Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cardiomyocytes from AKAP7 knockout mice respond normally to adrenergic stimulation.

doi: 10.1073/pnas.1215219109

Figure Lengend Snippet: Fig. 2. AKAP7α is expressed in brain but not in cardiomyocytes or skeletal muscle. (A) Lysates of various tissues from WT or AKAP7 KO mice were probed by immunoblot using rabbit polyclonal anti-AKAP7 antibody 12591- 1. Long isoforms, ∼37–42 kDa, are faintly detected in all WT tissues. Some nonspecific bands (e.g., ∼55 kDa) are observed in tissue from WT and KO animals. Total protein was measured by BCA assay and 80 μg were loaded per lane; GAPDH and β-actin are shown as additional loading controls. (B) AKAP7 was immunoprecipitated from lysates using guinea pig anti-AKAP7 YO869 antibody to enrich protein before probing via immunoblot as above. The single band at 15 kDa corresponds to AKAP7α and is only weakly detected in nonbrain tissues. RAtr, right atrium; Vent, ventricle; CMyo, iso- lated cardiomyocytes; SkM, skeletal muscle (quadriceps); IMed, kidney inner medulla. (C and D) Quantitative RT-PCR using Taq-Man MGB probes detec- ted AKAP7 long isoforms in all tissues, but short isoforms only appreciably in the brain.

Article Snippet: For immunoprecipitation, 2 mg of lysate was diluted in 1 mL of buffer and rotated at 4 °C overnight with 4 μg rabbit anti-CaV1.2 antibody CNC1 (46), 4 μg rabbit anti-RIIα (sc-909; Santa Cruz), or 6 μg of guinea pig anti-AKAP7 antibody YO869.

Techniques: Western Blot, BIA-KA, Immunoprecipitation, Quantitative RT-PCR

Journal: STAR Protocols

Article Title: Volumetric super-resolution imaging by serial ultrasectioning and stochastic optical reconstruction microscopy in mouse neural tissue

doi: 10.1016/j.xpro.2021.100971

Figure Lengend Snippet:

Article Snippet: Guinea pig polyclonal anti-Calbindin (1:100) , Synaptic Systems , Cat#214 005.

Techniques: Recombinant, Saline, Electron Microscopy, Plasmid Preparation, Software, Imaging, Control, Mass Measurement, Capsules, Microscopy, Staining, Adhesive, Spectrophotometry

Journal: Neuron

Article Title: Targeted proteoform mapping uncovers specific Neurexin-3 variants required for dendritic inhibition

doi: 10.1016/j.neuron.2022.04.017

Figure Lengend Snippet:

