Effect of an inhibitor of serotonin synthesis--para-chlorophenylalanine on the structure of primary sleep in the frog Rana temporaria

On the basis of statistical analysis of the EEG of the forebrain in frogs, new data have been obtained concerning neurophysiological nature of the resting forms of the primary sleep. It was found that natural rest occurring with the increase in the rigid muscle tonue (P-2) is identical to the condition which results from injections of parachlorphenylalanine, an inhibitor of serotonin synthesis. New data were obtained showing participation of the posterior hypothalamus in regulation of one of the resting forms of the primary sleep (P-2). Possible homology of the rest of catatonic type (P-2) and anabiosis of frogs is discussed.     

Histochemical determination of RNA and protein in the brain during primary sleep in the frog Rana temporaria and during intermediate sleep in the turtle Emys orbicularis

It has been shown that resting forms of the primary sleep in the frog and intermediate sleep in the tortoise (P-1, P-2 and P-3) differ from each other with respect to neurochemical characteristics. Cytospectrophotometric studies reveal the increase in the content (per one cell) and concentration of total protein and RNA in cells of the preoptic nuclei of the frog only during one of the resting forms of the primary sleep (P-3), which represents functional homologue of the intermediate sleep of reptiles and sleep of homoiothermic animals. These anabolic changes were not observed during two other resting forms of the primary sleep, this finding being consistent with divergent development of resting forms of the primary sleep in evolution of vertebrates.     

Neurophysiological characteristics of the resting forms of primary sleep and hypobiosis in the frog Rana temporaria

Neurophysiological studies have been made on natural resting forms of the primary sleep, wakefulness and hypobiosis in the frog. New method of presentation of the distribution density of EEG signal by a frequency spectrum was used. New data were presented concerning seasonal changes in the frequency parameters of one of the resting forms of the primary sleep, SLS-2. The role of this form of rest in the origin of hypobiosis in the frog Rana temporaria is discussed. Besides, new data were obtained which confirm that the resting form, SLS-3, of the primary sleep is an evolutionary precursor of the intermediate form of the sleep in reptiles and slow wave and activated phase of the sleep in warm-blooded vertebrates.     

Blood flow and pO2 in the posterior hypothalamus of cats during paradoxical sleep

In chronic experiments on cats it has been found by recording of the brain local blood flow (BLBF) and of oxygen tension (pO2) in the posterior and anterior hypothalamus, that at sleep phases alternation, the changes of these parameters are differently directed: during the paradoxical sleep the level of BLBF and pO2 oscillations frequency increased in the posterior hypothalamus and decreased in the anterior one. During slow-wave sleep opposite relations were observed. Opposite directions of changes of BLBF level and pO2 oscillations frequency in one and the same phase of sleep show that they are of local origin and must be determined by functional-metabolic shifts. In particular, the increase of BLBF level and frequency of pO2 oscillations must reflect a rise of posterior hypothalamus functional-metabolic activity during paradoxical sleep.     

Morphodynamics of the nervous system in the regenerative processes of Convoluta convoluta (Turbellaria, Acoela)]

By means of the method for revealing cholinesterases the alterations in the nervous system of Convoluta convoluta after mechanical lesion were studied. After a transversal transection of the animal into two pieces the anterior part of the body acquired the normal form within 3-4 days; the nervous system underwent inconsiderable transformations associated with a formation of a plexus in the injured area. The posterior parts of the body regenerated slowly. During the first days disintegration of the resting parts of the nervous system was observed. Later on the diffuse cholinesterase activity suggesting the beginning of the brain formation was manifested in the wound field. Plexus strands "grow" in the caudal direction from the brain in parallel with its development. Within three weeks the nervous system of the animals which had restored their mouth opening, body shape and normal behaviour consists of the brain and plexus not arranged in nervous trunks. The reaction of the nervous system to the ablation of a part of the brain and to a continuous mechanical injury were studied. The data obtained show a high morphological lability of the nervous system of Convoluta convoluta, its ability to radical structural rearrangements.     

Mesoderm induction in the future tail region of Xenopus

Summary We have used interspecific grafts between Xenopus borealis and Xenopus laevis to study the signalling system that produces tail mesoderm. Early gastrula ectoderm grafted into the posterior neural plate region of neurulae responds to a mesodermal inducing signal in this region and forms mainly tail somites; this signal persists until at least the early tail bud stage. Ventral ectoderm grafted into the posterior neural plate loses its competence to respond to this signal after stage 10 1/2. We have established the specification of anterior and posterior neural plate ectoderm. In ectodermal sandwiches or when grafted into unusual positions, anterior regions gave rise to mainly nervous system and posterior regions to large amounts of muscle, together with some nervous system. Thus it was impossible to assess the competence of posterior neural plate ectoderm to form further mesoderm and hence to establish if mesodermal induction continues during neurulation in unmanipulated embryos.     

Primary sleep in vertebrates and its role in the genesis of hypobiosis in poikilotherms and hibernation in mammals

New data are presented on the homology between winter hibernation and hypobiosis in poikilotherms, as well as one of the resting forms of the primary sleep in vertebrates. Different stages of formation of a cycle "awakefulness-sleep" in evolution of vertebrates are discussed. On the basis of universal behavioural, somato-vegetative, neurophysiological, neurochemical correlates of resting forms, hypobiosis and winter hibernation, a discussion is made of the problem of genetic fixation in the genotype and phenotype of heterothermic mammals (hibernating ones) of those characters which are typical of the sleep in homoiothermic and poikilothermic vertebrates.     

Functional similarity of the P-2 form of primary sleep and hypobiosis in the frog Rana temporaria (based on data from luminescent microspectral analysis of the cytoplasm of neurosecretory neurons)

Using luminescent microspectral analysis of preparations stained by acridine orange, studies have been made on the ratio between single- and double-stranded parts in ribosomal RNA from the cytoplasm of neurosecretory neurones of the preoptic nucleus of the frog Rana temporaria. The animals were investigated in active period, during P-2 form of the primary sleep, in hypobiosis, and after injection of the active factor (peptide fraction with a molecular mass 1-10,000 Da) extracted from the small intestine of the ground squirrel Citellus undulatus during winter hibernation. It was shown that unlike actively awake frogs, animals from other experimental series exhibited similar changes, i.e. the decrease in the affinity of acridine orange to single- and double-stranded parts of rRNA and the decrease in the value of alpha which reflects the ratio of single- and double-stranded parts. It was also demonstrated that injection of the active factor from hibernating ground squirrels to frogs results in a condition which is rather similar to a natural resting form (P-2) of the primary sleep in cold-blooded vertebrates.     

Changes in cardiac rhythm of rats in dehydration

80 mongrel rats were studied for peculiarities of the heart activity regulation in dehydration, conditions of the vegetative homeostasis being different. Data of the variation pulsometry were used. A sympathetic shift of the vegetative homeostasis was a common dehydration-caused response. The shift was significant in rats with an initial equilibrium of the vegetative homeostasis and prevailing parasympathetic effects. The survival rate of this group of rats was high. In the group of rats with the initial prevalence of a sympathetic tonus a short-term sympathetic shift was replaced by an increase of parasympathetic effects. The survival rate of this group was much lower. Therefore, rats with initial prevalence of the sympathetic compartment tonus of the vegetative nervous system are more labile to the effect of the dehydration stress.     

Neuropeptides in the gastrointestinal canal of Necturus maculosus

Summary The presence and distribution of regulatory peptides in nerves and endocrine cells of the stomach, intestine and rectum of a urodele amphibian, the mudpuppy, Necturus maculosus, was studied immunohistochemically in sections or whole-mount preparations of the gut wall. The effect of the occurring peptides on gut motility was studied in isolated strip preparations of circular and longitudinal smooth muscle from different parts of the gut.Bombesin-, neurotensin-, substance P- and VIP-like immunoreactivity was present in abundant nerve fibres in the myenteric plexus of both stomach, intestine and rectum. Single fibres or bundles were present in the circular muscle layer and in a well-developed deep muscular plexus in the intestine and rectum. Immunoreactive nerve cells were found in the myenteric plexus of the stomach, intestine (neurotensin only) and rectum. Gastrin/CCK-like immunoreactivity was observed only in a few fibres in stomach and rectum.Endocrine cells containing bombesin-, met-enkephalin-, gastrin/CCK-, neurotensin-, somatostatin- or substance P- like immunoreactivity were present in the mucosa.The effect of bombesin was an inhibition of the rhythmic activity in circular muscle preparations and in longitudinal muscle from the rectum, while longitudinal muscle from the stomach usually responded with a weak increase in tonus. Neurotensin, like bombesin, was inhibitory on the spontaneous rhythmic activity of circular muscle throughout the gut, while the effect on longitudinal muscle was an increase in tonus. Met-enkephalin and substance P increased the tonus of all types of preparations, and often, in addition, initiated a rhythmic activity superimposed on this maintained tonus. VIP had a general inhibitory effect on the preparations, decreasing tonus and/or abolishing rhythmic activity.It is concluded that bombesin-, neurotensin-, substance P- and VIP-like peptides are present in nerves throughout the urodele gut and may have physiological functions in regulating the motility of the gut. The gastrin/CCK-like peptide present in nerves of the stomach and rectum may affect the function of these parts of the gut. The regulatory peptides present in endocrine cells may, perhaps with the exception of the somatostatin-like peptide, affect the motility humorally.     

Distribution of immunoreactive mesotocin and vasotocin in the brain and pituitary of chickens

The distribution of vasotocin and mesotocin in the pituitary and central nervous system in male chickens was determined using radioimmunoassays. Neither peptide was detected in the pineal. Mesotocin, but not vasotocin, was detected in the cerebellum. Both peptides were found in the septal area, archistriatum, paleostriatum, optic lobe, anterior, medial and posterior hypothalamus, midbrain, pons, medulla oblongata, and the anterior and posterior pituitary. Equal amounts of the 2 peptides were present in the septal area, archistriatum and anterior hypothalamus whereas vasotocin was more abundant (2- to 10-fold) in the paleostriatum, optic lobe, midbrain, and pituitary. The amount of mesotocin was about twice that of vasotocin in the medulla oblongata and the medial and posterior hypothalamus. The wide distribution of vasotocin and mesotocin in extrahypothalamic sites in the central nervous system suggests that the peptides may, as in mammals, have a role in a variety of autonomic and endocrine regulatory processes in chickens.     

Vasopressin and developmental onset of flank marking behavior in golden hamsters

Golden hamsters start displaying flank marking behavior (a form of scent marking) around postnatal day 20 (P-20). Because the behavior is dependent upon the central activity of arginine vasopressin (AVP), the present study was conducted to correlate this activation with changes in the vasopressinergic system. A first set of experiments was performed to compare flank marking activity between P-18 and P-22. A second set of experiments was performed to compare the density of AVP receptors between the age periods and assess responsiveness to AVP microinjection. Finally, a third set of experiments incorporated immunocytochemistry, radioimmunoassay, in situ hybridization, and Northern blot analysis to determine the location and numbers of AVP immunoreactive neurons and the level of mRNA correlating with the developmental onset of flank marking behavior. Our results show that flank marking develops between P-18 and P-22. Male and female hamsters do not display odor-induced flank marking anytime before P-19. However, all animals show odor-induced flank marking by P-22. The onset of flank marking does not appear to be associated with any change in AVP receptor binding in the anterior hypothalamus. Indeed, flank marking can be triggered in hamsters on P-18 by the microinjection of AVP in the anterior hypothalamus. This would suggest that the postsynaptic mechanisms contributing to the transduction of the AVP signal and the motor control of flank marking are intact prior to the onset of odor-induced flank marking. In contrast, AVP levels in the hypothalamus and pituitary increase by two to threefold between P-18 and P-22, suggesting that changes in AVP synthesis and release from presynaptic sites may contribute to the onset of flank marking. Interestingly, there is no change in AVP mRNA between P-18 and P-22, which raises questions about posttranslational processing during this developmental period. These results suggest that heightened synthesis and release of AVP between P-18 and P-22 may contribute to the developmental onset of flank marking. © 1996 John Wiley & Sons, Inc.     

