Journal: Cell structure and function

Article Title: The antipsychotic olanzapine induces apoptosis in insulin-secreting pancreatic β cells by blocking PERK-mediated translational attenuation.

doi: 10.1247/csf.13012

Figure Lengend Snippet: Fig. 1. Effects of olanzapine and risperidone on viability of various cells. (A) HIT-T15 cells were untreated (CON) or treated with 100 μM olanzapine (OLA) or risperidone (RIS) for 7 h, and then fixed. Nuclei were stained with DAPI (blue), whereas fragmented chromosomal DNA was labeled with FITC (green) using the TUNEL method. The number of TUNEL-positive, apoptotic cells was measured in 500–600 cells, and their percentages are presented as the average±standard error of four independent experiments at bottom. ** P<0.01. (B) HIT-T15 cells were treated with 100 μM olanzapine (OLA), 100 μM risperidone (RIS) or 2 μg/ml tunicamycin (TM) for the indicated periods. Chromosomal DNA was extracted and analyzed by 2% agarose gel electrophoresis. DNA was stained with Gel Red. The migration positions of DNA size markers are shown at left. (C) HEK293 cells were treated and analyzed as in (A). (D) HepG2 cells were treated and analyzed as in (A). (E) Twenty-four hours after transfection with the reporter plasmid pGL3-GRP78P- luc and the reference plasmid pRLUC, HIT-T15 cells were treated with 100 μM olanzapine (OLA), 100 μM risperidone (RIS) or 2 μg/ml tunicamycin (TM) for 24 h. Firefly luciferase activity relative to Renilla luciferase activity in transfected cells was determined and presented as the average±standard error of two independent experiments.

Article Snippet: Olanzapine and risperidone were purchased from Toronto Research Chemicals.

Techniques: Staining, Labeling, TUNEL Assay, Agarose Gel Electrophoresis, Migration, Transfection, Plasmid Preparation, Luciferase, Activity Assay

Journal: Cell structure and function

Article Title: The antipsychotic olanzapine induces apoptosis in insulin-secreting pancreatic β cells by blocking PERK-mediated translational attenuation.

doi: 10.1247/csf.13012

Figure Lengend Snippet: Fig. 2. Effects of olanzapine and risperidone on the PERK pathway. (A) HIT-T15 cells were treated with 100 μM olanzapine (OLA), 100 μM risperidone (RIS) or 2 μg/ml tunicamycin (TM) for the indicated periods. Cell lysates were prepared and analyzed by immuno- blotting using anti-phosphorylated PERK and anti- ribophorin antibody. (B) Cell lysates prepared as in (A) were analyzed by immunoblotting using anti-PERK, anti-eIF2α, anti-phosphorylated eIF2α and anti-GAPDH antibodies. (C) Cell lysates prepared as in (A) were analyzed by immunoblotting using anti-ATF4, anti- CHOP and anti-GAPDH antibodies. (D) HEK293 cells were treated with 100 μM olanzapine (OLA) or 100 μM risperidone (RIS) for the indicated periods. Total RNA was extracted and subjected to RT-PCR analysis to detect splicing of XBP1 mRNA. PCR products from pcDNA-XBP1(unspliced) and pcDNA-XBP1(spliced) were electrophoresed for comparison. (E) Lysates were prepared from HEK293 cells treated as in (D) and analyzed by immunoblotting using anti-phosphorylated eIF2α and anti-GAPDH antibodies.

Article Snippet: Olanzapine and risperidone were purchased from Toronto Research Chemicals.

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Comparison

Journal: Cell structure and function

Article Title: The antipsychotic olanzapine induces apoptosis in insulin-secreting pancreatic β cells by blocking PERK-mediated translational attenuation.

doi: 10.1247/csf.13012

Figure Lengend Snippet: Fig. 3. Effect of olanzapine on phosphorylation of eIF2α by PERK and GCN2. (A) HIT-T15 cells were treated with 100 μM olanzapine (OLA), 100 μM risperidone (RIS) or 2 μg/ml tunicamycin (TM) for the indicated periods. Cell lysates were prepared and analyzed by immunoblotting using anti-GADD34 antibody. (B) HIT-T15 cells were cultured in the absence of leucine [DMEM/F12(-leu)] for the indicated periods. The status of XBP1 mRNA splicing was determined as in Fig. 2D. (C) HIT-T15 cells were cultured in the presence [DMEM/F12(+leu)] or absence [DMEM/F12(–leu)] of leucine with (+) or without (–) simultaneous treatment with 100 μM olanzapine for the indicated periods. Cell lysates were prepared and analyzed by immunoblotting using anti-phosphorylated eIF2α and anti-GAPDH antibodies. (D) Five micrograms of bacterially expressed and purified eIF2α (1–182 fragment) were incubated for 30 min with various concentrations of bacterially expressed and purified cytosolic portion of mouse PERK fused to GST in the presence of 1 mM ATP as well as in the absence or presence of 100 μM olanzapine (OLA) or 100 μM risperidone (RIS), and then subjected to phos-tag gel electrophoresis together with a molecular weight marker (25 kDa). The percentages of phosphorylated eIF2α in total eIF2α were quantified and presented at bottom. (E) HIT-T15 cells were pretreated with or without 100 μM olanzapine (OLA) for 30 min, washed with PBS and then treated with 2 μg/ml tunicamycin (TM) for the indicated periods. Cell lysates were prepared and analyzed by immunoblotting using anti-eIF2α and anti-phosphorylated eIF2α antibodies.

Article Snippet: Olanzapine and risperidone were purchased from Toronto Research Chemicals.

Techniques: Phospho-proteomics, Western Blot, Cell Culture, Purification, Incubation, Nucleic Acid Electrophoresis, Molecular Weight, Marker

Journal: Cell structure and function

Article Title: The antipsychotic olanzapine induces apoptosis in insulin-secreting pancreatic β cells by blocking PERK-mediated translational attenuation.

doi: 10.1247/csf.13012

Figure Lengend Snippet: Fig. 4. Effect of cycloheximide on olanzapine-induced apoptosis. (A) HIT-T15 cells were untreated (UNT), mock-treated (CON) or treated with 100 μM olanzapine (OLA) or 100 μM risperidone (RIS) for the indicated periods, and then pulse labeled for 10 min with 35S-methionine and cysteine without medium change. Cell lysates were prepared, subjected to SDS-PAGE and autoradiographed. Total radioactivity in each sample was determined, normalized with the value of UNT and presented as the average±standard error of five independent experiments on right. * P<0.05. (B) HIT-T15 cells were treated with 100 μM olanzapine (OLA), 100 μM risperidone (RIS) or 2 μg/ml tunicamycin (TM) for the indicated periods. The status of XBP1 mRNA splicing was determined as in Fig. 2D. (C) HIT-T15 cells were treated with 100 μM olanzapine (OLA) together with (+) or without (–) 20 μg/ml cycloheximide. The status of XBP1 mRNA splicing was determined as in Fig. 2D. (D) HIT-T15 cells were pretreated with (+) or without (–) 20 μg/ml cycloheximide for 15 min, washed out and then incubated in the absence (–) or presence (+) of 100 μM olanzapine (OLA) or 2 μg/ml tunicamycin (TM) for 24 h. DNA fragmentation assay was carried out as in Fig. 1B. (E) HIT-T15 cells treated as in (D) were subjected to TUNEL assay. The number of apoptotic cells was determined as in Fig. 1A, and their percentages are presented as the average±standard error of three independent experiments on right. * P<0.05.

Article Snippet: Olanzapine and risperidone were purchased from Toronto Research Chemicals.

Techniques: Labeling, SDS Page, Radioactivity, Incubation, DNA Fragmentation Assay, TUNEL Assay

Journal: Cell structure and function

Article Title: The antipsychotic olanzapine induces apoptosis in insulin-secreting pancreatic β cells by blocking PERK-mediated translational attenuation.

doi: 10.1247/csf.13012

Figure Lengend Snippet: Fig. 5. Effect of olanzapine on insulin. (A) HIT-T15 cells were initially fed with fresh medium and then incubated in the absence (CON) or presence of 100 μM olanzapine (OLA) or 100 μM risperidone (RIS) for the indicated periods. The amount of insulin secreted into the medium was determined by ELISA and is presented after normalization with the amounts of total cellular proteins. (B) HIT-T15 cells were untreated (CON) or treated with 100 μM olanzapine (OLA) or 100 μM risperidone (RIS) for 7 h (left). HIT-T15 cells were transfected with pcDNA (vector) or pcDNA- insulin (O/E) one day before (right). Total RNA was prepared and analyzed by northern blot hybridization with a probe specific to insulin mRNA. (C) Lysates were prepared from HIT-T15 cells treated as in (B, left) or HIT- T15 cells transfected with pcDNA-insulin one day before as in (B, right) and analyzed by immunoblotting using anti-insulin antibody.

Article Snippet: Olanzapine and risperidone were purchased from Toronto Research Chemicals.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Northern Blot, Hybridization, Western Blot

Journal: Cell structure and function

Article Title: The antipsychotic olanzapine induces apoptosis in insulin-secreting pancreatic β cells by blocking PERK-mediated translational attenuation.

doi: 10.1247/csf.13012

Figure Lengend Snippet: Fig. 6. Effect of insulin knockdown on olanzapine-induced apoptosis. (A) HIT-T15 cells were transfected with vector pSUPER alone or shRNA- producing plasmid (KD-1, KD-2). Two days after transfection, total RNA was prepared and analyzed by northern blot hybridization as in Fig. 5B. Band intensity was determined and normalized with the value of vector alone and is presented below. (B) Two days after transfection of HIT-T15 cells with pSUPER or KD-1, cells were treated with 100 μM olanzapine (OLA) for the indicated periods. The status of XBP1 mRNA splicing was determined as in Fig. 2D. (C) Two days after transfection of HIT-T15 cells with pSUPER or KD-1, cells were untreated (CON) or treated with 100 μM olanzapine (OLA), 100 μM risperidone (RIS) or 2 μg/ml tunicamycin (TM) for 24 h. DNA fragmentation assay was carried out as in Fig. 1B. (D) HIT-T15 cells were treated as in (C). TUNEL assay was carried out as in Fig. 1A. The data are presented as the average±standard error of two independent experiments. ** P<0.01.