Article Snippet: The following antibodies were used in this study: rabbit polyclonal anti-β-actin (Abcam; Cat# ab8227; RRID: AB_2305186; LOT# GR3314266-1), mouse monoclonal anti-calbindin (Swant; Cat# 300; RRID: AB_10000347; LOT# 17 (F)), goat polyclonal anti-calretinin (Swant; Cat# CG1; RRID: AB_10000342; LOT# 1§.1), mouse monoclonal anti-CamKII alpha (Thermo Fisher Scientific; 6G9; Cat# Ma1-048; RRID: AB_325403; LOT# TH269517), mouse monoclonal anti-cannabinoid receptor 1 (Immunogene; IMG-3C2; Cat# IMG-CB1R-mAb001; LOT# CJ03), guinea pig polyclonal anti-cholecystokinin (Synaptic Systems, Cat# 438004; RRID: AB_2814938; LOT# 1-1), mouse monoclonal anti-GAD67 (Millipore; 1G10.2; Cat# MAB5406; RRID: AB_2278725; LOT# 3015328), rabbit polyclonal anti-GAPDH (Enogene; Cat# E1C604; LOT# R14Q12), mouse monoclonal anti-gephyrin (Synaptic Systems; mAb7a; Cat# 147021; RRID: AB_2232546; LOT# 147021/15), mouse monoclonal anti-gephyrin (Synaptic Systems; mAb7a; Cat# 147011; RRID: AB_887717; LOT# 147011/54), rat monoclonal anti-HA (Roche; 3F10; Cat# 11867431001; RRID: AB_390919; LOT# 34502100), rabbit monoclonal anti-HA (Cell Signaling; 3724; Cat# 3724; RRID: AB_1549585; LOT# 9), guinea pig polyclonal anti-MAP2 (Synaptic Systems; Cat# 188004; RRID: AB_2138181; LOT# 2-26), mouse monoclonal anti-MAP2 (Synaptic Systems; 198A5; Cat# 188011; RRID: AB_2147096; LOT# 1-10), rabbit polyclonal anti-neurexin , chicken anti-neurexin , rabbit anti-neuroligin , rabbit monoclonal anti-nNOS (Cell Signaling; C7D7; Cat# 4231; RRID: AB_2152485; LOT# 2), goat polyclonal anti-parvalbumin (Swant, Cat# PVG214; RRID: AB_10000345), mouse monoclonal anti-PSD95 (Santa Cruz; 7E3; Cat# sc32290; RRID: AB_628114; LOT# J1509), goat polyclonal anti-somatostatin (Santa Cruz; Cat# sc7819; RRID: AB_2302603; LOT# L1611), mouse monoclonal anti-Synaptotagmin 2 (Zebrafish International Resource Center; Cat# znp-1; RRID: AB_10013783), mouse monoclonal anti-V5 (Biorad; SV5-PK1; Cat# MCA1360; RRID: AB_322378; LOT# 148239), guinea pig polyclonal anti-vGAT (Synaptic Systems; Cat# 131004; RRID: AB_887873; LOT# 2-42), guinea pig polyclonal anti-vGlut1 (Millipore; Cat# AB5905; RRID: AB_2301751; LOT# 3308226), rabbit polyclonal anti-VIP (Immunostar; Cat# 20077; RRID: AB_572270; LOT# 1513001), guinea pig anti-somatostatin (raised in the present study against amino acid residues 35-88 of mouse pro-somatostatin, GenBank: #BC010770.1), goat anti-cannabinoid receptor 1 (Nittobo Medical, MSFR100600; RRID: AB_2571592), guinea pig anti-GABAA receptor alpha 1 (Nittobo Medical, MSFR101540; RRID: AB_2571572), goat anti-vGAT (Nittobo Medical, MSFR106130; RRID: AB_2571623), guinea pig anti-PSD95 (Nittobo Medical, MSFR105180; RRID: AB_2571612).

Techniques: Recombinant, Reporter Assay, Multiplex Assay, Plasmid Preparation, Software, Microscopy

Journal: Research

Article Title: Lateral Hypothalamus Calcium/Calmodulin-Dependent Protein Kinase II α Neurons Encode Novelty-Seeking Signals to Promote Predatory Eating

doi: 10.34133/2022/9802382

Figure Lengend Snippet: Antibodies used in this research.

Article Snippet: Anti-Fos in guinea pig , Synaptic Systems , 226308 , Bolós M et al. J Neurosci . 39 (9):1605-1620 (2019) , 30651327 , AB_2619946 , 1 : 2000.

Techniques:

Journal: Frontiers in Synaptic Neuroscience

Article Title: Selective Enrichment of Munc13-2 in Presynaptic Active Zones of Hippocampal Pyramidal Cells That Innervate mGluR1α Expressing Interneurons

