Journal: Nature Communications

Article Title: Loss of embryonic neural crest derived cardiomyocytes causes adult onset hypertrophic cardiomyopathy in zebrafish

doi: 10.1038/s41467-018-07054-8

Figure Lengend Snippet: NC-Cm cells regulate Notch signaling during trabeculation. a – f Ventricle sections of a 3 dpf embryo from a sox10:tagRFP transgenic line crossed with the Notch reporter Tp1:GFP line, immunostained for the cardiomyocyte marker MF20. NC-derived lineages (tagRFP) did not show high levels of Notch response (GFP). Arrows indicate trabeculating NC-Cms that were next to a Notch-activated cardiomyocyte (arrowheads). Scale bar = 10 µm. g qPCR gene expression of isolated 4 dpf hearts from NC-Cm-ablated and control siblings. Values are delta delta Ct computations. Bars represent mean of four biological replicates. Points are individual experiments. Delta Ct values used to compute standard T test significance ( P -values shown) between NC-Cm ablated and control delta Ct values. h – k Jag2b fluorescent in situ hybridization in a magnified ventricle section of 3 dpf NC-Cm lineage-labeled embryos. Scale bar = 5 µm. Image is representative of N = 17 embryos imaged and analyzed in probe positive in situs compared with probe negative embryos imaged and analyzed. l Quantification of jag2b fluorescent in situ signal in individual NC-Cm and Cm ventricle cells. Relative intensity was computed by the average intensity of all jag2b expression in the heart cells and then comparing with individual NC-Cm tRFP + and Cm GFP + cell intensities. Circles are individual cells from N = 3 embryos from three independent in situ experiments. Bar = mean. Standard T test was used for P -value. m Schematic model in which NC-Cms provide spatial patterning of trabeculation using Notch signaling components. During the transition to initiate trabeculae, NC-Cms provide a significant source of jag2b expression that triggers its neighbor Cm to repress protrusion and trabeculation initiation. This results in evenly, spatially distinct trabeculae branches at 5dpf. However, without NC-Cms and their major source of jag2b expression (although not exclusive as indicated by the remaining Cm expressing jag2b at 3dpf), more Cms are poised to protrude and create trabeculae, yielding conjoined branches and poor trabeculae spacing by 5 dpf

Article Snippet: Antibody staining for MF20 (DSHB) was carried out as described .

Techniques: Transgenic Assay, Marker, Derivative Assay, Gene Expression, Isolation, Control, In Situ Hybridization, Labeling, In Situ, Expressing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Human Organotypic Airway and Lung Organoid Cells of Bronchiolar and Alveolar Differentiation Are Permissive to Infection by Influenza and SARS-CoV-2 Respiratory Virus

doi: 10.3389/fcimb.2022.841447

Figure Lengend Snippet: The product information and dilution of primary used to detect the various target epitopes in this study.

Article Snippet: Clara cell 10 protein (CC10) , Mouse anti-CC10 (E-11) #sc-365992 , 1:200 , Santa Cruz.

Techniques: Luciferase

Journal:

Article Title: Anandamide is a partial agonist at native vanilloid receptors in acutely isolated mouse trigeminal sensory neurons

doi: 10.1038/sj.bjp.0704904

Figure Lengend Snippet: Anandamide activates outward currents in capsaicin-sensitive mouse trigeminal sensory neurons. Selected trigeminal neurons were voltage clamped at +40 mV, as described in Methods. Superfusion of AEA (100 μM) produced a slowly developing outward current that reversed slowly on wash. Subsequent superfusion of capsaicin produced a large, rapidly developing outward current in the same neuron. This experiment is typical of 56 small trigeminal neurons challenged with both AEA and capsaicin, five additional cells did not respond to either agonist.

Article Snippet: Capsaicin, capsazepine and anandamide (in water soluble emulsion) were from Tocris Cookson (Bristol, U.K.).

Techniques: Produced

Journal:

Article Title: Anandamide is a partial agonist at native vanilloid receptors in acutely isolated mouse trigeminal sensory neurons

doi: 10.1038/sj.bjp.0704904

Figure Lengend Snippet: The activity of anandamide, but not capsaicin, is antagonized by bovine serum albumin. Trigeminal sensory neurons were voltage clamped at +40 mV and superfused with anandamide and capsaicin in the presence or absence of 0.05% BSA. (a) Concentration-response relationships for AEA in the absence and presence of BSA (0.5 mg ml−1). BSA strongly inhibited the outward currents produced by submaximally effective concentrations of AEA. Each point represents the mean±s.e.mean of six cells, the currents produced by AEA were normalized to that produced by 10 μM capsaicin. (b) Concentration-response relationships for capsaicin in the absence and presence of BSA (0.5 mg ml−1). Capsaicin currents were not affected by the inclusion of BSA. Each point represents the mean±s.e.mean of six cells.

Article Snippet: Capsaicin, capsazepine and anandamide (in water soluble emulsion) were from Tocris Cookson (Bristol, U.K.).

Techniques: Activity Assay, Concentration Assay, Produced

Journal:

Article Title: Anandamide is a partial agonist at native vanilloid receptors in acutely isolated mouse trigeminal sensory neurons

doi: 10.1038/sj.bjp.0704904

Figure Lengend Snippet: Anandamide acts via vanilloid and not cannabinoid receptors to produce outward currents in mouse trigeminal neurons. Trigeminal sensory neurons were voltage clamped at +40 mV and superfused with AEA and capsaicin alone or in the presence of the VR1 antagonist capsazepine or the CB1 receptor antagonist SR141716. (a) Representative trace showing that capsazepine (30 μM) blocks AEA-induced outward currents in a AEA and capsaicin-sensitive neuron. Typical of six similar experiments. (b) Summary data of the effect of a 2-min preincubation of SR141716 on currents induced by AEA (10 μM). The currents were normalized to those produced by 10 μM capsaicin, each bar represents the mean±s.e.mean of six cells. (c) Summary data of the effect of a 2-min preincubation of SR141716 on currents induced by capsaicin. The same cells were challenged with capsaicin, then superfused with SR141716 and rechallenged with SR141716+capsaicin. The bars represent the mean±s.e.mean of six cells.

Article Snippet: Capsaicin, capsazepine and anandamide (in water soluble emulsion) were from Tocris Cookson (Bristol, U.K.).

Techniques: Produced

Journal:

Article Title: Anandamide is a partial agonist at native vanilloid receptors in acutely isolated mouse trigeminal sensory neurons

doi: 10.1038/sj.bjp.0704904

Figure Lengend Snippet: Anandamide activation of VR1 is concentration-dependent, but less efficacious than capsaicin. Selected trigeminal neurons were voltage clamped at +40 mV, as described in Methods. Concentration-response curves for capsaicin and AEA to activate VR1 in trigeminal neurons. In each cell, capsaicin or AEA responses were normalized to that produced by 10–30 μM capsaicin in that cell. The EC50 for capsaicin was 300 nM, for AEA 3 μM. AEA produced maximal currents 38±2% of maximal capsaicin currents. Each point represents the mean±s.e.mean of at least six cells, error bars are contained within the symbol for several points.

Article Snippet: Capsaicin, capsazepine and anandamide (in water soluble emulsion) were from Tocris Cookson (Bristol, U.K.).

Techniques: Activation Assay, Concentration Assay, Produced

Journal:

Article Title: Anandamide is a partial agonist at native vanilloid receptors in acutely isolated mouse trigeminal sensory neurons

doi: 10.1038/sj.bjp.0704904

Figure Lengend Snippet: Anandamide analogs methanandamide and AM404 activates VR1 in sensory neurons. Trigeminal sensory neurons were voltage clamped at +40 mV and superfused with methanandamide or AM404 and the currents produced compared with capsaicin (10 μM). (a) Concentration-response curves for methanandamide and AM404 to activate outward currents in trigeminal neurons, each response was normalized to that produced by 10–30 μM capsaicin in the same cell. Methanandamide produced maximum currents of 37±6% of maximal capsaicin currents, at the highest concentration of AM404 tested the currents were 55±9% of the capsaicin maximum. Each point represents the mean±s.e.mean of at least six cells. (b) Representative trace showing that capsazepine (30 μM) blocks most of the methanandamide-induced outward currents in a capsaicin-sensitive neuron. (c) Summary data of the effect of a 2-min preincubation of capsazepine (30 μM) on currents produced by methanandamide (100 μM) and AM404 (100 μM), determined as illustrated in (b). Capsazepine significantly (P<0.01) blocked the currents produced by both agonists. Outward currents were normalized to those produced by 10 μM capsaicin, each bar represents the mean±s.e.mean of six cells.

Article Snippet: Capsaicin, capsazepine and anandamide (in water soluble emulsion) were from Tocris Cookson (Bristol, U.K.).

Techniques: Produced, Concentration Assay

Journal:

Article Title: Anandamide is a partial agonist at native vanilloid receptors in acutely isolated mouse trigeminal sensory neurons

doi: 10.1038/sj.bjp.0704904

Figure Lengend Snippet: Anandamide, methanandamide and AM404 inhibit capsaicin-stimulated VR1 currents. Trigeminal sensory neurons were voltage clamped at +40 mV and superfused with capsaicin (3 μM) alone until the outward current reached a steady state and then AEA (30 μM), methanandamide (100 μM) or AM404 (100 μM) were co-superfused with capsaicin. (a) An example trace illustrating the effects of co-superfusion of AEA (30 μM) with capsaicin (3 μM). Approximately 4 min of wash have been removed for clarity. This is typical of six cells. (b) Bar chart summarizing the effect of co-superfusion AEA, methanandamide and AM404 on the currents induced by capsaicin. Each drug significantly (P<0.01) reduced the capsaicin (3 μM) current. Each bar represents the mean±s.e.mean of 5–6 cells.

Article Snippet: Capsaicin, capsazepine and anandamide (in water soluble emulsion) were from Tocris Cookson (Bristol, U.K.).

Techniques: