Today, ART cycles using a donor egg cell are used more often, but this group of pregnancies has not been sufficiently studied in terms of detecting structural predictors of pregnancy complications caused by a special immunological relationship between the recipient mother and allogeneic blastocytes. A comprehensive clinical and morphological study of 89 allogeneic pregnancies was carried out. The morphological study was performed on placental sites biopsies taken at caesarean section (H & E, Orcein), for immunohistochemical analysis PanCK (AE1 / AE3), SMA (1A4), CD56 (123C3.D5), CD138 (B-A38), CD4 (SP35), CD25 (SP176), CD8 (SP16) were used as primary antibodies. A defect of the remodeling of the spiral arteries, including in subgroups without preeclampsia, the formation of perivascular foci of chronic inflammation were found (the accumulation of CD8 + T lymphocytes, CD56 + NK cells, CD138 + plasma cells, HLA-DR + dendritic cells). In subgroups with PE, significant changes were found for T lymphocytes (51.04 ± 2.3, 55.36 ± 11, 7 versus 37.12 ± 2.3), plasma cells (19.5 ± 3.2, 21.43 ± 8.4 versus 8.94 ± 1.2), Kruskal-Wallace test. Structural and immunohistochemical features of the allogenic placental bed may reflect the complexity of cytotrophoblast invasion and induction of maternal immunity with the formation of immune inflammation.
ВО «Ярославский государственный медицинский университет» Минздрава России, г. Ярославль, Россия Цель исследования -изучить возрастные морфологические характеристики обонятельных луковиц по распределению даблкортина (DCX) у крыс от 3 до 180 суток жизни.Материал и методы. DCX, маркер иммунореактивных нейрональных предшественников, выявля-ли на парасагиттальных срезах с учетом слоев, определяли численную плотность тел DCX-позитивных клеток (шт/мм 2 ) и их площадь (мкм 2 ).Результаты. Активность DCX отмечалась в телах и отростках части клеток, плотность распределе-ния зависела от слоя луковицы и возраста животных. У новорожденных, плотность позитивных нейронов в слоях луковиц не велика, затем нарастала, достигая максимума в митральном слое на 7-е, а в грануляр-ном -на 14-е сутки жизни. У крыс 2-месячного возраста плотность нейронов снижалась, но по-прежнему превышала показатели новорожденных, только у шестимесячных крыс плотность DCX+ клеток снижалась до минимума.Выводы. Снижение активности нейрогенеза по экспрессии даблкортина в обонятельной луковице у крыс происходило только к периоду полового созревания. Увеличение DCX иммунореактивности в перио-де новорожденности, вероятно, обусловлено перинатальным стрессом, процессами адаптации к внеутроб-ным условиям существования, расширением спектра и интенсивности сенсорных стимулов. Ключевые слова: обонятельные луковицы, нейрогенез, даблкортин, онтогенез, маркер, нейробласт, ростральный миграционный поток. © V. E. Varentsov, T. A. Rumyantseva, 2017Yaroslavl State Medical University, Yaroslavl, Russia Age-Related Differences of Doublecortin Expression in the Rat's Olfactory BulbsThe aim was to study age characteristics of doublecortin (DCX) distribution in olfactory bulbs of rats ranging from 3 to 180 days of life.Material and methods. DCX, a marker of immunoreactive neuronal precursors, was detected on parasagittal sections with distribution for layers. The numerical density of the positive cells bodies (pcs/mm 2 ) and their sizes were determined.Results. It was found that the DCX activity is noted in the bodies and in the processes of a part of the cells. The distribution density depends on the bulb layer and on the animal's age. In newborns the density of positive neurons in the bulbs layers is small, then it increases, reaching a maximum in the mitral layer by 7 th and in the granular layer -by the 14 th day. In 2-month aged rats the density of neurons decreases but still exceeds the rates of newborns. The density of DCX+ cells falls to the minimum only in six-month rats. Thus, a decrease in the activity of neurogenesis according to the expression of doublecortin in the olfactory bulb in rats occurs only to the period of puberty.
Целью работы было установление нормативных показателей распределения астроцитов и нейробластов в ростральном миграционном потоке и в различных слоях обонятельной луковицы у крыс в эволютивном периоде по позитивности специфичных маркеров GFAP и DCX, а также выявление возможной корреляции между уровнями экспрессии GFAP и DCX. Материал и методы. Материалом для исследования служили обонятельные луковицы двадцати самцов крыс линии Wistar 30-, 60-, 90-, 180-суточного возраста, что соответствует инфантильному, ювенильному и зрелому возрастным периодам. Изучено распределение GFAP, маркера астроцитов, и DCX, маркера незрелых нейронов, в ростральном потоке и обонятельных луковицах в различных возрастных группах. Данные маркеры выявляли на парасагиттальных срезах с учетом слоев обонятельной луковицы. Определяли численную плотность GFAP+ астроцитов (шт/мм 2), площадь распределения их отростков, среднюю площадь тел, а также численную плотность DCX+ нейронов (шт/мм 2). Результаты. В результате исследования установлены возрастные нормативные показатели численной плотности астроцитов, медианной площади распределения их отростков и численной плотности незрелых нейронов, установлено наличие корреляции между экспрессией DCX и GFAP в ростральном потоке и гранулярном слое обонятельной луковицы. Согласно полученным данным, пик интенсивности постнатального нейрогенеза приходится на ювенильный возраст (60-90 суток), численная плотность нейробластов зависит от слоя луковицы и возраста животного. Численная плотность астроцитов также имеет топографические и возрастные особенности, площадь распределения отростков астроцитов изменяется противоположно их численной плотности. Полученные результаты являются основой для оценки реакции глии и стволовых ниш на разные виды воздействий. Ключевые слова: нейрогенез, обонятельная луковица, ростральный миграционный поток, астроциты, нейробласты, даблкортин, DCX, GFAP.
The aim of the study was to evaluate the morphological features of nNOS-positive (nNOS-IR) neurons in the dorsolateral cortex of the frontal lobe of the cerebral hemispheres in albino rats during 180 days of postnatal development.Material and methods. The study was performed on 40 outbred white Wistar rats of different ages, from 1 to 180 days. The object of the study was an area of the right cerebral hemisphere on the dorsolateral surface near the frontal pole (neocortex). On paraffin serial sections of the frontal lobe, an immunohistochemical reaction was performed with antibodies to nNOS and a detection system with horseradish peroxidase. Neuronal morphometry was performed by microphotographs using the ImageJ-Fiji (NIH) 1.51h program, measuring the sectional area of the neuron body, the area of the nucleus, the nuclear-cytoplasmic ratio, and the intensity of the reaction.The significance of differences was assessed by paired Student's t-test.Results. It was found that in mature rats in the frontal lobe cortex nNOS-IR was detected in large multi-polar cells with high activity of the enzyme located in the supragranular layers, spindle-shaped cells with long positive processes at the border with the white matter (type 1), and two varieties of low-positive neurons – accumulations in the VI layer and single ones in other layers (type 2). Polymorphism of nNOS-IR neurons manifests from the birth, but it was possible to distinguish all subpopulations only from the 21st day. Each subpopulation is distinguished by its own age dynamics of the studied parameters and the nature of the distribution of positivity. In addition, in 3–7 day old rat pups, numerous small neurons at the border of the cortex and white matter have transient immunoreactivity.Conclusion. Thus, the division of nNOS-IR neurons into two morphological types proposed in the works of predecessors does not correspond to the number of subpopulations that could be described in the dorsolateral region of the prefrontal cortex in rats. This diversity of nNOS-IR neurons is consistent with the numerous functions described for nitric oxide. For an objective characterization of various classes of nNOS-IR cortical interneurons, it is necessary to use additional data obtained from transcriptomic, histological, electrophysiological, and functional experiments, taking into account species, topographic, and age features. Only an extended approach will make it possible to selectively influence different types of cells and reasonably interpret the results of experimental studies.
The aim of the study was to investigate changes of neuroblasts’ numeral destiny in rats’ olfactory bulb and rostral migratory stream after neurotoxic action in infant age.Material and methods. The distribution of DCX, a marker of immature neurons, was studied to reveal the dynamics of its expression in the evolutionary period of ontogenesis in olfactory bulbs and rostral migratory stream in 30, 60, 90, and 180-day-old rats. Modeling of neurotoxic effects was carried out by three times injections of capsaicin on the 30–32 day of life in a total dose of 120 mg/kg. The material for the study was sampled on the 15-, 30- and 60th days of the experiment. The marker was detected on parasagittal paraffin sections, taking into account the layers of the olfactory bulb. The numerical density of DCX+ neurons (pcs/mm2) and their part in the total number of cells were determined.Results. The standard age indicators of the numerical density of immature neurons and their share in the cell population, as well as similar indicators for two months after toxic effects, were established. The administration of toxic doses of capsaicin causes a massive death of mature neurons. Compensatory reaction manifested by activation of neurogenesis in the brain stem niches and an increase in the numerical density neuroblasts in the rostral stream.Conclusions. Activation of neurogenesis during neurodegeneration causes an increase in the number of DCX + neuronal progenitors in the olfactory bulb after 30 days after exposure and maintaining high levels until the end of observation.
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