Journal:

Article Title: Induction of Striatal Pre- and Postsynaptic Damage by Methamphetamine Requires the Dopamine Receptors

doi: 10.1002/syn.20185

Figure Lengend Snippet: D1R antagonist protects against METH-induced cell death. Pretreatment with D1R antagonist, SCH 23390 (SCH [0.05 mg/kg, 0.1 mg/kg, 0.25 mg/kg, 0.5 mg/kg, or 1 mg/kg of body weight]), 30 min before METH (30 mg/kg of body weight, i.p.) abrogated TUNEL-staining in the mouse striata in a dose-dependent fashion. Antagonist alone had no effect. (A) Epifluorescent images of TUNEL-stained mouse striata. Scale bar = 50 µm. (B) Figure shows mean ± SEM percentage of TUNEL-positive staining relative to total neuronal cell counts done previously (data not shown) with neuron-specific nuclear protein NeuN. *P < 0.01 compared with vehicle + saline, !P < 0.005 compared with vehicle + METH 30 mg/kg. No significance is found between regions of the same experimental group.

Article Snippet: Images were taken with a Nikon Eclipse E400 epifluorescent scope attached to a Hamamatsu digital camera C4742-95 using FITC filters.

Techniques: TUNEL Assay, Staining, Saline

Journal:

Article Title: Induction of Striatal Pre- and Postsynaptic Damage by Methamphetamine Requires the Dopamine Receptors

doi: 10.1002/syn.20185

Figure Lengend Snippet: D2R antagonist protects against METH-induced cell death. Pretreatment with D2R antagonist, raclopride (RAC [0.025 mg/kg, 0.5 mg/kg, or 1 mg/kg of body weight]), 30 min before METH (30 mg/kg of body weight, i.p.) attenuated TUNEL-staining in the mouse striata in a dose-dependent fashion. Antagonist alone had no effect. (A) Epifluorescent images of TUNEL-stained mouse striata. Scale bar = 50 µm. (B) Figure shows mean ± SEM percentage of TUNEL-positive staining relative to total neuronal cell counts done previously (data not shown) with neuron-specific nuclear protein NeuN. *P < 0.01 compared with vehicle + saline, !P < 0.05 compared with vehicle + METH. No significance is found between regions of the same experimental group.

Article Snippet: Images were taken with a Nikon Eclipse E400 epifluorescent scope attached to a Hamamatsu digital camera C4742-95 using FITC filters.

Techniques: TUNEL Assay, Staining, Saline

Journal:

Article Title: Induction of Striatal Pre- and Postsynaptic Damage by Methamphetamine Requires the Dopamine Receptors

doi: 10.1002/syn.20185

Figure Lengend Snippet: D1R and D2R antagonists protect against METH-induced reactive astrocytosis. (A) Figure shows epifluorescent images of GFAP-stained mouse striata. Arrows point to astrocytes stained with anti-GFAP conjugated to Cy3. Scale bar = 10 µm. (B) Pretreatment with D1R (SCH 23390 [0.1 mg/kg of body weight] i.p.) or D2R antagonist, raclopride (RAC [1 mg/kg of body weight] i.p.), 30 min before METH (30 mg/kg of body weight, i.p.) exposure attenuated the induction of reactive astrocysts in the mouse striata. Antagonists alone had no effect. *P < 0.0001 compared with vehicle + saline, !P < 0.0001 compared with vehicle + METH 30 mg/kg. No significance is found between regions. n = 6 per group.

Article Snippet: Images were taken with a Nikon Eclipse E400 epifluorescent scope attached to a Hamamatsu digital camera C4742-95 using FITC filters.

Techniques: Staining, Saline

Journal: Viruses

Article Title: PluriBAC: A Versatile Baculovirus-Based Modular System to Express Heterologous Genes in Different Biotechnological Platforms

doi: 10.3390/v15101984

Figure Lengend Snippet: Per os infection of Spodoptera frugiperda larvae with PluriBAC-derived recAcMNPV. ( A ) A representation of the pathway followed in PluriBAC scheme to construct the virus, and the final pGGL2Bac donor vector obtained to generate the recAcMNPV. CMV represents cytomegalovirus immediate-early gene 1 (CMV-IE1) promoter and SV40 refers to simian virus 40 polyadenylation signal (SV40 polyA). ( B ) PCR screening of the insert: CMV + dTomato + SV40 (1650 bp) and fluorescent infected High Five ™ cells. ( C ) Experimental protocol (created with BioRender.com , accessed on 14 September 2023) and epifluorescence microscopy of infected and uninfected Spodoptera frugiperda larvae with recAcMNPV.

Article Snippet: Recombinant protein expression was verified with epifluorescence microscopy Nikon Eclipse Ti-S (Nikon Instruments Inc. Melville, NY, USA) and photographed with Nikon DS-Ri2 camera.

Techniques: Infection, Derivative Assay, Construct, Virus, Plasmid Preparation, Epifluorescence Microscopy

Journal: Viruses

Article Title: PluriBAC: A Versatile Baculovirus-Based Modular System to Express Heterologous Genes in Different Biotechnological Platforms

doi: 10.3390/v15101984

Figure Lengend Snippet: CNS cells transduction with PluriBAC-derived recAcMNPV. ( A ) A representation of the pathway followed in PluriBAC workflow to generate the virus, where CMV stands for cytomegalovirus immediate-early gene 1 (CMV-IE1) promoter and SV40, for simian virus 40 polyA signal (SV40 polyA). ( B ) PCR screening of the expression cassette (1485 bp) and fluorescent High Five ™ cells infected with the recAcMNPV obtained. ( C ) Experimental protocol (created with BioRender.com , accessed on 14 September 2023) and epifluorescence microscopy of mice astrocytes and patient-derived GBM cells transduced with recombinant baculovirus. Citrine expression was observed 48 h after transduction. ( D ) Experimental protocol (created with BioRender.com , accessed on 14 September 2023) of the in vivo assay and citrine expression observation of mice normal astrocytes. C57Bl/6 mice were inoculated into the right striatum with BV (5 × 10 8 PFU). After 7 days, the expression of the reporter protein citrine (green) was evaluated with fluorescent microscopy. ( E ) Experimental protocol (created with BioRender.com , accessed on 14 September 2023) of the in vivo assay and citrine expression observation of mouse neoplastic astrocytes. C57Bl/6 mice were inoculated with mouse glioma neurospheres (NS) into the brain; 3 weeks later, tumors were injected with BV (5 × 10 8 PFU) and mice were euthanized after 7 days. The expression of the reporter protein citrine (green) in the tumor was evaluated with fluorescent microscopy. In all cases, DAPI was used for nuclear staining.

Article Snippet: Recombinant protein expression was verified with epifluorescence microscopy Nikon Eclipse Ti-S (Nikon Instruments Inc. Melville, NY, USA) and photographed with Nikon DS-Ri2 camera.

Techniques: Transduction, Derivative Assay, Virus, Expressing, Infection, Epifluorescence Microscopy, Recombinant, In Vivo, Microscopy, Injection, Staining

Journal: Viruses

Article Title: PluriBAC: A Versatile Baculovirus-Based Modular System to Express Heterologous Genes in Different Biotechnological Platforms

doi: 10.3390/v15101984

Figure Lengend Snippet: Characterization of the induction of pMDR1 with cisplatin using a PluriBAC-derived AcMNPV encoding dTomato fluorescent protein. ( A ) A representation of the pathway followed in PluriBAC workflow to generate the baculovirus. SV40: simian virus 40 polyA signal. ( B ) PCR screening of the inserts: pMDR1 (1393 bp), dTomato + SV40 (864 bp), and obtained fluorescent High Five™ cells infected with the recAcMNPV. ( C ) Experimental protocol (created with BioRender.com , accessed on 14 September 2023) and epifluorescence microscopy of transduced A549 cells treated with cisplatin at different times (for practical purposes, only two cisplatin concentrations are represented). Percentages of positive cells were calculated in the different cisplatin concentration assayed at 72 h post-transduction. A Student’s t test was performed between groups, * p < 0.001.

Article Snippet: Recombinant protein expression was verified with epifluorescence microscopy Nikon Eclipse Ti-S (Nikon Instruments Inc. Melville, NY, USA) and photographed with Nikon DS-Ri2 camera.

Techniques: Derivative Assay, Virus, Infection, Epifluorescence Microscopy, Concentration Assay, Transduction

Journal: Viruses

Article Title: PluriBAC: A Versatile Baculovirus-Based Modular System to Express Heterologous Genes in Different Biotechnological Platforms

doi: 10.3390/v15101984

Figure Lengend Snippet: Multiple insert PluriBAC-derived recBV constructed using Bac-to-Bac ® expression system. ( A ) A representation of the pathway followed in PluriBAC scheme to construct the virus, and the final pGGL1-Bac donor vector obtained to generate the recAcMNPV with Bac-toBac TM expression system. Tn7L and Tn7R: transposition sites, pSpect: Spectinomycin resistance promoter, Spect R: Spectinomycin resistance, pSfU6: small nucleolar U6 Spodoptera frugiperda promoter, ncRNA: synthetic non-coding RNA, CMV: cytomegalovirus immediate-early gene 1 (CMV-IE1) promoter, SV40: simian virus 40 polyA signal. ( B ) PCR screening of the multiple inserts. Insert 1: Tn7R (260bp), insert 3: pSfU6 + ncRNA + term (520 bp), insert 4: CMV (507 bp), insert 5: Tn7L (211 bp). Spectinomycin resistance and its promoter were screened by using Spectinomycin for clonal selection. ( C ) Experimental protocol (created with BioRender.com , accessed on 14 September 2023) of recAcMNPV generation using Bac-to-Bac TM system. Blue–white transposition screening in E. coli DH 10Bac TM cells and epifluorescence microscopy of infected High Five TM cells with recBV. Yellow arrows show some of the white colonies obtained.

Article Snippet: Recombinant protein expression was verified with epifluorescence microscopy Nikon Eclipse Ti-S (Nikon Instruments Inc. Melville, NY, USA) and photographed with Nikon DS-Ri2 camera.

Techniques: Derivative Assay, Construct, Expressing, Virus, Plasmid Preparation, Single Photon Emission Computed Tomography, Selection, Epifluorescence Microscopy, Infection