Article Snippet: Olanzapine and risperidone were purchased from Toronto Research Chemicals.

Techniques: Knockdown, Transfection, Plasmid Preparation, shRNA, Northern Blot, Hybridization, DNA Fragmentation Assay, TUNEL Assay

Journal: Journal of affective disorders

Article Title: Augmentative therapies do not potentiate the antidepressant-like effects of deep brain stimulation in rats

doi: 10.1016/j.jad.2014.03.007

Figure Lengend Snippet: Interaction between ventromedial prefrontal cortex deep brain stimulation (vmPFC DBS) and augmentative medications in the forced swim test (FST). DBS treated groups had a significant reduction in immobility scores when compared to saline treated controls. These effects, however, were not potentiated by buspirone, risperidone or pindolol. None of these drugs induced an antidepressant-like effect on their own. Data represent mean ± standard error. *p ≤ 0.05 when DBS + medication groups were compared saline treated controls. Sixteen animals were treated in each group.

Article Snippet: Drug administration The following medications and doses were selected based on their ability to potentiate the antidepressant-like effects of selective serotonin reuptake inhibitors in the FST ( Cryan et al., 2005 ; Detke et al., 1995b ; Wieland and Lucki, 1990 ): Buspirone (1 mg/kg; Tocris Bioscience), risperidone (0.1 mg/kg; Tocris Bioscience), and pindolol (6 mg/kg; Tocris Bioscience).

Techniques: Medications, Saline

Journal: Journal of affective disorders

Article Title: Augmentative therapies do not potentiate the antidepressant-like effects of deep brain stimulation in rats

doi: 10.1016/j.jad.2014.03.007

Figure Lengend Snippet: Antidepressant-like response in animals receiving ventromedial prefrontal cortex deep brain stimulation (vmPFC DBS) and augmentative medications in the forced swim test (FST). No significant differences were found when the percentage of improvement (i.e. reduction in immobility) recorded in animals from each DBS group was compared to that of rats given medications alone (i.e. DBS buspirone vs. buspirone; DBS risperidone vs. risperidone; DBS pindolol vs. pindolol). Data represent mean ± standard error. Sixteen animals were treated in each group.

Article Snippet: Drug administration The following medications and doses were selected based on their ability to potentiate the antidepressant-like effects of selective serotonin reuptake inhibitors in the FST ( Cryan et al., 2005 ; Detke et al., 1995b ; Wieland and Lucki, 1990 ): Buspirone (1 mg/kg; Tocris Bioscience), risperidone (0.1 mg/kg; Tocris Bioscience), and pindolol (6 mg/kg; Tocris Bioscience).

Techniques: Medications

Journal: Journal of analytical toxicology

Article Title: Quantification of 16 QT-prolonging Drugs and Metabolites in Human Postmortem Blood and Cardiac Tissue Using UPLC-MS-MS.

doi: 10.1093/jat/bkw014

Figure Lengend Snippet: Figure 1. Chromatography for the 16 compounds in whole blood and cardiac homogenate. The peaks are as follows: (1) O-desmethylvenlafaxine, (2) 9-hydroxyrisperidone, (3) risperidone, (4) E-10-hydroxynortriptyline, (5) venlafaxine, (6) N-desmethylvenlafaxine, (7) citalopram, (8) desmethylcitalopram, (9) EDDP, (10) Z-10-hydroxynortriptyline, (11) N-desalkylquetiapine, (12) quetiapine, (13) methadone, (14) amitriptyline, (15) nortriptyline, (16) chlorprothixene.

Article Snippet: Risperidone was obtained from EDQM (Strasbourg, France).

Techniques: Chromatography

Journal: Journal of pharmacological sciences

Article Title: Kamiuntanto increases prefrontal extracellular serotonin levels and ameliorates depression-like behaviors in mice.

doi: 10.1016/j.jphs.2018.11.008

Figure Lengend Snippet: Fig. 1. Effects of KUT on abnormal behaviors in isolation-reared mice. (A) Social encounter stimulation-induced hyperactivity was attenuated by oral administration of KUT (1000 mg/kg), fluoxetine (30 mg/kg), and risperidone (0.03 mg/kg). (B) Hyper- activity in a novel environment was attenuated by oral administration of risperidone (0.03 mg/kg), but not by KUT (300, 1000 mg/kg) or fluoxetine (30 mg/kg). (C) Increased immobility time in the forced swim test was reversed by oral administration of KUT (1000 mg/kg) and fluoxetine (30 mg/kg), but not by risperidone (0.03 mg/kg). Values are expressed as the mean ± SEM of 4e11 (A), 6e13 (B) and 6e11 (C) mice. **P < 0.01 vs. group-reared mice. yyP < 0.01 vs. vehicle-treated mice.

Article Snippet: Fluoxetine and risperidone were purchased from LKT Laboratories Inc. (St Paul, MN, USA).

Techniques: Isolation, Activity Assay