Subject(s)
Brain/embryology , Space Flight , Weightlessness , Animals , Cerebral Cortex/cytology , Cerebral Cortex/embryology , Corpus Striatum/cytology , Corpus Striatum/embryology , Embryo, Mammalian/physiology , Female , Histocytochemistry , Mitosis , Pregnancy , Rats , Rhombencephalon/embryology , Rhombencephalon/enzymologyABSTRACT
Beginning from the 13th day of pregnancy the rats were under conditions of weightlessness of spaceflight for 6 days. After landing in 18-day-old fetuses the state of their brain development is investigated comparing to that in control animals, that are on the Earth. As demonstrates analysis of a number of morphological processes: reproduction, migration, neuronal differentiation, growth of processes, establishment of nervous connections, neuroglial interconnections and vascularization--all they under conditions of weightlessness develop rather fully. Certain deviations in vascularization (as examples the medulla oblongata and the striated tuber are taken) are observed--the amount of vessels is greater and they are thinner--and changes in migration rate of cells is demonstrated by the example of the cortical plate formation. These changes are quickly levelled during their subsequent development on the Earth.
Subject(s)
Brain/embryology , Space Flight , Weightlessness/adverse effects , Animals , Brain/enzymology , Female , Gestational Age , Histocytochemistry , Pregnancy , Rats , USSRABSTRACT
Macroscopic and light microscopic examinations of the brain of 18-day fetuses, and neonate 15-, 30- and 100- day rats whose embryonic development from day 13 to day 18 occurred in space flight on Cosmos-1514 did not reveal any changes in the brain structures of the fetuses or pups of the three ages. However, the brain of the 18-day fetuses that developed in flight showed signs of insufficient tissue oxygenation, trends toward delayed cell migration in the cortex and delayed differentiation of neurosecretory cells of the hypothalamic supraoptic nuclei. The cell differentiation rate returned to normal during continued embryogenesis after flight at 1 g.
Subject(s)
Brain/embryology , Space Flight , Weightlessness , Animals , Brain/growth & development , Female , Morphogenesis , Pregnancy , RatsABSTRACT
There are 2 alternative procedures for cytophotometrical determination of 2 histochemical ingredients (DNA, proteins, and enzymes) in the same cell. The 1st possibility consists of the sequential measurement of 2 ingredients, when the 2nd chromophore is revealed after the determination and extraction of the 1st chromophore. Another one is the simultaneous measurement, when the 2 histochemical reactions (at 1st with the 1st chromophore and then with the other one) are performed on the fixed sections. So each of the sections includes 2 chromophores simultaneously. Next after photometry in 2 wavelengths, the quantity of each chromophore is calculated. This method gives a characteristic of mutual distribution of 2 ingredients in one and the same cell by coefficient of correlation. This article accentuates the causes of the limitations, and the estimation of mistakes. In the appendix there are described methodics and programmes of calculations.
Subject(s)
DNA/analysis , Ganglia, Spinal/analysis , Histocytochemistry/methods , Neurons/analysis , Proteins/analysis , Acetylcholinesterase/analysis , Animals , Chick Embryo , Chickens , Enzymes/analysis , Ganglia/analysis , Oxidoreductases/analysis , Rats , Spectrophotometry/methodsABSTRACT
Changes in the activity of 13 enzymes are described in the process of cytodifferentiation of the nerve cells of spinal ganglion, the motor neurons of spinal cord and large nerve cells of the III layer of tectum opticum in 7, 10 and 21 day old chick embryos. Cytophotometry was performed with MZFV-1 (LOMO) by means of plug-method. A relatively high activity of glucose-6-phosphat dehydrogenase, diaphorase, alpha-glycerophosphate dehydrogenase and, partially, acetylcholine esterase was found already in the 7 days old embryo. The activity of monoamine oxidase, aldolase-glyceroaldehyde phosphate dehydrogenase, isocitrate dehydrogenase, glutamate dehydrogenase increased markedly on the 21st day. When studying the reciprocal distribution of two enzymes in separate cells, pairs of enzymes with a high value of correlation coefficient were found. The cytodifferentiation was found to be accompanied by changes in the coefficient of correlation of the same pair of enzymes.
Subject(s)
Ganglia, Spinal/enzymology , Neurons/enzymology , Spinal Cord/enzymology , Superior Colliculi/enzymology , Animals , Cell Differentiation , Chick Embryo , Energy Metabolism , Ganglia, Spinal/embryology , Histocytochemistry , Neurons/cytology , Photometry , Spinal Cord/embryology , Superior Colliculi/embryology , Time FactorsABSTRACT
By means of doublewave cytophotometry of the histological sections and smears performed with the device mtsfv-1 (lomo) it has been demonstrated that differentiation of some neurons of the spinal ganglia and the spinal cord is accompanied with hyperreplication of DNA amount exceeding the diploid contents, while the large neurons in layer III of the tectum opticum remain diploid. In the process of differentiation an increased amount of RNA and histones, and decreasing portion of histones sented for the microcalculator b3-21.
Subject(s)
DNA/analysis , Ganglia, Spinal/embryology , Histones/analysis , RNA/analysis , Spinal Cord/embryology , Tectum Mesencephali/embryology , Animals , Cell Differentiation , Chick Embryo , Ganglia, Spinal/analysis , Microcomputers , Spinal Cord/analysis , Tectum Mesencephali/analysisABSTRACT
By means of cytophotometry and a computer in the cardiac intramural neural plexus of 17 experimental and 17 control rabbits acetilcholine esterase (AChE) activity (revealed by Karnovsky--Roots' method) was investigated. It was stated that a prolonged electrostimulation of the area hypothalamica posterior (AHP) produced statistically significant 2-fold decrease of AChE activity in the cardiac neural cells and fibres. When the stimulation of AHP was stopped, the decreased enzymatic activity was observed for the first two weeks, then by the end of the 4th week it again reached its control level. The results of the work were discussed in view of possible disorders of regulating vegetative-trophic effects of the posterior hypothalamus on innervative and executive tissue elements of the heart.
Subject(s)
Acetylcholinesterase/metabolism , Heart/innervation , Hypothalamus, Posterior/physiology , Hypothalamus/physiology , Myocardium/enzymology , Nerve Fibers/enzymology , Neurons/enzymology , Animals , Electric Stimulation , Enzyme Activation , Histocytochemistry , Male , Photometry , RabbitsABSTRACT
A method and device for obtaining embryo brain slices are described. Results of cultivation of slices of rhombencephalon and mesencephalon are presented obtained in accord with a proposed technique of the "swimming float" (Olenev, 1979). With this cultivation, original topographical relations between brain nuclei and fibrillar tracts remain unchanged that enables us to compare these cultures with histological sections of brain to identify strictly the structures being cultivated. Cultured neurons of nuclei of XII and VIII pairs of cranial nerves have been first described in addition to various divisions of the reticular formation commissura cochlearis dorsalis. Compared to cells examined in the in vivo conditions, neurons of the nucleus cochlearis laminaris, grown in tissue culture conditions, do not make plate-forming structures; dendrites of the neurons of III layer of tectum opticum do not develop. This may be explained by the absence in vitro of any stimulating influence from the acustic ganglion fibres and ganglion cells of the retina on these morphogenetic processes.
Subject(s)
Brain Stem/anatomy & histology , Culture Techniques/methods , Mesencephalon/anatomy & histology , Animals , Brain Stem/embryology , Chick Embryo , Culture Techniques/instrumentation , Mesencephalon/embryology , Time FactorsABSTRACT
Enzymatic activity was investigated in spinal ganglia of 10-day-old chick embryo and in 10-day-old cultures of these ganglia (the method of "flying glasses"). Histochemical preparations were photometrically treated with the one-wave plug-method in the device MCFB-1 (LOMO) with the diameter of the probe 8 mkm. As the result of the investigations on optic density, a definite conclusion was made about relative activity of 15 enzymes and a general scheme of neuronal metabolism was graphically presented under the conditions of cultivation, enzymatic activity of krebs cycle was demonstrated to decrease, while that of diaphorase and lactate dehydrogenase to increase. The data on topography of enzymatic distribution in neurons, satellites, lemmoncytes and fibroblasts were also presented.
Subject(s)
Dihydrolipoamide Dehydrogenase/metabolism , Ganglia, Spinal/enzymology , Oxidoreductases/metabolism , Animals , Cells, Cultured , Chick Embryo , Fibroblasts/enzymology , Ganglia, Spinal/cytology , Histocytochemistry , NADPH Dehydrogenase/metabolism , Neuroglia/enzymology , Neurons/enzymologySubject(s)
Micropore Filters , Organ Culture Techniques/methods , Animals , Chick Embryo , Culture Media , CzechoslovakiaABSTRACT
In order to reveal dehydrogenase and diaphorase in spinal ganglia neurons of 12-day-old chick embryos in cryostat sections, the following modifications of the medium were used: for dehydrogenase - sodium salt substrate 50 mM, NAD or NADPh 0.75 mM, nitro-BT 0.61 mM, phosphate buffer pH 7.2 15 mM, NaCl 50 mM, MgCL2 5 mM, for diaphorase - NAD-N2 or NADHh-N2 0.78-0.66 mM, NaCl 100 mM. To compare relative activity of the enzymes (optic density of histochemical preparations determined cytophotometrically) it is suggested to calculate the values obtained during proportional development of the staining regarding the time unit (hour). The possibility to compare the data obtained with the results of biochemical investigations is discussed, as well as an attempt is made to represent graphically metabolic peculiarities of various cell types.
Subject(s)
Dihydrolipoamide Dehydrogenase/metabolism , Oxidoreductases/metabolism , Animals , Chick Embryo , Ganglia, Spinal/enzymology , Histocytochemistry/methodsABSTRACT
The distribution and relative activities of some dehydrogenases and diaphorases in the rat's carotid body and ganglion cervicale superior were studied histochemically and cytophotometrically. The treatment of rats by the medium with decreased oxygen and increased carbon dioxide contents, a reliable rise of NAD--diaphorase activities was found in the interlobe tissue and (less expressed and late appearing) in the principal cells. In contrast, such changes were not observed in the ganglion cervicale superioir, that have not a special chemoreceptory function. It is supposed that these changes in enzyme activities in the carotid body is due to the chemoreceptory function of the intralobe tissue containing the IInd type cells and "special" cells of sensitive nerve terminals.
Subject(s)
Carotid Body/enzymology , Ganglia, Autonomic/enzymology , Oxidoreductases/metabolism , Animals , Dihydrolipoamide Dehydrogenase/metabolism , Glycerolphosphate Dehydrogenase/metabolism , Histocytochemistry , Isocitrate Dehydrogenase/metabolism , Ketoglutarate Dehydrogenase Complex/metabolism , L-Lactate Dehydrogenase/metabolism , Malate Dehydrogenase/metabolism , Rats , Succinate Dehydrogenase/metabolismABSTRACT
Spinal ganlia of a 9-day chick embryo were cultivated by the method of "floating rafts" in common medium (control) and in the medium containing amizyl (100 microgram/ml) or a neuregrowth factor (50 microgram/ml). With the action of amizyl there proved to be an increase in the number of surviving neurons; the majority of these neurons contained monoaminoxidase; there was a rise of NAD-diaphorase activity, and, to a lesser extent, of lactic dehydrogenase and isocitric dehydrogenase activities. The neurogrowth factor caused an increase in the number of nerve cells with acetylcholinesterase; there was an elevation of NAD-diaphorase and some rise of malic dehydrogenase activities; the activity of lactic dehydrogenase became maximal; as to succinic dehydrogenase--its activity was somewhat suppressed.
Subject(s)
Benactyzine/pharmacology , Ganglia, Spinal/drug effects , Nerve Growth Factors/pharmacology , Animals , Chick Embryo , Cholinesterases/metabolism , Dihydrolipoamide Dehydrogenase/metabolism , Ganglia, Spinal/enzymology , Histocytochemistry , L-Lactate Dehydrogenase/metabolism , Monoamine Oxidase/metabolism , Organ Culture TechniquesABSTRACT
By the aid of histochemical methods the authors sutdied the activity and distribution of acetylcholinesterase, monoamineoxidase, glutamatedehydrogynase and gammaaminobutyric acid -- transferase in the dorsal hypocampus of rats. It was shown that the activity of all the studied enzymes are significantly changed following 0.5 and 1.5 hours and are restituted following 48 hours after the formation of a pathological focus. It is concluded that a drop in the activity of glutamatedehydrogenase and gammaaminobutyric acid may be directed toward an accumulation of gammaaminobutyric acid in the epileptogenic focus and an inhibition of its activity. A drop in the histochemical reaction to acetylcholinesterase may indicate a decrease in the functions of the cholinergic systems with an active epileptogenic focus in the hypocampus.