Відмінності між версіями «Got An Thiazovivin Paradox ? If So Review This Guidance»
(Створена сторінка: Slices were cut to a thickness of 150?��m using a tissue slicer, and transferred to the recording chamber while remaining submerged. The recording chamber w...)
Поточна версія на 06:31, 18 липня 2017
Slices were cut to a thickness of 150?��m using a tissue slicer, and transferred to the recording chamber while remaining submerged. The recording chamber was immediately attached to a perfusion system, and the slices were perfused at a rate of 5?ml/min with Ames media bubbled with 95% O2 and 5% CO2. The Ames media was supplemented with 100?��M picrotoxin and 50?��M 1, 2, 5, Icotinib 6-Tetrahydropyridin-4yl methylphosphinic acid (TPMPA) to block GABAA and GABAC receptors, 10?��M strychnine to block glycine receptors, and 4?��M L-AP4 to hyperpolarize On bipolar cells. The standard recording solution for RBCs was composed of (in mM): 108 gluconic acid, 2 EGTA, 10 CsCl, 10 TEA, 4 MgATP, 1LiGTP. For FALI experiments, 1?mM of the free radical scavenger glutathione was included in the recording solution. For experiments looking at asynchronous release following step depolarizations to ?10?mV, the EGTA concentration was reduced to 0.5?mM. The Amacrine cell internal solution was composed of (in mM): 100 CsCH3SO3, 10 EGTA, 20 TEA, 10 HEPES, 4 MgATP. 2?mM QX314 was added to block action potentials. The pH was adjusted to 7.4 with CsOH. The osmolarity of both extracellular and intracellular solutions was 289-297, with a pH of 7.35�C7.40. All chemicals were obtained from Sigma, except for L-AP4, TPMPA (Tocris) and ribbon-binding peptides (Genscript). Glutathione was stored as a powder at 4��C, and was added to the pipette solution immediately before use. L-AP4 was stored as a YES1 stock solution at 4��C and was added to the bath solution at the day of experimentation. All other drugs were aliquoted, stored at ?20��C, and dissolved in the pipette solution on the day of use. All animal procedures were approved by the Animal Care and Use Committee of the National Eye Institute. The procedures for making ground squirrel retinal slices have Cell Cycle inhibitor been described previously (Li and DeVries, 2006). During recording, the tissue was superfused with bicarbonate buffered Ames media (Sigma-Aldrich) containing picrotoxin (50?��M) and Strychnine (10?��M). The pipette solution contained (in mM): CsCH3SO, 120; MgSO4, 2; HEPES 10; TEA 20; EGTA 5; ATP 3; GTP 1; and Neurobiotin (Vector Laboratories), 0.1. pH was adjusted to 7.4 with CsOH. Slices were mounted on a Zeiss Examiner 1D microscope and superfused continuously at room temperature. Recordings were obtained with Axopatch 200B amplifiers (Molecular Devices), and currents low-pass filtered at 1 or 5 kHz using the 4 pole Bessel filter on the amplifier. The input/series resistances of the cone and bipolar cell recordings were approximately 1 G��/?