Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: Ethanol’s Effects on Transient Receptor Potential Channel Expression in Brain Microvascular Endothelial Cells

doi: 10.1007/s11481-018-9796-3

Figure Lengend Snippet: Endothelial cell barrier permeability following treatment with a) different concentrations of EtOH or b) the TRPM7 antagonists, NS8593, CyPPA, or SKA-31. *p < 0.05; **p < 0.01. a Mouse BMVECs at Passage 7 were seeded into Corning transwell inserts seated in 24-well plates to construct an in-vitro Blood-Brain Barrier (BBB) model. The cells were allowed to grow in the inserts for three days to form a barrier. The same number of wells without cells were used as background. The wells were then treated with 0 mM, 17.4 mM, 52.2 mM and 104.4 mM EtOH for 24 h, and to determine permeability, 50 ng/μl Evans Blue Dye (EBD) was added into the inserts at the same time and the model was incubated at 37 °C for 24 h. The permeability coefficient was then calculated. P-value was calculated using student’s t test. b Mouse BMVECs at Passage 5 were seeded into Corning transwell inserts seated in 24-well plates to construct an in-vitro Blood-Brain Barrier (BBB) model. The cells were allowed to grow in the inserts for three days to form a barrier (BBB). Wells were then treated with three different Trpm 7 Channel Blockers, NS 8593, CyPPA and SKA-31 at 50 μM for 24 h, and the permeability assay was then performed to determine the effect of TRPM7 blockers on the BBB. P-values were calculated using student’s t test

Article Snippet: For Trpm 7 inhibition, three different Trpm 7 Channel Blockers, NS 8593, CyPPA and SKA-31 (Alomone Labs, Jerusalem, Israel) were added to the inserts at 50 μM, together with a tracer Sodium Fluorescein (NaF) at 10 μg/mL, and incubated in a humidified cell culture incubator at 37 °C with 5% CO 2 for 24 h. One hundred microliter samples from both the upper insert and the lower chamber were transferred to a clear 96-well plate.

Techniques: Permeability, Construct, In Vitro, Incubation

Journal: Vascular pharmacology

Article Title: GPR55 agonist lysophosphatidylinositol and lysophosphatidylcholine inhibit endothelial cell hyperpolarization via GPR-independent suppression of Na + -Ca 2+ exchanger and endoplasmic reticulum Ca 2+ refilling

doi: 10.1016/j.vph.2017.01.002

Figure Lengend Snippet: (A) Effect of 3 μM LPI on endothelial hyperpolarization to 2 μM Ach (n = 4). (B) Effect of 10 μM LPI on endothelial hyperpolarization to two consecutive administrations of 2 μM Ach (n = 3). The hyperpolarization to SKA-31 (10 μM) remained unaffected by LPI pre-exposure (n = 4). (C) Representative membrane potential recording from in situ mice aortic endothelium showing a failure of LPI (10 μM) to inhibit the hyperpolarization evoked by 10 μM SKA-31 (n = 3).

Article Snippet: LPC16:0 (1-palmitoyl-2-hydroxy- sn - glycero -3-phosphocholine) was purchased from Avanti Polar Lipids, LPI from Sigma Aldrich, paxilline and SKA-31 were purchased from Alomone Labs, KB-R7943 was purchased from TCI Chemicals.

Techniques: In Situ