RESUMO
Expression of choline acetyltransferase, 200-kDa neurofilament protein, 28-kDa calbindin, neuronal NO synthase, caspase 3, and Ki-67 in the motor neurons of spinal cord segments T3-T5 in male C57Bl/6 mice after 30-day space flight in the Bion-M1 biosatellite was studied by immunohistochemical methods. Under conditions space flight, the size of motoneurons increased, the number of neurons containing choline acetyltransferase and neurofilaments, decreased, and the number of calbindin-positive neurons increased; motoneurons, expressing neuronal NO synthase and caspase 3 appeared, while Ki-67 was not detected. Fragmentation of neurons with the formation structures similar to apoptotic (residual) bodies was observed in individual caspase 3-positive motoneurons.
Assuntos
Neurônios Motores/metabolismo , Voo Espacial , Medula Espinal/metabolismo , Ausência de Peso , Animais , Apoptose/genética , Calbindinas/genética , Calbindinas/metabolismo , Caspase 3/genética , Caspase 3/metabolismo , Contagem de Células , Tamanho Celular , Colina O-Acetiltransferase/genética , Colina O-Acetiltransferase/metabolismo , Regulação da Expressão Gênica , Imuno-Histoquímica , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Masculino , Camundongos , Neurônios Motores/química , Neurônios Motores/patologia , Proteínas de Neurofilamentos/genética , Proteínas de Neurofilamentos/metabolismo , Óxido Nítrico Sintase Tipo I/genética , Óxido Nítrico Sintase Tipo I/metabolismo , Medula Espinal/química , Medula Espinal/patologiaRESUMO
The expression of choline acetyltransferase (ChAT), neurofilament (NF) protein 200 kDa, calbindin (CAB) 28 kDa, neuronal NO-synthase (nNOS), caspase 3, Ki-67 was studied in motor neurons from TIIITV segments of the spinal cord in C57/ BL6 male mice by immunohistochemical methods 12 h after a 30 days-long space flight on the Bion-M1 biosatellite. Mice living under standard vivarium conditions served as a control. The motoneurons of experimental animals demonstrated the reactive changes that were manifested by the increase of their size, decrease in the number of subpopulations expressing ChAT and NF, increase of subpopulations containing CAB, appearance of motor neurons expressing nNOS, caspase-3, and the absence of Ki-67. The lack of proliferating gliocytes, the preservation of nucleolar component indicate the absence of necrobiotic changes characteristic of necrosis. The cytoplasmic vacuolization was observed in thionine-stained CAB-immunoreactive neurons. The disintegration of motor neurons into fragments resembling the apoptotic bodies was observed in single cells that contained caspase-3.
Assuntos
Neurônios Motores , Proteínas do Tecido Nervoso/metabolismo , Voo Espacial , Medula Espinal , Animais , Masculino , Camundongos , Neurônios Motores/metabolismo , Neurônios Motores/patologia , Medula Espinal/metabolismo , Medula Espinal/patologiaRESUMO
UNLABELLED: Mice were exposed to 1 month of space flight on the Russian biosatellite BION-M1 to determine its effect on the expression of genes involved in the maintenance of the mouse brain dopamine system. The current article focuses on the genes encoding glial cell line-derived neurotrophic factor (GDNF) and cerebral dopamine neurotrophic factor (CDNF). Space flight reduced expression of the GDNF gene in the striatum and hypothalamus but increased it in the frontal cortex and raphe nuclei area. At the same time, actual space flight reduced expression of the gene encoding CDNF in the substantia nigra but increased it in the raphe nuclei area. To separate the effects of space flight from environmental stress contribution, we analyzed expression of the investigated genes in mice housed for 1 month on Earth in the same shuttle cabins that were used for space flight and in mice of the vivarium control group. Shuttle cabin housing failed to alter the expression of the GDNF and CDNF genes in the brain structures investigated. Thus, actual long-term space flight produced dysregulation in genetic control of GDNF and CDNF genes. These changes may be related to downregulation of the dopamine system after space flight, which we have shown earlier. © 2015 Wiley Periodicals, Inc. SIGNIFICANCE: Our results provide the first evidence of microgravity effects on expression of the GDNF and CDNF neurotrophic factor genes. A considerable decrease in mRNA level of GDNF and CDNF in the nigrostriatal dopamine system was found. Because both GDNF and CDNF play a significant role in maintenance and survival of brain dopaminergic neurons, we can assume that this dysregulation in genetic control of GDNF and CDNF genes in substantia nigra could be among the reasons for the deleterious effects of space flight on the dopamine system.
Assuntos
Encéfalo/metabolismo , Regulação da Expressão Gênica/fisiologia , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Fatores de Crescimento Neural/metabolismo , Ausência de Peso , Animais , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fatores de Crescimento Neural/genética , RNA Mensageiro/metabolismo , Voo Espacial , Fatores de TempoRESUMO
Electron microscopic study of left ventricle cardiomyocytes and quantitative analysis of their mitochondriom was performed in rats exposed to tail-suspension, as a model of weightlessness effects, to artificial gravity produced by intermittent 2G centrifugation and a combination of these effects. It was found that the cardiomyocytes ultrastructure changed slightly after tail-suspension and after intermittent 2G influence, as well as under a combination of these effects. However, the number of intermitochondrial junctions increased significantly in the interfibrillar zone of cardiomyocytes under a combination of tail-suspension and intermittent 2G influence, which agrees with the cell hypertrophy described earlier.
Assuntos
Gravitação , Ventrículos do Coração/citologia , Miócitos Cardíacos/fisiologia , Miócitos Cardíacos/ultraestrutura , Simulação de Ausência de Peso , Animais , Centrifugação , Masculino , Microscopia Eletrônica , Mitocôndrias/ultraestrutura , Ratos , Ratos WistarRESUMO
Chronic action of gravity force of 2 G on pre- and postnatal rats up to day 75 of postnatal development gave rise to ultrastructural deviations in the somatosensory cortex (SSC) suggesting higher afferentation to SSC as compared with their counterparts at 1 g, more intensive synaptic transmission, afferent input by dendrites and neurons functioning with more frequent interneuronic contacts, glia activation and increased blood supply that altogether pointed to a permanent impact of elevated extero- and proprioceptive input throughout the time of SSC formation.
Assuntos
Dendritos/ultraestrutura , Hipergravidade , Neuroglia/ultraestrutura , Neurônios Aferentes/ultraestrutura , Córtex Somatossensorial/ultraestrutura , Animais , Animais Recém-Nascidos , Comunicação Celular , Feminino , Desenvolvimento Fetal , Feto , Masculino , Gravidez , Ratos , Córtex Somatossensorial/irrigação sanguínea , Transmissão SinápticaRESUMO
Electron microscopy examination of layers III-IV in the somatosensory cortex of rat's brain following 5- and 33-day and repeated 5-day exposure to 2 G revealed ultrastructural changes in dendritic spines, synapses, axon terminals and nervous cell body suggesting their functional activation and increase of the afferent input from hindlimb mechanoreceptors Hypertrophy of the acicular apparatus (AA) due to hypergravity, growth of AA destruction as a result of 33-day exposure and its ample evidence after repeated 5-day exposure lead to the conclusion that AA ultrastructure can be used for assessing afferent input into the brain cortex of animals exposed in a changed field of gravity.
Assuntos
Espinhas Dendríticas/ultraestrutura , Hipergravidade/efeitos adversos , Terminações Pré-Sinápticas/ultraestrutura , Córtex Somatossensorial/patologia , Sinapses/ultraestrutura , Vias Aferentes/patologia , Animais , Peso Corporal , Masculino , Mecanorreceptores/patologia , Microscopia Eletrônica , Ratos , Ratos Wistar , Fatores de TempoRESUMO
Quantitative cytochemical and morphometric methods were used to investigate cytochrome oxidase activity and sizes of bodies and nuclei of Purkinje's cells in the medical nodulus and upper central lobule of the vermis obtained from rats sacrificed in 5-6 hours of landing after the 14-day SLS-2 mission of NASA space "shuttle" Columbia (STS-58). The reduced cytochrome oxidase activity was explained by suppression of the functional activity of Purkinje's cells in microgravity. Results of the investigations suggest weakening of the regulatory effect of the vermis Purkinje's cells on giant neurons of the dorsocaudal segment of Deiters nucleus. They also strengthen the earlier hypothesis that space flight decays the inhibitory effect of nodulus Purkinje's cells on the medial vestibular nucleus for the reason of change in the "velocity storage" in mammals during and after flight.
Assuntos
Cerebelo/ultraestrutura , Propriocepção/fisiologia , Células de Purkinje/ultraestrutura , Distúrbios Somatossensoriais/patologia , Voo Espacial , Núcleo Vestibular Lateral/ultraestrutura , Ausência de Peso/efeitos adversos , Animais , Cerebelo/enzimologia , Cerebelo/fisiopatologia , Modelos Animais de Doenças , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Seguimentos , Imuno-Histoquímica , Masculino , Microscopia Eletrônica , Células de Purkinje/enzimologia , Ratos , Ratos Sprague-Dawley , Distúrbios Somatossensoriais/enzimologia , Distúrbios Somatossensoriais/fisiopatologia , Núcleo Vestibular Lateral/metabolismo , Núcleo Vestibular Lateral/fisiopatologiaRESUMO
The quantitative cytochemical and morphometrical techniques applied in the experiment with 24-d tail-suspension of rats demonstrated a decrease of cytochrome oxidase activity and downsizing of Purkinje's cell bodies in the proprioceptive cerebellum (cerebellar vermis upper central lobule), as well as a decrease of cytochrome oxidase activity in Purkinje's cells in the vestibular cerebellum (nodulus). The observed suppression of the Purkinje's cells functional activity indicates reductions in the proprioceptive and vestibular afferent inputs to these cells during simulation of the microgravity effects in rats. Intermittent exposure to hypergravity (2 G, one hour a day) prevented the decreases in cytochome oxidase activity in Purkinje's cells of both the proprioceptive and vestibular cerebella of tail-suspended rats suggesting a nearly same level of functioning as in the vivarium controls.
Assuntos
Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Hipergravidade/efeitos adversos , Propriocepção/fisiologia , Células de Purkinje/patologia , Animais , Modelos Animais de Doenças , Elevação dos Membros Posteriores , Masculino , Células de Purkinje/enzimologia , Ratos , Ratos WistarRESUMO
Cytochrome oxidase activity as well as neurons' body and nuclei sizes were measured in the spinal L5 ganglia receiving the hindlimb afferent input during primary and repeated 14-d tail-suspension of rats. The measurements were performed using the quantitative cytochemical and morphometric techniques. The animals were suspended 30 days and returned to vivarium. Thirty day later, they were suspended again, together with a group of intact rats, this time for 14 days. Primary 14-day suspension reduced significantly the cytochrome oxidase activity in small, middle and large neurons (cross-section area of bodies less than 800, 800-2000 microm2 and more than 2000 microm2, respectively) and yielded a trend-like diminution of the nerve cells bodies and nuclei. The postponed repeated suspension, unlike the primary 14-d one, resulted in a substantial downsizing of bodies and nuclei of middle neurons suggesting a more dramatic hypofunction and a more dramatic decrease of afferent input. However, the cytochrome oxidase activity was not suppressed considerably in small and middle neurons which might be explained by initiation of the afferent input recovery.
Assuntos
Gânglios Espinais/fisiologia , Vértebras Lombares , Simulação de Ausência de Peso , Animais , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Seguimentos , Gânglios Espinais/citologia , Masculino , Ratos , Ratos WistarRESUMO
The cytochrome oxidase activity, sizes of bodies, nuclei, and nucleoli of alpha-motoneurons from the L5 anterior horns were investigated in rats once or repeatedly suspended by tails to model the effects of microgravity. Rats were suspended 30 days, then held in vivarium 30 days w/o any behavior restriction and suspended once again 14 days simultaneously with rats that had not been suspended before. The first-time 14-d suspension reduced the cytochrome oxidase activity as well as sizes of bodies and nuclei in alpha-motoneurons which pointed to hypofunctioning of these nervous cells. Repeated (14-d) suspension after the 30-day maintenance in vivarium decreased the alpha-motoneurons parameters much less significantly implying mild hypofunction and a faster adaptation of L5 alpha-motoneurons to microgravity modeled again after a long period of recess.
Assuntos
Neurônios Motores/patologia , Nervos Espinhais/patologia , Simulação de Ausência de Peso , Ausência de Peso/efeitos adversos , Animais , Modelos Animais de Doenças , Seguimentos , Elevação dos Membros Posteriores/métodos , Masculino , Ratos , Ratos Wistar , Nervos Espinhais/fisiopatologiaRESUMO
Electron microscopy was used to study structural elements of the vestibular afferent input to the cerebellar nodulus Purkinje's cells--terminals of mossy fibers and granular cells in the granular layer and parallel fibers and Purkinje's cell dendrites in the molecular layer in rats decapitated in 2-3 hours and 9 days after the 9-day space flight aboard NASA shuttle Columbia (STS 40, SLS-1 mission). Analysis of the revealed ultrastructural changes on the base of morphofunctional correlations leads to the following conclusions: 1) space flight induced a prolonged reduction in vestibular input to most of the mossy fiber terminals and nodulus Purkinje's cells; 2) within the initial hours of recovery the vestibular input to a part of mossy fiber terminals and granular cells was increasing due to elevation of the sensitivity of vestibular receptors in microgravity; 3) regain of the vestibular input to Purkinje's cells after space flight is hampered by structural, as a result of microgravity effects, and also functional, developing shortly after space flight, impediments, and 4) in 9 d after landing the vestibular input to Purkinje's cells was almost normal. The observed reduction in the vestibular input to the nodulus Purkinje's cells during and after the spaceflight microgravity is presumably the key to the mechanism altering the velocity storage in mammals in microgravity and on return from space flight.
Assuntos
Dendritos/ultraestrutura , Microscopia Eletrônica/métodos , Neurônios Aferentes/ultraestrutura , Células de Purkinje/ultraestrutura , Voo Espacial , Nervo Vestibular/ultraestrutura , Animais , Seguimentos , Masculino , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Ausência de Peso/efeitos adversosRESUMO
The neuromuscular junctions were investigated in m. soleus of rats subjected to a 3-month tail suspension simulating the microgravity effects. Electron microscopy analysis revealed some ultrastructural signs of atrophy, degeneration and adipose dystrophy of muscle fibers. The aggregate of these findings points to progressive atrophy in m. soleus, while ultrastructural changes in the neuromuscular synapses testify a reduced functional activity of the synapses and partial denervation of the muscle fibers which, probably, underlay the atrophic process in the muscle. Increases in the number of axon terminals found in some neuromuscular synapses as well as of synaptic vesicles in individual axon terminals are likely to reflect formation of a particularly active pool of spinal motoneurons at L5, possibly associated with the growth in the number of fast fibers resulted from transformation.
Assuntos
Neurônios Motores/ultraestrutura , Músculo Esquelético/inervação , Junção Neuromuscular/ultraestrutura , Simulação de Ausência de Peso/efeitos adversos , Animais , Modelos Animais de Doenças , Progressão da Doença , Seguimentos , Elevação dos Membros Posteriores/efeitos adversos , Elevação dos Membros Posteriores/métodos , Masculino , Microscopia Eletrônica , Músculo Esquelético/ultraestrutura , Distrofia Muscular Animal/etiologia , Distrofia Muscular Animal/patologia , Terminações Pré-Sinápticas/ultraestrutura , Ratos , Ratos Wistar , Vesículas Sinápticas/ultraestrutura , Fatores de Tempo , Simulação de Ausência de Peso/veterináriaRESUMO
Nineteen-day centrifugation of rats at 2 G and ensuing 30-d readaptation to 1 g caused changes in the somatosensory cortex (SSC) ultrastructure pointing to intensification of the afferent input and functional activity of SSC neurons during hypergravity, and recovery of these processes in the course of readaptation. Repeated 5-d centrifugation of the rats at 2 G produced changes in ultrastructure of SSC layers 3 and 4 evidencing a more effective cortex functioning aimed at facilitation of adaptation to hypergravity repeated after the lengthy delay.
Assuntos
Hipergravidade , Córtex Somatossensorial/ultraestrutura , Voo Espacial , Adaptação Fisiológica , Animais , Axônios/ultraestrutura , Dendritos/ultraestrutura , Masculino , Neurônios/ultraestrutura , Ratos , Ratos WistarRESUMO
Scanning electron and light microscopy were applied to study the third ventricle ependyma in rat's brain after 30-d tail-suspension, 30-d readaptation to the horizontal position, and repeated 14-d suspension in parallel with another group of rats exposed to a single 14-d suspension. Despite the repeated blood redistribution toward the head, the second tail-suspension produced significantly less grave destruction of the ependymal cell structure and cilia ultrastructure than the single 14-d or previous 30-d one, which suggests that simulation of microgravity repeated after substantial delay imparts much more persistence to the cerebrospinal fluid outflow from the brain ventricles into the saggital sinus.
Assuntos
Epêndima/ultraestrutura , Ausência de Peso/efeitos adversos , Animais , Seguimentos , Elevação dos Membros Posteriores , Masculino , Microscopia Eletrônica , Ratos , Ratos WistarRESUMO
The ultrastructure of muscle spindles (incapsulated mechanoreceptors of stretch of extrafusal muscle fibres) of m. soleus in adult Wistar rats after repeated unloading of support on hind limbs with preservation of support loading on fore limbs has been studied by transmissing electron microscopy. It was shown that, along with muscle spindles with the ordinary number of intrafusal muscle fibres (four), m. soleus contains spindles with an increased number of intrafusal fibers (five to six). It was assumed that the increase in the number of intrafusal muscle fibers is due to the proliferation of their satellite cells.
Assuntos
Fibras Musculares Esqueléticas/ultraestrutura , Fusos Musculares/ultraestrutura , Animais , Contagem de Células , Masculino , Microscopia Eletrônica de Transmissão , Fibras Musculares Esqueléticas/citologia , Fusos Musculares/citologia , Ratos , Ratos Wistar , Suporte de CargaRESUMO
Postponed for a long time repeated simulation of the weightlessness effects in rats by tail-suspension increases in the amount of intrafusal muscle fibers in m. soleus spindles, reflecting presumably elevation of the sensitivity of mechanoreceptors.
Assuntos
Fusos Musculares/ultraestrutura , Músculo Esquelético/inervação , Simulação de Ausência de Peso , Ausência de Peso , Animais , Elevação dos Membros Posteriores , Masculino , Microscopia Eletrônica , Contração Muscular , Fusos Musculares/fisiologia , Músculo Esquelético/fisiologia , Músculo Esquelético/ultraestrutura , Estimulação Física , Ratos , Ratos WistarRESUMO
The electronmicroscopica/study of brain somatosensory cortex in the rats, exposed to postponed for a long time repeated lack of support load on hindlimbs by means of tail-suspension, has been revealed in hindlimb area of cortex the changes of ultrastructure of axodendritic synapses and neurons pointing out the acceleration of decrease in functional activity of synapses and neurons in 3-4 layers at postponed for a long time repeated simulation of weightlessness effects.
Assuntos
Córtex Somatossensorial/ultraestrutura , Sinapses/ultraestrutura , Ausência de Peso/efeitos adversos , Animais , Axônios/ultraestrutura , Dendritos/ultraestrutura , Modelos Animais de Doenças , Elevação dos Membros Posteriores , Masculino , Microscopia Eletrônica , Ratos , Ratos WistarRESUMO
Cytologic and morphometric investigations of the hypothesis, thyroid, thyroid and pancreas of rats after 19-d centrifugation at 2G revealed suppression of somatotropic hormone production by hypophyseal somatotrophs and insulin by pancreatic beta-cells, and intensification of thyrotrophic hormone production by hypophyseal thyreotrophs, thyroxin by thyreoid parenchyma and glucagon by pancreatic alpha-cells. These change in hormone production are viewed as an endocrine mechanism of the divergence of plastic processes and energy metabolism in animals subjected to chronic exposure to hypergravity.
Assuntos
Metabolismo Energético/fisiologia , Hipergravidade/efeitos adversos , Pâncreas/citologia , Hipófise/citologia , Glândula Tireoide/citologia , Animais , Insulina/metabolismo , Secreção de Insulina , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/metabolismo , Masculino , Pâncreas/metabolismo , Hipófise/metabolismo , Prognóstico , Ratos , Ratos Wistar , Somatotrofos/metabolismo , Glândula Tireoide/metabolismo , Tireotrofos/metabolismo , Tiroxina/metabolismoRESUMO
Results of the histological analysis of the brain of rats subjected to 93-d tail-suspension reveal morphological changes occurred to the arteries, veins and capillaries, ependymal cells of the ventricles and epithelium of the vascular plexus, and nervous and glial cells. On the one hand, these suggest venostatis and brain edema, impaired liquor production and liquor dynamics and increase in intracranial pressure and, on the other hand, adaptive processes in the arterial walls preventing the excess blood flow to the brain.
Assuntos
Encéfalo/irrigação sanguínea , Encéfalo/citologia , Artérias Cerebrais/citologia , Artérias Cerebrais/fisiologia , Simulação de Ausência de Peso , Glândulas Suprarrenais/fisiologia , Animais , Elevação dos Membros Posteriores , Masculino , Ratos , Ratos Wistar , Estresse FisiológicoRESUMO
Histological methods were employed to investigate brain vessels of rats subjected to 30-d tail-suspension and 30-d readaptation in the horizontal position afterwards, and suspended again for 14 days simultaneously with other rats that had no such experience in the past. Repeated suspension produced, although there was blood redistribution to the cranial end, much less profound changes in brain vessels as compared with the previous 30-d suspension and the first-time suspended mates. This suggests an unknown arterial mechanism triggered in the region forward of the extracerebral arteries of the brain surface which hampered an excess blood flow to the brain during repeated suspension.