Effects of Alcohol Intoxication
Investigation of Alcohol Intoxication Impact on Development of Limbic System and Correction of Neurogenesis, and some Clinical Indices in a Course of Hypoxen Utilization
Tech Area / Field
3 Approved without Funding
Ministry of Education and Science of Georgia / Beritashvili Institute of Physiology, Georgia, Tbilisi
- School of Medicine, Keio University, Japan, Tokyo\nUniversity of California, San Francisco / San Francisco / Laboratory of Neurobiology, USA, CA, San Francisco
Project summaryThe aim of the Project – investigation of disorders underlying development of the alcoholic embryopathy and normalization of the embryogenesis processes; Investigation of some clinical-laboratory-, electroencephalographic-, and tomographic indices with an aid of the Hypoxen.
Synthesized for the first time polymer from the class of redox polymers – Hypoxen, the sodium poly(-2,5-dihydroxiphenylen)-4-thiosulfate in respect to pharmacodynamics appears to be close to cytochrome-C and coenzyme-Q. However, due to the small size of its molecule (molecular mass – 568 ± 216), efficiency of Hypoxen exceeds that of cytochrome-C and coenzyme-Q by ten times. It shunts the electrone transport in the mitochondrial respiration chain, which is especially important in the organs with high rate of metabolism, the brain included (Popov W.G, In: New biotechnologies for practical health care, Moscow, 1999). Hypoxen possesses an electrone-accepting property, which determines its anti-radical and anti-oxidant action.
The Project consists of experimental and clinical parts.
Experimental part suggests to study the changes underlying development of the alcohol embryopathy (consumption of ethanol by females before the mating, for one month, and during pregnancy), which results in alteration of development in the cortical and subcortical structures of the limbic system; The hypoxen preapation will be used with an aim to normalize the limbic system structures in progeny and adult animals.
Morphological, morphometric, biochemical, and physiological investigations will be made. Such an approach will provide for early revealing of differentiation disorders of the limbic system, which result in functional alterations and correction of these alterations following hypoxen administration.
Experimental Series I: Considering the specificity of development of the cortical and subcortical structures of the limbic system, investigation of the ethanol impact on the major stages of histogenesis is suggested. According to the reference data, alcohol inhibits the protein synthesis, affects the S-period of the mitotic cycle, inhibiting thus the DNA replication, which, in its turn, influences participation of cyclins and mitogenic factors in development of the mitotic cycle. Ethanol exerts an inhibitory impact on the migration of the nervous cells as well.
Therefore, we suggest to investigate, in intact- (control) and experimental animals, mitotic index (MI) and index of the labeled cells (ILC) of the germinative zones at various stages of embryogenesis, which provide development of the limbic system (matrix cells of the lateral and III ventricles) with the neuroblasts. Proliferative activity and labeled granular cells of the hippocampal fascia dentata will be studied as well at the early stages of postnatal development.
Insofar the next important stage of embryogenesis is migration of the neuroblasts and glial cells into the respective structures of the system, an in vivo investigation of the ethanol impact on the cellular navigation will be carried out. The cellular migration, axonal development, and intercellular contacts’ formation will be studies as well in the model tissue culture experiments.
According to the reference data systematic consumption of alcohol by the adult animals leads to decrease of the neurons’ number in various cortical zones of the brain (Popova E.N. Bull.Exp.Biol.,Med.,1993, 116:318-321). Disorders of the proliferative activity and migration processes as well as death of the neurons, as a result of direct action of ethanol, may be a cause of decreased number of neurons in the cortical and subcortical structures of the limbic system. As a consequence, decrease of those neurits’ number may occur, which take part in formation of the afferent and efferent pathways.
According to the reference data systematic consumption of alcohol by the adult animals leads to decrease of the neurons’ number in various cortical zones of the brain (Popova, 1993). Disorders of the proliferative activity and migration processes as well as death of the neurons, as a result of direct action of ethanol, may be a cause of decreased number of neurons in the cortical and subcortical structures of the limbic system. As a consequence, decrease of those neurits’ number may occur, which take part in formation of the afferent and efferent pathways.
With an aid of morphometric methods a number of neurons in the limbic system as well as development of intercellular contacts and myelinization processes will be studied. Special attention will be payed to the ultrastructure of synaptic endings as well as of the system of neuron-glia-capillary, because ethanol exerts a cytotoxic influence on development of the intercellular contacts, contacts between the neurons and glial cells and capillary walls, which leads to disorders of the normal functioning of hemato-encephalic barrier and death of the neurones.
Migration processes and initiation of the neuroblasts’ differentiation in the CNS embryogenesis are closely related to the apoptosis process, which plays a major role in the final development of the limbic system’s structures. Assessment of an amount of free radicals is considered, by means of electron paramagnetic resonance method. With an aid of the high pressure chromatography, we consider to study effects of ethanol on the concentration of aspartate, gulamate, and nitric oxide, which being of high concentration exert a toxic influence on the neurons.
Due to exceptional sensitivity of DNA to ethanol, it is considered to investigate manifestation of cytotoxic effect in change of thermodynamic parameters of neuronal and glial DNA following alcohol intoxication of the organism. Conformational alterations of different types of chromatin in the neuronal and glial cell suspension will be determined by means of differential scanning microcflorimetry.
The data to be obtained will allow to determine effect of ethanol on active- and inactive chromatine and intranucleosomal DNA, and to assess the peculiarities of transfer of neural and glial cells into apoptotic cellular forms.
Disorders in development of cortical and subcortical structures of the limbic system in the progeny, resulting of alcohol intoxication of the maternal organism, leads to alterations of behavioral reactions of the animals. General and motivational-emotional behaviors as well as learning and memory processes will be studied in the offspring of alcoholized females with an aim to assess an impact of alcohol intoxication.
Experimental Series II: All the above-mentioned morphological, biochemical, and physiological parameters will be studied in the offspring of the females, which along with ethanol were given the Hypoxen.
Clinical part of the Project implies clinical-laboratory, electroencephalographic, and tomographic investigation in the alcohol-dependent- and in Hypoxen-treated patients. These investigation will reveal shifts in metabolism, functional activity, and structural organization of the brain regions. The method of non-invasive diagnostics – magnetic-resonance tomography – will provide for determination of the zones with disordered structural organization of the nervous tissue, including demyelinization, gliosis and atrophy as well as changes of paravascullar spaces and perfusion, which deteriorate the blood supply of the cortical and subcortical structures of the limbic system.
Expected results of investigations will have both basic and applied importance:
1. The role of maternal organism in formation of the Fetal Alcohol Syndrome will be revealed.
2. Peculiarities of disorders in those processes, which fulfill the histogenesis of the limbic system – proliferation, migration, synaptogenesis, and myelinization, - will be determined, as well; Number of those neurons will be evaluated, which take part in formation of cortical and subcortical structures of the limbic system.
3. The most alcohol-sensitive structures will be determined.
4. The role of glutamate and aspartate in cytotoxic influence on the neurons as well as impact of ethanol on the thermodynamic parameters of hetero- and euchromatine, and number of apoptotic neurons, will be studied; The free radicals’ dynamics will be evaluated.
5. Considering the specificity of the limbic system, character of the behavioral acts’ alterations will be studied in the offspring both at early stages of postnatal development and in adult inpiduals.
6. Probable anti-alcohol action of hypoxen in normalization of the limbic system histogenesis processes, metabolism, and behavior of the progeny, will be assessed.
7. Implementation of hypoxen in the clinical practice may reveal its feasibility to normalize clinical-laboratory, electroencephalographic, and tomographic indices and to correct clinical indices in alcohol intoxication.
Proposed Project will enable to re-orient activities of the arms-scientists towards the peaceful investigations.
The Project realization will be fulfilled in: Laboratory of Neuromorphology, I.Beritashvili Institute of Physiology, as well as in Institute of Physics and Institute of Cybernetics, Georgian Academy of Sciences; Research Institute of Narcology, Georgian Ministry of Health; Institute of Radiational and Interventional Diagnostics, Georgian Academy of Sciences. Under the auspices of the Laboratory of Neuromorphology an International Symposium – Cytochemical Aspects of Development of the Nervous System – was held. The data obtained by the Laboratory workers have been summarized and published in scientific journals, collections of works, and monographs. The laboratory workers frequently take part in various International symposia and conferences.
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