuromelanin-containing neurons, small neurons lacking neuromelanin, and glial cells. Levels of nNOS immunoreactivity were significantly lower in the LC (- 44%, p < 0.05), but not in the cerebellum, when comparing depressed with control subjects. nNOS levels were positively correlated with brain pH values in depressed, but not control, subjects in both brain regions. Low levels of nNOS in the LC may reflect altered excitatory input to this nucleus in major depression. However, pH appears to effect preservation of nNOS immunoreactivity in subjects with depression. This factor may contribute, in part, to low levels of nNOS in depression.     

Effect of experimental subarachnoid hemorrhage on calcium incorporation and adrenergic innervation of the cerebral arteries of the cat

The effect of experimental subarachnoid hemorrhage (SAH) on the adrenergic innervation and on the 45Ca2+ uptake of cat cerebral arteries was analyzed. Intracisternal injections of autologous blood reduced the noradrenaline content of perivascular nerve endings and 3H-noradrenaline uptake. These values returned to normal levels in a period of two weeks after SAH. The activity of dopamine-beta-hydroxylase was also reduced 3 and 7 days after SAH. Superior cervical gangliectomy and intracisternal injection of 6-hydroxydopamine also reduced these three parameters. The uptake of 45Ca2+ by arteries from animals submitted to SAH was greater than if the blood vessels were from untreated cats. Lantanum brought about a less 45Ca2+ displacement in the arterial segments from untreated animals than in those from cats after SAH. These results suggest that SAH induces a transient adrenergic denervation as well as changes in the membrane of smooth muscle cell which increase the quantity of Ca2+ bound to it. All these modifications might be involved in the cause of chronic cerebral vasospasm that appears after SAH.     

Open-field behavior of rats following bilateral coagulation of the locus coeruleus

A modified open-field test was used in the experiments on rats. It was shown that animals 6 days after bilateral coagulation of locus coeruleus displayed weight changes and some behavioural parameters distinct from those of sham-operated and control animals. The most pronounced differences observed in animals after locus coeruleus coagulation, as compared to the control, were inversion and adaptive responses to a sudden brief stimulus, locomotor dysfunction and absence of response to handling.     

Morphofunctional changes in the myocardium after extensive bilateral resection of the lungs

In the experiment, performed on 89 dogs, after 67-75% resection of the lungs, functional morphology of the myocardium has been studied by means of histological, electron microscopical and morphometrical methods. Bilateral extensive resections of the lungs are accompanied with hypertrophy of the cardiac muscle, that is revealed at all the levels of its structural organization. Simultaneously, the hypertrophy develops against the background of dystrophic and destructive changes of the myocardium, their depth and extensiveness correlate with duration of the postoperative observations. If an extrapulmonary anastomosis is formed between the inferior lobular pulmonary artery and the corresponding vein before the resection of 67-75% of the lungs, it eliminates the acute overloading of the right ventricle, makes better conditions for development of compensatory-adaptive processes in the myocardium, prevents advancement of the cardio-pulmonary deficiency during the postoperative period.     

Development of the adrenergic innervation in the myocardium and coronary arteries of the dog

The development of the adrenergic cardiac innervation was studied in premature dog fetuses, puppies and adult dogs by means of the formalin-induced fluorescence technique. A point-counting technique was used to evaluate the density of innervation. Two types of fluorescent profiles can be observed in the heart during development: (1) sprouting axons, and (2) beaded terminals. The axonal fluorescence disappears in adult neurons. A different morphology and a different time course of development enable to study separately the innervation of the myocardium (cardiomotor innervation) and that of the vessels (vasomotor innervation). The late prenatal innervation is very poor (0.1 hit). The first but very scant cardiomotor terminals appear in this period. A mature cardiomotor innervation is found in 4-month-old puppies [1.5 +/- 0.3 (SD) hits]. The vasomotor innervation is shifted to the right. The development of beaded vascular terminals begins and matures 1-2 weeks later. The growing fluorescent axons reveal that the myocardium is supplied by axons of the cardiac plexus and of the perivascular nerves; the vascular wall, on the other hand, is supplied by the perivascular nerves only. The developmental, spatial and morphological differences in innervation suggest that two different types of neurons exist in the sympathetic ganglia: (1) neurons innervating the vessels (coronaromotor neurons), and (2) neurons innervating the myocardium (cardiomotor neurons).     

