Journal: Neuropeptides

Article Title: Anti-CGRP antibodies block CGRP-induced diarrhea in mice

doi: 10.1016/j.npep.2016.11.004

Figure Lengend Snippet: Prevention of CGRP-induced diarrhea by anti-CGRP antibodies.

Article Snippet: Two humanized anti-CGRP antibodies (Ab3 and Ab6), vehicle, and control antibody (anti-digoxin, isotype human IgG1 lacking N-glycosylation) were provided by Alder Biopharmaceuticals Inc. (Bothell, WA).

Techniques:

Journal: Neuropeptides

Article Title: Anti-CGRP antibodies block CGRP-induced diarrhea in mice

doi: 10.1016/j.npep.2016.11.004

Figure Lengend Snippet: Anti-CGRP antibody blocks CGRP-induced diarrhea in mice. CGRP-induced diarrhea was completely blocked by ip anti-CGRP (anti-C) (30 mg/kg, Ab3) antibody in both wildtype mice given human CGRP and nestin/hRAMP1 mice given rat CGRP, which was significantly less (*p < 0.001) than mice pre-treated by ip antibody vehicle (Veh) or ip control (Con) antibody (n = 2 trials). Anti-CGRP antibody also blocked subsequent CGRP-induced diarrhea at the 0.05 mg/kg ip human CGRP dose (n = 1 trial). All trials included 9–12 mice per trial except ip Veh + icv CGRP trials, which included 5–10 mice per trial. Means ± range are given.

Article Snippet: Two humanized anti-CGRP antibodies (Ab3 and Ab6), vehicle, and control antibody (anti-digoxin, isotype human IgG1 lacking N-glycosylation) were provided by Alder Biopharmaceuticals Inc. (Bothell, WA).

Techniques: Control

Journal: Neuropeptides

Article Title: Anti-CGRP antibodies block CGRP-induced diarrhea in mice

doi: 10.1016/j.npep.2016.11.004

Figure Lengend Snippet: Anti-CGRP antibody reduced CGRP-induced excrement. Mass of urine and non-formed stool excreted by individual nestin/hRAMP1 mice following icv CGRP or vehicle. Prophylactic ip administration of anti-CGRP antibody (50 mg/kg, Ab6) 24 h prior prevented rat CGRP-induced diarrhea as compared to control antibody (*p < 0.0001). Both antibody and CGRP injection had significant effects (F1,44 = 38.44, p < 0.0001; F1,44 = 80.88, p < 0.0001, respectively). For all groups, n = 12. Data for individual mice with means ± SEM are given.

Article Snippet: Two humanized anti-CGRP antibodies (Ab3 and Ab6), vehicle, and control antibody (anti-digoxin, isotype human IgG1 lacking N-glycosylation) were provided by Alder Biopharmaceuticals Inc. (Bothell, WA).

Techniques: Control, Injection

Journal: Molecular Pain

Article Title: Sigma-1 receptor expression in sensory neurons and the effect of painful peripheral nerve injury

doi: 10.1186/1744-8069-9-47

Figure Lengend Snippet: σ1 Receptor (σ1R) in various Control dorsal root ganglion (DRG) cell populations. (A) Staining for σ1R was uniformly distributed within cytoplasmic areas of cells identified as neuronal profiles through co-staining with NeuN. No specific σ1R staining was found in neuronal nuclei. (B) σ1R was also found in cellular components identified as satellite glial cells by their expression of glutamine synthase (GS). Identically placed arrows in the left and middle panels indicate cellular areas clearly identifiable as satellite glial cell cytoplasm that expresses σ1R. (C) Classification of control DRG neurons by size (<700, 700-1300 and >1300 μm 3 ) showed greater σ1R intensity (arbitrary units, recorded using standardized image acquisition) in large neurons compared to others. (D) σ1Rs expression is compared in neuronal subtypes identified by their expression of calcitonin-gene related peptide (CGRP), isolectin B-4 (IB4), and neurofilament-200 (NF-200). (E) Higher fluorescent intensity for σ1Rs was found in the subpopulation of control neurons expressing NF-200. Mean ± SEM; number in bars represents the sample size; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: After four washes with PBST, sections were incubated with Alexa Fluor 568 Goat anti-rabbit antibody (1:500; Invitrogen, Camarillo, CA) for 2 h. To determine co-localization of σ1R with the neuron-specific nuclear protein (NeuN), neurofilament 200 (NF-200), CGRP, and glutamine synthetase, the sections were washed four times with PBST followed by incubated with anti-NeuN mouse monoclonal antibody (1:100, Millipore, Billerica, MA, catalog number MAB377, Lot No. LV1616015, monoclonal antibody raised against purified cell nuclei from mouse brain) [ ], anti-NF-200 mouse monoclonal antibody (1:1000, Abcam, Cambridge, UK, catalog number ab28029, monoclonal antibody raised against a non phosphorylated epitope from 200kD Neurofilament Heavy of most mammalian species) [ ], anti-CGRP mouse polyclonal antibody (1:50, Santa Cruz Biotechnology, Santa Cruz, CA, catalog number SC-28920, Lot No. H1407, epitope corresponding to amino acid 81-128 mapping at the C-terminus of CGRP of human origin) [ ], or anti-glutamine synthetase rabbit polyclonal antibody (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, catalog number SC-6640, Lot No. H1407, polyclonal affinity purified antibody raised against a peptide mapping at the C-terminus of glutamine synthetase of human origin) [ ] for 2 h. Following four washes with PBST, sections were incubated with Alexa Fluor 488 goat anti-mouse antibody or Alexa Fluor 488 goat anti-rabbit antibody (1:1000; Molecular Probes, Camarillo, CA) for 1 h. To determine co-localization of σ1R with IB4, sections were incubated with Alexa Fluor 488 conjugate IB 4 (1:50, Invitrogen, Camarillo, CA, catalog number I21411, Lot No. 743633,) [ ] after four washes with PBST.

Techniques: Control, Staining, Expressing

Journal: Molecular Pain

Article Title: Sigma-1 receptor expression in sensory neurons and the effect of painful peripheral nerve injury

doi: 10.1186/1744-8069-9-47

Figure Lengend Snippet: Effect of injury on σ1 receptor (σ1R) expression in sensory neurons identified by calcitonin-gene related peptide (CGRP) staining. (A) Staining of σ1R was distributed within cytoplasmic areas of all CGRP co-expressing neurons. σ1R immunofluorescence intensity was decreased in both spinal nerve ligation (SNL) 4th lumbar (L4) and 5th lumbar (L5) neurons compared with Control. (B) Summary data showed decrease of intensity of σ1R staining in CGRP-positive SNL L4 and L5 neurons. Mean ± SEM; number in bars represents the sample size; *** p < 0.001.

Article Snippet: After four washes with PBST, sections were incubated with Alexa Fluor 568 Goat anti-rabbit antibody (1:500; Invitrogen, Camarillo, CA) for 2 h. To determine co-localization of σ1R with the neuron-specific nuclear protein (NeuN), neurofilament 200 (NF-200), CGRP, and glutamine synthetase, the sections were washed four times with PBST followed by incubated with anti-NeuN mouse monoclonal antibody (1:100, Millipore, Billerica, MA, catalog number MAB377, Lot No. LV1616015, monoclonal antibody raised against purified cell nuclei from mouse brain) [ ], anti-NF-200 mouse monoclonal antibody (1:1000, Abcam, Cambridge, UK, catalog number ab28029, monoclonal antibody raised against a non phosphorylated epitope from 200kD Neurofilament Heavy of most mammalian species) [ ], anti-CGRP mouse polyclonal antibody (1:50, Santa Cruz Biotechnology, Santa Cruz, CA, catalog number SC-28920, Lot No. H1407, epitope corresponding to amino acid 81-128 mapping at the C-terminus of CGRP of human origin) [ ], or anti-glutamine synthetase rabbit polyclonal antibody (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, catalog number SC-6640, Lot No. H1407, polyclonal affinity purified antibody raised against a peptide mapping at the C-terminus of glutamine synthetase of human origin) [ ] for 2 h. Following four washes with PBST, sections were incubated with Alexa Fluor 488 goat anti-mouse antibody or Alexa Fluor 488 goat anti-rabbit antibody (1:1000; Molecular Probes, Camarillo, CA) for 1 h. To determine co-localization of σ1R with IB4, sections were incubated with Alexa Fluor 488 conjugate IB 4 (1:50, Invitrogen, Camarillo, CA, catalog number I21411, Lot No. 743633,) [ ] after four washes with PBST.

Techniques: Expressing, Staining, Immunofluorescence, Ligation, Control