rabbit anti post synaptic density 95  (Alomone Labs)

Journal: Heliyon

Article Title: Melatonin attenuates spatial learning and memory dysfunction in developing rats by suppressing isoflurane-induced endoplasmic reticulum stress via the SIRT1/Mfn2/PERK signaling pathway

doi: 10.1016/j.heliyon.2022.e10326

Figure Lengend Snippet: Melatonin mitigated isoflurane-induced hippocampal neuro apoptosis in neonatal rats. (A) Western blot bands of apoptosis signaling in the hippocampus. (B) The treatment of melatonin significantly reduced isoflurane induced increase of pro-apoptosis factor Bax expression in hippocampus of neonatal rats ( F (3, 20) = 71.360, P < 0.001). (C) The treatment of melatonin significantly reduced isoflurane induced decrease of anti-apoptosis factor Bcl-2 expression in hippocampus of neonatal rats ( F (3, 20) = 9.573, P < 0.001). (D) The treatment of melatonin significantly reduced isoflurane induced increase of pro-apoptosis factor cleaved-caspase 3 expression in hippocampus of neonatal rats ( F (3, 20) = 19.910, P < 0.001). (E) The treatment of melatonin significantly increase isoflurane induced decrease of synaptic protein PSD95 expression in hippocampus of neonatal rats ( F (3, 20) = 73.790, P < 0.001) (F) Immunofluorescence results of cleaved-caspase 3 and caspase 12 in hippocampus of all four groups rats. Expressions of cleaved-caspase 3 and caspase 12 in all four groups were presented through bar graphs: (G) for cleaved-caspase 3 ( F (3, 20) = 109.100, P < 0.001) and (H) for caspase 12 ( F (3, 20) = 105.200, P < 0.001). Data were presented as mean ± SEM (n = 6). ∗denotes p < 0.05, ∗∗denotes p < 0.01, ∗∗∗denotes p < 0.001 when comparing the two groups under each end of the capped line. Scale bar = 500 μm.

Article Snippet: The primary antibodies included the following: mouse anti-β-actin (1:2000; Qidongzi, Wuhan, China), mouse anti-SIRT1 (1:500; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-mitofusin 2 (Mfn2) (1:1000; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-protein kinase RNA-like ER kinase (PERK) (1:500; Affinity Biosciences, OH, USA), rabbit anti- C/EBP homologues protein (CHOP) (1:500; Affinity Biosciences, OH, USA), rabbit anti-78 kDa glucose regulated protein (GRP78) (1:500; Affinity Biosciences, OH, USA), rabbit anti-activating transcription factor 4 (ATF4) (1:500; Affinity Biosciences, OH, USA), rabbit anti-cysteine containing aspartate specific protease 12 (caspase 12) (1:500; Proteintech North America, IL, USA), rabbit anti-Bcl-2-associated X protein (Bax) (1:500; Proteintech North America, IL, USA), rabbit anti-post-synaptic density protein 95 (PSD95) (1:1000; Cell Signaling Technology, Danvers, MA, USA), rabbit anti-B-cell lymphoma-2 (Bcl-2) (1:500; Cell Signaling Technology, Danvers, MA, USA), and rabbit anti-cleaved-cysteine containing aspartate specific protease 3 (cleaved-caspase 3).

Techniques: Western Blot, Expressing, Immunofluorescence

Journal: Frontiers in Pediatrics

Article Title: Alterations in Excitatory and Inhibitory Synaptic Development Within the Mesolimbic Dopamine Pathway in a Mouse Model of Prenatal Drug Exposure

doi: 10.3389/fped.2021.794544

Figure Lengend Snippet: (A) Diagram of mouse brain regions associated with mesolimbic dopamine pathway (PFC, prefrontal cortex; ACC, anterior cingulate cortex; NAC, nucleus accumbens; VTA, ventral tegmental area; Hippo, hippocampus; Amyg, amygdala). (B–D) Representative fluorescent confocal microscopic images of excitatory synaptic staining (presynaptic VGlut1, green; postsynaptic PSD95, red) at 10X and 63X magnification of the regions analyzed in this study: (B) ACC, (C) NAC, and (D) PFC.

Article Snippet: Guinea pig anti-vesicular glutamate transporter 1 (VGlut1; Cat. #AB5905, EMD Millipore, Burlington, MA) at 1:2,000 dilution was used to identify excitatory glutamatergic presynaptic axonal regions while rabbit anti-post synaptic density protein 95 (PSD95; Cat. #51-6900, Invitrogen, Carlsbad, CA) at 1:300 dilution was used to identify postsynaptic dendritic regions.

