Preparative methods for brain slices: a discussion

Aitken, Peter G.; Breese, George R.; Dudek, F. F.; Edwards, FA; Espanol, Maryceline T.; Larkman, P.M.; Lipton, Peter; Newman, George C.; Nowak, Thaddeus S.; Panizzon, Kimberly L.; Raley‐Susman, Kathleen M.; Reid, Kenneth H.; Rice, Margaret E.; Sarvey, John M.; Schoepp, Darryle D.; Segal, Malcolm B.; Taylor, Charles P.; Teyler, Timothy J.; Voulalas, Pamela J.

doi:10.1016/0165-0270(94)00204-t

Cited by 134 publications

(92 citation statements)

References 2 publications

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“…This solution was preequilibrated with 95% O 2 plus 5% CO 2 . Immediately after the slicing procedure, slices were transferred to a chamber containing artificial cerebrospinal fluid (ACSF) and allowed to recover at 36°C for 45 min, followed by storage at room temperature in the same ACSF (Aitken et al, 1995). ACSF contained 126 mM NaCl, 3 mM KCl, 1.25 mM NaH 2 PO 4 , 1 mM MgSO 4 , 26 mM NaHCO 3 , 2 mM CaCl 2 , and 10 mM glucose equilibrated with 95% O 2 plus 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Ethanol Sensitivity of GABAergic Currents in Cerebellar Granule Neurons Is Not Increased by a Single Amino Acid Change (R100Q) in the α₆ GABA_A Receptor Subunit

Botta

Mameli²,

Floyd³

et al. 2007

J Pharmacol Exp Ther

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Cerebellar granule neurons (CGNs) extrasynaptically express GABA A receptors containing ␣ 6 ␤ x ␦ subunits, which mediate tonic inhibitory currents. Although it has been shown that the function of these receptors is potently and directly enhanced by ethanol, this finding has not been reproducible across different laboratories. In outbred Sprague-Dawley rats, a naturally occurring arginine (R) to glutamine (Q) mutation in position 100 of the ␣ 6 subunit was reported to increase the ethanol sensitivity of these receptors. However, we did not detect an action of this mutation in selectively bred rats (alcohol-tolerant and alcoholnontolerant). Consequently, we reexamined the effect of the mutation on ethanol sensitivity in Sprague-Dawley rats. Using patch-clamp electrophysiological techniques in cerebellar vermis parasagittal slices, we found that 25 mM ethanol increases the tonic current amplitude, tonic current noise, and spontaneous inhibitory postsynaptic current (sIPSC) frequency to a similar extent in ␣ 6 -100R/100R and ␣ 6 -100Q/100Q CGNs. Exposure to 80 mM ethanol increased the tonic current amplitude to a significantly greater extent in ␣ 6 -100R/100R than in ␣ 6 -100Q/ 100Q CGNs; however, the effects of 80 mM ethanol on the tonic current noise and sIPSC frequency were not significantly different between these groups. In the presence of tetrodotoxin, a non-N-methyl-D-aspartate receptor antagonist, exogenous GABA, and a GABA transporter inhibitor, neither 8 nor 40 mM ethanol consistently affected tonic current amplitude or noise in ␣ 6 -100R/100R or ␣ 6 -100Q/100Q CGNs. Thus, the ␣ 6 -R100Q GABA A receptor subunit polymorphism does not increase the acute ethanol sensitivity of extrasynaptic receptors, lending further support to the hypothesis that ethanol modulates these currents indirectly via a presynaptic mechanism.Studies indicate that ethanol enhances GABAergic transmission in several brain regions via presynaptic and postsynaptic mechanisms (reviewed in Siggins et al., 2005;

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Section: Methodsmentioning

confidence: 99%

Ethanol Sensitivity of GABAergic Currents in Cerebellar Granule Neurons Is Not Increased by a Single Amino Acid Change (R100Q) in the α₆ GABA_A Receptor Subunit

Botta

Mameli²,

Floyd³

et al. 2007

J Pharmacol Exp Ther

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“…During the recovery process, slices are thought to be exposed to increased extracellular adenosine, 6 in addition to suffering from ATP depletion due to the resumption of internal cell metabolism. 43 Takahashi et al 6 reported that extracellular adenosine release during OGD in the isolated spinal cord is highly dependent on temperature: the OGD-induced increase in adenosine levels at 35 C is about 10-fold higher than that at 25 C. In the acute slice tissue, it is probable that the levels of ADA 1,2 decrease due to a lack of ADA supply from blood circulation 2 and diffusive loss of ect-ADA. 2 Pre-incubation with ADA ( Figure 5(a)) increased the eFP detection ratio 3-6 h after the incubation ( Figure 5(b), CTRL: n ¼ 100, 18%; ADA: n ¼ 100, 40%.…”

Section: Ada Preserves Striatal Cell Morphology and Electrical Activitymentioning

confidence: 99%

Neuroprotective effects of adenosine deaminase in the striatum

Tamura

Ohta

Satoh

et al. 2016

J Cereb Blood Flow Metab

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Adenosine deaminase (ADA) is a ubiquitous enzyme that catabolizes adenosine and deoxyadenosine. During cerebral ischemia, extracellular adenosine levels increase acutely and adenosine deaminase catabolizes the increased levels of adenosine. Since adenosine is a known neuroprotective agent, adenosine deaminase was thought to have a negative effect during ischemia. In this study, however, we demonstrate that adenosine deaminase has substantial neuroprotective effects in the striatum, which is especially vulnerable during cerebral ischemia. We used temporary oxygen/glucose deprivation (OGD) to simulate ischemia in rat corticostriatal brain slices. We used field potentials as the primary measure of neuronal damage. For stable and efficient electrophysiological assessment, we used transgenic rats expressing channelrhodopsin-2, which depolarizes neurons in response to blue light. Time courses of electrically evoked striatal field potential (eFP) and optogenetically evoked striatal field potential (optFP) were recorded during and after oxygen/glucose deprivation. The levels of both eFP and optFP decreased after 10 min of oxygen/glucose deprivation. Bath-application of 10 mg/ml adenosine deaminase during oxygen/glucose deprivation significantly attenuated the oxygen/glucose deprivation-induced reduction in levels of eFP and optFP. The number of injured cells decreased significantly, and western blot analysis indicated a significant decrease of autophagic signaling in the adenosine deaminase-treated oxygen/glucose deprivation slices. These results indicate that adenosine deaminase has protective effects in the striatum.

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“…This control level of PO2, which is typically used for brain slice studies (1,23), is actually very hyperoxic at normobaric pressure compared with the intact CNS (Refs. 8, 28, 51; and see Table 1).…”

Section: Control Conditionsmentioning

confidence: 99%

Hyperbaric oxygen and chemical oxidants stimulate CO₂/H⁺-sensitive neurons in rat brain stem slices

Mulkey

Henderson²,

Putnam³

et al. 2003

Journal of Applied Physiology

153

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. Hyperbaric oxygen and chemical oxidants stimulate CO2/H ϩ -sensitive neurons in rat brain stem slices. J Appl Physiol 95: 910-921, 2003. First published April 18, 2003 10.1152/japplphysiol.00864. 2002-Hyperoxia, a model of oxidative stress, can disrupt brain stem function, presumably by an increase in O2 free radicals. Breathing hyperbaric oxygen (HBO 2) initially causes hyperoxic hyperventilation, whereas extended exposure to HBO 2 disrupts cardiorespiratory control. Presently, it is unknown how hyperoxia affects brain stem neurons. We have tested the hypothesis that hyperoxia increases excitability of neurons of the solitary complex neurons, which is an important region for cardiorespiratory control and central CO2/H ϩ chemoreception. Intracellular recordings were made in rat medullary slices during exposure to 2-3 atm of HBO 2, HBO 2 plus antioxidant (Trolox C), and chemical oxidants (N-chlorosuccinimide, chloramine-T). HBO 2 increased input resistance and stimulated firing rate in 38% of neurons; both effects of HBO 2 were blocked by antioxidant and mimicked by chemical oxidants. Hypercapnia stimulated 32 of 60 (53%) neurons. Remarkably, these CO 2/H ϩ -chemosensitive neurons were preferentially sensitive to HBO2; 90% of neurons sensitive to HBO2 and/or chemical oxidants were also CO2/H ϩ chemosensitive. Conversely, only 19% of HBO2-insensitive neurons were CO2/H ϩ chemosensitive. We conclude that hyperoxia decreases membrane conductance and stimulates firing of putative central CO2/H ϩ -chemoreceptor neurons by an O2 free radical mechanism. These findings may explain why hyperoxia, paradoxically, stimulates ventilation. central chemoreception; reactive oxygen species; cardiorespiratory control; intracellular recording; hyperoxia THE CENTRAL NERVOUS SYSTEM (CNS) is especially sensitive to oxidative stress. For example, hyperoxia, which is a popular model of oxidative stress, induced by breathing high levels of oxygen at hyperbaric pressure [i.e., hyperbaric oxygen (HBO 2 )], can rapidly disrupt neural function and result in CNS O 2 toxicity (16). Neurological responses to hyperoxia vary, depending on the oxygen tension in the brain and the duration of exposure. For example, the CNS response to hyperoxia can range from moderate, but reversible, changes in neural activity (7,49), to violent and reversible seizures at higher levels of oxygen (16), to irreversible motor deficits and ultimately death at the highest dosages of hyperoxia (16). In each of these instances, the effects of hyperoxia on the CNS are thought to result from increased production and accumulation of O 2 free radicals and subsequent oxidation of cellular components vital to maintaining normal mechanisms of neuronal excitability (16).The cardiorespiratory centers of the brain stem, similarly, are sensitive to a broad range of inspired oxygen. Hypoxia increases alveolar ventilation, primarily by stimulation of the peripheral chemoreceptors (18), although a central stimulatory effect on ventilation has also been reported (47, 64). Paradoxica...

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Preparative methods for brain slices: a discussion

Cited by 134 publications

References 2 publications

Ethanol Sensitivity of GABAergic Currents in Cerebellar Granule Neurons Is Not Increased by a Single Amino Acid Change (R100Q) in the α₆ GABA_A Receptor Subunit

Ethanol Sensitivity of GABAergic Currents in Cerebellar Granule Neurons Is Not Increased by a Single Amino Acid Change (R100Q) in the α₆ GABA_A Receptor Subunit

Neuroprotective effects of adenosine deaminase in the striatum

Hyperbaric oxygen and chemical oxidants stimulate CO₂/H⁺-sensitive neurons in rat brain stem slices

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Preparative methods for brain slices: a discussion

Cited by 134 publications

References 2 publications

Ethanol Sensitivity of GABAergic Currents in Cerebellar Granule Neurons Is Not Increased by a Single Amino Acid Change (R100Q) in the α6 GABAA Receptor Subunit

Ethanol Sensitivity of GABAergic Currents in Cerebellar Granule Neurons Is Not Increased by a Single Amino Acid Change (R100Q) in the α6 GABAA Receptor Subunit

Neuroprotective effects of adenosine deaminase in the striatum

Hyperbaric oxygen and chemical oxidants stimulate CO2/H+-sensitive neurons in rat brain stem slices

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Product

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Ethanol Sensitivity of GABAergic Currents in Cerebellar Granule Neurons Is Not Increased by a Single Amino Acid Change (R100Q) in the α₆ GABA_A Receptor Subunit

Ethanol Sensitivity of GABAergic Currents in Cerebellar Granule Neurons Is Not Increased by a Single Amino Acid Change (R100Q) in the α₆ GABA_A Receptor Subunit

Hyperbaric oxygen and chemical oxidants stimulate CO₂/H⁺-sensitive neurons in rat brain stem slices