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Glutamate-gated currents were evoked by pressure ejection of 1 mM glutamate onto NMJs using small glass pipettes and a picospritzer II, as previously described [68].
Exocytosis was initiated by pressure ejection of permeabilisation/stimulation buffer (139 mM potassium glutamate, 5 mM EGTA, 30 mM PIPES, 2 mM ATP, 2 mM MgCl2, 0.02 mM digitonin, 10 mM free calcium, pH 6.5) from a glass pipette situated on the opposite side of the cell.
A PCA training set was collected by pressure ejection of adenosine, hydrogen peroxide, and ATP onto brain slices by a Parker Hannifin picospritzer (Picospritzer III, Cleveland, OH).
To test this prediction, we measured the amplitude of glutamate-gated currents triggered by pressure ejection of 1 mM glutamate onto postsynaptic muscles.
In a previous study, sub-saturating D2 receptor-dependent currents repeatedly produced by pressure ejection of dopamine are augmented by bath application of CPA for 10 20 min (Perra et al., 2011).
Currents triggered by pressure ejection of glutamate were significantly reduced in sec8Δ1 mutants compared to controls (Fig. 4B C; Control = 1858 ± 222.4 pA, n = 13; sec8Δ1 = 1181 ± 105.5 pA, n = 11, p = 0.016).
Similar(53)
Local application of DA from the micropipette was performed by pressure ejection using a pneumatic pump (BH2, Medical System).
Fluorescent Aβ1 42 was applied locally by pressure ejection from a pipette.
Second, we can assay postsynaptic glutamate receptors directly, by immunohistochemistry and pressure ejection of glutamate onto voltage-clamped postsynaptic muscle cells [ 25].
Hence pressure ejection of Aβ1 42 approximates release of Aβ1 42 from a point source.
Pressure ejection of glutamate provides a means of measuring postsynaptic receptor function independent of presynaptic glutamate release.
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