doi: 10.3389/fnsyn.2021.773209

Figure Lengend Snippet: Munc13-2 immunolabeling is enriched on mGluR1α immunopositive dendrites. (A) Double immunolabeling for Munc13-2 (left, cyan) and mGluR1α (right, red) in the dorsal hippocampal CA1 region of the mouse (cartoon indicates the location of the region) shows similar distribution in the stratum oriens. Maximum intensity projection of six confocal images separated by 1 μm. (B) A dendritic segment of an mGluR1α immunopositive IN (white boxes on A ) is shown at a higher magnification, which is decorated by Munc13-2 immunopositive puncta. Maximum intensity projection of three confocal images separated by 1 μm. (C) 500 nm thick epoxy resin embedded section with preembedding immunolabeling for Munc13-2 (cyan) and mGluR1α (red) shows that Munc13-2 immunopositive puncta preferentially located on the small diameter of mGluR1α+ (distal) dendrites (dd), and mainly avoid the soma (s) and a proximal dendrite (pd) in the hippocampus of the mouse. Boxed area is enlarged on panel (D) . (D) Multiplexed postembedding immunolabeling carried out on the section shown in panel (G) . Munc13-2 immunopositive puncta marked by circles (representing ROIs for quantification) along the mGluR1α immunolabeled dendrite are immunopositive for Munc13-1, PSD95, AMPA receptors, Bassoon, Cav2.1,and Rim1/2 (all pseudo colored to green). Note that the intensity of Munc13-2 immunolabeling varies substantially. Alignment of sections after each round was based on mGluR1α immunolabeling (red). Numbers represent the labeling rounds during the multiplexed labeling. (E) All of the Munc13-2 immunopositive puncta contain PSD95 immunosignal. Their amount shows positive correlations (Spearman correlation r = 0.48 and 0.55, n = 40 in mouse #1 and n = 80 in mouse #2). (F) Correlations between the density of the Munc13-2 and Munc13-1 in individual AZs (each data point represents an AZ, n = 114 in mouse #1 and n = 80 in mouse #2; Spearman correlation r = 0.16 and 0.34). (G) mGluR1α IN targeting synapses have significantly larger (*) Munc13-1, AMPA receptors, Bassoon, Cav2.1 and Rim1/2 densities than those found in randomly selected glutamatergic synapses in the str. oriens ( p = 1.5 × 10 –5 , 5.5 × 10 –24 , 1.5 × 10 –4 , 6.6 × 10 –16 , 1.3 × 10 –5 , respectively, MW- U -test). Box plots represent median and 25/75 percentiles, square represent the mean value, whiskers represent SD. All immunolabelings were normalized to PSD95 intensity on panels (F,G) . (H) The Munc13-2 content of randomly selected synapses is only 4 ± 7% and 4 ± 10% (in two mice; p = 0 for both MW- U -test) of that of synapses on mGluR1α+ dendrites. (I–L) Munc13-2 immunolabeling (cyan) does not colocalize either with vesicular inhibitory amino acid transporter (VIAAT, red) ( I,J , n = 152 Munc13-2 and 222 VIAAT positive profiles in 2 mice) or with vesicular glutamate transporter-2 (vGluT2, red) (K,L) , n = 43 Munc13-2 and 33 vGluT2 positive profiles in 1 mouse). Single confocal images (I,K) . str. oriens, stratum oriens.

Article Snippet: Immunoglobulins were removed with a 5-min incubation in TBS containing 1% SDS (pH = 7.7) at 80 ° C. After 5 min of washing in TBST, new rounds of immunolabeling were performed using a guinea pig anti-PSD95 Ab (1:500, Synaptic Systems, Göttingen, Germany; Cat# 124014, RRID: AB_2619800 ), a guinea pig anti-panAMPA receptor Ab (1:100 Frontiers Cat#Af580; RRID: AB_257161 ), a chicken anti-Bassoon Ab (1:200 Synaptic Systems, Göttingen, Germany; Cat#141-016; RRID: AB_2661779 ), a guinea pig anti-Cav2.1 Ab (1:100 Synaptic Systems, Göttingen, Germany; Cat#152 205; RRID: AB_2619842 ), and finally with a guinea pig anti-Rim1/2 Ab (1:100 Synaptic Systems, Göttingen, Germany; Cat#140-205; RRID:AB_2631216 ) with the appropriate Alexa488-coupled secondary Abs (Alexa488-coupled donkey anti-guinea pig, 1:200; Jackson ImmunoResearch Laboratories, PA, United States; as above or Alexa488-coupled goat anti-chicken, 1:200; Jackson ImmunoResearch Laboratories, PA, United States, Cat# 103-545-155, RRID: AB_2337390 ).

Techniques: Immunolabeling, Labeling

Journal: eLife

Article Title: Intact synapse structure and function after combined knockout of PTPδ, PTPσ, and LAR

doi: 10.7554/eLife.66638

Figure Lengend Snippet:

Article Snippet: Antibody , Guinea pig anti-Synaptophysin , SySy , RRID: AB_1210382 , A106 , ICC (1:500).

Techniques: Mutagenesis, Recombinant, Software