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Fig. 2 | Molecular Brain

Fig. 2

From: The novel protein kinase C epsilon isoform modulates acetylcholine release in the rat neuromuscular junction

Fig. 2

Effect of εV1-2 in high Ca2+ or PMA medium and during electrical stimulation on transmitter release in diaphragm muscle. a Histogram shows the effect of high Ca2+ (5 mM), high magnesium (5 mM) and the P/Q-type channel blocker ω-Agatoxin-IVA (100 nM) on the evoked transmitter release in basal conditions and after CaC incubation (2: CaC; 10 μM). The histogram in (b) compares the effect of CaC or εV1-2 in high Ca2+ medium and during continuous electrical stimulation (1Hz, 1 h) on the transmitter release. Diaphragm muscles were preincubated (1 h) with high Ca2+ (5 mM; 1: Ca2+) and then evaluated the effect of εV1-2 (10 μM, 1 h of incubation; 2: εV1-2). We also evaluated the εV1-2 effect during electrical stimulation at 1 Hz (1: 1 Hz, 2: εV1-2). To evaluate the effect of the unspecific blocker CaC when the peptide εV1-2 is present, we performed a pretreatment with εV1-2 and a second incubation with CaC (10 μM, an additional hour; 2: CaC). This was done in conditions of both high Ca2+ (10 μM, 1 h of incubation, 1: εV1-2, Ca2+) and continuous electrical stimulation (1: εV1-2, 1Hz). c Changes in ACh release after PMA (10 nM) and PMA in presence of continuous electrical stimulation (PMA, 1 Hz, 1 h). We also evaluated the changes in ACh release when a PMA or a CaC (10 μM) preincubated muscle was then incubated with the other drug (1: CaC, 2: PMA; 1: PMA, 2: CaC). To determine whether nPKCε affects the PMA-induced enhancing of neurotransmission, we preincubated the neuromuscular preparation (1 h) with the εV1-2 peptide (1: εV1-2, 1 μM, 10 μM, 100 μM) and then evaluated the effect of PMA (2: PMA). We also studied the link between electrical stimulation and PMA effects in presence of electrical stimulation at 1Hz (1: εV1-2, 10 μM, 1 Hz; 2: PMA). * p < 0.05 vs. the corresponding control

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