Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: Schemes for Experiments and post hoc analyses. (A) Scheme of the ketamine injection paradigm for Experiment 1 during development of LID (daily L-DOPA injections). FAS = Forelimb adjusting steps test. (B) Scheme of the injection paradigm in PD rats for Experiment 2 . AR = amphetamine-rotation test; RR = RotaRod test. (C) Scheme of the injection paradigm for Experiment 3 during development of LID (daily L-DOPA injections). (D) Verification of unilateral 6-OHDA lesion and evaluation of striatal dopamine (DA) levels after ketamine in the rats from the study shown in (A). Electrochemical detection of striatal DA content (mean ± SEM) is reduced by>95% in the lesioned side. Striatal DA content was unchanged by a 10-h-treatment of either ketamine (K; n =9), R -ketamine ( R -K; n =9) vs. vehicle (V; n = 9) 1-h before rats were euthanized, showing no effect on overall striatal DA levels by ketamine or R -ketamine-treatment compared to vehicle in either the lesioned (Lx) or the intact hemisphere. (E) Verification of unilateral 6-OHDA lesion from the study shown in (B) using semi-quantitative TH western analysis in striatal tissue plotting % loss (mean ± SEM) in Lx vs. intact (In) hemisphere ( n =9). Two-tailed t -test, *** p < .001. (F) Verification of unilateral 6-OHDA lesions in the rats from the study depicted in (C). The graph shows the quantification of the TH-ir plotting the % loss (mean ± SEM) in the Lx vs. intact SN hemispheres ( n =10/group; V =vehicle, K =ketamine, K +A = ketamine+ ANA-12). (G) Example photomicrograph of a SN in Experiment 3 shows the unilateral reduction in TH-ir post-lesion. Two-way ANOVAs, Bonferroni post hoc tests, *** p < .001. (H) Verification of unilateral 6-OHDA lesion from the ANA-12-only control study, the negative control for Experiment 3 , using semi-quantitative TH western analysis in striatal tissue, plotting % loss (mean ± SEM) in Lx vs. intact hemisphere ( n = 10). Two-tailed t -test, *** p < .001.

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection, Western Blot, Two Tailed Test, Negative Control