Techniques: Staining

Journal: Frontiers in Pediatrics

Article Title: Alterations in Excitatory and Inhibitory Synaptic Development Within the Mesolimbic Dopamine Pathway in a Mouse Model of Prenatal Drug Exposure

doi: 10.3389/fped.2021.794544

Figure Lengend Snippet: Increased excitatory synapses with early life buprenorphine exposure. (A) Diagram illustrating co-localization of puncta representing presynaptic (VGlut1, green) and postsynaptic (PSD95, red) fluorescent antibody label pairs for quantifying excitatory glutamatergic synapses. (B–D) Representative IHC images (left) and quantification (right) of co-localized VGlut1 (green) and PSD95 (red) excitatory synaptic puncta (yellow arrowheads) from prenatal drug-exposed WT C57Bl/6J P21 mouse brain within (B) the anterior cingulate cortex (ACC), (C) nucleus accumbens (NAC), and (D) prefrontal cortex (PFC). n = 7 (vehicle control), 6 (GBP), 7 (Bup+GBP), 6 (Bup); ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Guinea pig anti-vesicular glutamate transporter 1 (VGlut1; Cat. #AB5905, EMD Millipore, Burlington, MA) at 1:2,000 dilution was used to identify excitatory glutamatergic presynaptic axonal regions while rabbit anti-post synaptic density protein 95 (PSD95; Cat. #51-6900, Invitrogen, Carlsbad, CA) at 1:300 dilution was used to identify postsynaptic dendritic regions.

Techniques:

Journal: Frontiers in Pediatrics

Article Title: Alterations in Excitatory and Inhibitory Synaptic Development Within the Mesolimbic Dopamine Pathway in a Mouse Model of Prenatal Drug Exposure

doi: 10.3389/fped.2021.794544

Figure Lengend Snippet: Dual exposure to buprenorphine and gabapentin decreased inhibitory synapses at P21. (A) Diagram illustrating co-localization of puncta representing presynaptic (VGAT, green) and postsynaptic (PSD95) fluorescent antibody label pairs for quantifying inhibitory GABAergic synapses. (B–D) Representative IHC images (left) and quantification (right) of co-localized VGAT (green) and gephyrin (red) inhibitory synaptic puncta (yellow arrowheads) from prenatal drug-exposed WT C57Bl/6J P21 mouse brain within (B) ACC, (C) NAC, and (D) PFC. n = 7 (vehicle control), 6 (GBP), 7 (Bup+GBP), 6 (Bup); ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Guinea pig anti-vesicular glutamate transporter 1 (VGlut1; Cat. #AB5905, EMD Millipore, Burlington, MA) at 1:2,000 dilution was used to identify excitatory glutamatergic presynaptic axonal regions while rabbit anti-post synaptic density protein 95 (PSD95; Cat. #51-6900, Invitrogen, Carlsbad, CA) at 1:300 dilution was used to identify postsynaptic dendritic regions.

Techniques:

Journal: Frontiers in Pediatrics

Article Title: Alterations in Excitatory and Inhibitory Synaptic Development Within the Mesolimbic Dopamine Pathway in a Mouse Model of Prenatal Drug Exposure

doi: 10.3389/fped.2021.794544

Figure Lengend Snippet: Altered excitatory synaptic connectivity following prenatal drug treatment in α2δ-1 haploinsufficient mice. Representative IHC images (left) of co-localized VGlut1 (green) and PSD95 (red) excitatory synaptic puncta (yellow arrowheads) from prenatal drug-exposed α2δ-1 +/- (Het) C57Bl/6J P21 mouse brain with quantification (right) compared to WT C57Bl/6J P21 mouse brains with the same prenatal treatment within (A) ACC, (B) NAC, and (C) PFC. Het: n = 11 (vehicle control), 7 (GBP), 7 (Bup+GBP), 7 (Bup); * p < 0.05; **** p < 0.0001.

Article Snippet: Guinea pig anti-vesicular glutamate transporter 1 (VGlut1; Cat. #AB5905, EMD Millipore, Burlington, MA) at 1:2,000 dilution was used to identify excitatory glutamatergic presynaptic axonal regions while rabbit anti-post synaptic density protein 95 (PSD95; Cat. #51-6900, Invitrogen, Carlsbad, CA) at 1:300 dilution was used to identify postsynaptic dendritic regions.

Techniques: