scholarly journals THE LOCALIZATION OF ENZYME ACTIVITIES IN THE RAT BRAIN

1960 ◽  
Vol 8 (3) ◽  
pp. 649-663 ◽  
Author(s):  
Norwin H. Becker ◽  
Sidney Goldfischer ◽  
Woo-Yung Shin ◽  
Alex B. Novikoff
Keyword(s):  
Rat Brain ◽  
Cell Membranes ◽  
Reductase Activity ◽  
Frozen Sections ◽  
Fixed Tissue ◽  
Quantitative Histochemistry ◽  
Capillary Endothelial Cells ◽  
Intracellular Organelles ◽  
The Brain ◽  
Nissl Substance

Studies with rat brain illustrate the usefulness of formol-calcium-fixed tissue for studying both enzymatic "chemoarchitectonics" and intracellular organelles. Unembedded frozen sections and polyvinyl alcohol-embedded sections may be used to demonstrate the activities of DPNH-tetrazolium reductase localized in mitochondria and ergastoplasm, TPNH-tetrazolium reductase localized in mitochondria, ATPase (and/or apyrase or ADPase) in cell membranes, and acid phosphatase in lysosomes.1 Among the observations recorded are: (1) the presence of lysosomes in all cells of the brain; (2) the presence of numerous large lysosomes near the nuclei of capillary endothelial cells; (3) a polarized arrangement of large lysosomes in epithelial cells of the ependyma and choroid plexus; (4) the presence of ATPase activity in the cell membranes of some neurons; (5) the presence of either an apyrase or combination of ATPase and ADPase in the cell membranes of neuroglia and capillaries; (6) the presence of both DPNH- and TPNH-tetrazolium reductase activities in neuroglia; (7) the presence of DPNH- and TPNH-tetrazolium reductase activities in mitochondria and of DPNH-tetrazolium reductase activity in Nissl substance. The possible functional significance of these localizations is briefly discussed, as is their relation to "quantitative histochemistry" data available in the literature.

Liquid Chromatography Analysis of Clozapine in Rat Brain Tissue, Using its Molecularly Imprinted Polymer after Administration of Toxic Dose of Drug and Lipid Emulsion Therapy

2019 ◽  
Vol 15 (3) ◽  
pp. 251-257
Author(s):  
Bahareh Sadat Yousefsani ◽  
Seyed Ahmad Mohajeri ◽  
Mohammad Moshiri ◽  
Hossein Hosseinzadeh
Keyword(s):  
Rat Brain ◽  
Brain Tissue ◽  
Molecularly Imprinted Polymer ◽  
Lipid Emulsion ◽  
Limit Of Detection ◽  
Imprinted Polymer ◽  
Toxic Dose ◽  
Molecularly Imprinted ◽  
The Brain ◽  
Rat Brain Tissue

Background:Molecularly imprinted polymers (MIPs) are synthetic polymers that have a selective site for a given analyte, or a group of structurally related compounds, that make them ideal polymers to be used in separation processes.Objective:An optimized molecularly imprinted polymer was selected and applied for selective extraction and analysis of clozapine in rat brain tissue.Methods:A molecularly imprinted solid-phase extraction (MISPE) method was developed for preconcentration and cleanup of clozapine in rat brain samples before HPLC-UV analysis. The extraction and analytical process was calibrated in the range of 0.025-100 ppm. Clozapine recovery in this MISPE process was calculated between 99.40 and 102.96%. The limit of detection (LOD) and the limit of quantification (LOQ) of the assay were 0.003 and 0.025 ppm, respectively. Intra-day precision values for clozapine concentrations of 0.125 and 0.025 ppm were 5.30 and 3.55%, whereas inter-day precision values of these concentrations were 9.23 and 6.15%, respectively. In this study, the effect of lipid emulsion infusion in reducing the brain concentration of drug was also evaluated.Results:The data indicated that calibrated method was successfully applied for the analysis of clozapine in the real rat brain samples after administration of a toxic dose to animal. Finally, the efficacy of lipid emulsion therapy in reducing the brain tissue concentration of clozapine after toxic administration of drug was determined.Conclusion:The proposed MISPE method could be applied in the extraction and preconcentration before HPLC-UV analysis of clozapine in rat brain tissue.

scholarly journals Age influences susceptibility of brain capillary endothelial cells to La Crosse virus infection and cell death

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Rahul Basu ◽  
Vinod Nair ◽  
Clayton W. Winkler ◽  
Tyson A. Woods ◽  
Iain D. C. Fraser ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Death ◽  
Virus Infection ◽  
La Crosse Virus ◽  
Brain Capillary ◽  
Adult Mice ◽  
Age Related ◽  
Capillary Endothelial Cells ◽  
The Brain

Abstract Background A key factor in the development of viral encephalitis is a virus crossing the blood-brain barrier (BBB). We have previously shown that age-related susceptibility of mice to the La Crosse virus (LACV), the leading cause of pediatric arbovirus encephalitis in the USA, was associated with the ability of the virus to cross the BBB. LACV infection in weanling mice (aged around 3 weeks) results in vascular leakage in the olfactory bulb/tract (OB/OT) region of the brain, which is not observed in adult mice aged > 6–8 weeks. Thus, we studied age-specific differences in the response of brain capillary endothelial cells (BCECs) to LACV infection. Methods To examine mechanisms of LACV-induced BBB breakdown and infection of the CNS, we analyzed BCECs directly isolated from weanling and adult mice as well as established a model where these cells were infected in vitro and cultured for a short period to determine susceptibility to virus infection and cell death. Additionally, we utilized correlative light electron microscopy (CLEM) to examine whether changes in cell morphology and function were also observed in BCECs in vivo. Results BCECs from weanling, but not adult mice, had detectable infection after several days in culture when taken ex vivo from infected mice suggesting that these cells could be infected in vitro. Further analysis of BCECs from uninfected mice, infected in vitro, showed that weanling BCECs were more susceptible to virus infection than adult BCECs, with higher levels of infected cells, released virus as well as cytopathic effects (CPE) and cell death. Although direct LACV infection is not detected in the weanling BCECs, CLEM analysis of brain tissue from weanling mice indicated that LACV infection induced significant cerebrovascular damage which allowed virus-sized particles to enter the brain parenchyma. Conclusions These findings indicate that BCECs isolated from adult and weanling mice have differential viral load, infectivity, and susceptibility to LACV. These age-related differences in susceptibility may strongly influence LACV-induced BBB leakage and neurovascular damage allowing virus invasion of the CNS and the development of neurological disease.

scholarly journals THE QUANTITATIVE HISTOCHEMISTRY OF THE BRAIN

1958 ◽  
Vol 232 (2) ◽  
pp. 979-993
Author(s):  
Mary V. Buell ◽  
Oliver H. Lowry ◽  
Nira R. Roberts ◽  
Mei-Ling W. Chang ◽  
Joyce I. Kapphahn
Keyword(s):  
Quantitative Histochemistry ◽  
The Brain

scholarly journals High-Cholesterol Diet Decreases the Level of Phosphatidylinositol 4,5-Bisphosphate by Enhancing the Expression of Phospholipase C (PLCβ1) in Rat Brain

2020 ◽  
Vol 21 (3) ◽  
pp. 1161 ◽  
Author(s):  
Yoon Sun Chun ◽  
Sungkwon Chung
Keyword(s):  
Neurodegenerative Diseases ◽  
Rat Brain ◽  
Phospholipase C ◽  
Cultured Cells ◽  
Cell Interaction ◽  
High Cholesterol Diet ◽  
Cholesterol Diet ◽  
High Cholesterol ◽  
Amyloid Aβ ◽  
The Brain

Cholesterol is a critical component of eukaryotic membranes, where it contributes to regulating transmembrane signaling, cell–cell interaction, and ion transport. Dysregulation of cholesterol levels in the brain may induce neurodegenerative diseases, such as Alzheimer’s disease, Parkinson disease, and Huntington disease. We previously reported that augmenting membrane cholesterol level regulates ion channels by decreasing the level of phosphatidylinositol 4,5-bisphosphate (PIP2), which is closely related to β-amyloid (Aβ) production. In addition, cholesterol enrichment decreased PIP2 levels by increasing the expression of the β1 isoform of phospholipase C (PLC) in cultured cells. In this study, we examined the effect of a high-cholesterol diet on phospholipase C (PLCβ1) expression and PIP2 levels in rat brain. PIP2 levels were decreased in the cerebral cortex in rats on a high-cholesterol diet. Levels of PLCβ1 expression correlated with PIP2 levels. However, cholesterol and PIP2 levels were not correlated, suggesting that PIP2 level is regulated by cholesterol via PLCβ1 expression in the brain. Thus, there exists cross talk between cholesterol and PIP2 that could contribute to the pathogenesis of neurodegenerative diseases.

Vital staining of cells of the brain vegetative centers

1926 ◽  
Vol 22 (5-6) ◽  
pp. 730-731
Author(s):  
G. P.
Keyword(s):  
Nervous System ◽  
Autonomic Nervous System ◽  
Vital Staining ◽  
Frozen Sections ◽  
Autonomic Nervous ◽  
The Brain ◽  
Dark Blue

V. Rakhmanov (Zhurn. Neurop. And Psych., 1925, No. 3-4) proposes to inject them with 1% Trypanblau solution in the amount of 1 cubic meter to study the vegetative centers in mice. with. weekly for 6-8 weeks. The brain is fixed in 10% formalin, frozen sections are stained with alum carmine or cochineal. In this case, dark blue dust-like grains appear in the plasma and nuclei of cells - selectively for the cells of the autonomic nervous system.

scholarly journals Superoxide dismutase, glutathione peroxidase and catalase in developing rat brain

1982 ◽  
Vol 204 (2) ◽  
pp. 535-540 ◽  
Author(s):  
I Mavelli ◽  
A Rigo ◽  
R Federico ◽  
M R Ciriolo ◽  
G Rotilio
Keyword(s):  
Superoxide Dismutase ◽  
Glutathione Peroxidase ◽  
Rat Brain ◽  
Rat Hepatocytes ◽  
Mitochondrial Fraction ◽  
Cytoplasmic Fraction ◽  
Mouse Ascites ◽  
Enzyme Pattern ◽  
Relative Deficiency ◽  
The Brain

The specific activities of Cu, Zn- and Mn-superoxide dismutases, of glutathione peroxidase and of catalase, the enzymes considered to be specifically involved in the defence of the cell against the partially reduced forms of oxygen, were determined as the function of postnatal age in the early (up to 60 days) period of rat brain development. The enzymes were assayed in the cytoplasmic fraction, in the crude mitochondrial fraction including peroxisomes, and in the mitochondria. The results show that the temporal changes of these enzymes cannot be correlated with each other, thus indicating that they do not concertedly parallel the increasing activity of aerobic brain metabolism during development. Specifically the cytoplasmic fraction shows a gradual increase of the Cu, Zn-superoxide dismutase activity with age, whereas the glutathione peroxidase activity is constant from birth. Furthermore the increase of the mitochondrial Mn-superoxide dismutase as a function of postnatal age is more remarkable than that of the cytoplasmic Cu, Zn-enzyme. Higher activities of catalase in adult animals are detectable only in the subcellular fraction containing peroxisomes, because of the modest catalase activity of the brain. These results indicate independent regulation of the expression of these enzyme activities in the process of brain differentiation and point to a relative deficiency of enzymic protection of the brain differentiation and point to a relative deficiency of enzymic protection of the brain against potentially toxic oxygen derivatives. This situation is similar to the pattern already described in the rat heart and in rat and mouse ascites-tumour cells, at variance with the much more efficient enzyme pattern present in rat hepatocytes.

scholarly journals Estimates of Michaelis-Menten Constants for the Two Membranes of the Brain Endothelium

1984 ◽  
Vol 4 (2) ◽  
pp. 241-249 ◽  
Author(s):  
Albert Gjedde ◽  
Ove Christensen
Keyword(s):  
Steady State ◽  
Endothelial Cell ◽  
Transport Properties ◽  
Glucose Transport ◽  
Cell Membranes ◽  
Experimental Studies ◽  
Facilitated Diffusion ◽  
Capillary Endothelial Cells ◽  
The Individual ◽  
Tracer Experiments

Tracer studies on facilitated diffusion across the blood–brain barrier lead to the calculation of Michaelis-Menten constants that describe the rate of transport. However, the barrier consists of two endothelial cell membranes, and the relevance of single Michaelis-Menten constants in relation to the two cell membranes is unknown. We have formulated a model of two endothelial cell membranes and show that the measured Michaelis-Menten constants are simple functions of the properties of the individual membranes when transport across the endothelium is rapid ( P1 > 10−6 cm s−1). We also show that the Michaelis-Menten constants determined in tracer experiments describe facilitated diffusion in the steady state only if the two membranes have similar transport properties. As an application of this observation, we have examined three experimental studies that measure glucose transport in the steady state and show that the Michaelis-Menten constants for glucose transport calculated from the tracer experiments are equal to the constants calculated from the steady-state experiments. We conclude that the luminal and abluminal membranes of brain capillary endothelial cells have equal glucose transport properties.

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