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: Low-dose racemic ketamine treatment once a week attenuates the development of LID in the preclinical model. In Experiment 1 6-OHDA-lesioned PD rats were injected daily with L-DOPA (days 0–13: 6 mg/kg; days 14–28: 12 mg/kg; i.p. ) to induce dyskinesia and tested for LAO-AIMs twice a week for 3 h by blinded investigators. (A) The mean LAO AIMs scores±SEM are plotted showing a 50% reduction after racemic low-dose ketamine treatments (K) when compared to the vehicle group (V) and a group treated with R -ketamine ( R -K), to test for contribution of the stereospecific ketamine isomer. The blue arrows point to the days of the 10-h racemic ketamine (20 mg/kg; i.p. ), R -ketamine (10 mg/kg; i.p. ) or vehicle treatment paradigm; n = 9 per group, * p < .05, ** p < .01, Kruskal-Wallis test with Dunn’s multiple comparisons post hoc tests. (B) Example time course of the LAO-AIMs data showed in (A) for day 11. (C) Example time course of the LAO-AIMs data showed in (A) for day 25. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: LAO-AIMs scores from Experiment 1 separated by sub-type of dyskinesia. Low-dose racemic ketamine treatment once a week attenuates the development of (A) limb (B) axial (C) orolingual AIMs in the preclinical model. 6-OHDA-lesioned PD rats were injected daily with L-DOPA (days 0–13: 6 mg/kg; days 14–28: 12 mg/kg; i.p. ) to induce dyskinesia and tested for LAO AIMs twice a week for 3 h by blinded investigators. The mean LAO AIMs scores ± SEM are plotted showing a 50% reduction after racemic low-dose ketamine treatments (K) when compared to the vehicle group (V) and a group treated with R -ketamine ( R -K), to test for contribution of the stereospecific ketamine isomer. The blue arrows point to the days of the 10-h racemic ketamine (20 mg/kg; i.p. ), R -ketamine (10 mg/kg; i.p. ) or vehicle treatment paradigm; n = 9 per group, * p < .05, ** p < .01, Kruskal-Wallis tests with Dunn’s post hoc tests. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: Ketamine does not interfere with the anti-PD effect of L-DOPA and reduces PD-motor behavior post-6-OHDA-lesion (Post-Lx) by itself. (A) Mean % contralateral/ipsilateral ratios of steps ± SEM using the FAS test paradigm in the LID cohort of Experiment 1 , are plotted after normalization to pre-lesion (Pre-Lx) indicate a significant anti-PD effect of ketamine * p < .05; repeated measures ANOVA. Ketamine does also not interfere with the anti-PD effect of L-DOPA, and a significant increase of stepping contralateral to the lesioned side after either L-DOPA alone or L-DOPA + Ketamine vs. all Post-Lx time points is seen: *** p < .001; one-way ANOVA on data prior to normalization, Tukey-Kramer corrected post hoc tests; n = 9 per group. (B) In Experiment 2 we tested ketamine treatment in a separate cohort of hemi-parkinsonian 6-OHDA-lesioned rats and used the RotaRod test to evaluate the deficit. The graph shows the mean latency to fall ± SEM, normalized to pre-lesion baseline (Pre-Lx). Post-lesion (Post-Lx) the latency to fall was reduced by 50% in these PD animals. This motor deficit was reversed by ketamine treatment (blue bars), already at the 1st injection, and the animals performed as good as at baseline. One-way ANOVA, with Tukey-Kramer corrected post hoc tests, on raw data before normalization. n = 9, * p < .05, ** p < .01. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: Ketamine’s long-term anti-dyskinetic activity was driven by BDNF signaling. (A) The mean LAO-AIMs scores ± SEM of Experiment 3 are plotted. The sustained anti-dyskinetic effect of low-dose ketamine is reduced by blocking the BDNF receptor TrkB, with co-injection of the TrkB antagonist ANA-12 (0.5 mg/kg; i.p. ) with ketamine (K + A). The blue arrows point to the days of the 10-h racemic (K) ketamine (20 mg/kg; i.p. ), or vehicle treatment paradigm (V). ANA-12 co-injection (green bars) did reduce the sustained anti-dyskinetic effect seen in ketamine-only injected LID (blue bars) leading to LAO AIMs comparable to those of the vehicle group (grey bars), indicating an involvement of BDNF in the sustained anti-dyskinetic effects of ketamine. n = 10 per group, * p < .05, ** p < .01, ANOVAs, Tukey-Kramer corrected post hoc tests. (B) Example time course of the LAO-AIMs for day 14 showed in (A). (C) A control study using 10-h ANA-12-only treatments on days 0 and 7 of daily L-DOPA-treatment (6 mg/kg; i.p .) verified that, while systemic TrkB antagonism does block the ketamine effect, it does not change development of LID in this model, and serves as an important negative control for the data shown in (A). The graph depicts the mean LAO-AIMs scores ± SEM from the vehicle control groups in Experiment 1 (V- E1 ; n = 9) and Experiment 3 (V- E3 ; n = 10), as well as the ANA-12-only control study (ANA-12; n = 10). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Activity Assay, Blocking Assay, Injection, Negative Control

Journal: Experimental neurology

Article Title: Preclinical evidence in support of repurposing sub-anesthetic ketamine as a treatment for L-DOPA-induced dyskinesia

doi: 10.1016/j.expneurol.2020.113413

Figure Lengend Snippet: LAO-AIMs scores from Experiment 3 separated by sub-type of dyskinesia. Low-dose ketamine treatment (blue arrows) once a week attenuates the development of individual AIMs scores. 6-OHDA-lesioned PD rats were injected daily with L-DOPA (days 0–14: 6 mg/kg; i.p. ) to induce dyskinesia and tested for LAO-AIMs twice a week for 3 h by blinded investigators. The anti-dyskinetic effect of low-dose ketamine (K, blue bars) reduced the individual (A) limb (B) axial (C) and orolingual AIMs (mean ± SEM) scores, compared to the vehicle (V, grey bars) group and a group treated with the TrkB antagonist, ANA-12 (K + A, green bars). The blue arrows point to the days of the 10-h racemic ketamine (20 mg/kg; i.p. ), R -ketamine (10 mg/kg; i.p. ) n = 10 per group, *p < .05, **p < .01, Kruskal-Wallis tests with Dunn’s post hoc tests. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: In Experiment 1 , ketamine (20 mg/kg; VetOne), R -ketamine (10 mg/kg; Cayman Chemicals, Ann Arbor, MI), and L-DOPA (6 and 12 mg/kg; Sigma-Aldrich) combined with benserazide hydrochloride (14 mg/kg; Sigma-Aldrich) were formulated in the vehicle solution, 0.9% USP grade sterile saline (VetOne).

Techniques: Injection

Journal: Translational Psychiatry

Article Title: R -ketamine: a rapid-onset and sustained antidepressant without psychotomimetic side effects

doi: 10.1038/tp.2015.136

Figure Lengend Snippet: Antidepressant effects of R -ketamine and S -ketamine in social defeat stress and LH models of depression. ( a ) The schedule of social defeat stress model and behavioral tests after treatment. ( b ) One percent SPT was performed 1 day after a single dose of saline, R -ketamine (10 mg kg −1 ) or S -ketamine (10 mg kg −1 ) (one-way ANOVA, F 3,42 =11.05, P =0.002). ( c – e ) Behavioral tests, including LMT (F 3,35 =0.038, P =0.99), TST (F 3,34 =12.046, P <0.001) and FST (F 3,34 =13.235, P <0.001), were performed 2 days after a single dose. ( f ) SPT was performed 6 days after a single dose (F 3,34 =9.974, P <0.001). ( g , h ) TST (F 3,32 =12.019, P <0.001) and FST (F 3,32 =14.479, P <0.001) were performed 7 days after a single dose. Data are shown as mean±s.e.m. ( n =8–11). * P <0.05, and *** P <0.001. ( i ) The schedule of learned helplessness (LH) model and behavioral tests after treatment. ( j , k ) The failure of LH (one-way ANOVA, F 2,15 =3.903, P =0.043) and the escape latency of LH (F 2,15 =4.317, P =0.033) were measured 5 days after a single dose of saline, R -ketamine (20 mg kg −1 ) or S -ketamine (20 mg kg −1 ). Data are shown as mean±s.e.m. ( n =6). * P <0.05. ANOVA, analysis of variance; FST, forced swimming test; IES, inescapable electric foot shock; LH, learned helplessness; LMT, locomotion test; NS, not significant; PS, post-shock test; SPT, sucrose preference test; TST, tail suspension test.

Article Snippet: R -ketamine hydrochloride and S -ketamine hydrochloride were prepared by recrystallization of RS -ketamine (Ketalar, ketamine hydrochloride, Daiichi Sankyo Pharmaceutical, Tokyo, Japan) and d -(−)-tartaric acid (or l - (+)-tartaric acid), as described previously.

Techniques:

Journal: Translational Psychiatry

Article Title: R -ketamine: a rapid-onset and sustained antidepressant without psychotomimetic side effects

doi: 10.1038/tp.2015.136

Figure Lengend Snippet: Effects of R -ketamine and S -ketamine on alterations in dendritic spine density in the brain regions after social defeat stress. Brain samples were collected 8 days after a single dose of saline, R -ketamine (10 mg kg −1 ) or S -ketamine (10 mg kg −1 ) for Golgi–Cox staining. Representative photomicrographs of Golgi–Cox-stained pyramidal neurons in the PrL of mPFC, IL of mPFC, NAc core, NAc shell, striatum, CA1, CA3 and DG of hippocampus from animals of each group. Scale bar=10 μm. ( a ) PrL (one-way analysis of variance, F 3,20 =8.123, P =0.001); ( b ) IL (F 3,20 =0.528, P =0.668); ( c ) NAc core (F 3,20 =6.318, P =0.003); ( d ) NAc shell (F 3,20 =6.332, P =0.003); ( e ) striatum (F 3,20 =0.381, P =0.768); ( f ) CA1 (F 3,20 =0.459, P =0.714); ( g ) CA3 (F 3,20 =8.448, P =0.001); ( h ) DG (F 3,20 =5.546, P =0.006). Values represent the mean±s.e.m. ( n =6). * P <0.05, ** P <0.01, *** P <0.001. DG, dentate gyrus; IL, infralimbic region; mPFC, medial prefrontal cortex; NAc, nucleus accumbens; N.S., not significant; PrL, prelimbic region.

Article Snippet: R -ketamine hydrochloride and S -ketamine hydrochloride were prepared by recrystallization of RS -ketamine (Ketalar, ketamine hydrochloride, Daiichi Sankyo Pharmaceutical, Tokyo, Japan) and d -(−)-tartaric acid (or l - (+)-tartaric acid), as described previously.

Techniques: Staining

Journal: Translational Psychiatry

Article Title: R -ketamine: a rapid-onset and sustained antidepressant without psychotomimetic side effects

doi: 10.1038/tp.2015.136

Figure Lengend Snippet: Role of AMPA receptor and BDNF–TrkB signaling in the mechanisms of antidepressant effect for R -ketamine and S -ketamine. ( a ) The schedule of social defeat stress model and behavioral tests after treatment. NBQX (10 mg kg −1 ), an AMPA receptor antagonist, was administered 30 min before saline, R -ketamine (10 mg kg −1 ) or S -ketamine (10 mg kg −1 ). Behavioral tests, including ( b ) LMT (one-way ANOVA, F 5,34 =0.06, P =0.997), ( c ) TST (F 5,39 =14.628, P <0.001) and ( d ) FST (F 5,42 =9.015, P <0.001), were performed 1 day after a single dose. ( e ) SPT was 7 days after a single dose of saline, R -ketamine (10 mg kg −1 ) or S -ketamine (10 mg kg −1 ) (F 5,40 =11.748, P <0.001). Values represent the mean±s.e.m. ( n =6–9). * P <0.05, ** P <0.01 and *** P <0.001. ( f ) The schedule of social defeat stress model and behavioral tests after treatment. ANA-12 (0.5 mg kg −1 ), a TrkB antagonist, was co-administered with saline or R -ketamine (10 mg kg −1 ) or S -ketamine (10 mg kg −1 ). Behavioral tests, including ( g ) LMT (one-way ANOVA, F 5,37 =0.414, P =0.836), ( h ) TST (F 5,33 =14.044, P <0.001) and ( i ) FST (F 5,32 =15.783, P <0.001), were performed 1 day after a single dose. ( j ) The SPT was 7 days after a single dose of saline, R -ketamine or S -ketamine (F 5,36 =11.825, P <0.001). Values represent the mean±s.e.m. ( n =6–8). * P <0.05, ** P <0.01 and *** P <0.001. AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor; ANOVA, analysis of variance; BDNF, brain-derived neurotrophic factor; FST, forced swimming test; LMT, locomotion test; N.S., not significant; SPT, sucrose preference test; TST, tail suspension test.

Article Snippet: R -ketamine hydrochloride and S -ketamine hydrochloride were prepared by recrystallization of RS -ketamine (Ketalar, ketamine hydrochloride, Daiichi Sankyo Pharmaceutical, Tokyo, Japan) and d -(−)-tartaric acid (or l - (+)-tartaric acid), as described previously.

Techniques: Derivative Assay

Journal: Translational Psychiatry

Article Title: R -ketamine: a rapid-onset and sustained antidepressant without psychotomimetic side effects

doi: 10.1038/tp.2015.136

Figure Lengend Snippet: Effect for R -ketamine and S -ketamine on the BDNF, TrkB phosphorylation and GluA1 in the brain regions. Brain samples were collected 7 days after a single dose of saline, R -ketamine (10 mg kg −1 ) or S -ketamine (10 mg kg −1 ) for Golgi–Cox staining. ( a ) Western blot analysis of BDNF in PFC, NAc, CA1, CA3 and DG. PFC (one-way ANOVA, F 3,13 =5.759, P =0.009); NAc (F 3,13 =3.544, P =0.045); CA1 (F 3,14 =0.065, P =0.978); CA3 (F 3,13 =8.324, P =0.002); DG (F 3,13 =7.444, P =0.003). The value was expressed as a percentage of that of control mice. Values represent the mean±s.e.m. ( n =5 or 6). * P <0.05, ** P <0.01, *** P <0.001. ( b , c ) Effects of R -ketamine and S -ketamine on changes in phosphorylation of TrkB in the mouse brain after social defeat stress. The ratio of p-TrkB to total TrkB in the brain regions was shown. PFC (one-way ANOVA, F 3,13 =3.962, P =0.033); NAc (F 3,13 =3.613, P =0.043); CA1 (F 3,14 =0.05, P =0.984); CA3 (F 3,14 =6.545, P =0.005); DG (F 3,13 =7.612, P =0.003). Total levels of TrkB protein in the all regions were not different among the four groups. Values represent the mean±s.e.m. ( n =5 or 6). * P <0.05, ** P <0.01. ( d ) Western blot analysis of GluA1 in PFC, NAc, CA1, CA3 and DG of the hippocampus. PFC (one-way ANOVA, F 3,13 =3.619, P =0.04); NAc (F 3,14 =7.834, P =0.003); CA1 (F 3,14 =0.060, P =0.98); CA3 (F 3,14 =5.207, P =0.013); DG (F 3,14 =5.108, P =0.014). The value was expressed as a percentage of that of control mice. Values represent the mean±s.e.m. ( n =5 or 6). * P <0.05, ** P <0.01, *** P <0.001. ANOVA, analysis of variance; BDNF, brain-derived neurotrophic factor; DG, dentate gyrus; NAc, nucleus accumbens; N.S., not significant; PFC, prefrontal cortex.

Article Snippet: R -ketamine hydrochloride and S -ketamine hydrochloride were prepared by recrystallization of RS -ketamine (Ketalar, ketamine hydrochloride, Daiichi Sankyo Pharmaceutical, Tokyo, Japan) and d -(−)-tartaric acid (or l - (+)-tartaric acid), as described previously.

Techniques: Staining, Western Blot, Derivative Assay

Journal: Translational Psychiatry

Article Title: R -ketamine: a rapid-onset and sustained antidepressant without psychotomimetic side effects

doi: 10.1038/tp.2015.136

Figure Lengend Snippet: Side-effect profiles for S -ketamine, but not R -ketamine, in mice. ( a ) Effects of R -ketamine and S -ketamine on locomotion in control mice. One hour after habituation, saline, R -ketamine (5, 10 or 20 mg kg −1 ) or S -ketamine (5, 10 or 20 mg kg −1 ) was administered intraperitoneally into mice. Two-way ANOVA analysis revealed significant interactions (drug: F 6,125 =6.441, P <0.001; time: F 17,125 =138.838, P <0.001; interaction (drug × time): F 102,1125 =2.814, P <0.001). Values represent the mean±s.e.m. ( n =8). ** P <0.01, *** P <0.001 compared with the saline-treated group. ( b ) Effects of R -ketamine (5, 10 or 20 mg kg −1 ) on PPI test in control mice. The MANOVA revealed no significant effect (Wilks' lambda=0.713, P =0.333). Values represent the mean±s.e.m. ( n =8). ( c ) Effects of S -ketamine (5, 10 or 20 mg kg −1 ) on the PPI test in control mice. The MANOVA revealed significant effect (Wilks' lambda=0.554, P =0.019). Values represent the mean±s.e.m. ( n =8). * P <0.05, ** P <0.01 compared with the saline-treated group. ( d ) The schedule of CPP model and behavioral tests after treatment. ( e ) RS -ketamine (10 mg kg −1 ) significantly ( P =0.0125) increased CPP scores in mice ( n =9). * P <0.05 compared with the saline-treated group. ( f ) R -ketamine (5, 10 or 20 mg kg −1 ) did not increase CPP score (F 3,35 =0.147, P =0.931). ( g ) S -Ketamine (5, 10 or 20 mg kg −1 ) significantly increased CPP score (F 3,34 =5.441, P =0.004). Values represent the mean±s.e.m. ( n =9–10). * P <0.05, ** P <0.01 compared with the saline-treated group. ANOVA, analysis of variance; CPP, conditioned place preference test; MANOVA, multivariate analysis of variance; PPI, prepulse inhibition.

Article Snippet: R -ketamine hydrochloride and S -ketamine hydrochloride were prepared by recrystallization of RS -ketamine (Ketalar, ketamine hydrochloride, Daiichi Sankyo Pharmaceutical, Tokyo, Japan) and d -(−)-tartaric acid (or l - (+)-tartaric acid), as described previously.

Techniques: Conditioned Place Preference, Inhibition

Journal: Translational Psychiatry

Article Title: R -ketamine: a rapid-onset and sustained antidepressant without psychotomimetic side effects

doi: 10.1038/tp.2015.136

Figure Lengend Snippet: Effect of R -ketamine and S -ketamine on PV-positive immunostaining in the brain. Mice were perfused 30 min after a single dose of saline, R -ketamine (10 mg kg −1 ) or S -ketamine (10 mg kg −1 ). Then PV immunohistochemistry was performed. Representative photomicrographs of PV immunohistochemistry in the PrL ( a ) and IL ( b ) of mPFC, NAc ( c ), CA1 ( d ), CA3 ( e ) and DG ( f ) of hippocampus from animals of each group. Scale bar=10 μm ( a – c ) or 200 μm ( d – f ). One-way analysis of variance revealed significant effects in the PrL (F 2,14 =7.57, P =0.016) of mPFC and DG (F 2,15 =13.834, P <0.001), but not IL of mPFC (F 2,14 =0.602, P =0.562), NAc (F 2,15 =0.019, P =0.981), CA3 (F 2,15 =0.015, P =0.985) and CA1 (F 2,14 =0.234, P =0.795). The data show the mean±s.e.m. ( n =5 or 6). * P <0.05, *** P <0.001 compared with the saline-treated group. DG, dentate gyrus; IL, infralimbic region; mPFC, medial prefrontal cortex; NAc, nucleus accumbens; N.S., not significant; PrL, prelimbic region; PV, parvalbumin.

Article Snippet: R -ketamine hydrochloride and S -ketamine hydrochloride were prepared by recrystallization of RS -ketamine (Ketalar, ketamine hydrochloride, Daiichi Sankyo Pharmaceutical, Tokyo, Japan) and d -(−)-tartaric acid (or l - (+)-tartaric acid), as described previously.

Techniques: Immunostaining, Immunohistochemistry

Journal: PLoS ONE

Article Title: Ligands Binding to Cell Surface Ganglioside GD2 Cause Src-Dependent Activation of N-Methyl-D-Aspartate Receptor Signaling and Changes in Cellular Morphology

doi: 10.1371/journal.pone.0134255

Figure Lengend Snippet: SH-SY5Y-TrkB cells were treated with mAb 3F8 (10 nM), control mIgG (10 nM) or positive control BDNF (4 nM) for the indicated times. Western blot analyses of whole cell lysates were done with specific anti-phospho-NR2B antibodies. Total NR2B levels did not change upon treatment (data not shown). (A) Anti-GD2 antibody 3F8 induces the phosphorylation of NR2B-Tyr 1472 . (B) Data from panel A were standardized to anti-β-actin or β-tubulin III levels, and were quantified versus untreated cells = 100%, n = 3–6 independent experiments each in triplicate ± SEM. ** p<0.01 versus control. (C) Pretreatment of SH-SY5Y-TrkB cells with the Src kinase inhibitor PP2 (20 μM) or the NMDA receptor antagonist ketamine (20 μM) inhibited 3F8-induced phosphorylation of NR2B-Tyr 1472 . (D) Data from panel C were standardized to anti-β-actin or β-tubulin III levels, and were quantified versus untreated cells = 100%, n = 3–6 independent experiments each in triplicate ± SEM. ** p<0.01 versus control.

Article Snippet: For pharmacological inhibition, the Src kinase inhibitor PP2 (20 μM) (Enzo Life Science), NMDA-R antagonist ketamine (Bioniche Animal Health) (20 μM) or the PKA inhibitor H-89 (Sigma) (20 ng/ml) were added to the resting cells 30 minutes before the indicated treatments.

