scholarly journals Metabolic changes in brain slices over time: a multiplatform metabolomics approach

2020 ◽  
Author(s):  
Carolina Gonzalez-Riano ◽  
Silvia Tapia-González ◽  
Gertrudis Perea ◽  
Candela González-Arias ◽  
Javier DeFelipe ◽  
...  
Keyword(s):  
Brain Slice ◽  
Ex Vivo ◽  
Brain Slices ◽  
Tca Cycle ◽  
Electrophysiological Recording ◽  
Slice Preparation ◽  
Experimental Conditions ◽  
The Status ◽  
The Brain ◽  
Brain Slice Preparation

ABSTRACTBrain slice preparations are widely used for research in neuroscience. However, a high-quality preparation is essential and there is no consensus regarding stable parameters that can be used to define the status of the brain slice preparation after its collection at different time points. Thus, it is critical to establish the best experimental conditions for ex-vivo studies using brain slices for electrophysiological recording. In this study, we used a multiplatform (LC-MS and GC-MS) untargeted metabolomics-based approach to shed light on the metabolome and lipidome changes induced by the brain slice preparation process. We have found significant modifications in the levels of 300 compounds, including several lipid classes and their derivatives, as well as metabolites involved in the GABAergic pathway and the TCA cycle. All these preparation-dependent changes in the brain biochemistry should be taken into consideration for future studies to facilitate non-biased interpretations of the experimental results.

scholarly journals Compartmentation of cerebral glutamate in situ as detected by 1H/13C n.m.r

1994 ◽  
Vol 298 (1) ◽  
pp. 121-127 ◽  
Author(s):  
R A Kauppinen ◽  
T R M Pirttilä ◽  
S O K Auriola ◽  
S R Williams
Keyword(s):  
Steady State ◽  
Brain Slice ◽  
Brain Slices ◽  
Slice Preparation ◽  
Total Acid ◽  
Significant Difference ◽  
13C Label ◽  
The Brain ◽  
Brain Slice Preparation

Incorporation of 13C label from either [1-13C]glucose to glutamate C-4 and lactate C-3 or from [2-13C]acetate to glutamate C-4 was monitored in situ in a superfused brain slice preparation by using 1H-detected/13C-edited (1H/13C) n.m.r. spectroscopy. The fractional enrichments of both metabolites were determined by this means in both brain slices and acid extracts of the preparations in order to assess their 1H-n.m.r. detectabilities. The 1H/13C satellite resonances from glutamate C-4 and lactate C-3 in brain tissue were followed from 4 min onwards in the presence of 5 mM [1-13C]glucose. Fractional enrichment of glutamate C-4 in the slice preparations was higher than in their acid extracts throughout the incubation of 100 min; at 30 min the enrichment was 15.9 +/- 0.6% in the slice preparations and 10.6 +/- 0.9% in extracts and at 100 min 24.5 +/- 1.7% compared with 19.7 +/- 0.4%, respectively. In contrast, lactate C-3 reached a steady-state fractional enrichment of approx. 43% by 15 min and there was no difference between the values determined in the slice preparations and the acid extracts. There was a significant difference between the glutamate C-4 fractional enrichments in the brain slices (7.4 +/- 0.6%) and extracts (5.1 +/- 0.3%) after 60 min of incubation with [2-13C]acetate. Thus 13C label from both glucose and exogenous acetate enters a pool of glutamate that is more amenable to 1H n.m.r. detection than total acid-extracted brain biochemical glutamate, whereas lactate is labelled with full 1H n.m.r. visibility. The results are discussed in the light of the biochemical factors that affect glutamate 1H-n.m.r. susceptibility and thus its n.m.r. visibility.

23. A new adult rat brain slice preparation for ex vivo NMR spectroscopy studies of stroke

1994 ◽  
Vol 52 (1) ◽  
pp. A11
Author(s):  
M.T. Espanol ◽  
L. Litt ◽  
L.-H. Chang ◽  
T.L. James ◽  
P.R. Weinstein ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Rat Brain ◽  
Brain Slice ◽  
Ex Vivo ◽  
Slice Preparation ◽  
Adult Rat ◽  
Adult Rat Brain ◽  
Spectroscopy Studies ◽  
Rat Brain Slice ◽  
Brain Slice Preparation

Halothane-induced Dilatation of Intraparenchymal Arterioles in Rat Brain Slices 

1997 ◽  
Vol 86 (4) ◽  
pp. 885-894 ◽  
Author(s):  
Christopher P. Harkin ◽  
Antal G. Hudetz ◽  
William T. Schmeling ◽  
John P. Kampine ◽  
Neil E. Farber
Keyword(s):  
Sodium Nitroprusside ◽  
Brain Slice ◽  
Brain Slices ◽  
Resistance Index ◽  
Nitric Oxide Donor ◽  
Slice Preparation ◽  
Cerebral Microvessels ◽  
Cerebrovascular Resistance ◽  
Prostaglandin F2 Alpha ◽  
Brain Slice Preparation

Background Halothane is a potent dilator of cerebral arteries. The predominant site of cerebrovascular resistance is thought to be intracerebral arterioles, and the effects of halothane on these vessels were not previously examined. This study compared the effects of halothane with those of the vasodilator and nitric oxide donor, sodium nitroprusside, on intraparenchymal microvessel responsiveness in a brain slice preparation. Methods Anesthetized Sprague-Dawley rats underwent thoracotomy and intracardiac perfusion and then were decapitated. Hippocampal brain slices were prepared and placed in a perfusion/recording chamber and superfused with artificial cerebrospinal fluid. An arteriole was located within the brain parenchyma and its diameter was monitored with videomicroscopy before, during, and after various concentrations of halothane or sodium nitroprusside were equilibrated in the perfusate. All vessels were preconstricted with prostaglandin F2 alpha before halothane or sodium nitroprusside treatment. An observer blinded to treatment analyzed vessel diameter changes with a computerized videomicrometer. Results Baseline microvessel diameter was 18 +/- 2 microns in the halothane group (n = 14) and 15 +/- 1 microns in the sodium nitroprusside group (n = 15). Prostaglandin F2 alpha (0.5 micron) preconstricted vessels by approximately 15% from resting diameter in both groups. Halothane significantly and dose dependently dilated intracerebral microvessels by 54% +/- 6%, 74% +/- 8%, 108% +/- 13%, and 132% +/- 7% (normalized to the preconstricted diameter) at 0.5%, 1.0%, and 2.5% halothane, respectively. This dilatation corresponds to a decrease in a calculated index of cerebrovascular resistance index of up to 117% +/- 2% at 2.5% halothane. Sodium nitroprusside, in concentrations ranging from 10(-8) to 10(-3)M, also dose dependently dilated these intraparenchymal vessels by 129% +/- 7% at the highest concentration. These alterations in microvessel diameter corresponded to a decrease in the cerebrovascular resistance index of up to 116 +/- 4% for the largest dose. Conclusions Halothane produces dose-dependent vasodilatation of intraparenchymal cerebral microvessels, thus predicting marked decreases in cerebrovascular resistance in this in vitro brain slice preparation. The effects of halothane on these cerebral microvessels are similar to those of the potent vasodilator sodium nitroprusside. These findings suggest that direct effects of halathane on cerebral microvessels diameter contribute substantially to alterations in cerebrovascular resistance and flow produced by this agent.

A dual chamber for comparative studies using the brain slice preparation

1985 ◽  
Vol 82 (3) ◽  
pp. 701-704 ◽  
Author(s):  
A Schurr ◽  
K.H Reid ◽  
M.T Tseng ◽  
H.L Edmonds ◽  
B.M Rigor
Keyword(s):  
Comparative Studies ◽  
Brain Slice ◽  
Slice Preparation ◽  
Dual Chamber ◽  
The Brain ◽  
Brain Slice Preparation

Fate of sperm membrane in fertilized ova

1993 ◽  
Vol 51 ◽  
pp. 40-41
Author(s):  
Frank J. Longo
Keyword(s):  
Electron Microscopy ◽  
Electrical Activity ◽  
Sea Urchins ◽  
Morphological Changes ◽  
Integrated System ◽  
Electrophysiological Recording ◽  
Experimental Conditions ◽  
The Status ◽  
Sperm Membrane ◽  
Temporal And Spatial

Measurement of the egg's electrical activity, the fertilization potential or the activation current (in voltage clamped eggs), provides a means of detecting the earliest perceivable response of the egg to the fertilizing sperm. By using the electrical physiological record as a “real time” indicator of the instant of electrical continuity between the gametes, eggs can be inseminated with sperm at lower, more physiological densities, thereby assuring that only one sperm interacts with the egg. Integrating techniques of intracellular electrophysiological recording, video-imaging, and electron microscopy, we are able to identify the fertilizing sperm precisely and correlate the status of gamete organelles with the first indication (fertilization potential/activation current) of the egg's response to the attached sperm. Hence, this integrated system provides improved temporal and spatial resolution of morphological changes at the site of gamete interaction, under a variety of experimental conditions. Using these integrated techniques, we have investigated when sperm-egg plasma membrane fusion occurs in sea urchins with respect to the onset of the egg's change in electrical activity.

TMOD-31. AN ORGANOTYPIC TISSUE PLATFORM TO BRIDGE IN VITRO AND IN VIVO ASSAYS FOR BRAIN CANCER TREATMENT

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi222-vi222
Author(s):  
Breanna Mann ◽  
Noah Bell ◽  
Denise Dunn ◽  
Scott Floyd ◽  
Shawn Hingtgen ◽  
...  
Keyword(s):  
Cell Lines ◽  
Brain Cancer ◽  
Brain Slice ◽  
Brain Slices ◽  
In Vitro Assays ◽  
Preclinical Model ◽  
Short Period ◽  
The Brain

Abstract Brain cancers remain one of the greatest medical challenges. The lack of experimentally tractable models that recapitulate brain structure/function represents a major impediment. Platforms that enable functional testing in high-fidelity models are urgently needed to accelerate the identification and translation of therapies to improve outcomes for patients suffering from brain cancer. In vitro assays are often too simple and artificial while in vivo studies can be time-intensive and complicated. Our live, organotypic brain slice platform can be used to seed and grow brain cancer cell lines, allowing us to bridge the existing gap in models. These tumors can rapidly establish within the brain slice microenvironment, and morphologic features of the tumor can be seen within a short period of time. The growth, migration, and treatment dynamics of tumors seen on the slices recapitulate what is observed in vivo yet is missed by in vitro models. Additionally, the brain slice platform allows for the dual seeding of different cell lines to simulate characteristics of heterogeneous tumors. Furthermore, live brain slices with embedded tumor can be generated from tumor-bearing mice. This method allows us to quantify tumor burden more effectively and allows for treatment and retreatment of the slices to understand treatment response and resistance that may occur in vivo. This brain slice platform lays the groundwork for a new clinically relevant preclinical model which provides physiologically relevant answers in a short amount of time leading to an acceleration of therapeutic translation.

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