Fine Structural Localization of Intravenously Injected Cytochrome C in the Porcine Choroid Plexus

1973 ◽  
Vol 31 ◽  
pp. 652-653
Author(s):  
D. A. Davis ◽  
T. H. Milhorat ◽  
B. J. Lloyd
Keyword(s):  
Electron Microscopy ◽  
Cerebrospinal Fluid ◽  
Cytochrome C ◽  
Horseradish Peroxidase ◽  
Choroid Plexus ◽  
Structural Localization

In the continuing search for suitable markers that can be used for examining mechanisms of cerebrospinal fluid (CSF) formation, we were recently led to a study of cytochrome c. This hemo-chromogen has two primary advantages as a marker: 1. It is g. considerably smaller protein (molecular wt. 13,000, radius 15 Å) than ferritin (molecular wt. 400,000, radius 5 Å) or horseradish peroxidase (molecular wt. 40,000, radius 25-30Å) ; 2. It is easily visualized by electron microscopy after reaction with 3,3'-diaminobenzidine (dab).Cytochrome c (Cal Biochem Grade A) was administered intravenously to pigs (7-9 kg. in wt.) in doses of 25-30 mg./100 g. wt.

scholarly journals Aquaporin-4 Expression In The Human Choroid Plexus

2021 ◽  
Author(s):  
Felix Deffner ◽  
Corinna Gleiser ◽  
Ulrich Mattheus ◽  
Andreas Wagner ◽  
Peter H Neckel ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Cerebrospinal Fluid ◽  
Choroid Plexus ◽  
Mrna Level ◽  
Age Groups ◽  
Freeze Fracture ◽  
Aquaporin 4 ◽  
Ependymal Cells ◽  
Aqp4 Expression ◽  
Age Related

Abstract Background: The choroid plexus (CP) consists of specialized ependymal cells and underlying stroma and blood vessels, producing the bulk of the cerebrospinal fluid (CSF). CP epithelial cells are the site of the internal blood-cerebrospinal fluid barrier, show epithelial characteristics (basal lamina, tight junctions), and express aquaporin-1 (AQP1) apically. In contrast, ventricle-lining ependymal cells express aquaporin-4 (AQP4) basolaterallly. The initial purpose of this study was to analyze the expression of aquaporins in the ependyma – CP transition zone in the human brain to gain insights in aquaporin regulation. The results prompted us to investigate aquaporin expression in the mouse CP of different age groups. Methods: We analyzed the CP from eight body donors (age 74-91) applying immunofluorescence, qPCR, and freeze-fracture electron microscopy. We used antibodies against AQP1, AQP4, NKCC1, and Na/K-ATPase. In addition, we compared the CP from young (2 months), adult (12 months) and old (30 months) mice by qPCR and immunofluorescence. Results: Unexpectedly, many cells in the human CP were positive not only for AQP1 but also for AQP4, normally restricted to ependymal cells and astrocytes. Expression of AQP1 and AQP4 was found in the CP of all eight body donors. These results were confirmed by qPCR, and by electron microscopy detecting AQP4-specific orthogonal arrays of particles. To find out whether AQP4 expression correlated with relevant transport-related proteins we investigated expression of NKCC1 and Na/K-ATPase. Immunostaining for NKCC1 was similar to AQP1 and revealed no particular pattern related to AQP4. Co-staining of AQP4 and Na/K-ATPase indicated a trend for an inverse correlation of their expression. To test for the possibility of age-related changes causing AQP4 expression in the CP, we analyzed mouse brains from different age groups and found a significant increase of AQP4 on the mRNA level in old mice compared to young and adult animals. Conclusions: We provide evidence for AQP4 expression in the human and murine CP related to aging which likely contributes to the water flow through the CP epithelium and CSF production. In two alternative hypotheses, we discuss this as a beneficial compensatory, or a detrimental mechanism influencing the previously observed CSF changes during aging.

scholarly journals FINE STRUCTURE OBSERVATIONS OF THE UPTAKE OF INTRAVENOUSLY INJECTED PEROXIDASE BY THE RAT CHOROID PLEXUS

1967 ◽  
Vol 15 (3) ◽  
pp. 160-165 ◽  
Author(s):  
NORWIN H. BECKER ◽  
ALEX B. NOVIKOFF ◽  
H. M. ZIMMERMAN
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Cerebrospinal Fluid ◽  
Choroid Plexus ◽  
Smooth Endoplasmic Reticulum ◽  
Multivesicular Bodies ◽  
Adult Rats ◽  
Dense Bodies ◽  
Pinocytotic Vesicles ◽  
Membrane Bound

The uptake by the choroid plexus of adult rats of intravenously injected horseradish peroxidase has been investigated by electron microscopy. Within 4 min, the injected protein passes the capillary and is rapidly distributed through extracellular space and choroidal cells. Peroxidase enters the choroidal cells within coated vesicles which act as pinocytotic vesicles. At 15 min, peroxidase activity is present in numerous membrane-bound vesicles, multivesicular bodies, dense bodies and what appear to be segments of smooth endoplasmic reticulum. None of the peroxidase-containing organelles is seen to empty to the ventricular surface. Egress of the extracellular peroxidase into the cerebrospinal fluid is apparently blocked by apical zonulae occludentes between the choroidal cells.

Nucleic Acid Molecules Prepared by Monolayer Techniques

1972 ◽  
Vol 30 ◽  
pp. 178-179
Author(s):  
Dimitrij Lang
Keyword(s):  
Electron Microscopy ◽  
Standard Deviation ◽  
Nucleic Acid ◽  
Cytochrome C ◽  
Nucleic Acids ◽  
Contour Length ◽  
Sample Standard Deviation ◽  
Molecular Weights ◽  
Length Measurements ◽  
Protein Monolayer

The success of the protein monolayer technique for electron microscopy of individual DNA molecules is based on the prevention of aggregation and orientation of the molecules during drying on specimen grids. DNA adsorbs first to a surface-denatured, insoluble cytochrome c monolayer which is then transferred to grids, without major distortion, by touching. Fig. 1 shows three basic procedures which, modified or not, permit the study of various important properties of nucleic acids, either in concert with other methods or exclusively:1) Molecular weights relative to DNA standards as well as number distributions of molecular weights can be obtained from contour length measurements with a sample standard deviation between 1 and 4%.

Comparative strengths and weaknesses of peroxidase and colloidal gold in pre- and post-embedding immunolabeling techniques

1987 ◽  
Vol 45 ◽  
pp. 1002-1005
Author(s):  
D. R. Abrahamson ◽  
P. L. St.John ◽  
E. W. Perry
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Light Microscopy ◽  
Colloidal Gold ◽  
Light Microscope ◽  
Ultrastructural Localization ◽  
The Other ◽  
Quantitative Studies ◽  
Dense Suspension ◽  
Antigen Localization

Antibodies coupled to tracers for electron microscopy have been instrumental in the ultrastructural localization of antigens within cells and tissues. Among the most popular tracers are horseradish peroxidase (HRP), an enzyme that yields an osmiophilic reaction product, and colloidal gold, an electron dense suspension of particles. Some advantages of IgG-HRP conjugates are that they are readily synthesized, relatively small, and the immunolabeling obtained in a given experiment can be evaluated in the light microscope. In contrast, colloidal gold conjugates are available in different size ranges and multiple labeling as well as quantitative studies can therefore be undertaken through particle counting. On the other hand, gold conjugates are generally larger than those of HRP but usually can not be visualized with light microscopy. Concern has been raised, however, that HRP reaction product, which is exquisitely sensitive when generated properly, may in some cases distribute to sites distant from the original binding of the conjugate and therefore result in spurious antigen localization.

scholarly journals 1616. Mechanism of Thrombocytopenia Induced by Oxazolidinones Antibiotics (Linezolid, Tedizolid): Demonstration of Impairment of Megakaryocyte Differentiation From Human Hematopoietic Stem Cells associated with Mitochondrial Toxicity

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S801-S801
Author(s):  
Paul M Tulkens ◽  
Tamara V Milosevic ◽  
Gaëlle Vertenoeil ◽  
William Vainchenker ◽  
Stefan N Constantinescu ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Stem Cells ◽  
Cytochrome C ◽  
Hematopoietic Stem Cells ◽  
Cytochrome C Oxidase ◽  
Panel Member ◽  
Human Model ◽  
Mitochondrial Toxicity ◽  
Review Panel ◽  
Hematopoietic Stem

Abstract Background Linezolid causes thrombocytopenia, which limits its use. In cell culture and in tissues from treated patients, linezolid impairs mitochondrial protein synthesis (due to structural similarities and common binding sites between bacterial and mitochondrial ribosomes). Recent studies have shown that mitochondria act as a key relay in the process leading from activation of the thrombopoietin receptor to megakaryocytes differentiation. Methods Validated ex-vivo human model of hematopoietic stem cells (HSC) differentiation for (i) measuring megakaryocytes, granulocyte-monocytes, and burst-forming unit-erythroids colony formation; (ii) differentiation into megakaryocytes (conversion of CD34+ into CD41+/CD42+ cells; morphology) and proplatelets formation, (iii) mitochondrial toxicity (electron microscopy; cytochrome c-oxidase activity [partly encoded by the mitochondrial genome]). Results We show that linezolid (and the recently approved tedizolid), both at concentrations corresponding to their human serum concentrations) inhibit the maturation of HSC into fully differentiated megakaryocytes (CD41 and CD42-positive cells) and the formation of proplatelets. Optic and Electron microscopy) showed an impairment of the formation of typical megakaryocytes (lack of large polylobulated nuclei and of intracellular demarcation membrane system [required for platelet formation]), together with disappearance of the internal structure of mitochondria. Biochemical studies showed a complete suppression of the activity of cytochrome c-oxidase (a key enzyme of the mitochondrial respiratory chain). Conclusion Our study provides for the first time insights in the mechanism of thrombocytopenia induced by linezolid and tedizolid, identifying mitochondria as their target and showing that the drugs will impair the differentiation of hematopoietic stem cells into mature platelets-releasing megakaryocytes. It illustrates how mitochondria dysfunction may play a key role in toxicology and diseases, while paving the way for rational approaches for the design and screening of less toxic derivatives for the benefit of future patients. Disclosures Paul M. Tulkens, MD, PhD, Bayer (Consultant, Advisor or Review Panel member, Speaker’s Bureau)Menarini (Speaker’s Bureau)Merck (Advisor or Review Panel member, Speaker’s Bureau)Trius (now part of Merck) (Advisor or Review Panel member, Research Grant or Support) Françoise Van Bambeke, PharmD, PhD, Bayer (Speaker’s Bureau)

Sex-Related Differences in Rat Choroid Plexus and Cerebrospinal Fluid: A cDNA Microarray and Proteomic Analysis

2016 ◽  
Vol 28 (1) ◽  
Author(s):  
T. Quintela ◽  
H. Marcelino ◽  
M. J. Deery ◽  
R. Feret ◽  
J. Howard ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Choroid Plexus ◽  
Cdna Microarray ◽  
Proteomic Analysis

Surgical removal of bilateral papillomas of the choroid plexus of the lateral ventricles with resolution of hydrocephalus

1979 ◽  
Vol 50 (5) ◽  
pp. 677-681 ◽  
Author(s):  
Steven K. Gudeman ◽  
Humbert G. Sullivan ◽  
Michael J. Rosner ◽  
Donald P. Becker
Keyword(s):  
Cerebrospinal Fluid ◽  
Choroid Plexus ◽  
Large Volume ◽  
Surgical Removal ◽  
Tumor Removal ◽  
Content Type ◽  
Lateral Ventricles ◽  
Csf Diversion ◽  
Csf Production ◽  
Fine Print

✓ The authors report a patient with bilateral papillomas of the choroid plexus of the lateral ventricles with documentation of cerebrospinal fluid (CSF) hypersecretion causing hydrocephalus. Special attention is given to the large volume of CSF produced by these tumors (removal of one tumor reduced CSF outflow by one-half) and to the fact that CSF diversion was not required after both tumors were removed. Since tumor removal alone was sufficient to stop the progression of hydrocephalus, we feel that this case supports the concept that elevated CSF production by itself is sufficient to cause hydrocephalus in patients with papillomas of the choroid plexus.

Cardiotrophin-1 in choroid plexus and the cerebrospinal fluid circulatory system

Neuroscience ◽  
2004 ◽  
Vol 127 (1) ◽  
pp. 43-52 ◽  
Author(s):  
A.L Gard ◽  
E Gavin ◽  
V Solodushko ◽  
D Pennica
Keyword(s):  
Cerebrospinal Fluid ◽  
Choroid Plexus ◽  
Circulatory System
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