microdialysis probe 6 kd cut-off molecular weight, membrane length; cma 11  (CMA Microdialysis)

Journal: Environmental Health Perspectives

Article Title: Role of OCT3 and DRP1 in the Transport of Paraquat in Astrocytes: A Mouse Study

doi: 10.1289/EHP9505

Figure Lengend Snippet: Measures of neurodegeneration and locomotor activity in WT mice exposed to PQ 2 + with and without DRP1 knockdown. (A,B) IHC staining of TH-positive cells in substantial nigra and terminals in striatum from mice that received (A) PBS or (B) PQ 2 + -CT . (C,D) Quantitative analysis of (C) TH-positive cells in SNpc and (D) striatal optical density (OD) of TH terminals. n = 10 mice per group. (E) Quantitative analysis of Nissl staining in SNpc. n = 10 mice per group. (F,G) In vivo microdialysis followed by HPLC tests for striatal DA release in mice that received (F) PBS or (G) PQ 2 + -CT . To evoke depolarization-induced release of DA, 240 nmol KCl in isotonic aCSF was perfused over a 15-min period (shaded box). (H) Peak areas under curves were analyzed. n = 5 mice per group. (I) The differences in stride length of mice before and after PQ 2 + -CT in mice exposed to either empty AAV control, neuronal DRP knockdown by AAV, astrocyte DRP1 knockdown by AAV, or non-selective DRP1 knockdown using mdivi-1. n = 10 mice per group (J) Rotarod tests of the mice in different groups. The value was measured before and after PQ 2 + -CT , and the differences were calculated by “after” minus “before.” Data are shown as mean ± SD . Two-way ANOVAs were followed by the Bonferroni multiple comparison test. *, p < 0.05 ; **, p < 0.01 ; *** /### , p < 0.001 ; ns, no significance. The numeric data are shown in Excel Table S3. In two-way ANOVAs, “*” was used to present the statistical difference between groups both treated by PQ 2 + , and “#” was used to present the statistical difference between the PBS- and the PQ 2 + -treated groups. Note: AAV, adeno-associated virus; aCSF, artificial cerebrospinal fluid; ANOVA, analysis of variance; Astro-DRP1 KD, DRP1 knockdown in astrocytes by AAV with gfaABC1D promoter; AUC, area under the curve; CT, chronic treatment; DA, dopamine; DRP1, dynamic related protein-1; HPLC, high-performance liquid chromatography; IHC, immunohistochemistry; KCl, potassium chloride; KD, knockdown; mdivi-1, mitochondrial division inhibitor-1; Neuro-DRP1 KD, DRP1 knockdown in neuron by AAV with hsyn promoter; OCT3;organic cation transporter-3; PBS, phosphate-buffered saline; PQ, paraquat; Scramble, empty AAV for control; SD, standard deviation; SNpc, substantia nigra pars compacta; TH, Tyrosine hydroxylase.

Article Snippet: The next day, a microdialysis probe (CMA8 with 2 mm membrane length; CMA; Harvard Bioscience, Inc.) was inserted into the guide cannula and connected to a low-torque dual channel swivel (Instech Laboratories, Inc.).

Techniques: Activity Assay, Knockdown, Immunohistochemistry, Staining, In Vivo, Control, Comparison, Virus, High Performance Liquid Chromatography, Saline, Standard Deviation

Journal: Environmental Health Perspectives

Article Title: Role of OCT3 and DRP1 in the Transport of Paraquat in Astrocytes: A Mouse Study

doi: 10.1289/EHP9505

Figure Lengend Snippet: Measures of neurodegeneration and locomotor activity in O c t 3 − / − mice exposed to PQ 2 + with and without DRP1 knockdown. (A,B) IHC staining of TH-positive cells in SNpc and striatum terminals in O c t 3 − / − mice that received (A) PBS and (B) PQ 2 + -CT . (C, D) Quantitative analysis of (C) TH-positive cells in SNpc and (D) striatal optical density (OD) of TH terminals. n = 10 mice per group. (E) Quantitative analysis of Nissl staining in SNpc. n = 10 mice per group. (F,G) In vivo microdialysis followed by HPLC tests for striatal DA release from the brains of mice that received (F) PBS or (G) PQ 2 + -CT . To evoke depolarization-induced release of DA, 240 nmol KCl in isotonic aCSF was delivered through the probe over a 15-min period (shaded box). (H) Peak areas under curves were analyzed. n = 5 mice per group. (I) The differences in stride length of mice before and after PQ 2 + -CT in mice exposed to either empty AAV control, neuronal DRP knockdown by AAV, astrocyte DRP1 knockdown by AAV, or non-selective DRP1 knockdown using mdivi-1. n = 10 mice per group (J) Rotarod tests of the mice in different groups. The value was measured before and after PQ 2 + -CT , and the differences were calculated by “after” minus “before.” n = 10 mice per group. Data are shown as mean ± SD . Two-way ANOVAs were followed by the Bonferroni multiple comparison test. *, p < 0.05 ; **, p < 0.01 *** /### , p < 0.001 . The numeric data are shown in Excel Table S4. In two-way ANOVAs, “*” was used to present the statistical difference between groups both treated by PQ 2 + , and “#” was used to present the statistical difference between PBS-treated group and PQ 2 + treated group. Note: AAV, adeno-associated virus; aCSF, artificial cerebrospinal fluid; ANOVA, analysis of variance; Astro-DRP1 KD, DRP1 knockdown in astrocytes by AAV with gfaABC1D promoter; AUC, area under the curve; CT, chronic treatment; DA, dopamine; DRP1, dynamic related protein-1; HPLC, high-performance liquid chromatography; IHC, immunohistochemistry; KCl, potassium chloride; KD, knockdown; Neuro-DRP1 KD, DRP1 knockdown in neuron by AAV with hsyn promoter; mdivi-1, mitochondrial division inhibitor-1; OCT3, organic cation transporter-3; PBS, phosphate-buffered saline; PQ, paraquat; Scramble, empty AAV for control; SD, standard deviation; SNpc, substantia nigra pars compacta; TH, Tyrosine hydroxylase.

Article Snippet: The next day, a microdialysis probe (CMA8 with 2 mm membrane length; CMA; Harvard Bioscience, Inc.) was inserted into the guide cannula and connected to a low-torque dual channel swivel (Instech Laboratories, Inc.).

Techniques: Activity Assay, Knockdown, Immunohistochemistry, Staining, In Vivo, Control, Comparison, Virus, High Performance Liquid Chromatography, Saline, Standard Deviation

Journal: Environmental Health Perspectives

Article Title: Role of OCT3 and DRP1 in the Transport of Paraquat in Astrocytes: A Mouse Study

doi: 10.1289/EHP9505

Figure Lengend Snippet: Evaluations of PQ 2 + or PQ + clearance in the brains of mice and in cultured astrocytes with or without DRP1 knockdown. (A,B) In vivo extracellular residue of the loading PQ 2 + ( PQ 2 + -LT ) was detected by microdialysis followed by HPLC in WT (A) and O c t 3 − / − (B) mice treated with either empty AAV control, neuronal DRP knockdown by AAV, astrocyte DRP1 knockdown by AAV, or non-selective DRP1 knockdown using mdivi-1. n = 5 mice per group. One-way ANOVA was followed by the Bonferroni multiple comparison test. Data are shown as mean ± SD . (C,D) PQ 2 + or PQ + uptake assays of GL261 astrocytes without or with SDT. PQ 2 + / PQ + concentration in the extracellular culture medium ranged from 0 to 800 μ M ( x -axis), and the uptake level of PQ 2 + / PQ + into astrocytes were detected in the cell lysate ( y -axis). Drp-1 siRNA was used to knockdown DRP1 in GL261 astrocytes. The average area under the curves were calculated and used for statistical analysis. n = 6 independent experiments per group. One-way ANOVA was followed by the Bonferroni multiple comparison test. Data are shown as mean ± SEM . **, p < 0.01 ; ***, p < 0.001 . The numeric data are shown in Excel Table S5. Note: AAV, adeno-associated virus; ANOVA, analysis of variance; Astro-DRP1 KD, DRP1 knockdown in astrocytes by AAV with gfaABC1D promoter; DRP1, dynamic related protein-1; HPLC, high-performance liquid chromatography; LT, loading treatment; mdivi-1, mitochondrial division inhibitor-1; Neuro-DRP1 KD, DRP1 knockdown in neuron by AAV with hsyn promoter; oct3, organic cation transporter-3; PQ, paraquat; Scramble, empty AAV for control; SD, standard deviation; SDT, sodium dithionite; SEM, standard error of the mean; siRNA, small interfering RNA; WT, wild-type.

Article Snippet: The next day, a microdialysis probe (CMA8 with 2 mm membrane length; CMA; Harvard Bioscience, Inc.) was inserted into the guide cannula and connected to a low-torque dual channel swivel (Instech Laboratories, Inc.).

Techniques: Cell Culture, Knockdown, In Vivo, Residue, Control, Comparison, Concentration Assay, Virus, High Performance Liquid Chromatography, Standard Deviation, Small Interfering RNA

Journal: Frontiers in Psychiatry

Article Title: Effects of Lisdexamfetamine, a Prodrug of D-Amphetamine, on Locomotion, Spatial Cognitive Processing and Neurochemical Profiles in Rats: A Comparison With Immediate-Release Amphetamine

doi: 10.3389/fpsyt.2022.885574

Figure Lengend Snippet: The effects of d-amphetamine and lisdexamfetamine on catecholamine neurotransmitters and the metabolites in mPFC. (A) The implantation location of the microdialysis guide cannula. (B) The effects of d-amphetamine and lisdexamfetamine on DA efflux. (C) The effects of d-amphetamine and lisdexamfetamine on DOPAC efflux. (D) The effects of d-amphetamine and lisdexamfetamine on HVA efflux. Amp: d-amphetamine; Lis: lisdexamfetamine. Data are presented by mean ± SEM, n = 5 in each group.

Article Snippet: A microdialysis probe (EICOM A-I: 0.22-mm OD, 4-mm membrane length with 50 kDa cutoff) was inserted into the guide cannula of awake rats.

Techniques: