Journal: Scientific Reports

Article Title: Ndufs4 ablation decreases synaptophysin expression in hippocampus

doi: 10.1038/s41598-021-90127-4

Figure Lengend Snippet: Ndufs4 deficiency shows altered synaptic protein expression without neuronal loss. ( A ) Representative image of co-immunofluorescence staining of NeuN and Caspase 3 in hippocampus and olfactory bulb (OB) of WT mouse and Ndufs4-KO. Insets enlarge the boxed area. Low magnification green indicates NeuN and red indicates caspase 3, scale bar: 400 µm. In high magnification red indicates caspase 3 with no green stain, scale: 20 µm. ( B ) Representative images of Western blot for synaptophysin, PSD95, Vglut1, EAAT1, EAAT2, EAAT3 in WT and Ndufs4-KO mouse hippocampus. ( C – H ) Analysis of band densities by ImageJ (n = 3).

Article Snippet: Rabbit polyclonal anti PSD95 , WB: 1:2000 IF: 1:200 , Cell Signaling, Cat. No. 3450) , Goat anti-rabbit IgG-HRP conjugated (1:2000) Goat anti-rabbit IgG-Alexa 568 (1:500).

Techniques: Expressing, Immunofluorescence, Staining, Western Blot

Journal: Scientific Reports

Article Title: Ndufs4 ablation decreases synaptophysin expression in hippocampus

doi: 10.1038/s41598-021-90127-4

Figure Lengend Snippet: List of primary and secondary antibodies.

Article Snippet: Rabbit polyclonal anti PSD95 , WB: 1:2000 IF: 1:200 , Cell Signaling, Cat. No. 3450) , Goat anti-rabbit IgG-HRP conjugated (1:2000) Goat anti-rabbit IgG-Alexa 568 (1:500).

Techniques: Diagnostic Assay

Journal: Aging (Albany NY)

Article Title: Activation of α7 nAChR by PNU-282987 improves synaptic and cognitive functions through restoring the expression of synaptic-associated proteins and the CaM-CaMKII-CREB signaling pathway

doi: 10.18632/aging.102640

Figure Lengend Snippet: Activation of α7 nAChR promotes the expression of synaptic-associated proteins in Aβ oligomer-treated neurons. The x-axis labels are the neurons isolated from the WT rat (control), the WT neuron cells treated with PNU (PNU), the WT neuron cells treated with Aβ (Aβ) and the WT neuron cells treated with PNU and Aβ (PNU+Aβ). The y-axis indicates the relative level of mRNA or protein (% of control). Detection of SYN ( A ) mRNA and ( B ) protein; PSD95 ( C ) mRNA and ( D ) protein; SNAP25 ( E ) mRNA and ( F ) protein; DYN1 ( G ) mRNA and ( H ) protein; AP180 ( I ) mRNA and ( J ) protein. The relative level in each group was measured by RT-qPCR and western blot analysis, and β-actin was used as an internal control. The results demonstrated that the protein expression levels of SYN, PSD95, SNAP25, DYN1 and AP180 were significantly decreased in Aβ oligomer-treated neurons, and this decrease was partially reversed by PNU treatment. Data are presented as the mean ± standard deviation. * P<0.05, ** P<0.01 vs. control group; # P<0.05, ## P<0.01 vs. Aβ.

Article Snippet: Rabbit anti-PSD95 polyclonal antibody (cat. no. GTX133091), rabbit anti-SNAP25 polyclonal antibody (cat. no. GTX113839), rabbit anti-DYN1 polyclonal antibody (cat. no. GTX110379), rabbit anti-CREB monoclonal antibody (cat. no. GTX61140) and rabbit anti-β-actin polyclonal antibody (cat. no. GTX109639) were purchased from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Activation Assay, Expressing, Isolation, Control, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Aging (Albany NY)

Article Title: Activation of α7 nAChR by PNU-282987 improves synaptic and cognitive functions through restoring the expression of synaptic-associated proteins and the CaM-CaMKII-CREB signaling pathway

doi: 10.18632/aging.102640

Figure Lengend Snippet: Activation of α7 nAChR increases the expression of synaptic-associated proteins in the hippocampus of APP/PS1_DT mice . The x-axes are the WT mice (control), the WT mice treated with PNU (WP), the APP/PS1_DT mice (APP/PS1) and the APP/PS1_DT mice treated with PNU (AP). The y-axes are the relative level of mRNA or protein (% of control group). Detection of SYN ( A ) mRNA and ( B ) protein; PSD95 ( C ) mRNA and ( D ) protein; SNAP25 ( E ) mRNA and ( F ) protein; DYN1 ( G ) mRNA and ( H ) protein; AP180 ( I ) mRNA and ( J ) protein by RT-qPCR and western blot analysis. Protein expression levels were detected by western blot analysis (β-actin was used as an internal control). RT-qPCR and western blot analysis demonstrated that the expression levels of SYN, PSD95, SNAP25, DYN1 and AP180 in the hippocampus of APP/PS1_DT mice were significantly decreased compared with the control group, and this decreasing trend was partially reversed by PNU treatment. Data are presented as the mean ± standard deviation. * P<0.05, ** P<0.01 vs. control group; # P<0.05, ## P<0.01 vs. APP/PS1 group.

Article Snippet: Rabbit anti-PSD95 polyclonal antibody (cat. no. GTX133091), rabbit anti-SNAP25 polyclonal antibody (cat. no. GTX113839), rabbit anti-DYN1 polyclonal antibody (cat. no. GTX110379), rabbit anti-CREB monoclonal antibody (cat. no. GTX61140) and rabbit anti-β-actin polyclonal antibody (cat. no. GTX109639) were purchased from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Activation Assay, Expressing, Control, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Aging (Albany NY)

Article Title: Activation of α7 nAChR by PNU-282987 improves synaptic and cognitive functions through restoring the expression of synaptic-associated proteins and the CaM-CaMKII-CREB signaling pathway

doi: 10.18632/aging.102640

Figure Lengend Snippet: Activation of α7 nAChRs increases the expression of PSD95 protein in the hippocampus of APP/PS1_DT mice. The expression of PSD95 protein in the hippocampus of the APP/PS1_DT mice at ( A ) 6-months and ( F ) 10-months old. The IOD of PSD95 in hippocampus ( B ) CA1 area, ( C ) CA2 area, ( D ) CA3 area and ( E ) CA4 area of 6-month-old APP/PS1_DT mice. The IOD of PSD95 in hippocampus ( G ) CA1 area, ( H ) CA2 area, ( I ) CA3 area and ( J ) CA4 area of 10-month-old APP/PS1_DT mice. Control, WT C57 mice injected with saline; WP, WT mice injected with PNU; APP/PS1, APP/PS1_DT mice injected with saline; and AP, APP/PS1_DT mice injected with PNU. Compared with the control group, the expression of PSD95 protein in the hippocampus of the APP/PS1_DT mice group was significantly decreased and was partially reversed by PNU. This result suggested that activation of α7 nAChR partially reversed the expression level of PSD95 protein in the hippocampus of APP/PS1_DT mice. Data are presented as the mean ± standard deviation. * P<0.05, ** P<0.01 vs. control group; # P<0.05, ## P<0.01 vs. APP/PS1 group.

Article Snippet: Rabbit anti-PSD95 polyclonal antibody (cat. no. GTX133091), rabbit anti-SNAP25 polyclonal antibody (cat. no. GTX113839), rabbit anti-DYN1 polyclonal antibody (cat. no. GTX110379), rabbit anti-CREB monoclonal antibody (cat. no. GTX61140) and rabbit anti-β-actin polyclonal antibody (cat. no. GTX109639) were purchased from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Activation Assay, Expressing, Control, Injection, Saline, Standard Deviation

Journal: Aging (Albany NY)

Article Title: Activation of α7 nAChR by PNU-282987 improves synaptic and cognitive functions through restoring the expression of synaptic-associated proteins and the CaM-CaMKII-CREB signaling pathway

doi: 10.18632/aging.102640

Figure Lengend Snippet: Activation of α7 nAChR by PNU-282987 improves synaptic and cognitive functions through restoring the expression of synaptic-associated proteins and the CaM-CaMKII-CREB signaling pathway. Synapses are the basic structure for the connection between neurons, and the function of a synapse is largely associated with synaptic-associated proteins. The present study found that activation of α7 nAChRs by PNU-282987 could largely restore the expression level of synaptic-associated proteins, including SYN, PSD95, SNAP25, DYN1 and AP180, which are downregulated by deposition of Aβ. α7 nAChRs belong to the ligand-gated ion channel coupled receptors, and form an ion channel in the center, which mainly regulates the flow of Ca 2+ plasma inside and outside the cell. As a second cytoplasmic messenger, Ca 2+ can flow into the cells through α7 nAChRs and bind to CaM to form an active CaM complex. The CaM complex can bind to CaMKII, which changes the CaMKII spatial conformation and phosphorylates CaMKII protein. Phosphorylated CaMKII enters the nucleus to catalyze CREB phosphorylation. CREB is located downstream of the α7 nAChR signal transduction pathway and acts as a transcription factor that regulates the growth and development of neurons and the formation of long-term memory.

Article Snippet: Rabbit anti-PSD95 polyclonal antibody (cat. no. GTX133091), rabbit anti-SNAP25 polyclonal antibody (cat. no. GTX113839), rabbit anti-DYN1 polyclonal antibody (cat. no. GTX110379), rabbit anti-CREB monoclonal antibody (cat. no. GTX61140) and rabbit anti-β-actin polyclonal antibody (cat. no. GTX109639) were purchased from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Activation Assay, Expressing, Clinical Proteomics, Phospho-proteomics, Transduction