Journal: Frontiers in Neuroanatomy

Article Title: Dopamine in the auditory brainstem and midbrain: co-localization with amino acid neurotransmitters and gene expression following cochlear trauma

doi: 10.3389/fnana.2015.00088

Figure Lengend Snippet: Antibodies.

Article Snippet: Sections containing the CN and IC were incubated for 48 h at 4°C with the primary antibody (TH rabbit polyclonal antibody 1:1500–1:2000, GAD67 mouse monoclonal antibody 1:1000, GLYT2 guinea pig polyclonal antibody 1:1000, VGLUT1 guinea pig polyclonal antibody 1:1000, VGLUT3 guinea pig polyclonal antibody 1:1000 (Chemicon, Temecula, CA, USA), VGLUT3 mouse monoclonal antibody 1:100 (NeuroMab, Davis, CA, USA), DBH mouse monoclonal antibody 1:400, and PNMT mouse monoclonal antibody 1:2000 (Chemicon, Temecula, CA, USA).

Techniques: Recombinant, Purification

Journal: Frontiers in Neuroanatomy

Article Title: Dopamine in the auditory brainstem and midbrain: co-localization with amino acid neurotransmitters and gene expression following cochlear trauma

doi: 10.3389/fnana.2015.00088

Figure Lengend Snippet: Dopamine and AAN related genes are differentially expressed in the inferior colliculus (IC). Changes were assessed in DA related (A) , inhibitory (B,C) , and excitatory (D) AAN related gene expression in the IC 3 days and 2 months following bilateral cochlear ablation. TH expression decreased significantly in the IC at both 3 days (31%) and 2 months (38%). In addition to the DA receptor subunit, Drd1A (15%-3 days; 23%-2 months; A ), four AAN related genes showed the same pattern of expression as TH: significant decreases at both time points. Of these, two were related to excitatory neurotransmission (VGLUT2: 60%-3 days and 32%-2 months; VGLUT3: 62%-3 days and 52%-2 months) and two related to inhibitory neurotransmission (GABAtp: 15%-3 days and 22%-2 months; GAD67: 29%-3 days and 36%-2 months). Interestingly, seven genes were significantly decreased (Drd2-30%; Drd5:-18%; GLYT1- 31%; GLYT2- 45%; GAD65- 22%; GABArb1- 26%) only at the 2 months time point (A,C,D) . Error bars: SD; asterisks: p ≤ 0.05.

Article Snippet: Sections containing the CN and IC were incubated for 48 h at 4°C with the primary antibody (TH rabbit polyclonal antibody 1:1500–1:2000, GAD67 mouse monoclonal antibody 1:1000, GLYT2 guinea pig polyclonal antibody 1:1000, VGLUT1 guinea pig polyclonal antibody 1:1000, VGLUT3 guinea pig polyclonal antibody 1:1000 (Chemicon, Temecula, CA, USA), VGLUT3 mouse monoclonal antibody 1:100 (NeuroMab, Davis, CA, USA), DBH mouse monoclonal antibody 1:400, and PNMT mouse monoclonal antibody 1:2000 (Chemicon, Temecula, CA, USA).

Techniques: Gene Expression, Expressing

Journal: Frontiers in Neuroanatomy

Article Title: Dopamine in the auditory brainstem and midbrain: co-localization with amino acid neurotransmitters and gene expression following cochlear trauma

doi: 10.3389/fnana.2015.00088

Figure Lengend Snippet: In the IC VGLUT3 is expressed and produced. Gene expression for VGLUT3 in the IC was demonstrated using PCR (A) , with a predicted band of approximately 1.9 kb. Western blotting for VGLUT3 protein in the IC resulted in a single band ∼65 kDa (B) .

Article Snippet: Sections containing the CN and IC were incubated for 48 h at 4°C with the primary antibody (TH rabbit polyclonal antibody 1:1500–1:2000, GAD67 mouse monoclonal antibody 1:1000, GLYT2 guinea pig polyclonal antibody 1:1000, VGLUT1 guinea pig polyclonal antibody 1:1000, VGLUT3 guinea pig polyclonal antibody 1:1000 (Chemicon, Temecula, CA, USA), VGLUT3 mouse monoclonal antibody 1:100 (NeuroMab, Davis, CA, USA), DBH mouse monoclonal antibody 1:400, and PNMT mouse monoclonal antibody 1:2000 (Chemicon, Temecula, CA, USA).

Techniques: Produced, Gene Expression, Western Blot

Journal: Frontiers in Neuroanatomy

Article Title: Dopamine in the auditory brainstem and midbrain: co-localization with amino acid neurotransmitters and gene expression following cochlear trauma

doi: 10.3389/fnana.2015.00088

Figure Lengend Snippet: In the IC VGLUT3 is distributed throughout each subdivision. Although some labeling is observed in terminals, the majority of labeling for VGLUT3 is contained within somata and dendrites (A–C) . In the CNIC large and small multipolar neurons with round somata and proximal dendrites are labeled for VGLUT3 (A,A’) . While prominent somatic labeling for VGLUT3 is found in the ECIC (B,B’) and DCIC (C,C’) localized to small elongated neurons with little to no labeling of proximal dendrites (arrowheads) as well as within larger neurons, including proximal dendrites (arrows). CNIC – central nucleus of the IC; ECIC – external cortex of the IC; DCIC – dorsal cortex of the IC; Scale bars: 20 μm.

Article Snippet: Sections containing the CN and IC were incubated for 48 h at 4°C with the primary antibody (TH rabbit polyclonal antibody 1:1500–1:2000, GAD67 mouse monoclonal antibody 1:1000, GLYT2 guinea pig polyclonal antibody 1:1000, VGLUT1 guinea pig polyclonal antibody 1:1000, VGLUT3 guinea pig polyclonal antibody 1:1000 (Chemicon, Temecula, CA, USA), VGLUT3 mouse monoclonal antibody 1:100 (NeuroMab, Davis, CA, USA), DBH mouse monoclonal antibody 1:400, and PNMT mouse monoclonal antibody 1:2000 (Chemicon, Temecula, CA, USA).

Techniques: Labeling

Journal: Frontiers in Neuroanatomy

Article Title: Dopamine in the auditory brainstem and midbrain: co-localization with amino acid neurotransmitters and gene expression following cochlear trauma

doi: 10.3389/fnana.2015.00088

Figure Lengend Snippet: In the DCIC VGLUT3 (green) and TH (red) are co-localized in somata and dendrites (A–F). In the DCIC double labeling for VGLUT3 and TH was primarily observed in somata and dendrites. Arrows indicate somatic and dendritic labeling for VGLUT3 and TH. The asterisks indicate neurons in the DCIC labeled only for VGLUT3. DCIC – dorsal cortex of the IC; Scale bars: 10 μm.

Article Snippet: Sections containing the CN and IC were incubated for 48 h at 4°C with the primary antibody (TH rabbit polyclonal antibody 1:1500–1:2000, GAD67 mouse monoclonal antibody 1:1000, GLYT2 guinea pig polyclonal antibody 1:1000, VGLUT1 guinea pig polyclonal antibody 1:1000, VGLUT3 guinea pig polyclonal antibody 1:1000 (Chemicon, Temecula, CA, USA), VGLUT3 mouse monoclonal antibody 1:100 (NeuroMab, Davis, CA, USA), DBH mouse monoclonal antibody 1:400, and PNMT mouse monoclonal antibody 1:2000 (Chemicon, Temecula, CA, USA).

Techniques: Labeling

Journal: Frontiers in Neuroanatomy

Article Title: Dopamine in the auditory brainstem and midbrain: co-localization with amino acid neurotransmitters and gene expression following cochlear trauma

doi: 10.3389/fnana.2015.00088

Figure Lengend Snippet: Gene expression and immunocytochemistry comparison in the cochlear nucleus (CN).

Article Snippet: Sections containing the CN and IC were incubated for 48 h at 4°C with the primary antibody (TH rabbit polyclonal antibody 1:1500–1:2000, GAD67 mouse monoclonal antibody 1:1000, GLYT2 guinea pig polyclonal antibody 1:1000, VGLUT1 guinea pig polyclonal antibody 1:1000, VGLUT3 guinea pig polyclonal antibody 1:1000 (Chemicon, Temecula, CA, USA), VGLUT3 mouse monoclonal antibody 1:100 (NeuroMab, Davis, CA, USA), DBH mouse monoclonal antibody 1:400, and PNMT mouse monoclonal antibody 1:2000 (Chemicon, Temecula, CA, USA).

Techniques: Gene Expression, Immunocytochemistry, Comparison

Journal: Frontiers in Neuroanatomy

Article Title: Dopamine in the auditory brainstem and midbrain: co-localization with amino acid neurotransmitters and gene expression following cochlear trauma

doi: 10.3389/fnana.2015.00088

Figure Lengend Snippet: Gene expression and immunocytochemistry summary in the inferior colliculus (IC).

Article Snippet: Sections containing the CN and IC were incubated for 48 h at 4°C with the primary antibody (TH rabbit polyclonal antibody 1:1500–1:2000, GAD67 mouse monoclonal antibody 1:1000, GLYT2 guinea pig polyclonal antibody 1:1000, VGLUT1 guinea pig polyclonal antibody 1:1000, VGLUT3 guinea pig polyclonal antibody 1:1000 (Chemicon, Temecula, CA, USA), VGLUT3 mouse monoclonal antibody 1:100 (NeuroMab, Davis, CA, USA), DBH mouse monoclonal antibody 1:400, and PNMT mouse monoclonal antibody 1:2000 (Chemicon, Temecula, CA, USA).

Techniques: Gene Expression, Immunocytochemistry