p q ca 2 channel  (Mochida Pharmaceutical)

Journal: Molecular Neurobiology

Article Title: Involvement of the Voltage-Gated Calcium Channels L- P/Q- and N-Types in Synapse Elimination During Neuromuscular Junction Development

doi: 10.1007/s12035-022-02818-2

Figure Lengend Snippet: a Representative confocal image of a nerve terminal arborization. Singly, dually, and innervated by three or more axons NMJs from YFP muscles and also images of the morphologic maturation (S1, the most inmature, and S4, almost fully differentiated, stages) of the postsynaptic clusters from P9 mice. The bar indicates 10 μm. b Confocal immunofluorescence location of α 1D L-, N-, and P/Q-type voltage-dependent calcium channels (VDCCs) at the NMJ. Triple labeling of VDCCs (green fluorescence) with syntaxin (blue fluorescence) and nAChR-α-bungarotoxin (red fluorescence) in merge images. Figure shows the presence of α 1D L-, N-, and P/Q-type-VDCC (in green) in the nerve terminal of P9 Levator auris longus (LAL) muscle endplates. The bar indicates 10 μm

Article Snippet: Muscles were incubated overnight at 4 °C with anti-Ca V 1.3 (CACNA1D) antibody voltage-dependent L-type calcium channel subunit α 1D (1/100; ACC-005, Alomone Labs, Jerusalem, Israel); anti-Ca V 2.1 (CACNA1A) antibody voltage-dependent P/Q-type calcium channel subunit α 1A (1/100; ACC-001, Alomone Labs, Jerusalem, Israel); anti-Ca V 2.2 (CACNA1B) antibody voltage-dependent N-type calcium channel subunit α 1B (1/100; ACC1-002, Alomone Labs, Jerusalem, Israel), and anti-mouse syntaxin (1/1000, S066, Sigma, St Louis, MO, USA).

Techniques: Immunofluorescence, Labeling, Fluorescence

Journal: Biomedicines

Article Title: Silymarin Inhibits Glutamate Release and Prevents against Kainic Acid-Induced Excitotoxic Injury in Rats

doi: 10.3390/biomedicines8110486

Figure Lengend Snippet: Silymarin inhibits 4-aminopyridine-evoked glutamate release and intracellular Ca 2+ elevation in rat cerebral cortex synaptosomes. ( A ) Glutamate release was evoked by the addition of 1 mM 4-aminopyridine in the absence (control) and presence of 30 µM silymarin added 10 min before depolarization. ( B ) Concentration-dependent inhibition of 4-aminopyridine-evoked glutamate release by silymarin. ( C ) Effect of silymarin on 4-aminopyridine-evoked glutamate release in extracellular Ca 2+ -free solution or in the presence of the vesicular transporter inhibitor bafilomycin A1. ( D ) Silymarin decreases 4-aminopyridine-evoked increase in [Ca 2+ ] i . Data are mean ± standard error of the mean (SEM) ( n = 5–6 per group). *** p < 0.001 (in comparison with the control).

Article Snippet: Silymarin (5–30 µM, purity >98%, ChemFaces), ethylene glycol bis(β-aminoethyl ether)-N,N,N1,N1-tetraacetic acid (EGTA, 300 µM, Sigma-Aldrich, MO, USA), the vesicular transporter inhibitor bafilomycin A1 (0.1 µM, Tocris, Bristol, UK), endoplasmic reticulum Ca 2+ release inhibitor dantrolene (10 µM, Tocris, Bristol, UK), mitochondrial Na + /Ca 2+ exchange inhibitor 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3 H )-one (GP37157, 10 µM, Tocris, Bristol, UK), N- and P/Q-type Ca 2+ channel inhibitor ω-conotoxin MVIIC (ω-CgTX MVIIC, 4 µM, Alomone lab, Jerusalem, Israel), inositol 1,4,5-trisphosphate (IP 3 )receptor antagonist xestospongin C (1 µM, Tocris, Bristol, UK), protein kinase A (PKA) inhibitor N-[2-( p -bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H89, 100 µM, Tocris, Bristol, UK), protein kinase C (PKC) inhibitor bisindolylmaleimide I (GF109203X, 10 µM, Tocris, Bristol, UK), mitogen-activated protein kinase (MAPK) inhibitor 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one (PD98059, 50 µM, Tocris, Bristol, UK), extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor FR180204 (10 µM, Tocris, Bristol, UK), Ca 2+ indicator fura-2-acetoxymethyl ester (Fura-2-AM, 5 µM, Life Technologies, Bengaluru, India), membrane potential-sensitive dye 3′,3′-dipropylthiadicarbocyanine iodide (DiSC 3 (5), 5 µM, Invitrogen, CA, USA), K + channel blocker 4-aminopyridine (1 mM, Sigma-Aldrich, MO, USA), and glutamate analog KA (15 mg/kg, Sigma-Aldrich, MO, USA) were used.

Techniques: Concentration Assay, Inhibition

Journal: Biomedicines

Article Title: Silymarin Inhibits Glutamate Release and Prevents against Kainic Acid-Induced Excitotoxic Injury in Rats

doi: 10.3390/biomedicines8110486

Figure Lengend Snippet: Silymarin-mediated inhibition of 4-aminopyridine-evoked glutamate release in the presence of N- and P/Q-type Ca 2+ channel blocker ω-CgTX MVIIC, ryanodine receptor inhibitor dantrolene, mitochondrial Na + /Ca 2+ exchanger inhibitor CGP37157, or IP3 receptor antagonist xestospongin C. Silymarin was added 10 min before the addition of 4-aminopyridine, and other drugs were added 10 min before this. Data are mean ± SEM ( n = 5 per group). ** p < 0.01, *** p < 0.001 (in comparison with the control), # p < 0.001 (compared with the dantrolene-, CGP37157-, or xestospongin C-treated group).

Article Snippet: Silymarin (5–30 µM, purity >98%, ChemFaces), ethylene glycol bis(β-aminoethyl ether)-N,N,N1,N1-tetraacetic acid (EGTA, 300 µM, Sigma-Aldrich, MO, USA), the vesicular transporter inhibitor bafilomycin A1 (0.1 µM, Tocris, Bristol, UK), endoplasmic reticulum Ca 2+ release inhibitor dantrolene (10 µM, Tocris, Bristol, UK), mitochondrial Na + /Ca 2+ exchange inhibitor 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3 H )-one (GP37157, 10 µM, Tocris, Bristol, UK), N- and P/Q-type Ca 2+ channel inhibitor ω-conotoxin MVIIC (ω-CgTX MVIIC, 4 µM, Alomone lab, Jerusalem, Israel), inositol 1,4,5-trisphosphate (IP 3 )receptor antagonist xestospongin C (1 µM, Tocris, Bristol, UK), protein kinase A (PKA) inhibitor N-[2-( p -bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H89, 100 µM, Tocris, Bristol, UK), protein kinase C (PKC) inhibitor bisindolylmaleimide I (GF109203X, 10 µM, Tocris, Bristol, UK), mitogen-activated protein kinase (MAPK) inhibitor 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one (PD98059, 50 µM, Tocris, Bristol, UK), extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor FR180204 (10 µM, Tocris, Bristol, UK), Ca 2+ indicator fura-2-acetoxymethyl ester (Fura-2-AM, 5 µM, Life Technologies, Bengaluru, India), membrane potential-sensitive dye 3′,3′-dipropylthiadicarbocyanine iodide (DiSC 3 (5), 5 µM, Invitrogen, CA, USA), K + channel blocker 4-aminopyridine (1 mM, Sigma-Aldrich, MO, USA), and glutamate analog KA (15 mg/kg, Sigma-Aldrich, MO, USA) were used.

Techniques: Inhibition

Journal: Biomedicines

Article Title: Silymarin Inhibits Glutamate Release and Prevents against Kainic Acid-Induced Excitotoxic Injury in Rats

doi: 10.3390/biomedicines8110486

Figure Lengend Snippet: The proposed working model of silymarin depressing glutamate release from synaptosomes and having a preventive effect against KA-induced neuronal death. In rat synaptosomes, silymarin inhibits glutamate release by suppressing N- and P/Q-type Ca 2+ channels and ERK1/2 activity. In a rat model of KA-induced excitotoxicity, silymarin pretreatment substantially attenuates KA-induced neuronal death by suppressing inflammatory processes and HSP70 expression and upregulating Akt activation. The red T-shaped lines represent inhibition.

Article Snippet: Silymarin (5–30 µM, purity >98%, ChemFaces), ethylene glycol bis(β-aminoethyl ether)-N,N,N1,N1-tetraacetic acid (EGTA, 300 µM, Sigma-Aldrich, MO, USA), the vesicular transporter inhibitor bafilomycin A1 (0.1 µM, Tocris, Bristol, UK), endoplasmic reticulum Ca 2+ release inhibitor dantrolene (10 µM, Tocris, Bristol, UK), mitochondrial Na + /Ca 2+ exchange inhibitor 7-chloro-5-(2-chlorophenyl)-1,5-dihydro-4,1-benzothiazepin-2(3 H )-one (GP37157, 10 µM, Tocris, Bristol, UK), N- and P/Q-type Ca 2+ channel inhibitor ω-conotoxin MVIIC (ω-CgTX MVIIC, 4 µM, Alomone lab, Jerusalem, Israel), inositol 1,4,5-trisphosphate (IP 3 )receptor antagonist xestospongin C (1 µM, Tocris, Bristol, UK), protein kinase A (PKA) inhibitor N-[2-( p -bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H89, 100 µM, Tocris, Bristol, UK), protein kinase C (PKC) inhibitor bisindolylmaleimide I (GF109203X, 10 µM, Tocris, Bristol, UK), mitogen-activated protein kinase (MAPK) inhibitor 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one (PD98059, 50 µM, Tocris, Bristol, UK), extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor FR180204 (10 µM, Tocris, Bristol, UK), Ca 2+ indicator fura-2-acetoxymethyl ester (Fura-2-AM, 5 µM, Life Technologies, Bengaluru, India), membrane potential-sensitive dye 3′,3′-dipropylthiadicarbocyanine iodide (DiSC 3 (5), 5 µM, Invitrogen, CA, USA), K + channel blocker 4-aminopyridine (1 mM, Sigma-Aldrich, MO, USA), and glutamate analog KA (15 mg/kg, Sigma-Aldrich, MO, USA) were used.

Techniques: Activity Assay, Expressing, Activation Assay, Inhibition

Journal: eLife

Article Title: Rapid purification and metabolomic profiling of synaptic vesicles from mammalian brain

doi: 10.7554/eLife.59699

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit polyclonal Ca 2+ channel P/Q-type alpha1A unit , Synaptic Systems , 152103, RRID: AB_887699 , (1:300) dilution.

Techniques: Recombinant, Plasmid Preparation, CRISPR, Construct, Glo Assay, Magnetic Beads

Journal: Diabetes

Article Title: Adrenaline stimulates glucagon secretion by Tpc2-dependent Ca 2+ mobilization from acidic stores in pancreatic α-cells

doi: 10.2337/db17-1102

Figure Lengend Snippet: (a) Glucagon secreted from isolated NMRI mouse islets in response to 3 mM glucose with(out) 5μM adrenaline ± isradipine or ω-agatoxin (as indicated). p<0.05 vs the effect of adrenaline + 3mM glucose (*) or vs the basal (3mM glucose + respective antagonist) group. (b) [Ca2+]i upon adrenergic (isoprenaline (n=27), or noradrenaline (n=19)) stimulation alone (n=29) or with (as indicated) isradipine (n=17, preincubated, n=29, acute), ω-agatoxin (n=13), propranolol (n=11), prazosin (n=49), diazoxide (n=20), EGTA (n=84). p<0.05 vs. the effect of adrenaline alone (*) or vs the basal (3 mM glucose+respective (ant)agonist) of the same recording. (c) Effect of adrenaline on α-cell action potential firing (representative of 11 experiments). Examples of action potentials recorded in the absence and presence of adrenaline (taken from the recording above as indicated) are shown.

Article Snippet: Chemicals The following substances were used (source given in parentheses): the L-type Ca 2+ channel blocker isradipine and the P/Q Ca 2+ channel blocker ω-agatoxin (Alomone Labs; Jerusalem, Israel); the α 1 -antagonist prazosin (Abcam, Cambridge, UK); the membrane-permeable PKA inhibitor myr-PKI, the IP 3 receptor inhibitor Xestospongin C, the NAADP antagonist Ned-19, the V-ATPase inhibitor bafilomycin, the sER ATPase inhibitor thapsigargin and noradrenaline (Tocris Bioscience, Bristol, UK); the EPAC2 inhibitor ESI-05 (BioLog, Bremen, Germany); the insulin receptor antagonist (S961, Novo-Nordisk, Denmark).

Techniques: Isolation