drugs sl327  (Tocris)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Name:
    SL 327
    Description:
    Selective inhibitor of MEK1 and MEK2 brain penetrant
    Catalog Number:
    1969
    Price:
    None
    Purity:
    ≥99% (HPLC)
    Category:
    MEK Inhibitors MEK Kinases Enzymes Pharmacology
    Formula:
    α-[Amino[(4-aminophenyl)thio]methylene]-2-(trifluoromethyl)benzeneacetonitrile
    Buy from Supplier


    Structured Review

    Tocris drugs sl327
    SL 327
    Selective inhibitor of MEK1 and MEK2 brain penetrant
    https://www.bioz.com/result/drugs sl327/product/Tocris
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    drugs sl327 - by Bioz Stars, 2020-02
    90/100 stars

    Related Products / Commonly Used Together

    am251 am

    Images

    1) Product Images from "Mechanism and treatment for the learning and memory deficits associated with mouse models of Noonan syndrome"

    Article Title: Mechanism and treatment for the learning and memory deficits associated with mouse models of Noonan syndrome

    Journal: Nature neuroscience

    doi: 10.1038/nn.3863

    PTPN11 D61G overexpression induces learning and memory and LTP deficits that can be reversed by MEK inhibition a. AAV– PTPN11 D61G infection results in overexpression of SHP2 D61G . Anti–SHP2 immunohistochemistry shows robust overexpression of SHP2 in the hippocampus of AAV– PTPN11 D61G –infused brains (left) compared with AAV– GFP infused brains (right). Full-length blots/gels are presented in Supplementary Figure 11 . b. PTPN11 D61G overexpression increases basal Erk activity (phospho–Erk level) and prevents further Erk activation in response to TBS. Left , Representative immunoblot showing p–Erk (upper) and total Erk (lower) in PTPN11 D61G –expressing slices and GFP –expressing slices. Slices were prepared 1 h after TBS. Right , Bar graph displays normalized p–Erk levels (mean ± s.e.m.). CTL, control without TBS. c. MEK inhibitor SL327 reverses spatial memory deficits in PTPN11 D61G –overexpressing mice in the Morris water maze. Quadrant occupancy analysis for the probe trial reveals that PTPN11 D61G /veh mice showed no preference for the target quadrant (target vs. other quadrants, Dunnett’s Multiple Comparison Test after one-way ANOVA, P > 0.05). PTPN11 D61G /veh mice also spent significantly less time in the target quadrant compared with GFP /veh mice. SL327 treatment significantly increased the time spent in the target quadrant in PTPN11 D61G -expressing mice compared with vehicle-treated PTPN11 D61G mice ( PTPN11 D61G /SL327, 37.25 ± 3.50 %, n=10, unpaired two-tailed t-test, t = 2.335, * P
    Figure Legend Snippet: PTPN11 D61G overexpression induces learning and memory and LTP deficits that can be reversed by MEK inhibition a. AAV– PTPN11 D61G infection results in overexpression of SHP2 D61G . Anti–SHP2 immunohistochemistry shows robust overexpression of SHP2 in the hippocampus of AAV– PTPN11 D61G –infused brains (left) compared with AAV– GFP infused brains (right). Full-length blots/gels are presented in Supplementary Figure 11 . b. PTPN11 D61G overexpression increases basal Erk activity (phospho–Erk level) and prevents further Erk activation in response to TBS. Left , Representative immunoblot showing p–Erk (upper) and total Erk (lower) in PTPN11 D61G –expressing slices and GFP –expressing slices. Slices were prepared 1 h after TBS. Right , Bar graph displays normalized p–Erk levels (mean ± s.e.m.). CTL, control without TBS. c. MEK inhibitor SL327 reverses spatial memory deficits in PTPN11 D61G –overexpressing mice in the Morris water maze. Quadrant occupancy analysis for the probe trial reveals that PTPN11 D61G /veh mice showed no preference for the target quadrant (target vs. other quadrants, Dunnett’s Multiple Comparison Test after one-way ANOVA, P > 0.05). PTPN11 D61G /veh mice also spent significantly less time in the target quadrant compared with GFP /veh mice. SL327 treatment significantly increased the time spent in the target quadrant in PTPN11 D61G -expressing mice compared with vehicle-treated PTPN11 D61G mice ( PTPN11 D61G /SL327, 37.25 ± 3.50 %, n=10, unpaired two-tailed t-test, t = 2.335, * P

    Techniques Used: Over Expression, Inhibition, Infection, Immunohistochemistry, Activity Assay, Activation Assay, Expressing, CTL Assay, Mouse Assay, Two Tailed Test

    PTPN11 D61G overexpression enhances excitatory synaptic function through increased Ras-Erk signaling a. AMPA receptor-mediated currents were measured at the peak of the currents at − 65 mV, and NMDA currents were measured 50 ms after onset at + 40 mV. The average of 15 traces is shown. Scale, 100 pA and 40 ms. b. Group data showing the increased AMPA:NMDA current ratio in AAV– PTPN11 D61G mice compared with AAV- GFP mice. SL327 treatment (1 μM, 1 h) significantly reversed the AMPA:NMDA current ratio in the PTPN11 D61G group without affecting GFP –expressing mice. Two-way ANOVA, interaction between viral treatment and drug, F 1, 31 = 10.53, ** P
    Figure Legend Snippet: PTPN11 D61G overexpression enhances excitatory synaptic function through increased Ras-Erk signaling a. AMPA receptor-mediated currents were measured at the peak of the currents at − 65 mV, and NMDA currents were measured 50 ms after onset at + 40 mV. The average of 15 traces is shown. Scale, 100 pA and 40 ms. b. Group data showing the increased AMPA:NMDA current ratio in AAV– PTPN11 D61G mice compared with AAV- GFP mice. SL327 treatment (1 μM, 1 h) significantly reversed the AMPA:NMDA current ratio in the PTPN11 D61G group without affecting GFP –expressing mice. Two-way ANOVA, interaction between viral treatment and drug, F 1, 31 = 10.53, ** P

    Techniques Used: Over Expression, Mass Spectrometry, Mouse Assay, Expressing

    2) Product Images from "The Neuroprotective Effect of Lithium in cannabinoid Dependence is Mediated through Modulation of Cyclic AMP, ERK1/2 and GSK-3β Phosphorylation in Cerebellar Granular Neurons of Rat"

    Article Title: The Neuroprotective Effect of Lithium in cannabinoid Dependence is Mediated through Modulation of Cyclic AMP, ERK1/2 and GSK-3β Phosphorylation in Cerebellar Granular Neurons of Rat

    Journal: Iranian Journal of Pharmaceutical Research : IJPR

    doi:

    The effect of SL327 (SL) as a specific inhibitor of ERK1/2 pathway and AM251 (AM, 1 µM) as a cannabinoid antagonist on the phosphorylated-ERK1/2 (p-ERK1/2), total ERK1/2, phosphorylated-GSK-3β (p-GSK-3β) and total GSK-3β expression in cerebellar granular cells (CGNs). CGNs were pre-treated with SL (10 µM), 30 min before added WIN 55,212-2 (WIN, 1 µM) and AM was co-treated with WIN in CGNs 12-well plate and the cell lysates were prepared after 15, 30, 60 and 180 minutes. The protein expression was analyzed by western blotting. The p-ERK1/2 and ERK1/2 expression (A and B), and the p-GSK-3β and GSK-3β expression (A and C), were evaluated. The protein expression was normalized to β-actin as internal control (CTRL). The band intensity was measured and presented as the percent of un-treated cells (CTRL). All data were presented as Mean ± SD. Statistically significant values are presented as follows: *** p
    Figure Legend Snippet: The effect of SL327 (SL) as a specific inhibitor of ERK1/2 pathway and AM251 (AM, 1 µM) as a cannabinoid antagonist on the phosphorylated-ERK1/2 (p-ERK1/2), total ERK1/2, phosphorylated-GSK-3β (p-GSK-3β) and total GSK-3β expression in cerebellar granular cells (CGNs). CGNs were pre-treated with SL (10 µM), 30 min before added WIN 55,212-2 (WIN, 1 µM) and AM was co-treated with WIN in CGNs 12-well plate and the cell lysates were prepared after 15, 30, 60 and 180 minutes. The protein expression was analyzed by western blotting. The p-ERK1/2 and ERK1/2 expression (A and B), and the p-GSK-3β and GSK-3β expression (A and C), were evaluated. The protein expression was normalized to β-actin as internal control (CTRL). The band intensity was measured and presented as the percent of un-treated cells (CTRL). All data were presented as Mean ± SD. Statistically significant values are presented as follows: *** p

    Techniques Used: Expressing, Western Blot

    The effect of SL327 (SL) pre-treated to WIN 55, 212-2 (WIN) and AM251 (AM) on the phosphorylated-ERK1/2 (p-ERK1/2), total ERK1/2, phosphorylated-GSK-3β (p-GSK-3β) and total GSK-3β expression in cerebellar granular cells (CGNs). CGNs were pre-treated with SL (10 µM), 30 min before added WIN (1 µM) and AM (1 µM), and the cell lysates were prepared after 15, 30, 60 and 180 minutes. The protein expression was analyzed by western blotting. The p-ERK1/2 and ERK1/2 expression (A and B), and the p-GSK-3β and GSK-3β expression (A and C), were evaluated. The protein expression was normalized to β-actin as internal control (CTRL). The band intensity was measured and presented as the percent of un-treated cells (CTRL). All data were presented as Mean ± SD. Statistically significant values are presented as follows: *** p
    Figure Legend Snippet: The effect of SL327 (SL) pre-treated to WIN 55, 212-2 (WIN) and AM251 (AM) on the phosphorylated-ERK1/2 (p-ERK1/2), total ERK1/2, phosphorylated-GSK-3β (p-GSK-3β) and total GSK-3β expression in cerebellar granular cells (CGNs). CGNs were pre-treated with SL (10 µM), 30 min before added WIN (1 µM) and AM (1 µM), and the cell lysates were prepared after 15, 30, 60 and 180 minutes. The protein expression was analyzed by western blotting. The p-ERK1/2 and ERK1/2 expression (A and B), and the p-GSK-3β and GSK-3β expression (A and C), were evaluated. The protein expression was normalized to β-actin as internal control (CTRL). The band intensity was measured and presented as the percent of un-treated cells (CTRL). All data were presented as Mean ± SD. Statistically significant values are presented as follows: *** p

    Techniques Used: Expressing, Western Blot

    Related Articles

    Fluorescence:

    Article Title: A metabolic perturbation by U0126 identifies a role for glutamine in resveratrol-induced cell death
    Article Snippet: SL327 was obtained from Tocris Bioscience. .. Vectashield mounting medium for fluorescence with DAPI was obtained from Vector Laboratories.

    Synthesized:

    Article Title: Concurrent activation of β2-adrenergic receptor and blockage of GPR55 disrupts pro-oncogenic signaling in glioma cells
    Article Snippet: .. ( R,R ′)-MNF and ( R,R ′)-fenoterol [( R,R ′)-Fen] were synthesized as described previously [ ]., API-2, SL327, U0126, AM251, H-89, protein kinase inhibitor-(14–22)-amide (PKI), O-1602, and Tocrifluor 1117 (T1117) were from Tocris Bioscience. .. LPI, isoproterenol (ISO) and ICI-118,551 were purchased from Sigma-Aldrich.

    Solubility:

    Article Title: mGluR1-Mediated Excitation of Cerebellar GABAergic Interneurons Requires Both G Protein-Dependent and Src–ERK1/2-Dependent Signaling Pathways
    Article Snippet: Drugs DHPG, 7-(hydroxyimino)cyclopropa-[b]chromen-1a-carboxylate ethyl ester (CPCCOEt), (S )-(+)-α-amino-4-carboxy-2-methylbenzeneacetic acid (LY367385), 1-[2-(4-methoxyphenyl)-2-[3-(4-methoxyphenyl)propoxy]ethyl-1H -imidazole (SKF96365), (E)-2-cyano-3-(3,4-dihydrophenyl)-N-(phenylmethyl)-2-propenamide (AG490), (-)-bicuculline methochloride, 3-(4-chlorophenyl) 1-(1,1-dimethylethyl)-1H -pyrazolo[3,4-d]pyrimidin-4-amine (PP2), 1-phenyl-1H-pyrazolo[3,4-d]pyrimidin -4-amine (PP3), SR95531 (gabazine), U73122, U73343, and SL327 were obtained from Tocris Bioscience (Bristol, UK); 2-aminoethoxydiphenylborane (2-APB) was obtained from Abcam Biochemicals (Cambridge, UK); A polyclonal TRPC1 antibody and its control antigen, amino acid residues 557-571 of human TRPC1 were obtained from Alomone Labs (Cat#: ACC-010, Jerusalem, Israel). .. Drugs were dissolved in dH2 O or DMSO, according to the solubility advice of the manufacturer.

    Mouse Assay:

    Article Title: Genetic targeting of ERK1 suggests a predominant role for ERK2 in murine pain models
    Article Snippet: .. In dose-finding experiments with SL327, mice were acclimated as above, injected with SL327 or vehicle (DMSO), returned to their cage for 30 minutes, injected with 3.5% formalin, and sacrificed 3 minutes after paw injection. .. Spinal cord levels lumbar 3–6 (L3–L6) were isolated, and ipsilateral and contralateral sides were separated.

    Article Title: Genetic targeting of ERK1 suggests a predominant role for ERK2 in murine pain models
    Article Snippet: .. In experiments with the MEK inhibitor, SL327 (Tocris Bioscience, Ellisville, MO), mice were allowed to acclimate as above, injected with SL327 or vehicle (DMSO), and then returned to the behavior chamber. .. Thirty minutes later, mice were injected into the plantar surface of the hindpaw with 3.5% formalin, returned to the behavior chamber, and video recorded from below with a webcam (Logitech) at a resolution of 960 × 720 for one hour without the experimenter present.

    Article Title: Genetic targeting of ERK1 suggests a predominant role for ERK2 in murine pain models
    Article Snippet: .. 50 mg/kg SL327 significantly attenuated formalin-induced spontaneous nociceptive behaviors in WT mice (2-way RM ANOVA F=8.09, DFn=1, DFd=198, P=0.0108). .. Importantly, no difference was detected between WT and ERK1 KO mice injected with SL327 (2-way RM ANOVA F=0.91, DFn=1, DFd=187, P=0.3528).

    Concentration Assay:

    Article Title: Concurrent activation of β2-adrenergic receptor and blockage of GPR55 disrupts pro-oncogenic signaling in glioma cells
    Article Snippet: ( R,R ′)-MNF and ( R,R ′)-fenoterol [( R,R ′)-Fen] were synthesized as described previously [ ]., API-2, SL327, U0126, AM251, H-89, protein kinase inhibitor-(14–22)-amide (PKI), O-1602, and Tocrifluor 1117 (T1117) were from Tocris Bioscience. .. All compounds were dissolved in DMSO and were applied to cells at a final DMSO concentration of 0.1%.

    other:

    Article Title: Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury
    Article Snippet: Due to the potentially traumatic nature of i.c.v. administration, we wished to examine the efficacy of our inhibitor, SL327, an analogue of UO126, when introduced via the intraperitoneal route.

    Article Title: Heme Oxygenase Activity and Hemoglobin Neurotoxicity Are Attenuated by Inhibitors of the MEK/ERK Pathway
    Article Snippet: These activities were not significantly altered by U0126, SL327, or .

    Activity Assay:

    Article Title: Genetic targeting of ERK1 suggests a predominant role for ERK2 in murine pain models
    Article Snippet: .. Moreover, our experiments with the systemically active MEK inhibitor, SL327, provide additional evidence that ERK1/2 activity is required for the behavioral response to formalin. .. Since SL327 exerts similar effects in WT and ERK1 KO mice, it seems likely that ERK2 is specifically required in this behavioral model, although this must be directly tested.

    Expressing:

    Article Title: The mitogen and stress-activated protein kinase 1 regulates the rapid epigenetic tagging of dorsal horn neurons and nocifensive behaviour
    Article Snippet: .. The reduction in both formalin-induced p-MSK1 and p-H3S10 after SL327, by 73% and 62.5%, respectively, indicated a strong requirement of ERK signalling for the full expression of p-MSK1 and p-H3S10, as previously shown., , , , , It is important to note that p-ERK, p-MSK1, and p-H3S10 peak at different time points, and therefore the percentages of exact overlap are most probably greater than those suggested here. .. To conclusively establish that p-H3S10 is entirely downstream of p-ERK would require the complete elimination of p-ERK—a condition that was not fulfilled with 50 mg/kg SL327, a dose previously used by others.

    Article Title: The mitogen and stress-activated protein kinase 1 regulates the rapid epigenetic tagging of dorsal horn neurons and nocifensive behaviour
    Article Snippet: .. In this study, SL327 was used as a pharmacological tool to investigate the expression of molecular targets downstream of ERK, specifically p-MSK1 and p-H3S10, after noxious stimulation. .. Formalin stimulation induced expression of p-ERK, p-H3S10, and p-MSK1 in laminae I and II of the ipsilateral dorsal horn at 30 minutes (26.3 ± 2.7, 10.4 ± 1.5, and 11.2 ± 1.5 cells per 40-μm section, respectively) (Fig. B, C, D and E).

    BIA-KA:

    Article Title: A metabolic perturbation by U0126 identifies a role for glutamine in resveratrol-induced cell death
    Article Snippet: BY4741 yeast and Protein BCA kit were obtained from Thermo Fisher Scientific. .. SL327 was obtained from Tocris Bioscience.

    Injection:

    Article Title: Genetic targeting of ERK1 suggests a predominant role for ERK2 in murine pain models
    Article Snippet: .. In dose-finding experiments with SL327, mice were acclimated as above, injected with SL327 or vehicle (DMSO), returned to their cage for 30 minutes, injected with 3.5% formalin, and sacrificed 3 minutes after paw injection. .. Spinal cord levels lumbar 3–6 (L3–L6) were isolated, and ipsilateral and contralateral sides were separated.

    Article Title: Activation of a novel p70 S6 kinase 1-dependent intracellular cascade in the basolateral nucleus of the amygdala is required for the acquisition of extinction memory
    Article Snippet: .. SL327 (Tocris) was dissolved in a vehicle solution of 5% DMSO, 5% Tween-80 and saline and injected at a dose of 30 mg/kg. .. Rapamycin and PF-4708671 were injected intraperitoneally (i.p.) to a volume of 5 ml/kg relative to body weight one hour before the behavioral manipulation began; SL327 was injected 45 minutes before behavior.

    Article Title: Genetic targeting of ERK1 suggests a predominant role for ERK2 in murine pain models
    Article Snippet: .. In experiments with the MEK inhibitor, SL327 (Tocris Bioscience, Ellisville, MO), mice were allowed to acclimate as above, injected with SL327 or vehicle (DMSO), and then returned to the behavior chamber. .. Thirty minutes later, mice were injected into the plantar surface of the hindpaw with 3.5% formalin, returned to the behavior chamber, and video recorded from below with a webcam (Logitech) at a resolution of 960 × 720 for one hour without the experimenter present.

    Article Title: Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury
    Article Snippet: .. In order to investigate the effect of MEK1/2 inhibition on the brain regions of interest – isocortex, pyriform cortex, hippocampus, striatum, thalamus and external capsule – P7 C57/Bl6 animals were injected with a single i.p. dose of SL327 (133 μg/g BW) either 20 min preceding or 1 h following both 30 and 60 min hypoxia ( n = 10 per group). ..

    Article Title: Activation of a novel p70 S6 kinase 1-dependent intracellular cascade in the basolateral nucleus of the amygdala is required for the acquisition of extinction memory
    Article Snippet: .. Rapamycin and PF-4708671 were injected intraperitoneally (i.p.) to a volume of 5 ml/kg relative to body weight one hour before the behavioral manipulation began; SL327 was injected 45 minutes before behavior. ..

    Recombinant:

    Article Title: A metabolic perturbation by U0126 identifies a role for glutamine in resveratrol-induced cell death
    Article Snippet: SL327 was obtained from Tocris Bioscience. .. Recombinant human EGF was obtained from R & D Systems.

    Inhibition:

    Article Title: Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury
    Article Snippet: .. In order to investigate the effect of MEK1/2 inhibition on the brain regions of interest – isocortex, pyriform cortex, hippocampus, striatum, thalamus and external capsule – P7 C57/Bl6 animals were injected with a single i.p. dose of SL327 (133 μg/g BW) either 20 min preceding or 1 h following both 30 and 60 min hypoxia ( n = 10 per group). ..

    Plasmid Preparation:

    Article Title: A metabolic perturbation by U0126 identifies a role for glutamine in resveratrol-induced cell death
    Article Snippet: SL327 was obtained from Tocris Bioscience. .. Vectashield mounting medium for fluorescence with DAPI was obtained from Vector Laboratories.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Tocris drugs sl327
    PTPN11 D61G overexpression induces learning and memory and LTP deficits that can be reversed by MEK inhibition a. AAV– PTPN11 D61G infection results in overexpression of SHP2 D61G . Anti–SHP2 immunohistochemistry shows robust overexpression of SHP2 in the hippocampus of AAV– PTPN11 D61G –infused brains (left) compared with AAV– GFP infused brains (right). Full-length blots/gels are presented in Supplementary Figure 11 . b. PTPN11 D61G overexpression increases basal Erk activity (phospho–Erk level) and prevents further Erk activation in response to TBS. Left , Representative immunoblot showing p–Erk (upper) and total Erk (lower) in PTPN11 D61G –expressing slices and GFP –expressing slices. Slices were prepared 1 h after TBS. Right , Bar graph displays normalized p–Erk levels (mean ± s.e.m.). CTL, control without TBS. c. MEK inhibitor <t>SL327</t> reverses spatial memory deficits in PTPN11 D61G –overexpressing mice in the Morris water maze. Quadrant occupancy analysis for the probe trial reveals that PTPN11 D61G /veh mice showed no preference for the target quadrant (target vs. other quadrants, Dunnett’s Multiple Comparison Test after one-way ANOVA, P > 0.05). PTPN11 D61G /veh mice also spent significantly less time in the target quadrant compared with GFP /veh mice. SL327 treatment significantly increased the time spent in the target quadrant in PTPN11 D61G -expressing mice compared with vehicle-treated PTPN11 D61G mice ( PTPN11 D61G /SL327, 37.25 ± 3.50 %, n=10, unpaired two-tailed t-test, t = 2.335, * P
    Drugs Sl327, supplied by Tocris, used in various techniques. Bioz Stars score: 90/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/drugs sl327/product/Tocris
    Average 90 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    drugs sl327 - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

    Image Search Results


    PTPN11 D61G overexpression induces learning and memory and LTP deficits that can be reversed by MEK inhibition a. AAV– PTPN11 D61G infection results in overexpression of SHP2 D61G . Anti–SHP2 immunohistochemistry shows robust overexpression of SHP2 in the hippocampus of AAV– PTPN11 D61G –infused brains (left) compared with AAV– GFP infused brains (right). Full-length blots/gels are presented in Supplementary Figure 11 . b. PTPN11 D61G overexpression increases basal Erk activity (phospho–Erk level) and prevents further Erk activation in response to TBS. Left , Representative immunoblot showing p–Erk (upper) and total Erk (lower) in PTPN11 D61G –expressing slices and GFP –expressing slices. Slices were prepared 1 h after TBS. Right , Bar graph displays normalized p–Erk levels (mean ± s.e.m.). CTL, control without TBS. c. MEK inhibitor SL327 reverses spatial memory deficits in PTPN11 D61G –overexpressing mice in the Morris water maze. Quadrant occupancy analysis for the probe trial reveals that PTPN11 D61G /veh mice showed no preference for the target quadrant (target vs. other quadrants, Dunnett’s Multiple Comparison Test after one-way ANOVA, P > 0.05). PTPN11 D61G /veh mice also spent significantly less time in the target quadrant compared with GFP /veh mice. SL327 treatment significantly increased the time spent in the target quadrant in PTPN11 D61G -expressing mice compared with vehicle-treated PTPN11 D61G mice ( PTPN11 D61G /SL327, 37.25 ± 3.50 %, n=10, unpaired two-tailed t-test, t = 2.335, * P

    Journal: Nature neuroscience

    Article Title: Mechanism and treatment for the learning and memory deficits associated with mouse models of Noonan syndrome

    doi: 10.1038/nn.3863

    Figure Lengend Snippet: PTPN11 D61G overexpression induces learning and memory and LTP deficits that can be reversed by MEK inhibition a. AAV– PTPN11 D61G infection results in overexpression of SHP2 D61G . Anti–SHP2 immunohistochemistry shows robust overexpression of SHP2 in the hippocampus of AAV– PTPN11 D61G –infused brains (left) compared with AAV– GFP infused brains (right). Full-length blots/gels are presented in Supplementary Figure 11 . b. PTPN11 D61G overexpression increases basal Erk activity (phospho–Erk level) and prevents further Erk activation in response to TBS. Left , Representative immunoblot showing p–Erk (upper) and total Erk (lower) in PTPN11 D61G –expressing slices and GFP –expressing slices. Slices were prepared 1 h after TBS. Right , Bar graph displays normalized p–Erk levels (mean ± s.e.m.). CTL, control without TBS. c. MEK inhibitor SL327 reverses spatial memory deficits in PTPN11 D61G –overexpressing mice in the Morris water maze. Quadrant occupancy analysis for the probe trial reveals that PTPN11 D61G /veh mice showed no preference for the target quadrant (target vs. other quadrants, Dunnett’s Multiple Comparison Test after one-way ANOVA, P > 0.05). PTPN11 D61G /veh mice also spent significantly less time in the target quadrant compared with GFP /veh mice. SL327 treatment significantly increased the time spent in the target quadrant in PTPN11 D61G -expressing mice compared with vehicle-treated PTPN11 D61G mice ( PTPN11 D61G /SL327, 37.25 ± 3.50 %, n=10, unpaired two-tailed t-test, t = 2.335, * P

    Article Snippet: Drugs SL327 (Tocris) was dissolved in DMSO (16 mg/ml) and was injected intraperitoneally once daily, 30 min before the water maze experiment at a dose of 32mg/kg.

    Techniques: Over Expression, Inhibition, Infection, Immunohistochemistry, Activity Assay, Activation Assay, Expressing, CTL Assay, Mouse Assay, Two Tailed Test

    PTPN11 D61G overexpression enhances excitatory synaptic function through increased Ras-Erk signaling a. AMPA receptor-mediated currents were measured at the peak of the currents at − 65 mV, and NMDA currents were measured 50 ms after onset at + 40 mV. The average of 15 traces is shown. Scale, 100 pA and 40 ms. b. Group data showing the increased AMPA:NMDA current ratio in AAV– PTPN11 D61G mice compared with AAV- GFP mice. SL327 treatment (1 μM, 1 h) significantly reversed the AMPA:NMDA current ratio in the PTPN11 D61G group without affecting GFP –expressing mice. Two-way ANOVA, interaction between viral treatment and drug, F 1, 31 = 10.53, ** P

    Journal: Nature neuroscience

    Article Title: Mechanism and treatment for the learning and memory deficits associated with mouse models of Noonan syndrome

    doi: 10.1038/nn.3863

    Figure Lengend Snippet: PTPN11 D61G overexpression enhances excitatory synaptic function through increased Ras-Erk signaling a. AMPA receptor-mediated currents were measured at the peak of the currents at − 65 mV, and NMDA currents were measured 50 ms after onset at + 40 mV. The average of 15 traces is shown. Scale, 100 pA and 40 ms. b. Group data showing the increased AMPA:NMDA current ratio in AAV– PTPN11 D61G mice compared with AAV- GFP mice. SL327 treatment (1 μM, 1 h) significantly reversed the AMPA:NMDA current ratio in the PTPN11 D61G group without affecting GFP –expressing mice. Two-way ANOVA, interaction between viral treatment and drug, F 1, 31 = 10.53, ** P

    Article Snippet: Drugs SL327 (Tocris) was dissolved in DMSO (16 mg/ml) and was injected intraperitoneally once daily, 30 min before the water maze experiment at a dose of 32mg/kg.

    Techniques: Over Expression, Mass Spectrometry, Mouse Assay, Expressing

    Histological and electron microscopy assesment of pERK expression in P7 mouse forebrain following 30 min HI A and B , distribution of normal pERK immunoreactivity in the forebrain of sham animal ( A ) and an animal with unilateral carotid occlusion ( B ). C and G , increased pERK immunoreactivity in untreated animals at 15 min following 30 min HI ( C ). Response was ablated with the application of MEK inhibitor SL327 (133 μg/g BW) ( G ). D and E , schematic summary of white matter pERK‐IR in naive ( D ) and after a 30 min HI insult ( E ). Light microscopy overview at the intersection between hippocampus (top), thalamus (left) and cerebral cortex (right), coronal section at mid‐parietal level ( D and E ). Note the faint pERK‐IR in control animal with carotid occlusion only ( D , Ctrl), and the strong increase of expression in fibre tracts in external capsule (ec), fornix (fx), cortico‐thalamic fibres (ct), and descending tracts of the internal capsule (ic) at 1 h recovery following HI ( E ). F and H , electron microscopy of the internal capsule, at 15 min recovery following HI. Early pERK reactivity is located to the axons only. Arrows point to pERK positive clusters within adjacent axons. Scale bar ( A–C, G ]

    Journal: The Journal of Physiology

    Article Title: Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury

    doi: 10.1113/JP275649

    Figure Lengend Snippet: Histological and electron microscopy assesment of pERK expression in P7 mouse forebrain following 30 min HI A and B , distribution of normal pERK immunoreactivity in the forebrain of sham animal ( A ) and an animal with unilateral carotid occlusion ( B ). C and G , increased pERK immunoreactivity in untreated animals at 15 min following 30 min HI ( C ). Response was ablated with the application of MEK inhibitor SL327 (133 μg/g BW) ( G ). D and E , schematic summary of white matter pERK‐IR in naive ( D ) and after a 30 min HI insult ( E ). Light microscopy overview at the intersection between hippocampus (top), thalamus (left) and cerebral cortex (right), coronal section at mid‐parietal level ( D and E ). Note the faint pERK‐IR in control animal with carotid occlusion only ( D , Ctrl), and the strong increase of expression in fibre tracts in external capsule (ec), fornix (fx), cortico‐thalamic fibres (ct), and descending tracts of the internal capsule (ic) at 1 h recovery following HI ( E ). F and H , electron microscopy of the internal capsule, at 15 min recovery following HI. Early pERK reactivity is located to the axons only. Arrows point to pERK positive clusters within adjacent axons. Scale bar ( A–C, G ]

    Article Snippet: In order to investigate the effect of MEK1/2 inhibition on the brain regions of interest – isocortex, pyriform cortex, hippocampus, striatum, thalamus and external capsule – P7 C57/Bl6 animals were injected with a single i.p. dose of SL327 (133 μg/g BW) either 20 min preceding or 1 h following both 30 and 60 min hypoxia ( n = 10 per group).

    Techniques: Electron Microscopy, Expressing, Hi-C, Light Microscopy

    Ipsilateral ERK phosphorylation Dose response for SL327 inhibition of pERK immunoreactivity (optical luminosity value (OLV)), applied 20 min before a 30 min HI insult in hemispheric regions ipsilateral to carotid occlusion. A , CTX 12–2 dorsal cerebral cortex (12 to 2 o'clock segment), CTX 2–4 middle cerebral cortex (2 to 4 o'clock segment). B , PYRI: pyriform cortex, HIP: hippocampus. C , THAL: thalamus, STR; striatum. Increasing the dose of SL327 from 15 to 30 μg/g BW correlates to an 80% reduction in immunoreactivity.

    Journal: The Journal of Physiology

    Article Title: Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury

    doi: 10.1113/JP275649

    Figure Lengend Snippet: Ipsilateral ERK phosphorylation Dose response for SL327 inhibition of pERK immunoreactivity (optical luminosity value (OLV)), applied 20 min before a 30 min HI insult in hemispheric regions ipsilateral to carotid occlusion. A , CTX 12–2 dorsal cerebral cortex (12 to 2 o'clock segment), CTX 2–4 middle cerebral cortex (2 to 4 o'clock segment). B , PYRI: pyriform cortex, HIP: hippocampus. C , THAL: thalamus, STR; striatum. Increasing the dose of SL327 from 15 to 30 μg/g BW correlates to an 80% reduction in immunoreactivity.

    Article Snippet: In order to investigate the effect of MEK1/2 inhibition on the brain regions of interest – isocortex, pyriform cortex, hippocampus, striatum, thalamus and external capsule – P7 C57/Bl6 animals were injected with a single i.p. dose of SL327 (133 μg/g BW) either 20 min preceding or 1 h following both 30 and 60 min hypoxia ( n = 10 per group).

    Techniques: Inhibition

    Schedule of experimental procedures A , WT (C57/Bl6) mice underwent 30 min hypoxic–ischaemic (HI) insult and were then killed at 15 min post‐hypoxia for pERK immunoreactivity evaluation. B , pERK immunoreactivity was assessed at multiple time points up to 48 h post‐insult to P7 WT mice. C , a dose response of SL327, controlled to vehicle alone, was administered 20 min prior to 30 min HI and pERK immunoreactivity was assessed at 15 min post‐insult. D , WT mice were subject to either 30 min or 60 min HI, with 133 μg/g SL327 or EtOH (vehicle) administered either 20 min prior to or 60 min post‐insult. Brain histology was assessed at 48 h. E , inhibition of neuronal pERK immunoreactivity was confirmed at 15 min post‐HI in synapsin‐cre driven ERK tg mutant mice compared to littermate WT controls. F , brain histology was assessed at 48 h after 30 min HI in synapsin‐cre driven ERK tg mutant mice and littermate WT controls. G , brain histology was assessed at 48 h after 30 min HI in GFAP‐cre driven ERK tg mutant mice and littermate WT controls. H , saline or LPS was injected at 12 h prior to 30 min HI in both synapsin‐cre and GFAP‐cre driven ERK tg mutant mice and littermate WT controls. Brain histology was assessed at 48 h.

    Journal: The Journal of Physiology

    Article Title: Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury

    doi: 10.1113/JP275649

    Figure Lengend Snippet: Schedule of experimental procedures A , WT (C57/Bl6) mice underwent 30 min hypoxic–ischaemic (HI) insult and were then killed at 15 min post‐hypoxia for pERK immunoreactivity evaluation. B , pERK immunoreactivity was assessed at multiple time points up to 48 h post‐insult to P7 WT mice. C , a dose response of SL327, controlled to vehicle alone, was administered 20 min prior to 30 min HI and pERK immunoreactivity was assessed at 15 min post‐insult. D , WT mice were subject to either 30 min or 60 min HI, with 133 μg/g SL327 or EtOH (vehicle) administered either 20 min prior to or 60 min post‐insult. Brain histology was assessed at 48 h. E , inhibition of neuronal pERK immunoreactivity was confirmed at 15 min post‐HI in synapsin‐cre driven ERK tg mutant mice compared to littermate WT controls. F , brain histology was assessed at 48 h after 30 min HI in synapsin‐cre driven ERK tg mutant mice and littermate WT controls. G , brain histology was assessed at 48 h after 30 min HI in GFAP‐cre driven ERK tg mutant mice and littermate WT controls. H , saline or LPS was injected at 12 h prior to 30 min HI in both synapsin‐cre and GFAP‐cre driven ERK tg mutant mice and littermate WT controls. Brain histology was assessed at 48 h.

    Article Snippet: In order to investigate the effect of MEK1/2 inhibition on the brain regions of interest – isocortex, pyriform cortex, hippocampus, striatum, thalamus and external capsule – P7 C57/Bl6 animals were injected with a single i.p. dose of SL327 (133 μg/g BW) either 20 min preceding or 1 h following both 30 and 60 min hypoxia ( n = 10 per group).

    Techniques: Mouse Assay, Inhibition, Mutagenesis, Injection

    Effect of SL327 on αMβ2+ microglial activation, astroglial activation, neuronal tissue loss (Nissl body presence) and TUNEL+ cell death, when applied 20 min before ( A , C , E , G ) or 1 h post ( B , D , F , H , I ) 30 or 60 min HI Assessment at ×20 microscopy field magnification (mean + SEM over 3 fields). A and B , the levels of CD11b+ microglia are significantly decreased in the SL327 group in white matter (EC) as well as in most grey matter regions (STR, CTX, HIP). E and F , Nissl score was decreased in pre‐treated animals. Cortex was particularly spared compared to vehicle alone. G and H , this trend to decrease is observed with number of TUNEL+ cells. SL327 treated pups have a reduction in dying cells compared to EtOH treated animals, significantly so in pre‐treated STR and HIP. C and D , extent of gliosis and reactive (GFAP+) astrocyte activation was unaffected by the application of SL327. * P

    Journal: The Journal of Physiology

    Article Title: Extracellular signal‐regulated kinase 2 has duality in function between neuronal and astrocyte expression following neonatal hypoxic–ischaemic cerebral injury

    doi: 10.1113/JP275649

    Figure Lengend Snippet: Effect of SL327 on αMβ2+ microglial activation, astroglial activation, neuronal tissue loss (Nissl body presence) and TUNEL+ cell death, when applied 20 min before ( A , C , E , G ) or 1 h post ( B , D , F , H , I ) 30 or 60 min HI Assessment at ×20 microscopy field magnification (mean + SEM over 3 fields). A and B , the levels of CD11b+ microglia are significantly decreased in the SL327 group in white matter (EC) as well as in most grey matter regions (STR, CTX, HIP). E and F , Nissl score was decreased in pre‐treated animals. Cortex was particularly spared compared to vehicle alone. G and H , this trend to decrease is observed with number of TUNEL+ cells. SL327 treated pups have a reduction in dying cells compared to EtOH treated animals, significantly so in pre‐treated STR and HIP. C and D , extent of gliosis and reactive (GFAP+) astrocyte activation was unaffected by the application of SL327. * P

    Article Snippet: In order to investigate the effect of MEK1/2 inhibition on the brain regions of interest – isocortex, pyriform cortex, hippocampus, striatum, thalamus and external capsule – P7 C57/Bl6 animals were injected with a single i.p. dose of SL327 (133 μg/g BW) either 20 min preceding or 1 h following both 30 and 60 min hypoxia ( n = 10 per group).

    Techniques: Activation Assay, TUNEL Assay, Microscopy

    Spinal extracellular signal-regulated kinase (ERK) regulates p-MSK1 and p-H3S10 expression after formalin injection. (A) Diagram of postulated signaling pathways upstream of p-H3S10. (B) Dorsal horn images of p-ERK and p-MSK1 in vehicle- and MEK inhibitor SL327-treated animals 30 minutes after formalin stimulation. (C) Dorsal horn images of p-ERK and p-H3S10 in vehicle- and MEK inhibitor SL327-treated animals 30 minutes after formalin stimulation. (D) Dorsal horn images of p-H3S10 and p-MSK1 in vehicle- and MEK inhibitor SL327-treated animals 30 minutes after formalin stimulation. (B-D): Pictures show a single focal plane. Final column images show a merge of the first 2 images; colocalisation is seen in yellow; arrowheads indicate examples of colocalisation. The white line indicates the medial border between lamina I and white matter. Scale bar, 50 μm. (E) Quantification of p-ERK, p-H3S10, and p-MSK1 single-labeled cells 30 minutes after formalin injection. n = 8 in each treatment group, 5 sections per animal. (F) Quantification of p-ERK, p-H3S10, and p-MSK1 double-labeled cells 30 minutes after formalin. n = 4 in each group. (E-F) Values presented as group mean ± SEM (per 40 μm section). * P

    Journal: Pain

    Article Title: The mitogen and stress-activated protein kinase 1 regulates the rapid epigenetic tagging of dorsal horn neurons and nocifensive behaviour

    doi: 10.1097/j.pain.0000000000000679

    Figure Lengend Snippet: Spinal extracellular signal-regulated kinase (ERK) regulates p-MSK1 and p-H3S10 expression after formalin injection. (A) Diagram of postulated signaling pathways upstream of p-H3S10. (B) Dorsal horn images of p-ERK and p-MSK1 in vehicle- and MEK inhibitor SL327-treated animals 30 minutes after formalin stimulation. (C) Dorsal horn images of p-ERK and p-H3S10 in vehicle- and MEK inhibitor SL327-treated animals 30 minutes after formalin stimulation. (D) Dorsal horn images of p-H3S10 and p-MSK1 in vehicle- and MEK inhibitor SL327-treated animals 30 minutes after formalin stimulation. (B-D): Pictures show a single focal plane. Final column images show a merge of the first 2 images; colocalisation is seen in yellow; arrowheads indicate examples of colocalisation. The white line indicates the medial border between lamina I and white matter. Scale bar, 50 μm. (E) Quantification of p-ERK, p-H3S10, and p-MSK1 single-labeled cells 30 minutes after formalin injection. n = 8 in each treatment group, 5 sections per animal. (F) Quantification of p-ERK, p-H3S10, and p-MSK1 double-labeled cells 30 minutes after formalin. n = 4 in each group. (E-F) Values presented as group mean ± SEM (per 40 μm section). * P

    Article Snippet: The reduction in both formalin-induced p-MSK1 and p-H3S10 after SL327, by 73% and 62.5%, respectively, indicated a strong requirement of ERK signalling for the full expression of p-MSK1 and p-H3S10, as previously shown., , , , , It is important to note that p-ERK, p-MSK1, and p-H3S10 peak at different time points, and therefore the percentages of exact overlap are most probably greater than those suggested here.

    Techniques: Expressing, Injection, Labeling

    NCS-Rapgef2-dependent ERK activation in mouse NAc following D1 dopamine receptor agonist SKF 81297 and psychostimulants D-amphetamine or cocaine administration. A , C57BL6N wild-type mice received either saline or D1 dopamine receptor agonist SKF 81297 (2 mg/kg, i.p.). Phosphorylation of ERK in the NAc, especially in the shell, was robustly induced 15 min after D1 agonist treatment. A systemic injection of MEK inhibitor SL327 (60 mg/kg, i.p.) 60 min before treatment with SKF 81297 significantly reduced phosphorylation of ERK in NAc. Lower panels are pictures with higher magnification from the shell areas in upper panels (indicated by the white frame). Scale bar: 200 µm (upper panel) and 50 μm (lower panel). B–D , AAV viral vector-directed ablation of Rapgef2 expression in the NAc impaired ERK activation induced by D1 dopamine receptor agonist SKF 81297. Rapgef2 cko/cko mice were unilaterally injected with AAV9.hSynap.HI.eGFP-Cre.WPRE.SV40 or control viral vector AAV9.hSynap.eGFP , which encodes eGFP-fused cre recombinase or eGFP alone under the control of human synapsin promoter. Four weeks later, animals were treated with saline or SKF81297 (2 mg/kg, i.p.) and perfused 15 min later. Phosphorylation of ERK ( B ) induced by SKF81297 in the NAc was absent on the side of the brain with cre viral injection (shown by nuclear eGFP signal) (upper panels), but clearly present in the NAc injected with control eGFP viral vector (lower panels). Phospho-CREB ( C ) induced by SKF81297 in the NAc on either the side of the brain with cre injection (upper panels) or on the side of the brain receiving control viral vector (lower panels) administration. Loss of NCS-Rapgef2 protein expression ( D ) was observed in the NAc of Rapgef2 cko/cko mice four weeks following unilateral injection of AAV9.hSynap.HI.eGFP-Cre.WPRE.SV40 (left panel), but not control viral vector (right panel). Scale bar: 50 μm. E , Rapgef2 cko/cko mice were unilaterally injected with AAV9.hSynap.HI.eGFP-Cre.WPRE.SV40 into the NAc. Neurons showing phospho-ERK after SKF 81297(2 mg/kg, i.p.), D-amphetamine (10 mg/kg, i.p.), or cocaine (30 mg/kg, i.p.) were counted using the NIH ImageJ software. Cell counts represent the average obtained from 3three to five animals per treatment group measured in a 318 × 318 μm area in the NAc. Student’s t test showed significant reduction of ERK phosphorylation on viral vector injected side for all drug treatment groups (* p

    Journal: eNeuro

    Article Title: NCS-Rapgef2, the Protein Product of the Neuronal Rapgef2 Gene, Is a Specific Activator of D1 Dopamine Receptor-Dependent ERK Phosphorylation in Mouse Brain

    doi: 10.1523/ENEURO.0248-17.2017

    Figure Lengend Snippet: NCS-Rapgef2-dependent ERK activation in mouse NAc following D1 dopamine receptor agonist SKF 81297 and psychostimulants D-amphetamine or cocaine administration. A , C57BL6N wild-type mice received either saline or D1 dopamine receptor agonist SKF 81297 (2 mg/kg, i.p.). Phosphorylation of ERK in the NAc, especially in the shell, was robustly induced 15 min after D1 agonist treatment. A systemic injection of MEK inhibitor SL327 (60 mg/kg, i.p.) 60 min before treatment with SKF 81297 significantly reduced phosphorylation of ERK in NAc. Lower panels are pictures with higher magnification from the shell areas in upper panels (indicated by the white frame). Scale bar: 200 µm (upper panel) and 50 μm (lower panel). B–D , AAV viral vector-directed ablation of Rapgef2 expression in the NAc impaired ERK activation induced by D1 dopamine receptor agonist SKF 81297. Rapgef2 cko/cko mice were unilaterally injected with AAV9.hSynap.HI.eGFP-Cre.WPRE.SV40 or control viral vector AAV9.hSynap.eGFP , which encodes eGFP-fused cre recombinase or eGFP alone under the control of human synapsin promoter. Four weeks later, animals were treated with saline or SKF81297 (2 mg/kg, i.p.) and perfused 15 min later. Phosphorylation of ERK ( B ) induced by SKF81297 in the NAc was absent on the side of the brain with cre viral injection (shown by nuclear eGFP signal) (upper panels), but clearly present in the NAc injected with control eGFP viral vector (lower panels). Phospho-CREB ( C ) induced by SKF81297 in the NAc on either the side of the brain with cre injection (upper panels) or on the side of the brain receiving control viral vector (lower panels) administration. Loss of NCS-Rapgef2 protein expression ( D ) was observed in the NAc of Rapgef2 cko/cko mice four weeks following unilateral injection of AAV9.hSynap.HI.eGFP-Cre.WPRE.SV40 (left panel), but not control viral vector (right panel). Scale bar: 50 μm. E , Rapgef2 cko/cko mice were unilaterally injected with AAV9.hSynap.HI.eGFP-Cre.WPRE.SV40 into the NAc. Neurons showing phospho-ERK after SKF 81297(2 mg/kg, i.p.), D-amphetamine (10 mg/kg, i.p.), or cocaine (30 mg/kg, i.p.) were counted using the NIH ImageJ software. Cell counts represent the average obtained from 3three to five animals per treatment group measured in a 318 × 318 μm area in the NAc. Student’s t test showed significant reduction of ERK phosphorylation on viral vector injected side for all drug treatment groups (* p

    Article Snippet: The MEK inhibitor SL327 (60 mg/kg; Tocris) was dissolved in DMSO and then diluted twice in sterile water.

    Techniques: Activation Assay, Mouse Assay, Injection, Plasmid Preparation, Expressing, Software