Characterization of a developmentally transient adrenergic depolarizing response in cultured locus coeruleus neurons

The effects of iontophoretically applied noradrenaline have been tested on intracellularly recorded locus coeruleus neurons grown in explant cultures from neonatal mice. In addition to hyperpolarizing responses mediated by alpha 2-adrenergic receptors, as observed in locus coeruleus neurons in vivo and in brain slices from adult animals, alpha 1-mediated depolarizations were observed to succeed the initial hyperpolarizations in some cultures. It was shown that the depolarizing responses were only present in younger cultures, i.e., less than 26 days in vitro. In cultures less than 20 days old, all cells displayed the biphasic hyperpolarizing-depolarizing responses. Both components of the response appear to be direct, since they were present when synaptic transmission was blocked by including tetrodotoxin or by altering divalent cations in the perfusate. The depolarizing responses were frequently reduced in solutions with altered divalent cation content, and this might reflect a calcium dependency of this response. The hyperpolarizing and depolarizing components of the responses to noradrenaline were progressively blocked by increasing concentrations of the selective antagonists yohimbine and prazosin, respectively, in the dose ranges of 100 mM - 1 microM (yohimbine) and 20-200 nM (prazosin). Recent results from electrophysiological studies of locus coeruleus neurons in brain slices suggest that similar changes occur in the animal as well as in culture. It is possible that the transient depolarizing responses reflect a developmentally important enhanced responsiveness of locus coeruleus neurons during the early postnatal period.     

Morphofunctional restructuring of the nervous system of the lymphoid organs during electrostimulation of the hypothalamus

The intramural nervous apparatus of rabbit lymphoid organs was examined after 15 and 30 sessions of electric stimulation of the posterior hypothalamus. The function of nerve cells was evaluated by cytophotometry according to the content of catecholamines and acetylcholinesterase (ACE) activity. In the mesenteric lymph nodes, stimulation led to a decrease in the activity of both adrenergic and ACE-containing nerve cells. The spleen demonstrated a reduction in the catecholamine content in the perivascular nerve plexuses, with a simultaneous rise of the catecholamine content in the fibers located in the parenchyma, which evidences inhibition of the neurogenous mechanisms of vasomotor control and possible selective activation of the sympathetic mechanisms involved in the regulation of the immunogenesis. Thymic preparations impregnated with silver salts manifested high argyrophilia of the nerve cells, characteristic of the reactive phase of the destructive process. The problems of the hypothalamic effects on the blood supply and functions of the immune system organs are discussed.     

Adrenergic and cholinergic innervation of rat cerebral arteries

Summary A consecutive demonstration of both aminergic and cholinergic nerves of rat cerebral arteries was carried out on whole mount preparations. For demonstration of aminergic nerves the glyoxylic acid method was used, while for cholinergic nerves Karnovsky's technique was utilized consecutively. This procedure provided a highly sensitive and reproducible demonstration of the two systems on the same specimen. The results obtained were as follows: 1) Aminergic and cholinergic nerves were distributed densely in the proximal portions of the major cerebral arteries examined. They were dense in the arteries of the anterior circulation and sparse in the posterior circulation. Each nerve had two different patterns, i.e. circular and longitudinal. Both circular and longitudinal patterns were observed in the proximal portions of the arteries, while in the distal portions, longitudinal fibres were predominantly present. 2) Superior cervical ganglionectomy produced no change in cholinergic nerve distribution, while it caused decreased density of aminergic nerves on the ipsilateral side on the arteries of the anterior circulation. The contralateral anterior cerebral arteries were partially affected. Bilateral sympathectomies abolished aminergic nerves in all arteries except the vertebral artery.     

Comparative characteristics of adrenergic innervation of the cerebrospinal arteries of vertebrates and man

Adrenergic innervation of the cerebrospinal arteries in redeye, frog, tortoise, hen, sea-gull, guinea-pig, rabbit, cat, dog and man has been studied by means of fluorescent-histochemical method. In all the animals there is a similar principle of innervation. The nervous apparatus is revealed as continuous integral plexuses. In the process of historical development the role of the nervous factor in regulation of the cerebrospinal circulation is increasing. As the innervational degree of the spinal arteries decreases, the vertebrates make the following line: mammals, birds, reptiles, amphibia, fishes. In all the animals, besides the fishes, the greatest concentration of the nervous fibers is revealed in the areas of the spinal cord thickenings.     

Morphofunctional state of the cholinergic innervation of the rat pancreas during vagotomy]

Cholinergic innervation of the albino rat pancreas was investigated under normal conditions and after bilateral subdiaphragmatic vagotomy (one, three, seven, twenty one days and two months after the operation). In control animals cholinergic nerve fibers were found in all the organ structures -- both in the vascular and in the gland-duct systems. Vagotomy resulted in a short-term increase in the nerve fibers revealed at the end of the first day, followed by their decrease on the seventh day; but in two months the number of detectable nerve fibers approached the initial level.     

Generators of synchronous activity of the locus coeruleus during development

The locus coeruleus is thought to play an important role in the development of the central nervous system through a coordinated release of noradrenaline. The influence of the locus coeruleus on its diverse targets is strongly synchronized by the existence of electrical and chemical coupling between neurones, afferent supply of the nucleus from restricted sources and diffuse innervation of target areas. Extensive coupling between neonatal locus coeruleus neurones produces rhythmic synchronized electrical activity and distributes afferent synaptic activity throughout the entire nucleus. The strength of electrical coupling declines with age but appears to persist to a limited extent in the adult.     

Increased sympathetic innervation in the cerebral and mesenteric arteries of hypertensive rats

The density of catecholamine-containing nerve fibers was studied in the cerebral and mesenteric arteries from normotensive Wistar-Kyoto rats (WKY), spontaneously hypertensive rats (SHR), and stroke-prone SHR (SHRSP) in the growing (SHR, WKY) and adult (SHR, SHRSP, WKY) animals. Cerebral arteries from SHR showed an increased adrenergic innervation from day 1. The nerve plexuses reached an adult pattern earlier in SHR than in WKY. The arteries from adult SHR and SHRSP (22 weeks old) showed a markedly higher nerve density than WKY. There was a positive linear correlation between blood pressure and nerve density for four cerebral arteries. The mesenteric arteries were not innervated at birth. However, hyperinnervation of these arteries in the SHR was already present at 10 days of age as compared with WKY. Sympathectomy with anti-nerve growth factor and guanethidine caused a complete disappearance of fluorescent fibers in the mesenteric arteries from SHR and WKY, and in the cerebral arteries of WKY. The same procedure caused only partial denervation of the cerebral arteries from hypertensive animals. We postulate that the increase in nerve density in the cerebral arteries from the hypertensive rats may contribute to the development of arterial hypertrophy in chronic hypertension through the trophic effect of the sympathetic innervation on vascular structure.     

The dynamic behavioral changes in rats after destruction of the locus coeruleus]

Destruction of locus coeruleus (LC) was conducted in adult rats (male rats of the Wistar line, 250-300 g) electrolytically or by means of 6-hydroxydopamine (6-OHDA). Behaviour of the intact animals sham-operated and of rats with destructed LC was evaluated before and every week after the operation during 5 weeks. Statistically significant alterations in the behaviour were revealed only in the initially low-active rats. These animals were characterized firstly, by low motor and exploratory activity in all tests (open field and automated devices), disturbance of the reactivity to novelty--only in one week after the operation with subsequent compensation of shifts; secondly, by low level of investigatory activity in the open field and absence of habituation to the repetitive tests--from the 21-st day after the operation.     

Morphofunctional changes in the pineal gland during adaptation to hypothermia

The influence of hypothermal stress (+4 degrees during 3 h) on the ways of serotonin metabolism in pineal gland and its structure has been studied in dynamics on adult male Wistar rats. It has been revealed that melatonin-producing epiphyseal function suffers from phase changes in dynamics of adaptation--significant rising during 15 min. after beginning of the experiment, rehabilitation up to normal--in 30 min, and fast suppressing--in 3 hrs. Suppressing of the functional pineal activity is not due to switched serotonin metabolism with melatonin and new indoles release, but to a partial pinealocytes breaking from their active function.