Journal: eLife

Article Title: Distinct transcriptomic profile of satellite cells contributes to preservation of neuromuscular junctions in extraocular muscles of ALS mice

doi: 10.7554/eLife.92644

Figure Lengend Snippet: ( A ) Representative compacted z-stack scan images of whole-mount EDL, soleus diaphragm extraocular muscles stained with antibodies against neurofilament medium chain (NF, labeling axons), synaptophysin (SYP, labeling axon terminals) and Alexa Fluor 488 conjugated α-Bungarotoxin (BTX, labeling AChRs on muscle membrane). scale bars, 50 μm. ( B ) Mean ratios of well innervated NMJs (SYP signals are present in >60% of BTX-positive area), partially innervated NMJs (SYP signals are present in 30–60% of BTX positive area) and poorly innervated NMJs (SYP signals are present in <30% of BTX-positive area) and in different types of muscles dissected from WT controls and end-stage G93A mice with or without NaBu feeding (see , and for NMJs measured per muscle type per gender. Briefly, EDL, soleus and diaphragm muscles were from four male and six female mice per group; WT EOM group was from four male and four female mice; G93A EOM group was from three male and four female mice; G93A EOM with NaBu feeding group was from six female mice). Each dot in the box-and-dot plots represents quantification result from a single mouse. * p<0.05; ** p<0.01; **** p<0.0001; ns, not significant (t-test). ANOVA p values are also shown. Figure 1—source data 1. Quantification results of the three types of NMJs in muscles of different origins and treatment conditions. Figure 1—source data 2. qRT-PCR results for Scn5a relative expression in whole muscles of different origins and treatment conditions. The calculation of ddCt used the averaged dCt value of Scn5a of EDL muscles derived from WT mice as the normalization control.

Article Snippet: Antibody , Anti-rat Synaptophysin (Rabbit polyclonal IgG) , Thermo Fisher Scientific , PA11043 , IF (1:400).

Techniques: Muscles, Staining, Labeling, Membrane, Quantitative RT-PCR, Expressing, Derivative Assay, Control

Journal: eLife

Article Title: Distinct transcriptomic profile of satellite cells contributes to preservation of neuromuscular junctions in extraocular muscles of ALS mice

doi: 10.7554/eLife.92644

Figure Lengend Snippet: ( A ) Representative images of well-innervated, partially innervated and poorly innervated NMJs in WT diaphragm muscles. BTX: Alexa Fluor 488 conjugated α-Bungarotoxin; SYP: synaptophysin; NF: neurofilament medium chain. ( B ) qRT-PCR based relative quantification (RQ, fold of change against WT muscles) of Scn5a expression using RNAs extracted from whole muscles of EDL, soleus, diaphragm and EOM. Each dot in the box-and-dot plots represent result from one mouse. Please also see . Briefly, muscles in WT and G93A groups were from three male and three female mice per group; G93A EDL with NaBu feeding group were from three male and three female mice. G93A soleus with NaBu feeding group were from two male and three female mice; G93A diaphragm with NaBu feeding group were from two male and four female mice; G93A EOM with NaBu feeding group were from four male and two female mice. * p<0.05; ** p<0.01; ns, not significant (t-test). ANOVA p values are also shown.

Article Snippet: Antibody , Anti-rat Synaptophysin (Rabbit polyclonal IgG) , Thermo Fisher Scientific , PA11043 , IF (1:400).

Techniques: Muscles, Quantitative RT-PCR, Quantitative Proteomics, Expressing

Journal: eLife

Article Title: Distinct transcriptomic profile of satellite cells contributes to preservation of neuromuscular junctions in extraocular muscles of ALS mice

doi: 10.7554/eLife.92644

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-rat Synaptophysin (Rabbit polyclonal IgG) , Thermo Fisher Scientific , PA11043 , IF (1:400).

Techniques: Cell Culture, Muscles, Isolation, Bacteria, Transfection, Construct, Plasmid Preparation, Virus, Recombinant, Concentration Assay, Staining, Sequencing

Journal: bioRxiv

Article Title: Postweaning development influences endogenous VPAC1 modulation of LTP induced by theta-burst stimulation: a link to maturation of the hippocampal GABAergic system?

doi: 10.1101/2024.02.18.580862

Figure Lengend Snippet: A., E ., and I. Western-blot immunodetection of PSD-95, VGlut1, Gephyrin, VGAT and synaptophysin obtained in one individual experiment in which total hippocampal membranes were subjected to WB analysis. Averaged total PSD-95 ( B. ), VGlut1 ( C. ), Gephyrin ( F. ), VGAT ( G. ) and synaptophysin ( J. ) immunoreactivities. VGlut1/PSD-95 ( D. ) ratio and VGAT/Gephyrin ratio ( H. ) for each animal were also plotted as an estimate of synaptic growth. VGlut1/synaptophysin ( K. ) ratio and VGAT/Synaptophysin ratio ( L. ) are also depicted and were used as an estimate of GABAergic vs glutamatergic synaptic enhancement. Values are mean ± S.E.M of five independent experiments performed in duplicate and were normalized to the immunoreactivity of α-tubulin, used as loading control. 100% - averaged target immunoreactivity obtained for 3-week-old animals. * Represents p < 0.05 ** represents p < 0.01 ( ANOVA , Tukey’s multiple comparison test) as compared to protein levels in 3-week-old animals or as otherwise indicated.

Article Snippet: These were then blocked for 1 h with either a 3% BSA solution or 5% milk solution in Tris-buffered saline (20mM Tris, 150mM NaCl) containing 0.1% Tween-20 (TBST), and incubated overnight at 4°C with and either rabbit polyclonal anti-VPAC1 (1:600, Alomone Labs #AVR-001, RRID: AB_2341081), rabbit polyclonal anti-VPAC2 (1:500, Alomone Labs #AVR-002, RRID: RRID: AB_2341082), rabbit polyclonal anti VIP (1:300, Proteintech, # 16233-1-AP, RRID: AB_2878233), mouse monoclonal anti-gephyrin (1:3000, #147011, Synaptic Systems, AB_2810214), rabbit polyclonal anti-PSD-95 (1:750, #CST-2507, Cell Signalling Tech., AB_561221), rabbit polyclonal anti-synaptophysin (1:7500, Synaptic Systems #101002, RRID:AB_887905), rabbit polyclonal anti-VGAT (1:2500, Synaptic Systems #131002, RRID: AB_887871), rabbit polyclonal anti-VGlut1 (1:3000, Synaptic Systems #135302, RRID: AB_887877), and either mouse monoclonal anti-β-actin (1:5000, Proteintech #60008-1-Ig, RRID: AB_2289225) or rabbit polyclonal anti-alpha-tubulin (1:5000, Proteintech #11224-1-AP; RRID: AB_2210206) primary antibodies.

Techniques: Western Blot, Immunodetection, Comparison

Journal: bioRxiv

Article Title: Postweaning development influences endogenous VPAC1 modulation of LTP induced by theta-burst stimulation: a link to maturation of the hippocampal GABAergic system?

doi: 10.1101/2024.02.18.580862

Figure Lengend Snippet: A. and D. Western-blot immunodetection of VIP VPAC 1 and VPAC 2 receptors obtained in one individual experiment in which hippocampal synaptosomes were subjected to WB analysis. Averaged total VPAC 1 ( B. ) and VPAC 2 ( C. ) immunoreactivity. Values are mean ± S.E.M of five independent experiments performed in duplicate and were normalized to the immunoreactivity of α-tubulin, used as loading control. 100% - averaged VPAC 1 or VPAC 2 immunoreactivity obtained for 3-week-old animals. VPAC 1 /synaptophysin ( K. ) ratio and VPAC 2 /Synaptophysin ratio ( L. ) are also depicted. * Represents p < 0.05 and *** represents p < 0.001 ( ANOVA , Tukey’s multiple comparison test) as compared to protein levels in 3-week-old animals or as otherwise indicated.

Article Snippet: These were then blocked for 1 h with either a 3% BSA solution or 5% milk solution in Tris-buffered saline (20mM Tris, 150mM NaCl) containing 0.1% Tween-20 (TBST), and incubated overnight at 4°C with and either rabbit polyclonal anti-VPAC1 (1:600, Alomone Labs #AVR-001, RRID: AB_2341081), rabbit polyclonal anti-VPAC2 (1:500, Alomone Labs #AVR-002, RRID: RRID: AB_2341082), rabbit polyclonal anti VIP (1:300, Proteintech, # 16233-1-AP, RRID: AB_2878233), mouse monoclonal anti-gephyrin (1:3000, #147011, Synaptic Systems, AB_2810214), rabbit polyclonal anti-PSD-95 (1:750, #CST-2507, Cell Signalling Tech., AB_561221), rabbit polyclonal anti-synaptophysin (1:7500, Synaptic Systems #101002, RRID:AB_887905), rabbit polyclonal anti-VGAT (1:2500, Synaptic Systems #131002, RRID: AB_887871), rabbit polyclonal anti-VGlut1 (1:3000, Synaptic Systems #135302, RRID: AB_887877), and either mouse monoclonal anti-β-actin (1:5000, Proteintech #60008-1-Ig, RRID: AB_2289225) or rabbit polyclonal anti-alpha-tubulin (1:5000, Proteintech #11224-1-AP; RRID: AB_2210206) primary antibodies.

Techniques: Western Blot, Immunodetection, Comparison