Techniques: Control, Positive Control, Western Blot, Phospho-proteomics

Journal: PLoS ONE

Article Title: Ligands Binding to Cell Surface Ganglioside GD2 Cause Src-Dependent Activation of N-Methyl-D-Aspartate Receptor Signaling and Changes in Cellular Morphology

doi: 10.1371/journal.pone.0134255

Figure Lengend Snippet: SH-SY5Y-TrkB cells loaded with Fluo-8NW dye solution were treated with ( A ) positive control ionomycin (5 μM), or BDNF (4 nM), (B) mAb 3F8 (10 nM) ( C, D ) and (E, F) SS58 (25 μM). Negative controls buffer or vehicle DMSO, ionomycin plus EDTA to quench Ca ++ , and irrelevant mIgG (10 nM) were also tested. For pharmacological inhibition, the Src kinase inhibitor PP2 (20 μM) or the NMDA-R antagonist ketamine (20 μM) were added 30 minutes before treatment (data shown in C, D, E and F , respectively). For Ca ++ assays, the data are a representative result of 4 independent experiments each done in quadruplicate ± SD.

Article Snippet: For pharmacological inhibition, the Src kinase inhibitor PP2 (20 μM) (Enzo Life Science), NMDA-R antagonist ketamine (Bioniche Animal Health) (20 μM) or the PKA inhibitor H-89 (Sigma) (20 ng/ml) were added to the resting cells 30 minutes before the indicated treatments.

Techniques: Positive Control, Inhibition

Journal: Frontiers in Pain Research

Article Title: S -Ketamine Oral Thin Film—Part 1: Population Pharmacokinetics of S -Ketamine, S -Norketamine and S -Hydroxynorketamine

doi: 10.3389/fpain.2022.946486

Figure Lengend Snippet: Mean measured plasma concentrations following application of the 50 and 100 mg S-k etamine oral thin film (OTF): (A) S -ketamine, (B) S -norketamine, and (C) S -hydroxynorketamine. Individual concentrations are given in panels (D–F) for the 50 mg oral thin film and (G–I) for the 100 mg oral thin film. In black the results of placement below the tongue, in red buccal placement. The OTF was administered at t = 0 min for 10 min (green bars); at t = 360 min, an intravenous dose of 20 mg S -ketamine was administered over 20 min (light orange bars).

Article Snippet: To calculate the bioavailability of the OTF, six hours after placement of the oral thin film, all subjects received an intravenous S -ketamine (Ketanest-S, Pfizer, the Netherland) infusion of 20 mg over 20 min.

Techniques: Clinical Proteomics

Journal: Frontiers in Pain Research

Article Title: S -Ketamine Oral Thin Film—Part 1: Population Pharmacokinetics of S -Ketamine, S -Norketamine and S -Hydroxynorketamine

doi: 10.3389/fpain.2022.946486

Figure Lengend Snippet: Peak concentration (CMAX), time of CMAX, and area-under-the time-concentration curve (AUC) of S -ketamine, S -norketamine and S -hydroxynorketamine following 50 and 100 mg S -ketamine oral thin film (OTF).

Article Snippet: To calculate the bioavailability of the OTF, six hours after placement of the oral thin film, all subjects received an intravenous S -ketamine (Ketanest-S, Pfizer, the Netherland) infusion of 20 mg over 20 min.

Techniques: Concentration Assay

Journal: Frontiers in Pain Research

Article Title: S -Ketamine Oral Thin Film—Part 1: Population Pharmacokinetics of S -Ketamine, S -Norketamine and S -Hydroxynorketamine

doi: 10.3389/fpain.2022.946486

Figure Lengend Snippet: Adverse effects.

Article Snippet: To calculate the bioavailability of the OTF, six hours after placement of the oral thin film, all subjects received an intravenous S -ketamine (Ketanest-S, Pfizer, the Netherland) infusion of 20 mg over 20 min.

Techniques:

Journal: Frontiers in Pain Research

Article Title: S -Ketamine Oral Thin Film—Part 1: Population Pharmacokinetics of S -Ketamine, S -Norketamine and S -Hydroxynorketamine

doi: 10.3389/fpain.2022.946486

Figure Lengend Snippet: Final pharmacokinetic model. K = S -ketamine, N = S -norketamine and H = S -hydroxynorketamine. KA1 and KA2 are S -ketamine rate constants. G.I. tract = gastrointestinal tract. Cp = plasma concentration. K1, N1 and H1 are the central compartments for S -ketamine, S -norketamine and S -hydroxynorketamine, respectively. VN1 and VK1 are the volumes of the central compartments of S -ketamine and S -norketamine, respectively. Kx, Nx and Hx are the peripheral compartments for S -ketamine, S -norketamine and S -hydroxynorketamine, respectively, with x = compartment 2 or 3. CL = clearance with CLK1 and CLN1 S -ketamine and S-norketamine clearances from the central compartment toward the metabolism compartment, respectively and CLK2, CLK3, CLN2 and CLH2 intercompartmental clearances. CLH1 is the terminal S -hydroxynorketamine clearance. MTT = mean transit (or delay) time with MTTG the mean transit time from the gut to the liver.

Article Snippet: To calculate the bioavailability of the OTF, six hours after placement of the oral thin film, all subjects received an intravenous S -ketamine (Ketanest-S, Pfizer, the Netherland) infusion of 20 mg over 20 min.

Techniques: Clinical Proteomics, Concentration Assay

Journal: Frontiers in Pain Research

Article Title: S -Ketamine Oral Thin Film—Part 1: Population Pharmacokinetics of S -Ketamine, S -Norketamine and S -Hydroxynorketamine

doi: 10.3389/fpain.2022.946486

Figure Lengend Snippet: S -ketamine OTF pharmacokinetics.

Article Snippet: To calculate the bioavailability of the OTF, six hours after placement of the oral thin film, all subjects received an intravenous S -ketamine (Ketanest-S, Pfizer, the Netherland) infusion of 20 mg over 20 min.

Techniques: Drug discovery

Journal: Frontiers in Pain Research

Article Title: S -Ketamine Oral Thin Film—Part 1: Population Pharmacokinetics of S -Ketamine, S -Norketamine and S -Hydroxynorketamine

doi: 10.3389/fpain.2022.946486

Figure Lengend Snippet: Goodness-of-fit plots for S -ketamine (A–C) , S -norketamine (D–F) and S -hydroxynorketamine (G–I) . (A , D,G) : measured concentration vs. individual predicted. (B,E,H) : individual weighted residuals vs. time. (C,F,I) : Normalized discrepancy errors vs. time.

Article Snippet: To calculate the bioavailability of the OTF, six hours after placement of the oral thin film, all subjects received an intravenous S -ketamine (Ketanest-S, Pfizer, the Netherland) infusion of 20 mg over 20 min.

Techniques: Concentration Assay

Journal: Frontiers in Pain Research

Article Title: S -Ketamine Oral Thin Film—Part 1: Population Pharmacokinetics of S -Ketamine, S -Norketamine and S -Hydroxynorketamine

doi: 10.3389/fpain.2022.946486

Figure Lengend Snippet: Simulations showing the effect of changing the duration of placement of the 50 mg oral thin file in the mouth by changing both F1 (bioavailability) and D (duration of absorption) on the plasma concentrations of S -ketamine (A) , S -norketamine (B) and S -hydroxynorketamine (C) . F is a factor by which D1 is adjusted and ranges from 0.5 (red lines) to 1 (blue lines) and 2 (green lines).

Article Snippet: To calculate the bioavailability of the OTF, six hours after placement of the oral thin film, all subjects received an intravenous S -ketamine (Ketanest-S, Pfizer, the Netherland) infusion of 20 mg over 20 min.

Techniques: Clinical Proteomics

Journal: Frontiers in Pain Research

Article Title: S -Ketamine Oral Thin Film—Part 1: Population Pharmacokinetics of S -Ketamine, S -Norketamine and S -Hydroxynorketamine

doi: 10.3389/fpain.2022.946486

Figure Lengend Snippet: Simulation showing the S -ketamine (continuous green line) and S -hydroxynorketamine (broken green line) concentration profiles following a 0.5 mg/kg S -ketamine infusion, given for 40 min in a 70-kg individual. As comparator the equivalent concentrations are given following the 100 mg S -ketamine oral thin film (blue continuous = S -ketamine, and broken blue line = S -hydroxynorketamine).

Article Snippet: To calculate the bioavailability of the OTF, six hours after placement of the oral thin film, all subjects received an intravenous S -ketamine (Ketanest-S, Pfizer, the Netherland) infusion of 20 mg over 20 min.

Techniques: Concentration Assay