scholarly journals Enhanced resolution of histochemical distribution of glucose-6-phosphate dehydrogenase activity in rat neural tissue by use of a semipermeable membrane.

1991 ◽  
Vol 39 (7) ◽  
pp. 937-943 ◽  
Author(s):  
M A Philbert ◽  
C M Beiswanger ◽  
T L Roscoe ◽  
D K Waters ◽  
H E Lowndes
Keyword(s):  
Vestibular Nucleus ◽  
Neural Tissue ◽  
Semipermeable Membrane ◽  
G6pd Activity ◽  
Ependymal Cells ◽  
Regional Localization ◽  
Enhanced Resolution ◽  
Membrane Technique ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
The Brain

We examined the histochemical distribution of glucose-6-phosphate dehydrogenase (G6PD) activity in neural tissue using different diffusion barriers. Although polyvinyl alcohol and agar overlays permitted regional localization of G6PD, a semipermeable membrane revealed cellular differences in G6PD activity within populations of neurons. Distribution of G6PD activity in selected regions of the nervous system was examined using the membrane technique. White matter usually exhibited strong G6PD activity. The neuronal somata of the dorsal root ganglia (L4-L6) and anterior horns of the spinal lumbar enlargement demonstrated a variation in activity which was independent of somal size. Satellite cells showed intense activity when the membrane technique was used. Hippocampal pyramidal and granular cells of the dentate gyrus exhibited moderate, uniform G6PD activity, but only weak activity was seen in hippocampal and dentate molecular layers. High levels of activity were observed in the vascular endothelial cells of the brain, spinal cord, and choroid plexus, and in the ependymal cells of the spinal central canal and ventricles of the brain. The superior vestibular nucleus appeared to have little G6PD activity in either the neuron cell bodies or the surrounding parenchyma. The use of a semipermeable membrane for localization of G6PD activity in neural tissues permits enhanced resolution of neuron elements and may provide a more accurate assessment of G6PD activity in histological preparations.

A study of the properties, morphogenetic potencies and prospective fate of outer and inner layers of ectodermal and chordamesodermal regions during gastrulation, in various Anuran amphibians

Development ◽  
1983 ◽  
Vol 75 (1) ◽  
pp. 67-86
Author(s):  
T. A. Dettlaff
Keyword(s):  
Outer Layer ◽  
Normal Development ◽  
Neural Plate ◽  
Ependymal Cells ◽  
Epithelial Layer ◽  
Bombina Bombina ◽  
Cell Layers ◽  
The Brain ◽  
Early Gastrula

In both the ectodermal and the chordamesodermal regions of Anuran embryos, the outer layer of cells possesses epithelial properties and has the same restricted morphogenetic potencies. It is thus interchangeable between the regions, capable of epiboly and, when underlain by notochord material, of the formation of bottle-shaped cells as at the blastoporal groove, and invagination. When taken from the chordamesoderm region, this outer layer has no inducing effect on the ectoderm of the early gastrula. In normal development the outer layer of the neural plate takes an active part in forming the neural tube cavity. It gives rise to the neuroepithelial roof of the diencephalon and medulla oblongata and, when underlain by neuroblasts that develop from the inner cell layers, to ependymal cells of the brain wall. The outer layer of the notochord material is included in the epithelial layer underlying the roof of the gastrocoel - the hypochordal plate. The inner layers of these regions consist of loosely arranged cells and normally have no epithelial properties although, when taken from the ectoderm region, they may acquire such properties upon long-term contact with the environment. However they have wide morphogenetic potencies; the differences in these potencies between cells taken from the various presumptive regions being less than the differences between outer and inner cell layers in each region. Maps are provided which show the arrangement of presumptive rudiments in the ectoderm and chordamesoderm on sagittal sections through Bombina bombina embryos in early and late gastrulation.

scholarly journals Selection of Ovine Oocytes by Brilliant Cresyl Blue Staining

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Liqin Wang ◽  
Jiapeng Lin ◽  
Juncheng Huang ◽  
Jing Wang ◽  
Yuncheng Zhao ◽  
...  
Keyword(s):  
Embryonic Stem ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
G6pd Activity ◽  
Blue Staining ◽  
Animal Cloning ◽  
Cresyl Blue ◽  
Deep Blue ◽  
Glucose 6 Phosphate Dehydrogenase

Sheep oocytes derived from the ovaries collected from the slaughterhouse are often used for research onin vitroembryo production, animal cloning, transgenesis, embryonic stem cells, and other embryo biotechnology aspects. Improving thein vitroculture efficiency of oocytes can provide more materials for similar studies. Generally, determination of oocyte quality is mostly based on the layers of cumulus cells and cytoplasm or cytoplasm uniformity and colors. This requires considerable experience to better identify oocyte quality because of the intense subjectivity involved (Gordon (2003), Madison et al. (1992) and De Loos et al. (1992)). BCB staining is a function of glucose-6-phosphate dehydrogenase (G6PD) activity, an enzyme synthesized in developing oocytes, which decreases in activity with maturation. Therefore, unstained oocytes (BCB−) are high in G6PD activity, while the less mature oocytes stains are deep blue (BCB+) due to insuffcient G6PD activity to decolorize the BCB dye.

scholarly journals Reovirus Neurotropism and Virulence Are Dictated by Sequences in the Head Domain of the Viral Attachment Protein

2018 ◽  
Vol 92 (23) ◽  
Author(s):  
Danica M. Sutherland ◽  
Pavithra Aravamudhan ◽  
Melanie H. Dietrich ◽  
Thilo Stehle ◽  
Terence S. Dermody
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Ependymal Cells ◽  
Attachment Protein ◽  
Newborn Mice ◽  
Viral Attachment ◽  
Head Domain ◽  
Serotype 1 ◽  
The Central Nervous System ◽  
The Brain

ABSTRACTViral capsid components that bind cellular receptors mediate critical functions in viral tropism and disease pathogenesis. Mammalian orthoreoviruses (reoviruses) spread systemically in newborn mice to cause serotype-specific disease in the central nervous system (CNS). Serotype 1 (T1) reovirus infects ependymal cells to cause nonlethal hydrocephalus, whereas serotype 3 (T3) reovirus infects neurons to cause fulminant and lethal encephalitis. This serotype-dependent difference in tropism and concomitant disease is attributed to the σ1 viral attachment protein, which is composed of head, body, and tail domains. To identify σ1 sequences that contribute to tropism for specific cell types in the CNS, we engineered a panel of viruses expressing chimeric σ1 proteins in which discrete σ1 domains have been reciprocally exchanged. Parental and chimeric σ1 viruses were compared for replication, tropism, and disease induction following intracranial inoculation of newborn mice. Viruses expressing T1 σ1 head sequences infect the ependyma, produce relatively lower titers in the brain, and do not cause significant disease. In contrast, viruses expressing T3 σ1 head sequences efficiently infect neurons, replicate to relatively higher titers in the brain, and cause a lethal encephalitis. Additionally, T3 σ1 head-expressing viruses display enhanced infectivity of cultured primary cortical neurons compared with T1 σ1 head-expressing viruses. These results indicate that T3 σ1 head domain sequences promote infection of neurons, likely by interaction with a neuron-specific receptor, and dictate tropism in the CNS and induction of encephalitis.IMPORTANCEViral encephalitis is a serious and often life-threatening inflammation of the brain. Mammalian orthoreoviruses are promising oncolytic therapeutics for humans but establish virulent, serotype-dependent disease in the central nervous system (CNS) of many young mammals. Serotype 1 reoviruses infect ependymal cells and produce hydrocephalus, whereas serotype 3 reoviruses infect neurons and cause encephalitis. Reovirus neurotropism is hypothesized to be dictated by the filamentous σ1 viral attachment protein. However, it is not apparent how this protein mediates disease. We discovered that sequences forming the most virion-distal domain of T1 and T3 σ1 coordinate infection of either ependyma or neurons, respectively, leading to mutually exclusive patterns of tropism and disease in the CNS. These studies contribute new knowledge about how reoviruses target cells for infection in the brain and inform the rational design of improved oncolytic therapies to mitigate difficult-to-treat tumors of the CNS.

General concepts of hypothalamus-pituitary anatomy

2011 ◽  
pp. 71-81
Author(s):  
Ignacio Bernabeu ◽  
Monica Marazuela ◽  
Felipe F. Casanueva
Keyword(s):  
Median Eminence ◽  
Third Ventricle ◽  
Optic Chiasm ◽  
Perivascular Space ◽  
Ependymal Cells ◽  
The Third ◽  
Long Time ◽  
Central Grey Matter ◽  
Imaginary Line ◽  
The Brain

The hypothalamus is the part of the diencephalon associated with visceral, autonomic, endocrine, affective, and emotional behaviour. It lies in the walls of the third ventricle, separated from the thalamus by the hypothalamic sulcus. The rostral boundary of the hypothalamus is roughly defined as a line through the optic chiasm, lamina terminalis, and anterior commissure, and an imaginary line extending from the posterior commissure to the caudal limit of the mamillary body represents the caudal boundary. Externally, the hypothalamus is bounded rostrally by the optic chiasm, laterally by the optic tract, and posteriorly by the mamillary bodies. Dorsolaterally, the hypothalamus extends to the medial edge of the internal capsule (Fig. 2.1.1) (1). The complicated anatomy of this area of the central nervous system (CNS) is the reason why, for a long time, little was known about its anatomical organization and functional significance. Even though the anatomy of the hypothalamus is well established it does not form a well-circumscribed region. On the contrary, it is continuous with the surrounding parts of the CNS: rostrally, with the septal area of the telencephalon and anterior perforating substance; anterolaterally with the substantia innominata; and caudally with the central grey matter and the tegmentum of the mesencephalon. The ventral portion of the hypothalamus and the third ventricular recess form the infundibulum, which represents the most proximal part of the neurohypophysis. A bulging region posterior to the infundibulum is the tuber cinereum, and the zone that forms the floor of the third ventricle is called the median eminence. The median eminence represents the final point of convergence of pathways from the CNS on the peripheral endocrine system and it is supplied by primary capillaries of the hypophyseal portal vessels. The median eminence is the anatomical interface between the brain and the anterior pituitary. Ependymal cells lining the floor of the third ventricle have processes that traverse the width of the median eminence and terminate near the portal perivascular space; these cells, called tanycytes, provide a structural and functional link between the cerebrospinal fluid (CSF) and the perivascular space of the pituitary portal vessels. The conspicuous landmarks of the ventral surface of the brain can be used to divide the hypothalamus into three parts: anterior (preoptic and supraoptic regions), middle (tuberal region), and caudal (mamillary region). Each half of the hypothalamus is also divided into a medial and lateral zone. The medial zone contains the so-called cell-rich areas with well-defined nuclei. The scattered cells of the lateral hypothalamic area have long overlapping dendrites, similar to the cells of the reticular formation. Some of these neurons send axons directly to the cerebral cortex and others project down into the brainstem and spinal cord.

Ventricular and Cerebrospinal Fluid System

2017 ◽  
pp. 409-434
Author(s):  
Eduardo E. Benarroch ◽  
Jeremy K. Cutsforth-Gregory ◽  
Kelly D. Flemming
Keyword(s):  
Cerebrospinal Fluid ◽  
Neural Tissue ◽  
Local Environment ◽  
System P ◽  
Neurologic Disorders ◽  
Unique System ◽  
The Central Nervous System ◽  
Protective Structures ◽  
And Function ◽  
The Brain

The meninges, ventricular system, subarachnoid space, and cerebrospinal fluid (CSF) constitute a functionally unique system that has an important role in maintaining a stable environment within which the central nervous system can function. The membranes that constitute the meninges serve as supportive and protective structures for neural tissue. The CSF itself provides a cushioning effect during rapid movement of the head and mechanical buoyancy to the brain. In addition to providing a pathway for the removal of brain metabolites, it functions as a chemical reservoir that protects the local environment of the brain from changes that may occur in the blood, thus ensuring the brain’s continued undisturbed performance. The CSF system is present at the supratentorial, posterior fossa, and spinal levels. Because of this extensive anatomical distribution and function, pathologic alterations of the CSF system can occur in many neurologic disorders.

Vitamin D3 and glucose-6-phosphate dehydrogenase in rat duodenal epithelial cells

1989 ◽  
Vol 257 (5) ◽  
pp. G760-G765
Author(s):  
L. B. Nasr ◽  
J. D. Monet ◽  
P. Lucas ◽  
C. A. Bader
Keyword(s):  
Vitamin D ◽  
Intraperitoneal Administration ◽  
Middle Part ◽  
G6pd Activity ◽  
Dihydroxyvitamin D3 ◽  
Rat Duodenum ◽  
High Level ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Stimulation Of

A microdensitometric method was employed to determine enzyme activities in situ in undisrupted tissue rat duodenum. The effect of 1 alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] on glucose-6-phosphate dehydrogenase (G6PD) activity and on the two utilization pathways of synthesized NADPH, H1 (mixed function oxidation) and H2 (biosynthesis), was studied. In normal animals, a crypt-to-villus gradient of G6PD activity and of both NADPH utilization pathways was observed. A high level of NADPH utilization occurred predominantly via the H2 pathway. In vitamin D-deficient rat animals, G6PD activity in the middle part of the villus was approximately 60% lower than in normal animals [10.05 +/- 0.35 vs. 3.95 +/- 0.26 (means +/- SE) A585.min-1.micron-3 X 10(5), P less than 0.001] with reduced NADPH utilization via the H2 pathway (8.39 +/- 0.49 vs. 2.73 +/- 0.43 A585.min-1.micron-3 X 10(5), P less than 0.001) but not the H1 pathway (1.65 +/- 0.17 vs. 1.22 +/- 0.19 A585.min-1.micron-3 X 10(5), P = NS). Intraperitoneal administration of 1,25(OH)2D3 (500 pmol) to vitamin D-deficient animals resulted in increased G6PD activity within 30 min (4.09 +/- 0.38 vs. 5.51 +/- 0.39 A585.min-1.micron-3 X 10(5), P less than 0.05), attaining normal levels within 2 h. The H2 but not the H1 pathway of NADPH utilization increased significantly in response to 1,25(OH)2D3. This increase is essentially located in the basal and middle parts of the villus. Thus 1,25(OH)2D3 may influence biosynthesis in the duodenum via stimulation of G6PD activity and the H2 pathway of NADPH utilization.

scholarly journals Multiple forms with glucose 6-phosphate dehydrogenase activity in Musca domestica L. as revealed by electrophoresis on cellulose acetate gel.

1978 ◽  
Vol 26 (10) ◽  
pp. 846-854 ◽  
Author(s):  
L Cima ◽  
A Malacrida ◽  
G Gasperi ◽  
L Sacchi ◽  
A Grigolo
Keyword(s):  
Musca Domestica ◽  
Nicotinamide Adenine Dinucleotide ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
G6pd Activity ◽  
Nicotinamide Adenine ◽  
Molecular Forms ◽  
Type I ◽  
Coenzyme Specificity ◽  
Low Sensitivity ◽  
Glucose 6 Phosphate Dehydrogenase

Single newly emerged males of Musca domestica, WHO strain, usually show five electrophoretic bands of glucose 6-phosphate dehydrogenase (G6PD) activity. Of these five molecular forms, designated with Roman numerals in order from the origin, we have considered the first three: these have been characterized with respect to their substrate and coenzyme specificity and to their sensitivity to some sulfhydryl inhibitors. The data show band III to be G6P specific, nicotinamide adenine dinucleotide phosphate dependent and to be a type I enzyme according to Kamada and Hori's classification. Bands I and II, on the other hand, show wide substrate specificity and low sensitivity to the sulfhydryl inhibitors assayed. In addition, in the absence of an exogenous substrate and in the presence of nicotinamide adenine dinucleotide as a coenzyme, fairly weak bands, which can be ascribed to the so called "nothing dehydrogenase" effect, are seen in the position I and II. Nevertheless, the data reported do not allow a clear definition of the enzymatic type corresponding to bands I and II of G6PD activity.

scholarly journals Distribution of the Cystine/Glutamate Antiporter System x−c in the Brain, Kidney, and Duodenum

2006 ◽  
Vol 54 (5) ◽  
pp. 549-557 ◽  
Author(s):  
Joseph Burdo ◽  
Richard Dargusch ◽  
David Schubert
Keyword(s):  
Oxidative Stress ◽  
Vascular Endothelial Cells ◽  
Ependymal Cells ◽  
Vascular Endothelial ◽  
Specific Antibodies ◽  
Glutathione Status ◽  
Border Areas ◽  
Low Levels ◽  
Rate Limiting ◽  
The Brain

System x−c, one of the main transporters responsible for central nervous system cystine transport, is comprised of two subunits, xCT and 4F2hc. The transport of cystine into cells is rate limiting for glutathione synthesis, the major antioxidant and redox cofactor in the brain. Alterations in glutathione status are prevalent in numerous neurodegenerative diseases, emphasizing the importance of proper cystine homeostasis. However, the distribution of xCT and 4F2hc within the brain and other areas has not been described. Using specific antibodies, both xCT and 4F2hc were localized predominantly to neurons in the mouse and human brain, but some glial cells were labeled as well. Border areas between the brain proper and periphery including the vascular endothelial cells, ependymal cells, choroid plexus, and leptomeninges were also highly positive for the system x−c components. xCT and 4F2hc are also present at the brush border membranes in the kidney and duodenum. These results indicate that system x−c is likely to play a role in cellular health throughout many areas of the brain as well as other organs by maintaining intracellular cystine levels, thereby resulting in low levels of oxidative stress. (J Histochem Cytochem 54: 549–557, 2006)

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