Vol 156, No 5 (2019)

Articles

MORPHO-FUNCTIONAL TRANSFORMATION OF EPENDYMAL CELLS IN THE DYNAMICS OF POSTNATAL DEVELOPMENT OF THE RAT BRAIN VENTRICLES

Pavlov A.V., Fokanova O.A., Korableva T.V.

Abstract

Objective - to study morphometric parameters and ciliary activity of ciliated ependymal cells in the dynamics of postnatal development of the brain ventricles. Material and Methods. With the help of a digital intravital video microscopy, histological techniques and morphometry, we studied vascular plexuses and structural and functional characteristics of ciliated ependymal cells on the biopsy samples of the 109 brain ventricles of Wistar rats during the first year of life. Results. In the rat brain ventricles, morpho-functional transformations of the vascular plexuses and ciliated ependymal cells were most pronounced in the first 20 days after birth. They included an accelerated increase in the size of the vascular plexuses, the maximal values of the morphometric parameters of ciliated ependymal cells and indicators of their ciliary activity, the highest rate of cerebrospinal fluid movement in the parietal layer compared with animals of reproductive age. By 4-12 months in all ventricles the studied indicators of ciliary clearance were reduced to 30-50 % of the maximum values. Conclusions. During the first 20 days of postnatal ontogenesis, we observed significant changes in the morphometric and functional parameters of rat brain ventricles ciliated ependymal cells, activity of which, apparently, is the main mechanism for the movement of cerebrospinal fluid. Later, the development of vascular plexuses provides a more efficient mechanism of cerebrospinal fluid circulation, therefore, the role of the transport function of ependymal cells decreases.
Morphology. 2019;156(5):9-16
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MORPHOLOGICAL PATTERN AND MORPHOMETRIC STUDY OF THE CEREBRAL CORTEX OF RATS WITH DIFFERENT TYPOLOGICAL CHARACTERISTICS OF HIGHER NERVOUS ACTIVITY

Krishtop V.V., Pozhilov D.A.

Abstract

Objective - to study the histological and immunohistochemical characteristics of the cerebral cortex in animals with different stress tolerance and level of cognitive abilities. Material and methods. The morphology of the cortex of the brain precentral gyrus was studied in 24 rats with different levels of stress tolerance, verified in open field test, and cognitive abilities, verified in the Morris water maze. The sections were stained according to Nissl method and with antibodies against GFAP. Results. In animals with low stress tolerance, a significant increase in the number of neurons with two nucleoli was found; large nuclei and the area of cytoplasm, as well as a larger number of neurons demonstrating irreversible changes were also detected. The average distance between the perikaryon of neurons and the satellite cells in this subgroup was also larger, and the number of perivascular gliocytes was smaller. In animals with a low level of cognitive abilities, the features revealed by Nissl staining were similar to those of animals with high stress tolerance but less pronounced. Animals with high stress tolerance were characterized by a significantly smaller distribution area of the processes and a smaller number of GFAP-positive cells per 1 mm2 of the section. Animals with high cognitive abilities typically had a significantly greater number of GFAP-positive cell bodies per 1 mm2 of section and a significantly smaller area occupied by processes. Conclusions. The level of the animal stress tolerance correlates with the nature of neuro-glial relationship; high level of stress tolerance and high cognitive abilities are associated with a smaller number of neurons with two nucleoli, a great number of dead satellite cells, low values of the average distribution of astrocyte processes. Other parameters associated with high stress tolerance and high cognitive abilities are of mutually exclusive or insignificant character.
Morphology. 2019;156(5):17-21
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IMMUNOHISTOCHEMICAL CHARACTERISTICS OF THE NEURONS OF RAT VENTROMEDIAL AND DORSOMEDIAL HYPOTHALAMIC NUCLEUS

Moiseev K.Y., Emanuylov A.I., Maslyukov P.M.

Abstract

Objective - to identify the localization, percentage and morphometric characteristics of neurons immunoreactive (IR) to neuronal nitric oxide synthase (nNOS) and calbindin (CB) in the ventromedial (VMH) and dorsomedial (DMH) rat hypothalamic nuclei. Material and methods. The work was performed on female Wistar rats aged 3-4 months using immunohistochemical methods. Results. nNOS-IR and CB-IR neurons were detected in the VMH and DMH. VMH and DMH had heterogeneous neurochemical composition. The largest percentage of intensely fluorescent CB-IR neurons was demonstrated in DMH in the ventral region of the nucleus, while in the VMH CB-IR neurons with weak fluorescence were detected in large numbers in the ventrolateral region. In all areas of DMH and VMH, nNOS-IR neurons with low fluorescence intensity were demonstrated. The average cross-sectional area of CB-IR and nNOS-IR cells in the VMH and DMH was not significantly different from immunonegative neurons. Conclusions. The neurons of the VMH and DMH hypothalamic nuclei, which are involved in the regulation of metabolism in the body, contain nNOS and CB. In turn, VMH and DMH have heterogeneous neurochemical composition.
Morphology. 2019;156(5):22-25
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THE PROLIFERATION ACTIVITY IN RAT OLFACTORY BULB IN A NEURODEGENERATION MODEL

Varentsov V.E., Rumyantseva T.A., Kiseleva E.V., Myasishcheva T.S.

Abstract

Objective - to evaluate the effect of neuronal death in the neonatal period on the proliferative activity in the rat olfactory bulb (OB). Material and methods. The study was conducted on 48 male Wistar 1 to 90 days of age. The object of study was the right OB. Neurodegeneration was induced by single subcutaneous injection of 10 % capsaicin solution to the rats on the first day of life. Ki-67 was detected using monoclonal rabbit antibodies (ab16667, UK, 1:100). Microphotographs of standard sections were used to determine the numerical density of positive nuclei (units/mm2). Results. The activity of cell proliferation in OB depended on the age and layer. In response to capsaicin administration, the proliferative activity increased during the first week. The intensity of the activation had topological characteristic - the maximum and earliest effect was observed in the center of the OB and then it spreaded to the outer layers. At the beginning of the observation, the main source of proliferating cells was the germinative zone of the OB, in the subsequent periods - the migrating neuronal precursors of the rostral migratory stream. Conclusion. Diffuse death of a part of neurons caused by the capsaicin administration leads to the development of a compensatory process manifested by the activation of the canonical stem niches and the germinative zone of the OB, as well as the prolongation of the proliferative activity of radially migrating cells to the 90th day of life in rats.
Morphology. 2019;156(5):26-31
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AGE-RELATED CHARACTERISTICS OF OLFACTORY BULB ASTROCYTES REACTION TO THE ADMINISTRATION OF CAPSAICIN

Pozhilov D.A., Moskalenko A.V., Pshenisnov K.K.

Abstract

Objective - to compare the reaction of rat olfactory bulb astrocytes to capsaicin administration at different stages of postnatal ontogenesis. Materials and methods. On serial paraffin sections of the olfactory bulbs of 70 male Wistar rats aged 30 to 240 days, astrocytes and mature neurons were detected using GFAP and NeuN markers. NeuN was demonstrated in the perinuclear cytoplasm and nucleus of mature neurons, glial fibrillary acidic protein (GFAP) of astrocytes - in the cytoplasm and astrocyte processes. The numerical density of mature neurons and astrocytes (pcs/ mm2), as well as the average area of astrocyte processes (μm2) were assessed in norm and after neurotoxic exposure to subcutaneous administration of capsaicin in rats of different age groups. Results. Postnatal changes in the studied microstructural parameters of the olfactory bulb in rats were normally observed from 30 to 180 days. In adulthood, stabilization of both the numerical density of neurons and astrocytes, and the average area of the processes of astrocytes were observed. It was found that neurotoxic exposure caused neuronal death and an increase in the numerical density of astrocytes in different layers of olfactory bulb, caused by a response to damage. More profound microstructural changes in the primary and secondary neurogenic niches were noted. Conclusions. Neuronal death was maximal at 15-30 days and reactive gliosis - at 30 days after neurotoxin administration. In infantile rats the intensity of gliosis decreased by the 60th day of the experiment, in adulthood gliosis did not tend to decrease.
Morphology. 2019;156(5):32-38
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CHANGES OF THE IMMUNOHISTOCHEMICAL CHARACTERISTICS OF NEURONS OF THE MIDDLE GROUP OF THE HYPOTHALAMIC NUCLEI DURING AGING

Porseva V.V., Korzina M.B., Spirichev A.A., Vyshnyakova P.A., Aryaeva D.A., Nozdrachev A.D., Masliukov P.M.

Abstract

The aim of the study - to identify differences in the immunohistochemical characteristics of the neurons of the arcuate (ARС), ventromedial (VMH) and dorsomedial (DMH) hypothalamus between young and old rats. Material and methods. The work was performed on male Wistar rats aged 3-4 months and 2-2,5 years using immunohistochemical methods. Results. In DMH, the percentage of calbindin (CB)immunoreactive (IR) of neurons is significantly reduced, and the percentage of calretinin (CR)-IR of neurons increases in DMH and VMH during aging. In old animals, the number of neuronal nitric oxide synthase (nNOS)-IR neurons and the degree of immunofluorescence to nNOS increased in all studied nuclei. In old rats, there is an increase in the density of neuropeptide Y-IR fibers in the VMH and a decrease in the DMH. Conclusions. The neurochemical composition of the nuclei of the middle group (mainly DMH and VMH) of the hypothalamus changes during aging.
Morphology. 2019;156(5):39-43
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THE STUDY OF THE DYNAMICS OF THE MEDIATOR CONTENT OF SYMPATHETIC NEURONS IN EARLY ONTOGENESIS OF CAT

Emanuilov A.I.

Abstract

Objective - to demonstrate the localization and immunohistochemical characteristics of cat sympathetic neurons in the superior cervical (SCG) and stellate (SG) ganglia. Material and methods. The work was performed on 1-, 10-, 20-, 30-day-old cats using retrograde axonal transport of Fast Blue (FB) which was injected in the neck muscle, and immunohistochemical methods. Results. FB-labeled neurons were detected in both studied ganglia in cats from the moment of birth. The largest number of labeled neurons in kittens of all age groups were immunoreactive (IR) to tyrosine hydroxylase (TG). The percentage of labeled neurons reactive both to TG and neuropeptide Y significantly increased in both ganglia during the first 10 days of life. The percentage of calbindin-IR neurons in the SCG decreased from birth by the 30th day of life, in the SG - increased in the first 10 days and then decreased by the 30th day of life. Immunoreactivity to choline acetyltransferase, vasointestinal polypeptide, and neuronal nitric oxide synthase were found in less than 1 % of labeled neurons in kittens of all age groups. Somatostatin-IR labeled neurons were not detected in the SCG and SG. Conclusions. In ontogenesis a reorganization of the mediator content of sympathetic neurons occurs, which in cats ends by the 20th day of life.
Morphology. 2019;156(5):44-48
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TOPOGRAPHY AND CYTOARCHITECTURE OF THE AREA X OF THE SPINAL CORD IN POSTNATAL ONTOGENESIS OF THE RAT

Porseva V.V., Nozdrachev A.D.

Abstract

Objective - to study age-related changes in the topography and cytoarchitecture of the area X of the rat spinal cord at different stages of postnatal ontogenesis. Materials and methods. The study was carried out on pieces of the spinal cord taken at the level of the second thoracic segment of 65 female Wistar rats aged 3, 5, 7, 10, 15, 20, 30, 60, 90, 120, 150, 180, and 360 days (5 animals in each age group). The age-related changes in the square area of the lamina X, its neurons number per section, the density of neurons in the area of central canal and dorsal and ventral gray commisures were studied histologically on 14-mm-thick tangential cryosections stained by thionine according to Nissl method. Results. The area X on the transverse section of the spinal cord thoracic segment was clearly identified in 3-day-old rats. In the rat postnatal development, the area of the lamina X decreased, mainly due to the decrease in size of the dorsal gray commissure, despite the increase in the area of the ventral commissure, which was observed in rats older than 20 days. Simultaneously with a decrease in the area of lamina X, a decrease in the total number of its neurons per section was observed. The density of neurons around the circumference of the central canal decreased by the 7th day, in the dorsal gray commissure - by the 10th day, and in the region of the ventral grey commissure - by the 20th day. In rats older than 20 days, the density of neurons in the zone of the central canal and ventral gray commissure did not change, and in the dorsal gray commissure it increased again by 240 days, reaching the same indices as in 7-day-old rats. Conclusions. The area X and its individual regions, as well as the total number and the density of neurons per section, vary in different ways at different stages of postnatal ontogenesis. The data are of interest for the study of changes of the organization of gray matter of the spinal cord in the postnatal ontogenesis.
Morphology. 2019;156(5):49-54
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NEUROPEPTIDE Y-ERGIC NEURONS OF THE SMALL INTESTINE IN POSTNATAL ONTOGENESIS

Maslyukov P.M., Vyshnyakova P.A., Aryaeva D.A., Budnik A.F.

Abstract

Objective - to demonstrate the localization, percentage and morphometric characteristics of NPY-immunoreactive (IR) neurons in the intramural ganglia of the myenteric (MP) and submucous neural plexus (SP) of the small intestine in rats of different age groups. Material and methods. The work was performed on 1-, 10-, 20-, 30-, 60-day-old and 2-year-old rats using immunohistochemical methods. Results. NPY-IR neurons were detected in the small intestine from the moment of birth and throughout all studied age periods. In the intramural ganglia of the MP, their number was very small and did not change significantly during ontogenesis. In the SP, NPY-IR neurons were detected in large numbers in newborns; the maximal number of the neurons was observed at 20-30 days. The average cross-sectional area of the NPY-IR cells in the SP was significantly larger than such area of immunonegative neurons in 10-day-old and older rats. Conclusions. Thus, neuropeptide Y is fairly common in various parts of the autonomic nervous system, including sympathetic, parasympathetic and metasympathetic, and plays an important role in the processes of age-related neuronal development.
Morphology. 2019;156(5):55-58
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AGE-RELATED CHANGES IN INTRAMURAL GANGLIONIC NEURONS OF THE LARGE INTESTINE EXPRESSING CALBINDIN AND CALRETININ

Budnik A.F., Maslyukov P.M., Nozdrachev A.D.

