Journal: PLoS ONE

Article Title: Increased Learning and Brain Long-Term Potentiation in Aged Mice Lacking DNA Polymerase μ

doi: 10.1371/journal.pone.0053243

Figure Lengend Snippet: ( A ) Hierarchical clustering of the differential mRNA expression between wild-type and Polµ −/− old (18 months) mice. ( B ) Schematic representation of the expression network detected using the software Ingenuity. ( C ) qRT-PCR analysis of genes belonging to the previously described network ( B ) and others found in DNA expression arrays carried out with brain of acutely treated animals . Black bars denote values for wild-type animals and white bars correspond to Polµ −/− animals. ( D ) Circulating levels of IGF1 and GH (ng/mL) were monitored by ELISA in old (21–24 months) animals. ( E, F ) qRT-PCR expression ( E ) and immunoconfocal analysis ( F ) of Igf1 and Igf1R in the hippocampal CA1area from wild-type and Polμ −/− young adult (4 months) mice. Igf1 immunoreactivity-labeled non-pyramidal cells (short arrows) of stratum oriens (so), stratum pyramidale (sp), and stratum radiatum (sr; not shown); no differences in labeling frequency or labeling intensity were found between Polμ −/− and wild-type mice. Igf1R-immunostaining delineated cell processes in the stratum radiatum (sr) of Polμ −/− but not of wild-type mice. As compared with the case of aged mice, unspecific punctate labeling (long arrows) was infrequent in these tissue sections. Slight differences were found, however, in the occurrence of Igf1-immunoreactive cells in the CA1 area of aged mice, and in the expression of Igf1R between young wild-type and Polμ −/− mice. Scale bar, 25 µm.

Article Snippet: Primary antibodies used were mouse monoclonal antibody M23 to IGF1 (Abcam, Cambridge, UK; 1∶300), mouse monoclonal antibody AP-20 to microtubule-associated protein 2 (map2, 2a+2b; Sigma; 1∶250), rabbit polyclonal antibody sc-713 to IGF1Rβ (Santa Cruz Biotechnology, CA, USA; 1∶150) and rabbit polyclonal antibody to the Zn 2+ vesicular transporter ZnT3 (Synaptic Systems, Göttingen, Germany; 1∶500).

Techniques: Expressing, Software, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Labeling, Immunostaining

Journal: Annals of Translational Medicine

Article Title: Gastrodin exerts protective effects in reactive TNC1 astrocytes via regulation of the Notch signaling pathway

doi: 10.21037/atm-21-5787

Figure Lengend Snippet: GAS reduced TNF-α expression and enhanced BDNF, IGF-1, and Sirt3 expression in reactive astrocytes after OGD-stimulation. (A-D) Immunofluorescence images show TNF-α, BDNF, IGF-1, and Sirt3 (red) in GFAP-positive (green) astrocytes. DAPI (blue) shows the nuclei. (E) The quantitative analysis reveals BDNF, IGF-1, and Sirt3 expressions were drastically increased following treatment with GAS. TNF-α expression increased sharply in the OGD group, but were drastically decreased by GAS treatment. Significant differences are expressed as: #, P<0.05 vs. CON group; *, P<0.05 vs. OGD group. Scale bars: 20 µm. The values represented as mean ± SEM in triplicate. BDNF, brain-derived neurotrophic factor; CON, control; GAS, gastrodin; GFAP, glial fibrillary acidic protein; IGF-1, insulin-like growth factor-1; OGD, oxygen-glucose deprivation; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α.

Article Snippet: The primary antibodies were as follows: glial fibrillary acidic protein (GFAP) (monoclonal, 1:1,000, MAB360, Millipore, USA), Notch-1 (rabbit polyclonal 1:1,000, ab52627, Abcam, UK), intracellular Notch receptor domain (NICD, rabbit polyclonal 1:1,000, ab8925, Abcam, UK), recombining binding protein suppressor of hairless (RBP-JK) (rabbit polyclonal 1:100, ab180588, Abcam, UK), transcription factor hairy and enhancer of split-1 (Hes-1) (mouse polyclone 1:500, SC-166410, Santa Cruz, UK); BDNF (rabbit monoclonal 1:1,000, ab108139, Abcam, UK), insulin-like growth factor-1 (IGF-1) (rabbit polyclonal 1:1,000, orb10886, Biorbyt, UK); Sirtuin 3 (Sirt3, mouse polyclone 1:500, SC-365175, Santa Cruz, USA); tumor necrosis factor-α (TNF-α) (rabbit polyclonal antibody, 1:1,000, AB1837P, Millipore, USA); and mouse anti-β-actin monoclonal antibody (1:5,000, Proteintech, China).

Techniques: Expressing, Immunofluorescence, Derivative Assay, Control

Journal: Annals of Translational Medicine

Article Title: Gastrodin exerts protective effects in reactive TNC1 astrocytes via regulation of the Notch signaling pathway

doi: 10.21037/atm-21-5787

Figure Lengend Snippet: GAS increased the protein expressions of BDNF, IGF-1, and Sirt3, but reduced the protein expressions of IL-1β and TNF-α in TNC1 astrocytes. (A) Western blotting results of BDNF, IGF-1, Sirt3, IL-1β and TNF-α. (B-F) The quantitative analysis of BDNF, IGF-1, Sirt3, IL-1β and TNF-α. The results showed that the expressions of BDNF, IGF-1, and Sirt3 in OGD groups had negligible increases, and increased obviously in groups with GAS. Meanwhile, IL-1β and TNF-α expressions were decreased in TNC1 astrocytes following treatment with GAS. Significant differences are expressed as: #, P<0.05 vs. CON group; *, P<0.05 vs. OGD group. The values represent the mean ± SEM in triplicate. BDNF, brain-derived neurotrophic factor; CON, control; GAS, gastrodin; IGF-1, insulin-like growth factor-1; IL-1β, interleukin 1β; OGD, oxygen-glucose deprivation; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α.

Article Snippet: The primary antibodies were as follows: glial fibrillary acidic protein (GFAP) (monoclonal, 1:1,000, MAB360, Millipore, USA), Notch-1 (rabbit polyclonal 1:1,000, ab52627, Abcam, UK), intracellular Notch receptor domain (NICD, rabbit polyclonal 1:1,000, ab8925, Abcam, UK), recombining binding protein suppressor of hairless (RBP-JK) (rabbit polyclonal 1:100, ab180588, Abcam, UK), transcription factor hairy and enhancer of split-1 (Hes-1) (mouse polyclone 1:500, SC-166410, Santa Cruz, UK); BDNF (rabbit monoclonal 1:1,000, ab108139, Abcam, UK), insulin-like growth factor-1 (IGF-1) (rabbit polyclonal 1:1,000, orb10886, Biorbyt, UK); Sirtuin 3 (Sirt3, mouse polyclone 1:500, SC-365175, Santa Cruz, USA); tumor necrosis factor-α (TNF-α) (rabbit polyclonal antibody, 1:1,000, AB1837P, Millipore, USA); and mouse anti-β-actin monoclonal antibody (1:5,000, Proteintech, China).

Techniques: Western Blot, Derivative Assay, Control

Journal: Annals of Translational Medicine

Article Title: Gastrodin exerts protective effects in reactive TNC1 astrocytes via regulation of the Notch signaling pathway

doi: 10.21037/atm-21-5787

Figure Lengend Snippet: DAPT further enhanced the effect of GAS on the expressions of Notch pathway proteins, neuroinflammatory factors, and neurotrophic factors. (A) Protein expression of NICD, RBP-JK and Hes-1. (B) The quantitative analysis showed the expression levels of NICD, RBP-JK, Hes-1 in OGD-activated TNC1 astrocytes were reduced significantly with DAPT treatment, in the GAS + OGD + DAPT (GAS + O + D) group, the expressions of NICD and HES-1 were further reduced. (C) Protein expression levels of BDNF, IGF-1, Sirt3, IL-1β, and TNF-α. (D) The quantitative analysis showed BDNF and IGF-1 level in OGD-activated TNC1 astrocytes were obviously increased with DAPT treatment. BDNF and IGF-1 expressions in OGD-activated TNC1 astrocytes administered a combined DAPT and GAS treatment reached a peak increase compared to the OGD + D or GAS + OGD (GAS + O) groups. Sirt3 expression in OGD-activated TNC1 astrocytes administered a combined DAPT and GAS treatment was reduced significantly compared with the OGD + D group. In OGD-activated TNC1 astrocytes, the IL-1β and TNF-α levels were reduced significantly with DAPT treatment. Combination of DAPT and GAS cumulatively decreased the expressions of IL-1β and TNF-α. Significant differences are expressed as: κ, P<0.05 between the control and DAPT group; *, P<0.05 compared with the OGD group; #, P<0.05 between the OGD + D and GAS + O + D group; ns, P>0.05 compared with the OGD + D group; Δ, P<0.05 compared with the GAS+ O group. The values represent the mean ± SEM in triplicate. DAPT, N-[N-(3,5-Difluorophenacetyl)-1-alany1]-S-phenyglycine t-butylester; GAS, gastrodin; Hes-1, transcription factor hairy and enhancer of split-1; IGF-1, insulin-like growth factor-1; IL-1β, interleukin 1β; NICD, intracellular Notch receptor domain; ns, not significant; OGD, oxygen-glucose deprivation; RBP-JK, recombining binding protein suppressor of hairless; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α; ns, no significance.

Article Snippet: The primary antibodies were as follows: glial fibrillary acidic protein (GFAP) (monoclonal, 1:1,000, MAB360, Millipore, USA), Notch-1 (rabbit polyclonal 1:1,000, ab52627, Abcam, UK), intracellular Notch receptor domain (NICD, rabbit polyclonal 1:1,000, ab8925, Abcam, UK), recombining binding protein suppressor of hairless (RBP-JK) (rabbit polyclonal 1:100, ab180588, Abcam, UK), transcription factor hairy and enhancer of split-1 (Hes-1) (mouse polyclone 1:500, SC-166410, Santa Cruz, UK); BDNF (rabbit monoclonal 1:1,000, ab108139, Abcam, UK), insulin-like growth factor-1 (IGF-1) (rabbit polyclonal 1:1,000, orb10886, Biorbyt, UK); Sirtuin 3 (Sirt3, mouse polyclone 1:500, SC-365175, Santa Cruz, USA); tumor necrosis factor-α (TNF-α) (rabbit polyclonal antibody, 1:1,000, AB1837P, Millipore, USA); and mouse anti-β-actin monoclonal antibody (1:5,000, Proteintech, China).

Techniques: Expressing, Control, Binding Assay