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At least 703 records · Page 39Linked to original sources
PubMed1990

Cell isolation and genotoxicity assessment in gastric mucosa.

It has been claimed that in vitro digestion of in vivo DNA-labeled gastric mucosa is suitable for evaluation of genotoxic effects of drugs or chemicals. This method was then used to show that omeprazole (a novel antiulcer drug) was potentially genotoxic. In this study we have examined the method used and the interaction of omeprazole and its derivatives with purified DNA. The method was shown to enrich for dividing cells (6.92 +/- 0.693%, N = 43, 2-hr labeling) in the digest from the intact tissue and was therefore unsuitable for estimating unscheduled DNA synthesis in the gastric mucosa induced by chemicals or drugs including omeprazole. It was further shown that neither omeprazole or its acid-activated product, a cationic sulfenamide, were able to react with isolated purified DNA from either a prokaryote (E. coli) or a eukaryote (salmon sperm). Hence any conclusions using this method attributing acute genotoxic effects to any chemical are based on unrecognized artifacts of the technique and are unsound. In addition, these results negate the suggestion that omeprazole or its gastric metabolites are genotoxic.

Animals↗
PubMed1982

Epithelial cell volume modulation and regulation.

Epithelial cell volume is a sensitive indicator of the balance between solute entry into the cell and solute exit. Solute accumulation in the cell leads to cell swelling because the water permeability of the cell membranes is high. Similarly, solute depletion leads to cell shrinkage. The rate of volume change under a variety of experimental conditions may be utilized to study the rate and direction of solute transport by an epithelial cell. The pathways of water movement across an epithelium may also be deduced from the changes in cellular volume. A technique for the measurement of the volume of living epithelial cells is described, and a number of experiments are discussed in which cell volume determination provided significant new information about the dynamic behavior of epithelia. The mechanism of volume regulation of epithelial cells exposed to anisotonic bathing solution is discussed and shown to involve the transient stimulation of normally dormant ion exchangers in the cell membrane.

Animals↗
PubMed1980

Cryoprotection of human bone marrow committed stem cells (CFU-c) by dextran, glycerol and dimethyl sulfoxide.

Dextran, glycerol and dimethyl sulfoxide (DMSO), alone or in combination, were used for cryoprotection of human bone marrow cells. The viability of cryopreserved cells was assessed by culture of myelopoiesis-committed stem cells (CFU-c) in vitro. A significantly better protection against freezing injury was obtained by 9% dextran in combination with 3 or 5% DMSO, and also with 5 or 10% DMSO alone, than with either 15% glycerol or 9% dextran with 1% DMSO.

Animals↗
PubMed1987

Maxi K+ channels in leaky epithelia are regulated by intracellular Ca2+, pH and membrane potential.

We have studied a Ca2+-activated K+ channel in the ventricular membrane of the epithelium of choroid plexus by means of the patch-clamp technique, using excised inside-out patches. The channel was highly K+ selective. It had a conductance of approximately 200 pS with 112 mM KCl on both sides of the membrane. The probability for the channel being open increased with intracellular Ca2+, pH and with membrane potential. The channel shows two gating modes. The primary gating mode has open and closed times which depend strongly on membrane potential, intracellular Ca2+ and pH. It accounts for the variation of the channel open probability. Lowering intracellular pH from 7.4 to 6.4 reduced the channel open probability mainly by increasing the channel closed time. It appears, that H+ can compete with Ca2+ in binding to the same site, thereby preventing channel opening. A second gating mode consisted of short-lived closures, or flickers. The open and closed time for this process were largely independent of membrane potential, intracellular Ca2+ and pH. The channel density was approximately 0.4 micron-2 corresponding to a K+-permeability of 2.2 10(-5) cm s-1 if the channels were fully open. In cell-attached patches we measured the open probability of the channel in the intact cell membrane. The channel is almost totally closed under normal cellular conditions. This type of channel is therefore not the membrane component that forms the electrodiffusive pathway for K+-ions.

Animals↗
PubMed1997

5-Lipoxygenase metabolites of arachidonic acid regulate volume decrease by mudpuppy red blood cells.

We examined whether metabolites of arachidonic acid (AA) regulate K+ efflux during regulatory volume decrease (RVD) by mudpuppy red blood cells (RBCs). Volume regulation was inhibited by the phospholipase A2 antagonists mepacrine (10 microM) and ONO-RS-082 (10 microM); the inhibitory effect of ONO-RS-082 was reversed by gramicidin (5 microM). Eicosatetraynoic acid (ETYA, 100 microM), a general antagonist of AA metabolism, also blocked RVD. In addition, volume regulation was inhibited by the lipoxygenase pathway antagonist nordihydroguaiaretic acid (NDGA, 10 microM), the 5 lipoxygenase antagonists AA-861 (5 microM) and curcumin (20 microM), and by the 5-lipoxygenase activating protein inhibitor L-655,298 (5 microM). Inhibition by all four of these agents was reversed with gramicidin. In contrast, the 12- and 15-lipoxygenase pathway inhibitor ethyl-3,4-dihydroxy-benzylidene-cyanoacetate (EDBCA, 1 microM) and the cytochrome P-450 monooxygenase pathway blocker ketoconazole (20 microM) had no effect. On the other hand, the cyclooxygenase pathway inhibitor aspirin (100 microM) slightly enhanced RVD. Consistent with these findings, a K(+)-selective whole cell conductance responsible for K+ efflux during cell swelling was inhibited by ONO-RS-082 (10 microM), NDGA (10 microM), AA-861 (5 microM), curcumin (20 microM), and L-655,298 (5 microM). In contrast, EDBCA (1 microM), ketoconazole (20 microM), and indomethacin (10 microM) did not block this whole cell conductance. These results indicate that a channel mediating K+ loss during RVD is regulated by a 5-lipoxygenase metabolite of arachidonic acid.

Animals↗
PubMed2003

Neural computations in the tiger salamander and mudpuppy outer retinae and an analysis of GABA action from horizontal cells.

A neural network architecture based on the neural anatomy and function of retinal neurons in tiger salamander and mudpuppy retinae is proposed to study basic aspects of early visual information processing. The model predictions for the main response characteristics of retinal neurons are found to be in agreement with neurophysiological data, including the antagonistic role of horizontal cells in the outer plexiform layer. The examination of possible gamma-aminobutyric acid (GABA) action from horizontal cells suggests that GABA(A) alone, GABA(B) alone, or their weighted combination can generate the response characteristics observed in bipolar cells.

Ambystoma↗
PubMed1986

Neurotensin actions in the retina: mechanisms and variability.

The effects of neurotensin on mudpuppy retinal cells were studied using extracellular and intracellular electrophysiological recording techniques and bath application of the peptide. Ganglion and amacrine cells (but not bipolar or horizontal cells) were reversibly depolarized by low micromolar concentrations of neurotensin. Depolarizations also occurred with neurotensin application during cobalt block of synaptic transmission and were accompanied by decreased input resistances. This suggests neurotensin may act directly on amacrine and ganglion cells as a conventional excitatory transmitter. However, in many retinas, cells responded to light stimuli and to other drugs but not to neurotensin. These negative results are important in considering the peptide's normal role in retinal function.

Action Potentials↗
PubMed1986

Kynurenic acid distinguishes kainate and quisqualate receptors in the vertebrate retina.

Excitatory amino acid receptors (EAARs) underlie major synaptic pathways in the brain, retina and spinal cord. Several subclasses of EAARs have been proposed, based on pharmacological studies using a variety of agonists and antagonists. Kynurenic acid (Kyn), a metabolite of tryptophan, has been recently proposed as a potent EAAR antagonist. In this report, we show that Kyn can be used to separate two distinct classes of EAAR in the vertebrate retina: it blocks kainic acid (KA) responses but has minimal effects on responses mediated by quisqualate (QQ). At concentrations which block the KA responses, Kyn also blocks the light-evoked synaptic responses of all types of third-order neurons in the retina. These results suggest that KA receptors are the major receptor subtypes which underlie synaptic transmission and that QQ receptors are minimally utilized by light-activated pathways under the conditions of our experiments.

Animals↗
PubMed1987

Subtle actions of 2-amino-4-phosphonobutyrate (APB) on the Off pathway in the mudpuppy retina.

The principal action of 2-amino-4-phosphonobutyrate (APB) applied to the retina is the elimination of the On-bipolar cell light response. In this study, we report that APB also has more subtle effects; it sometimes enhances Off responses of Off bipolar cells and On, Off, and On-Off inner retina neurons. Possible mechanisms for this enhancement are considered.

Action Potentials↗
PubMed1991

Rods and cones activate different excitatory amino acid receptors on the mudpuppy retinal horizontal cell.

We have studied rod and cone neurotransmission onto horizontal cells (HCs) of the mudpuppy retina by: (1) dual whole-cell recordings from synaptically connected pairs in a superfused retinal slice preparation, and (2) intracellular recordings from HCs in a superfused retina-eyecup preparation. In the presence of relatively weak concentrations of broad spectrum excitatory amino acid antagonists, HC recordings show that cone-mediated responses were significantly more attenuated than those mediated by rods. These observations suggest that the HC postsynaptic receptors for rods and cones are pharmacologically different. One advantage of having distinct rod- and cone-activated receptors on second-order neurons may be to preserve different rod and cone functions under varying levels of light stimulation and provide separate means of regulating rod and cone neurotransmission onto the second-order cells.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1994

Pharmacology of the GABAB receptor in amphibian retina.

Amacrine and ganglion cells in the amphibian retina contain GABAB, as well as GABAA, receptors. Baclofen, a GABAB agonist, hyperpolarizes the dark membrane potential of these third order neurons and makes their light responses more transient. GABAB receptors in the retina have a similar agonist profile to GABAB receptors described at other sites in the brain. Namely, preferential activation by the R-enantiomer of baclofen, and agonist sensitivity in the order 3-aminopropylphosphinic acid > baclofen >> 3-aminopropylphosphonic acid. The GABAB receptor was not activated by 4-aminobutylphosphonic acid. Several antagonists, such as phaclofen, saclofen, and 2-hydroxysaclofen, were ineffective in the amphibian retina. However, CGP35348 blocked the action of applied baclofen and produced effects on the light response that were opposite to those of baclofen. Applied agonists and antagonists support the hypothesis that GABAB receptors serve to regulate the balance of sustained and transient signals to the inner retina.

Ambystoma↗
PubMed1995

Pharmacology of selective and non-selective metabotropic glutamate receptor agonists at L-AP4 receptors in retinal ON bipolar cells.

Retinal ON bipolar cells possess metabotropic glutamate receptors (mGluRs) which are sensitive to L-2-amino-4-phosphonobutyric acid (L-AP4). Recent studies suggest there are multiple subtypes of L-AP4 receptors. In order to provide a more complete description of the pharmacology of the retinal L-AP4 receptor, we examined the actions of a number of compounds which are active at L-AP4 receptors and other mGluRs. Four groups of compounds were studied: (1) AP4 analogues (e.g. L-AP5, L-SOP, cyclobutylene AP5, and N-Me-AP4), (2) non-selective mGluR agonists (ibotenate and quisqualate), (3) selective mGluR agonists (L-CCG-I), and (4) agonists proposed to be selective for specific mGluR subtypes (DCG-IV and t-ADA). Concentration-response curves were obtained using the b-wave of the electroretinogram (ERG) as an assay for L-AP4 receptor activation. Whole cell voltage clamp recordings from ON bipolar cells in the retinal slice preparation of the mudpuppy were used to determine whether the compounds acted as L-AP4 receptor agonists. All compounds were L-AP4 receptor agonists, except t-ADA which was ineffective. The results reveal pharmacological differences between L-AP4 receptors in mudpuppy ON bipolar cells and those in other systems, consistent with the proposal that there are multiple L-AP4 receptor subtypes. For example, retinal L-AP4 receptors are more potently activated by L-AP5 than L-SOP, whereas L-SOP has been shown to be more potent than L-AP5 in L-AP4 receptors in the lateral perforant path (LPP) of the rat hippocampus. L-SOP is also relatively more potent at the cloned L-AP4 receptors mGluR4, 6, and 7 than in mudpuppy ON bipolar cells in situ. The different potencies of these compounds in retina and LPP is ascribed to both steric and charge factors. The results with DCG-IV and t-ADA are consistent with the proposal that these are subtype-selective agonists, but DCG-IV is likely to be selective only at very low concentrations (< or = 1 microM).

Aminobutyrates↗
PubMed1995

Galantide distinguishes putative subtypes of galanin receptors in mudpuppy parasympathetic neurons.

The effect of the chimeric ligand galantide on the galanin-induced activation of membrane K+ conductance and inhibition of voltage-dependent Ca2+ conductance has been studied using voltage-clamped dissociated mudpuppy parasympathetic neurons. Galantide did not activate the K+ conductance but produced a concentration-dependent antagonism (IC50 = 4 nM) of the galanin-induced increase in K+ conductance. Galantide acted like galanin and inhibited the voltage-dependent Ba2+ current (IBa). The inhibition of IBa also was concentration dependent (IC50 = 16 nM) and the maximum inhibition produced by galantide was approximately 40%. We also demonstrate that the galanin-(1-16) fragment increased the membrane K+ conductance and decreased IBa, suggesting that the NH2 portion of the galanin molecule is sufficient to mediate both actions. One interpretation of these observations is that different galanin receptors mediate the different effects of galanin on the mudpuppy parasympathetic neurons.

Animals↗