Abstract

Objective - to identify the localization, percentage and morphometric characteristics of calbindin (CB) and calretinin (CR)-immunoreactive (IR) neurons in the methasympathetic intramural ganglia of the myenteric (MP) and submucous plexus (SP) of the large intestine of rats of different age groups. Material and methods. Thick sections of the transverse colon of Wistar rats aged 1, 10, 20, 30, 60 days and 2 years were studied by immunohistochemical and morphometric methods. Results. CB-IR and CR-IR neurons were demonstrated in rats of all studied age groups. In the MP, the percentage of CB-IR and CR-IR neurons on the 10th day of life was significantly greater than after birth, and did not change in the older groups. In the SP of newborn rats, CB was not detected (it was demonstrated only from the 10th day), and CR was detected in 72,0±2,52 % of neurons. A significantly higher percentage of CB-IR and CR-IR neurons was detected in 2-month-old and older rats. The average cross-sectional area of CB-IR and CR-IR cells was significantly larger than the average cross-sectional area of the immunonegative neurons in all age groups. Conclusions. In early postnatal ontogenesis, there was an increase in the proportion of CB-IR and CR-IR neurons in the intramural ganglia of the intestine.
Morphology. 2019;156(5):59-64
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STRUCTURE OF THE THYROID GLAND IN RATS WITH DIFFERENT STRESS TOLERANCE DURING CEREBRAL HYPOPERFUSION

Krishtop V.V., Rumyantseva T.A., Agadzhanova L.S.

Abstract

Objective - to study the structural features of the thyroid gland in animals with different stress tolerance during cerebral hypoperfusion. Material and methods. The microstructure of the thyroid gland was studied in 104 rats with different levels of stress tolerance verified in open field test. Thyrocytes height, follicle diameter, areas of single islands of the interfollicular epithelium, relative proportion of perifollicular blood capillaries and average area of mast cell cytoplasm were measured on the thyroid gland slides. Results. Animals with high stress tolerance, compared with animals with low stress tolerance, had significant differences in the structure of the thyroid gland: a greater height of the thyroid epithelium and a greater specific area of blood capillaries as well as larger average area of mast cells of the thyroid gland. Cerebral hypoperfusion was accompanied by the activation of folliculogenesis, a decrease of mast cell area and an increase in the perifollicular capillary blood volume. In animals with a high level of stress tolerance a decrease in the height of thyrocytes in the early periods switched to its growth on the 35th day of the experiment, and activation of folliculogenesis was registered. Conclusions. The animals stress tolerance level is associated with the structural parameters of the thyroid gland. During cerebral hypoperfusion, groups with high and low stress tolerance have different dynamics and severity of compensatory changes.
Morphology. 2019;156(5):65-69
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COMPLEX ASSESSMENT OF THE ADRENAL MEDULLA RESPONSE TO PARTIAL LASER DESTRUCTION OF A PAIRED ORGAN USING THE METHOD OF NATURAL PERIODIZATION OF BIOLOGICAL PROCESSES

Kemoklidze K.G.

Abstract

Objective - complex assessment of morphological changes in the adrenal medulla (AM) after partial laser destruction of a paired organ, based on the calculation of variation coefficients and the identification of natural periods Material and methods. On sections of the adrenal glands of adult rats (n=41), with a differentiating staining of adrenalocytes and noradrenalocytes, morphometry of 19 AM parameters was performed during 2 months after a surgical laser injury to the paired organ. The data were processed according to the natural periodization method. Results. According to the degree of variation of the parameters in each time point from their average values for the entire observation period and from control we calculated the coefficients of variation Cv and Cvc. Their dynamics allowed us to reveal the natural periods: I period - enhanced response (1-14 days) with phases I.1 - a quick, but weaker reaction (days 1-7) and I.2 - a slow, but stronger reaction (7-14 days); and II period - normalization (days 14-56). Passing the curves of Cvc and Cv through the time point of the 21st day almost in a straight-line indicates a possibility of optimizing similar experiments taking into account this time point. The convergence of all parameters with the control on the 56th day shows that it is impractical to prolong the experiment. Conclusions. 1) The crucial point of the systemic reaction of AM as a paired structure to laser damage is the 14th day. 2) When planning similar experiments, it is possible to optimize them taking into account the 21st day. 3) Extension of similar experiments longer than 2 months is not rational.
Morphology. 2019;156(5):70-74
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SOVERShENSTVOVANIE PREPODAVANIYa TOPOGRAFIChESKOY ANATOMII S ISPOL'ZOVANIEM SREDSTV VIRTUAL'NOY REAL'NOSTI

Filimonov V.I., Novikov Y.V., Gagarin V.V., Kochergin A.F., Abakshina M.N.
Morphology. 2019;156(5):85-89
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