[Development of the habenulointerpeduncular tract in rats].

Development of the habenulointerpeduncular tract has been carried out on fixed brain preparations obtained from 21 day rat embryos and from neonatal animals on the 0 and 9 days of postnatal development by diffusion oflipophilic fluorescent carbocyanine dye DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate) through neuron membranes. The marker was introduced into the nuclei of the habenula, the interpeduncular nucleus, and into the area of raphe nuclei. Neurons and fibers that contained Dil were identified on vibratome sections by fluorescent and confocal microscopy. we have found that reciprocal links between the lateral habenula nucleus and raphe nuclei are formed in the prenatal period by stage E21. Raphe nuclei innervating neurons were located in dorso- and ventrocaudal parts of the lateral habenula nucleus. Projections of the medial habenula nucleus onto interpeduncular nucleus were found only in the postnatal P2 period. Neurons that provide a source of these projections form characteristic assemblies inside the medial habenula nucleus. Therefore, the present study for the first time describes heterogenic formation of different projection systems that are involved in the habenulointerpeduncular tract of rats at perinatal ontogenesis.

Perinatal development of the mammillothalamic tract and innervation of the anterior thalamic nuclei.

Axonal projections originating from the mammillary bodies represent important pathways that are essential for spatial information processing. Mammillothalamic tract is one of the main efferent projection systems of the mammillary body belonging to the limbic "Papez circuit". This study was aimed to describe the schedule of the mammillothalamic tract development in the rat using carbocyanine dye tracing. It was shown for the first time that fibers of the mammillothalamic tract being the collaterals of the mammillotegmental tract axons start bifurcating from the mammillotegmental tract on E17. The axons of the mammillothalamic tract grow simultaneously and reach the ventral region of the anterior thalamus where they form first terminal arborizations on E20-E21. Ipsilateral projections from the medial mammillary nucleus to the anteromedial and anteroventral thalamic nuclei develop from E20 to P6. Bilateral projections from the lateral mammillary nucleus to the anterodorsal thalamic nuclei develop later, on P3-P6, after the formation of the thalamic decussation of the mammillary body axons. Unique spatial and temporal pattern of the perinatal development of ascending mammillary body projections to the anterior thalamic nuclei may reflect the importance of these connections within the limbic circuitry.

[Perinatal development of mammillotegmental connections in rats].

Development of direct axonal connections of the hypothalamic mammillary bodies with ventral and dorsal tegmental nuclei of Gudden was studied on fixed rat brains from day 14 of embryonic development until day 10 of postnatal development using the method of diffusion of the lipophilic fluorescent carbocyanine tracer 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate along the neuronal membranes. The tracer was inserted into the mammillary bodies or into the tegmentum and after incubation in a fixative fluorescent nerve cells and nerve fibers were visualized in the brain tissue. The mammillotegmental tract was found to start developing earlier than other conducting systems of the mammillary bodies. On days 14-15 of embryonic development, it was visualized as a bundle of axons running from the mammillary bodies caudally to the midbrain. A group of neurons in the midbrain tegmentum and their axons going to the mammillary bodies via the mammillary peduncle were first visualized on day 19 of embryonic development. The mammillotegmental tract and mammillary peduncle developed progressively from the moment of birth. Ventral and dorsal tegmental nuclei were formed in the midbrain by day 10 of the postnatal development. Thus, the formation of reciprocal connections of the mammillary bodies with midbrain tegmental nuclei was first described during perinatal development in rats.

[Effect of catecholamines on migration and differentiation of gonadotropin releasing hormone-neurons in rats]. / Vliianie kateholaminov na migratsiiu i differentsirovku gonadotropin-relizing gormon-neironov u krys.

The GnRH producing neurons are the key link of neuroendocrine regulation of the adult reproductive system. Synthesis and secretion of GnRH are, in turn, under the afferent catecholaminergic control. Taking into account that catecholamines exert morphogenetic effects on target cells during ontogenesis, this study was aimed at investigation of the effects of catecholamines on development of GnRH neurons in rats during ontogenesis. We carried out comparative quantitative and semiquantitative analyses of differentiation and migration of GnRH neurons in fetuses of both sexes under the conditions of normal metabolism of catecholamines (administration of saline) or their pharmacologically induced deficiency (administration of alpha-methylparatyrosine). The inhibition of catecholamine synthesis from day 11 of embryogenesis led to an increasing number of GnRH neurons in rostral regions of the trajectory of their migration over the brain: in the area of olfactory tubercles on day 17 and in the area of olfactory bulb on days 18 and 21. In addition, the optical density of GnRH neurons located in the rostral regions of migration was higher in the fetuses after administration of alpha-methylparatyrosine during embryogenesis, as compared to the control. It has been concluded that catecholamines stimulate the migration of GnRH neurons and affect their differentiation.

Distribution of serotonin 5-hydroxytriptamine 1B (5-HT(1B)) receptors in the normal rat hypothalamus.

This is the study first attempting to evaluate distribution of neurons expressing serotonin 5-hydroxytriptamine 1B (5-HT(1B)) receptors in hypothalamus by using immunocytochemistry. The 5-HT(1B)-immunoreactive neurons were widely distributed in hypothalamus. Accumulations of 5-HT(1B) neurons occurred in magnocellular nuclei, supraoptic nucleus, paraventricular nucleus (dorsolateral part) and accessory perifornical, circular and retrochiasmatic nuclei. Magnocellular neurons manifested an intense immunostaining suggesting a high level of 5-HT(1B) receptors. Large and middle-sized neurons with different 5-HT(1B) staining patterns were scattered throughout lateral hypothalamus, periventricular nucleus and lateral preoptic area. Immunofluorescent double-labeling revealed a great overlapping of the distribution 5-HT(1B) neurons and dense network of neuropeptide Y-immunoreactive fibers in paraventricular, supraoptic and arcuate nuclei. The potential functional significance of 5-HT(1B) receptors in the 5-HT control of endocrine functions and feeding are discussed.

[The source of transitory innervation of suprachiasmatic nucleus by tyrosine hydroxylase-immunoreactive fibers during the postnatal period in rats]. / Istochnik tranzitornoi innervatsii suprakhiazmaticheskogo iadra tirozingidroksilaza-immunoreaktivnymi voloknami v postnatal'nom periode u krys.

Suprachiasmatic nucleus in the rats during early postnatal development is transitorily innervated by tyrosine hydroxylase-immunoreactive fibers that are neither catecholamine- nor serotoninergic. The goal of this immunocytochemical investigation was to find out if tyrosine hydroxylase-immunoreactive neurons of anterior hypothalamus could be the source of this innervation. According to the obtained immunocytochemical data, multiple multipolar tyrosine hydroxylase-immunoreactive neurons are localized around the suprachiasmatic nucleus in the rats at days 2 and 10 of postnatal development. Most of them were observed ventrally and laterally to the nucleus. The axons of the neurons are oriented towards the suprachiasmatic nucleus. Further investigation demonstrated considerably decreased number of tyrosine hydroxylase-immunoreactive neurons surrounding the suprachiasmatic nucleus in the adult animals as compared to early postnatal period, which correlates to the number of tyrosine hydroxylase-immunoreactive fibers in this nucleus. Hence, tyrosine hydroxylase-immunoreactive neurons in the ventral region of anterior hypothalamus can be considered as a potential source of transitory innervation of suprachiasmatic nucleus by tyrosine hydroxylase-immunoreactive fibers during early postnatal development.

[Involvement of accessory neurosecretory nuclei of hypothalamus in the formation of hypothalamohypophyseal system during prenatal and postnatal development in rats]. / Uchastie dopolnitel'nykh neirosekretnykh iader gipotalamo-gipofizarnoi sistemy v pre- i postnatal'nom ontogeneze u krys.

We studied the development of direct axonal connections of the accessory neurosecretory hypothalamic nuclei with the posterior pituitary lobe on the fixed rat brain from day 15 of embryogenesis until day 10 of postnatal development using the retrograde diffusion method of the lipophilic fluorescent carbocyanine dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate along the neuronal membranes. The marker was applied onto the posterior pituitary lobe and, after incubation in a fixative, fluorescing bodies of nerve cells were visualized in the hypothalamus. Neuronal axons of the retrochiasmatic nucleus were the first of the accessory nuclei to ingrow in the posterior pituitary lobe (on days 16-17 of embryogenesis). Neurons of the circular and dorsolateral nuclei and the nuclei of the median bundle of the forebrain sent their axons to the posterior pituitary lobe starting from the first days of postnatal development. No direct connections of the anterior commissure and perifornical accessory nuclei with the posterior pituitary lobe were found in perinatal development. These facts are discussed in the light of concepts about the different functional role of accessory peptidergic hypothalamic nuclei in rats.

Axonal projections from the hypothalamus to the median eminence in rats during ontogenesis: DiI tracing study.

This study has determined the ontogenetic schedule of the arrival of the axons from the hypothalamus and the diagonal band in the median eminence in rats by using the fluorescent lipophilic carbocyanine dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) as a retrograde tracer. After fixation of the brain, the crystals of the dye were implanted in the median eminence on the 13th, 14th, 15th, 16th, 17th, 20th embryonic days, and on the 2nd postnatal day. This was followed by fluorescent staining of the neuronal cell bodies in the hypothalamus. According to our data, the axons of rare hypothalamic neurons first reached the primordium of the median eminence on the 14th embryonic day. For two subsequent days, the number of neurons projecting the axons to the median eminence appeared to increase considerably. They were widely distributed through the hypothalamus and in the ventromedial region of the more rostral forebrain. Till the 20th embryonic day, the majority of the fluorescent neurons were concentrated mainly in the paraventricular nucleus (dorsal and medial parts) and the arcuate nucleus, and to a lesser extent in the medial preoptic nucleus, the supraoptic nucleus, the diagonal band, and the retrochiasmatic nucleus. In neonates, DiI-labelled neurons appeared additionally in the accessory dorsolateral nucleus, medial preoptic area lateral to the diagonal band, anterior hypothalamic area, and in the anterior periventricular nucleus. Thus, the axons of differentiating neurons arrive in the median eminence from the 14th embryonic day till the neonatal period, providing the pathway for the neurohormone transfer to the hypophysial portal circulation.

Projections from the hypothalamus to the posterior lobe in rats during ontogenesis: 1,1'-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate tracing study.

The objective of this study was to determine the schedule of the arrival of the axons from the hypothalamus to the posterior lobe of the pituitary (PL) in rats during ontogenesis by using the fluorescent lipophilic carbocyanine dye 1,1'-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate (DiI) as a retrograde tracer. After preliminary fixation of the brain, DiI crystals were implanted in the PL on embryonic day 15 (E15), E16, E17, and E19 as well as on postnatal day 2 (P2) and P9. This was followed by a DiI retrograde diffusion along the plasma membrane and subsequent staining of hypothalamic neuronal cell bodies. The supraoptic nucleus (SO) contained an accumulation of fluorescent cells that extended toward the diamond-like swelling of the third ventricle as early as E15. These data suggest that the magnocellular neurons of the SO send their axons to the PL at the very beginning of differentiation, perhaps even before reaching their final position. The initial axons of the neurons of the paraventricular nucleus proper (PV) appeared to reach the PL significantly later, at E17. In addition to the SO and the PV, accessory magnocellular nuclei contributed to the innervation of the PL in perinatal rats. The neurons of the retrochiasmatic accessory nucleus first sent their axons to the PL on E16-E17. Axons that originated from other accessory hypothalamic nuclei reached the PL after birth, suggesting a delay in their involvement in the regulation of visceral functions compared with other magnocellular nuclei. Thus, the axons of magnocellular neurons reach the PL unexpectedly early in embryogenesis, raising the possibility of the functional significance of vasopressin and oxytocin as fetal neurohormones.

[The development of connections of the magnocellular hypothalamic nuclei with the posterior hypophyseal lobe in rat prenatal ontogeny]. / Razvitie sviazei krupnokletochnykh iader gipotalamusa s zadnei dolei gipofiza v prenatal'nom ontogeneze u krys.

The development of projections of the hypothalamic nuclei into the posterior lobe of the pituitary was studied on the fixed brain of rat fetuses from day 15 until day 19 of embryogenesis using retrograde staining with the fluorescent carbocyanine dye DiI. The formation of connections of the supraoptic and retrochiasmatic nuclei of the hypothalamus with the posterior lobe of the pituitary takes place during prenatal development on days 15 and 16-17, respectively, while only an insignificant number of the paraventricular nucleus neurons send their axons to the posterior lobe of the pituitary in rat fetuses. These facts suggest different temporal involvement of the above nuclei in formation of the hypothalamic-hypophysial neurosecretory system in rat fetuses.

[A comparative study of the osmoregulating system in the hamsters Phodopus roborovskii and Phodopus sungorus]. / Sravnitel'noe izuchenie osmoreguliruiushchei sistemy u mokhnonogikh khomiachkov Phodopus roborovskii i Phodopus sungorus.

The hamster Phodopus roborovskii is adapted to live in desert, Phodopus sungorus in less arid regions. Hamsters of both species have rather close values of blood serum osmolality and ion concentration, after dehydration the value of urine osmolality is found to be equally high. The microdissection of the kidney did not reveal any difference in the structure and arrangement of nephrons in cortex and medulla. According to the results of immunocytochemical analysis of the hypothalamo-hypophyseal neurosecretory system of hamster Phodopus roborovskii, compared to Phodopus sungorus, has a smaller quantity of vasopressin-positive neurons, which is likely to be a result of a release of larger quantities of vasopressin in long-term dehydration. It is suggested that of importance in the animal adaptation to life in arid condition may be not only morphofunctional reorganization of kidney but also of the hypothalamo-hypophyseal system which is involved in permanent secretion of vasopressin.

[The detection of cytoplasmic creatinine phosphokinase BB in nerve cell nuclei under normal conditions, in schizophrenia, and in Alzheimer's disease patients]. / Vyiavlenie tsitoplzmaticheskoi KFK BB v iadrakh nervnykh kletok v norme, pri shizofrenii i bolezni Al'tsgeimera.

This work is devoted to the study of CPK BB content in nuclear fraction of nervous cells in normal brain and in brains of mental (schizophrenia and Alzheimer disease) patients. With the help of the immunocytochemistry and immunoblotting was detected, that the nuclear membrane fraction of brain contains significant amount of CPK BB. On the contrary, in the nuclear membrane fractions of schizophrenic and Alzheimer disease brains the content of this isoenzyme decreased. Therefore in the brain of schizophrenic and Alzheimer disease patients the content of CPK BB decreased not only in cytosolic fractions, but also in the fractions of nuclear membranes. We demonstrated also, that cytosolic CPK BB associates not only with nuclear membranes, but with synaptosomal and microsomal fractions and mitochondrias of normal brain cells.

[Neuromorphology and neurochemistry of senile dementias in the light of studies on glial response]. / Neiromorfologiia i neirokhimiia senil'nykh dementsii v svete izucheniia glial'nykh reaktsii.

The present study describes a few morphological and neurochemical disturbances in astroglial cells in senile dementia of the Alzheimer type (SDAT). The greatly elevated number of fibrous astrocytes in brain samples with SDAT is observed. The content of glial fibrillar acid protein is elevated in these cells. Glial filaments are detected to surround lipid centers. Moreover, there are deviations in energy metabolism: a profound decrease in the levels of the brain creatine kinase CKBB that is one of the principle ATP-regulation enzyme. It is suggested that there is a relationship between all the disturbances observed in glial cells of SDAT patients. These disturbances are likely to be links of the same process which leads to abnormal brain ageing.

[Immunocytochemical detection of creatine kinase isoenzymes (CK BB) in human cerebral cortex]. / Immunotsitokhimicheskoe vyiavlenie kreatinfosfokinazy BB v kore golovnogo mozga cheloveka.

The localization of creatine phosphokinase-BB (CPK-BB) in the neocortex of persons without mental pathology, who died of cardiovascular diseases (n-15) was investigated immunocytochemically. It has been found that there are several differences in the visualization of CPK-BB in neurons and astrocytes under various conditions of autopsy material treatment. The data obtained may attest to the existence of the immunochemically different forms of CPK-BB in the human brain. The appropriate conditions were defined for satisfactory visualization of CPK-BB on material of pathomorphologic collections.

[Decreased level of creatine phosphokinase BB in the brain of patients with mental disorders (complex immunochemical and immunocytochemical studies)]. / Snizhenie soderzhaniia kreatinfosfokinazy BB v mozgu bol'nykh psikhicheskimi zabolevaniiami (kompleksnoe immunokhimicheskoe i immunotsitokhimicheskoe issledovanie).

Immunochemical and immunocytochemical studies were carried out to reveal the cerebral isozyme creatine phosphokinase (CPK BB) in autopsy brain of patients with different mental diseases. The studies covered schizophrenic patients (n-19), patients with senile dementia (n-9), with Alzheimer's disease (n-13) and controls without any mental pathology (n-17). It has been demonstrated that in mental pathology in the frontal cortex (field 10), there was an appreciable decrease in the content of immunoreactive CPK BB. That decrease was significantly more pronounced in Alzheimer's disease than in schizophrenia or senile dementia (p less than 0.01). Apparently, the decrease of the content of immunoreactive CPK BB determines to a considerable measure the early detected decrease of CPK BB activity in the patients' brain.

[Effect of astasilid on the stimulation of peritoneal macrophages]. / Vliianie astazilida na stimuliatsiiu peritoneal'nykh makrofagov.

The effect of astasilid, a sucrose monoester and the effect of mainly unsaturated fatty acids from the lipid fraction of Astasia longa on immunocompetent cells--macrophages of the mouse peritoneal cavity were studied. It was shown that astasilid increased 7.5-8.5-fold expression of Fc-receptors on the macrophage plasmic membranes and stimulated 5.5-6.5-fold the macrophage capacity for Fc-dependent phagocytosis of sheep red blood cells. Astasilid had no effect on migration of the macrophages into the abdominal cavity.

[Astasian: A new stimulator of peritoneal macrophages]. / Astazian--novyi stimuliator peritoneal'nykh makrofagov.

Immunological properties of astasin, a novel substance isolated from Astasia longa was studied. 3-4 days after intraperitoneal administration of astasin in a dose of 1000 micrograms per a mouse (50 mg/kg) in the form of suspension in physiological solution or gel in dimethylsulfoxide there was observed maximum increase in the number of the Fc-receptor cells of the macrophage plasmatic membranes and intensity of sheep red blood cell phagocytosis (3.6-4 and 3.2-4-fold, respectively). Dimethylsulfoxide solution of astasin increased migration of the cells into the abdominal cavity by 5.8 times. The respective figure for the astasin suspension was 28. times. Therefore, differences in the nature of the effect and quantitative indices after the exposure to astasin in the above two forms were observed. Sodium chloride isotonic solution of astasin had a retarded but more prolonged effect on the macrophages which was probably due to gradual digestion of the polysaccharide phagocyted granules by the macrophages.

[Immunoperoxidase study of the localization of alpha-endorphin in the brain]. / Immunoperoksidaznoe issledovanie lokalizatsii al'fa-éndorfina v mozge.

Location of alpha-endorphin in the rat brain was studied by means of the indirect immunoperoxidase method (peroxidase-antiperoxidase). alpha-endorphin immunoreactivity was detected in neural fibers but not in neuronal bodies. Immunoreactive fibers were located predominantly in the hypothalamus: in the preoptic area, periventricular zone, dorsal and ventrolateral portions of the gray tuber. Individual fibers containing alpha-endorphin were also discovered in the cerebral portions directly adjacent to the hypothalamus (the ventral part of the striate body, septum and thalamus). The questions related to immunocytochemical detection of peptides in the perikaryons of neurons and their processes are discussed.