A reappraisal of the anatomy of the levator ani muscle in man

A study of the attachments of the musculotendinous fibres of the levator ani muscle shows that it is made of two portions: a thick anterior portion which is mostly fleshy and a thin posterior portion which is mostly aponeurotic. The anterior portion consists of two layers: a superficial perineal layer and a deep pelvic layer. Both layers have a common origin from the back of the body of the pubic bone and the anterior part of the tendinous arch. In addition both layers make a U-shaped loop around the recto-anal junction. The posterior fibres of the deep pelvic layer received nerve supply only from the third and fourth sacral nerves. The rest of the muscle was supplied from the sacral nerves as well as the perineal branches of the pudendal nerve. The role of the anterior fibres in reinforcing the sphincters of the anal canal and fixation of the pelvic viscera is stressed. The close anatomical relation between the posterior portion of the muscle and the obturator internus suggests that the latter may play a role in supporting the weak posterior portion of the levator ani, especially during straining positions associated with lateral rotation at both hips.     

Stomodeal and neurohypophysial placodes in Ciona intestinalis: insights into the origin of the pituitary gland

The ascidian larva has a central nervous system which shares basic characteristics with craniates, such as tripartite organisation and many developmental genes. One difference, at metamorphosis, is that this chordate-like nervous system regresses and the adult's neural complex, composed of the cerebral ganglion and associated neural gland, forms. It is known that neural complex differentiation involves two ectodermal structures, the neurohypophysial duct, derived from the embryonic neural tube, and the stomodeum, i.e. the rudiment of the oral siphon; nevertheless, their precise role remains to be clarified. We have shown that in Ciona intestinalis, the neural complex primordium is the neurohypophysial duct, which in the early larva is a short tube, blind anteriorly, with its lumen in continuity with that of the central nervous system, i.e. the sensory vesicle. The tube grows forwards and fuses with the posterior wall of the stomodeum, a dorsal ectodermal invagination of the larva. The duct then loses posterior communication with the sensory vesicle and begins to grow on the roof of the vesicle itself. The neurohypophysial duct differentiates into the neural gland rudiment; its dorsal wall begins to proliferate neuroblasts, which migrate and converge to build up the cerebral ganglion. The most anterior part of the neural gland organizes into the ciliated duct and funnel, whereas the most posterior part elongates and gives rise to the dorsal strand. The hypothesis that the neurohypophysial duct/stomodeum complex possesses cell populations homologous to the craniate olfactory and adenohypophysial placodes and hypothalamus is discussed.     

Effect of blue-green alga (Cyanobacteria) and their exometabolites on formation of resting forms and variability of Yersinia pseudotuberculosis

Research, carried out with the use of bacteriological methods and polymerase chain reaction, revealed that the transformation of Y. pseudotuberculosis, associated with blue-green algae Anabaena variabilis, into resting (noncultivable) forms took shorter time than in soil extract containing no algae. The exometabolites of "old" cultures of these algae sharply accelerated the formation of resting Y. pseudotuberculosis forms. The influence of the algae and the products of their metabolism was manifested far more intensively at 22 degrees C than at 4 degrees C. After passage through infusoria resting Y. pseudotuberculosis forms, preserved in the mucous covering of cyanobacteria, partially reverted into vegetative forms, capable of growing on solid culture media. The revertants essentially differed from the initial vegetative forms by having lower enzymatic activity, agglutinability and cytopathogenicity, as well as by the loss of plasmid p45. The probable role of blue-green algae, widely spread in soils and water reservoirs, in the processes of reversible transformation of Y. pseudotuberculosis vegetative and resting forms, closely connected with seasonal changes of temperature conditions.     

Further consideration of phenobarbital effects on cytochrome P-450 activity in the killifish, Fundulus heteroclitus

1. Ethoxyresorufin O-deethylase (EROD) activity, aldrin epoxidase (AE) activity, cytochrome P-450 content, and levels of cytochrome P-450E (the major BNF-inducible P-450 form and primary EROD catalyst in scup) or its homologues were measured in hepatic microsomes isolated from Fundulus heteroclitus, scup (Stenotomus chrysops) and brook trout (Salvelinus fontinalis) treated with beta-naphthoflavone (BNF) or phenobarbital (PB). 2. In all three teleost species, BNF treatment caused expected increases in P-450 content, EROD activity and P-450E level; but either no change or a slight decrease in AE turnover rate (nmol/min/nmol P-450). 3. Polyclonal antibodies to P-450E did not inhibit AE activity in microsomes from BNF-treated scup, confirming that this major BNF-inducible P-450 form does not catalyze AE activity in fish. 4. In contrast, PB treatment did not affect hepatic AE activity, P-450 content or levels of "P-450E" in F. heteroclitus, but did variably affect EROD activity which was suppressed in one experiment and elevated in another. 5. The results indicate that (i) contrary to previous reports, neither PB nor MC-type inducers increase AE activity in F. heteroclitus, (ii) MC-type inducers do not affect AE activity in the other teleost species examined, and (iii) AE activity is not a reliable indicator of P-450 induction by environmental chemicals. 6. We emphasize the need to establish the mechanism of PB action, and the nature of any fish P-450 forms analogous to PB-inducible forms in mammals in order to conclusively evaluate PB-responses in fish.     

The role of the posterior hypothalamus in the regulation of paradoxical sleep

Posterior hypothalamus was found to take part in the inhibitory control of the paradoxical sleep executive mechanisms responsible for the ECoG desynchronisation and phasic events. Functional activity of the posterior hypothalamus seems to be at its lowest during the paradoxical sleep stage as characterised by phasic events and the ECoG desynchronisation, and increases during the stage with alpha-like activity in the ECoG and absence of phasic events, the latter having, probably, a "sentinel" function.     

Adrenovascular lesions from neurogenic stress modified by suppression of nervous centres in rat

Adrenovascular lesions (hemorrhages and/or edema) caused by neurogenic stress, are modified by suppression of different nervous centres. Spinal transection at Th5 abolishes the adrenovascular response to neurogenic stress. Destruction of the anterior or posterior hypothalamus decreases this vascular response; the greatest effect was obtained by destruction of the hypothalamus medius or by hemidecortication. This effect was bilateral but more manifest contralaterally in cases with unilateral destruction. These results show that the adrenovascular reaction to neurogenic stress is regulated by the central nervous system, mainly by the hypothalamus and cerebral cortex.     

Influence of lead exposure on catecholamine metabolism in discrete rat brain nuclei

1. Using the rat exposed both acutely and chronically to lead as a model of lead neurotoxicity, various parameters of catecholamine metabolism were investigated. 2. The steady-state concentrations of noradrenaline, adrenaline and dopamine together with the activities of tyrosine hydroxylase and phenylethanolamine N-methyl transferase were measured in discrete brain nuclei--periventricular, paraventricular, median eminence, posterior and anterior hypothalamus, caudate putamen and globus pallidus. 3. Lead exposure resulted in significant fall in the activity of the rate-limiting enzyme of catecholamine synthesis, tyrosine hydroxylase which was associated with alterations in concentrations of catecholamines in the median eminence, periventricular nucleus and anterior hypothalamus. 4. No other brain nuclei investigated exhibited any effect of lead on the catecholaminergic nervous system and, therefore, the effect of lead on rat brain can be considered to be regionally specific.     

The mechanisms of the influence of the anterior and posterior hypothalamus on the electrical activity of the large hemispheres of the forebrain and on autonomic reactions in hens

Studies have been made of the effect of stimulation of the anterior and posterior hypothalamus on the electrical activity of the hemispheres, arterial blood pressure, heart and respiration rates. Mainly desynchronizing mechanisms were revealed in the ascending influences from both the anterior and posterior hypothalamus. Concerning the descending influences, it was found that stimulation of the anterior hypothalamus evokes depressor reactions, whereas stimulation of the posterior hypothalamus results in pressor reactions. Peculiarities of evolutionary development of the ascending and descending mechanisms of the posterior and anterior hypothalamus are discussed.