rabbit polyclonal vesicular glutamate transporter 1 (vglut1  (Bio-Techne corporation)

Journal: Brain Sciences

Article Title: Hindbrain Stimulation Modulates Object Recognition Discrimination Efficiency and Hippocampal Synaptic Connections

doi: 10.3390/brainsci13101425

Figure Lengend Snippet: ( A ) percentage of total puncta which had colocalized Synaptic vesicle glycoprotein 2A (SV2A) and Postsynaptic density protein 95 (PSD95) in the Dorsal CA1 region of the hippocampus. ( B ) representation of colocalized SV2A/PSD95 puncta in the CA1 shown with white arrows. ( C ) vesicular GABA Amino Acid Transporter (vGAT) normalized to total number of cells in the frontal cortex. ( D ) vesicular glutamate transporter 1 (vGLUT) normalized to the total number of cells in the frontal cortex. ( E ) example of vGAT staining in frontal cortex. ( F ) example of vGLUT staining in frontal cortex. * p < 0.05, bars represent 10 µm.

Article Snippet: Free-floating sections were blocked and permeabilized with 10% goat donkey serum in 0.1% Triton in 1 M PBS and then incubated for 48 h with the rabbit polyclonal Postsynaptic density protein 95 (PSD95, 1:250, Thermo Scientific, Waltham, MA, USA) and mouse monoclonal Synaptic vesicle glycoprotein 2A (SV2A, 1:250, Santa Cruz Biotechnology, Dallas, TX, USA), or for 24 h with rabbit polyclonal Vesicular glutamate transporter 1 (vGLUT1, 1:500, Novus Biotechne, Littleton, CO, USA) and mouse monoclonal Vesicular gamma-aminobutyric acid (GABA) Amino Acid Transporter (vGAT, 1:250, Invitrogen, Carlsbad, CA, USA).

Techniques: Staining

Journal: Alzheimer's Research & Therapy

Article Title: Aβ42 oligomer-specific antibody ALZ-201 reduces the neurotoxicity of Alzheimer’s disease brain extracts

doi: 10.1186/s13195-022-01141-1

Figure Lengend Snippet: Effect of immunodepletion on the neurotoxicity of AD brain extracts. Primary mouse neurons were treated for 24 h with artificial cerebrospinal fluid (aCSF), extracts of non-AD controls (grey) or extracts of AD patients (black) that were either untreated (-) or immunodepleted with ALZ-201, 4G8 or IgG3. High-content automated microscopy was employed for the quantification of the number of A neuronal nuclei: B morphology (as determined by segments, branches and extremities), and C number of vGlut1-positive presynapses. Morphological and synaptic parameters were normalised against the number of neurons. Values are displayed as the % change to cultures treated with aCSF. Bar graphs show the mean ± SD and data points represent the individual values for control (grey) and (immunodepleted) AD brain extracts (black). Shapiro-Wilk normality testing was followed by parametric (ordinary) or non-parametric (Kruskall-Wallis) one-way ANOVA, and groups were compared to pre-immunodepleted conditions using Dunn’s or Dunnet’s post hoc test. Significance values: ( B : segments) AD vs Ctrl <0.0001, vs 4G8 <0.0001, vs ALZ-201 =0.006; ( B : branches) AD vs Ctrl = 0.0008, vs 4G8 <0.0001, vs ALZ-201 <0.0001; ( B : extremities) AD vs Ctrl = 0.0053, vs 4G8 = 0.0004, vs ALZ-201 = 0.0145; ( C : presynapses) AD vs Ctrl <0.0001, vs 4G8 <0.0001, vs ALZ-201 = 0.0011. ns = not significant

Article Snippet: The primary antibodies used were vesicular glutamate transporter 1 (vGlut1) rabbit polyclonal (1:1000) (Synaptic Systems), neurofilament H mouse monoclonal (1:1000) (SMI-32P, Eurogentec) and microtubule-associated protein 2 (Map2) chicken polyclonal (1:250) (Abcam).

Techniques: Immunodepletion, Microscopy, Control

Journal: The Journal of Neuroscience

Article Title: Induction of BDNF Expression in Layer II/III and Layer V Neurons of the Motor Cortex Is Essential for Motor Learning

doi: 10.1523/JNEUROSCI.0288-20.2020

Figure Lengend Snippet: BDNF is enriched in glutamatergic corticostriatal presynaptic terminals. A , Confocal (top) and SIM (bottom) microscopic images showing BDNF-IR in the same section in glutamatergic (left) versus dopaminergic terminals (right) in the dorsal striatum. B , BDNF-IR is present in VGluT1-positive terminals (magenta arrows). Single BDNF-IR signals overlap with TH (white arrows). VGluT1- and TH-positive terminals reside in direct regional proximity but do not overlap. C , Quantification of BDNF signals in VGluT1-positive terminals and TH-positive terminals. True colocalization between BDNF/VGluT1 was confirmed by Costes p value (Costes p > 0.95) but not between BDNF/TH (Costes p ≪ 0.95). D , Representative Western blots showing recombinant BDNF (lanes 1, 2) versus endogenous BDNF derived from anterior cortex or striatum of P21 NFL-Cre BDNF fl/ko mice (lane 3), P21 sedentary mice (lanes 4, 5), and P21 runners after 72 h voluntary running-wheel exercise (lanes 6, 7); 30 µg of protein lysate was loaded for each sample. BDNF levels were normalized to cytochrome C. Band intensities were determined from extracts of 9 independent mice and presented in % of P21 sedentary mice. Statistical analysis reveals significant increase in BDNF protein levels in both brain areas after running-wheel exercise. Statistical analysis: unpaired t test (anterior CTX: t = 5,312, p < 0.0001; striatum: t = 2,784, p = 0.0133). E , SIM images showing BDNF-IR in VGluT1-positive terminals in the dorsal striatum in sedentary mice (top row) and after 72 h of voluntary running-wheel exercise (bottom row). Data are presented as box and whiskers (Tukey). +, Mean. Vertical line indicates median. Black dots indicate outliers. n , number indicated below. Raw data are provided in Extended Data and . Scale bars: A , 2.5 µm; B , 1.5 µm; E , Overview, 2 µm; Detail, 1 µm. * p < 0.05; **** p < 0.0001.

Article Snippet: Presynaptic corticostriatal terminals were labeled with rabbit polyclonal antibodies against vesicular glutamate transporter 1 (VGluT1) (Synaptic Systems, #135302, RRID: AB_887877 ).

Techniques: Western Blot, Recombinant, Derivative Assay

Journal: The Journal of Neuroscience

Article Title: Induction of BDNF Expression in Layer II/III and Layer V Neurons of the Motor Cortex Is Essential for Motor Learning

doi: 10.1523/JNEUROSCI.0288-20.2020

Figure Lengend Snippet: Image preparation

Article Snippet: Presynaptic corticostriatal terminals were labeled with rabbit polyclonal antibodies against vesicular glutamate transporter 1 (VGluT1) (Synaptic Systems, #135302, RRID: AB_887877 ).

Techniques: