Journal: British Journal of Pharmacology

Article Title: Distinct effects of CGRP on typical and atypical smooth muscle cells involved in generating spontaneous contractions in the mouse renal pelvis

doi: 10.1111/j.1476-5381.2009.00514.x

Figure Lengend Snippet: Distribution of PGP9.5-, calcitonin gene-related peptide (CGRP)- and transient receptor potential cation channel, subfamily V, member 1 (TRPV1) immunoreactive nerves in the mouse renal pelvis. In the proximal renal pelvis, nerve fibres were stained with antibodies against the neural marker PGP 9.5 (Aa). CGRP-immunoreactive nerve fibres were abundant in the same preparation (Ab). A merged image indicates that most of thick and thin nerve bundles contain CGRP-immunoreactive fibres (Ac). Double staining of the mid renal pelvis with TRPV1 (Ba) and CGRP (Bb) antibodies showed that many CGRP-positive fibres also expressed TRPV1-immnoreactivity (Bc). Scale bars: 80 µm.

Article Snippet: To examine colocalization of CGRP and transient receptor potential cation channels (TRPV1; vanilloid receptors), whole mounts were incubated with rabbit anti-CGRP antibody (1:50, 11189, Progen Biotechnik, Heidelberg, Germany) and guinea pig anti-TRPV1 antibody (1:1000, GP14100, Neuromics, Edina, MN, USA) for 2 days at 4°C.

Techniques: Staining, Marker, Double Staining

Journal: Neurology International

Article Title: Semi-Automated Recording of Facial Sensitivity in Rat Demonstrates Antinociceptive Effects of the Anti-CGRP Antibody Fremanezumab

doi: 10.3390/neurolint15020039

Figure Lengend Snippet: Schematic workflow of the experiments. The first week was used for priming the animals to the attractive source, in the second week baseline measurements without barrier, and with the mechanical and thermal barrier were performed, finished by injection of antibodies (either anti-CGRP antibody fremanezumab or control antibody). In the third and fourth week, the measurements were repeated with the same sequence at days 4–6 and days 11–13 after antibody injection.

Article Snippet: The animals were divided into two groups, one group that received isotype control antibody and the other anti-CGRP antibody, fremanezumab (Teva Pharmaceuticals, Redwood City, CA, USA).

Techniques: Injection, Control, Sequencing

Journal: Neurology International

Article Title: Semi-Automated Recording of Facial Sensitivity in Rat Demonstrates Antinociceptive Effects of the Anti-CGRP Antibody Fremanezumab

doi: 10.3390/neurolint15020039

Figure Lengend Snippet: Gain of body weight in animals injected with fremanezumab or control antibody from baseline to the two test sequences at 4 and 11 days. Females ( A ) gained less weight than males ( B ), in which the increase was clearly significant (repeated measures ANOVA with factor antibody, F 2,40 = 18.9, p < 0.0005). There was no significant difference between the animals that received fremanezumab or control antibody in any group at any time of measurement. White digits in the bars represent the number of animals tested. Differences between days: # p < 0.05, ## p < 0.005, ### p < 0.0005 (LSD post hoc test).

Article Snippet: The animals were divided into two groups, one group that received isotype control antibody and the other anti-CGRP antibody, fremanezumab (Teva Pharmaceuticals, Redwood City, CA, USA).

Techniques: Injection, Control

Journal: Neurology International

Article Title: Semi-Automated Recording of Facial Sensitivity in Rat Demonstrates Antinociceptive Effects of the Anti-CGRP Antibody Fremanezumab

doi: 10.3390/neurolint15020039

Figure Lengend Snippet: Comparison of baselines for approaches to the source ( A ), drinking time ( B ), and drinking volume ( C ) before the injection of antibodies. Groups designated for the control antibody and fremanezumab, respectively, showed some variance but no significant differences. After inserting the mechanical barrier, the number of trials to approach the source tended to increase ( A ), which was significant in the group designated for fremanezumab (repeated measures ANOVA with factor antibody, F 2,44 = 3.99, p < 0.05 and LSD post hoc test, p < 0.05). On the contrary, the mechanical and thermal barriers dramatically reduced both the drinking time ( B ) and drinking volume ( C ) (repeated measures ANOVA, F 2,44 = 39.17 and 74.03, respectively, p < 0.0005). White digits in the bars represent the number of animals tested. Difference between no barrier situation and mechanical and thermal barrier, respectively: # p < 0.05, ### p < 0.0005 (LSD post hoc test).

Article Snippet: The animals were divided into two groups, one group that received isotype control antibody and the other anti-CGRP antibody, fremanezumab (Teva Pharmaceuticals, Redwood City, CA, USA).

Techniques: Comparison, Injection, Control

Journal: Neurology International

Article Title: Semi-Automated Recording of Facial Sensitivity in Rat Demonstrates Antinociceptive Effects of the Anti-CGRP Antibody Fremanezumab

doi: 10.3390/neurolint15020039

Figure Lengend Snippet: Tests without the barrier before (baseline) and at days 4 and 11 after antibody injection (1st and 2nd test sequence). ( A ) Animals injected with the control antibody tended to approach the drinking source more frequently than animals injected with fremanezumab (repeated measures ANOVA with factor antibody, F 2,44 = 6.52, p < 0.05), which was significant at day 4 after antibody injection (LSD post hoc test, p < 0.05). ( B ) The cumulated time, during which the animals stayed at the source to drink, increased at days 4 and 11 compared to baseline, independent of the type of antibody injected (repeated measures ANOVA with factor antibody, F 2,44 = 8.90, p < 0.005). ( C ) Similarly, the cumulated volume consumed by the animals increased at days 4 and 11 (repeated measures ANOVA, F 2,44 = 17.17, p < 0.0005). White digits in bars represent the number of animals tested. Difference between antibodies: * p < 0.05; difference in the course of repeated tests: # p < 0.05; ## p < 0.005 (LSD post hoc tests).

Article Snippet: The animals were divided into two groups, one group that received isotype control antibody and the other anti-CGRP antibody, fremanezumab (Teva Pharmaceuticals, Redwood City, CA, USA).

Techniques: Injection, Sequencing, Control

Journal: Neurology International

Article Title: Semi-Automated Recording of Facial Sensitivity in Rat Demonstrates Antinociceptive Effects of the Anti-CGRP Antibody Fremanezumab

doi: 10.3390/neurolint15020039

Figure Lengend Snippet: Tests with the thermal barrier before (baseline) and at days 6 and 13 (1st and 2nd test sequence) after antibody injection. At days 6 and 13, the animals partly approached the drinking source more often ( A ) and stayed longer at the source ( B ) compared to baseline, independently of the type of injected antibody (repeated measures ANOVA with factor antibody, F 2,44 = 3.92, p < 0.05 for A and F 2,44 = 3.60, p < 0.05 for B; F 2,44 = 10.44, p < 0.0005.81 for ( C )). At day 12, the drinking time ( B ) and the consumed volume ( C ) were significantly higher in animals injected with fremanezumab compared to the control antibody (repeated measures ANOVA with combined factors repetition and antibody extended by the LSD post hoc test, F 2,44 = 45.10, p < 0.05). The separated analysis of the sexes showed that this was particularly due to the female animals, which drank longer and significantly more when they were injected with fremanezumab (( B , C ), right insets). White digits in bars represent the number of animals tested. Difference between antibodies: * p < 0.05, ** p < 0.005; difference in the course of repeated tests: # p < 0.05; ## p < 0.005 (LSD post hoc test).

Article Snippet: The animals were divided into two groups, one group that received isotype control antibody and the other anti-CGRP antibody, fremanezumab (Teva Pharmaceuticals, Redwood City, CA, USA).

Techniques: Sequencing, Injection, Control

Journal: The Journal of Physiological Sciences : JPS

Article Title: Immobilization-induced hypersensitivity associated with spinal cord sensitization during cast immobilization and after cast removal in rats

doi: 10.1007/s12576-013-0277-4

Figure Lengend Snippet: Intensity of calcitonin gene-related peptide (CGRP) expression in the ipsilateral dorsal horn of the spinal cord. Representative photomicrographs of CGRP immunohistochemistry in the ipsilateral dorsal horn from the Im-4 weeks, Im-8 weeks, and the control groups (age-matched with the Im-4 weeks group) are shown (a). The CGRP-positive neural fibers were clearly observed in the deep layer of the dorsal horn only in the Im-8 weeks group (arrowheads). Percentage control of fluorescence intensity of CGRP expression in the superficial layer (laminae I-II) (b, d) and deep layers (laminae III-VI) were calculated (c, e) in the ipsilateral and contralateral dorsal horn. *P < 0.05, significantly different compared to the age-matched control group. # P < 0.05, significantly different compared to the Im-4 weeks group. Scale bar = 100 μm

Article Snippet: Next, sections were blocked for 20 min with 5 % bovine albumin dissolved in PBS, followed by incubation with an anti-CGRP polyclonal antibody (1:500 rabbit; ImmunoStar Inc. Hudson, WI, USA) overnight at 4 °C.

Techniques: Expressing, Immunohistochemistry, Control, Fluorescence

Journal: BMC Musculoskeletal Disorders

Article Title: Persistent synovial inflammation plays important roles in persistent pain development in the rat knee before cartilage degradation reaches the subchondral bone

doi: 10.1186/s12891-018-2221-5

Figure Lengend Snippet: Distribution of CGRP-positive nerve fibers in IFP and L4 DRG. a Representative immunohistochemical images of the knee joint at 28 days after the injection of MIA. Arrowheads in red indicate CGRP-positive nerve fibers. b Representative images of L4 DRG at day 28 post-MIA injection. The red signal indicates neuronal cell bodies projected from the knee joint (left column, FG). Neural cells positive for CGRP are indicated in green (middle column). Merged images are shown in the right column. Arrows in white indicate the CGRP-positive nerve cells projected from the knee joint. c , d Differences in innervation density between the MIA and control sides. Ten different areas of 0.01 mm 2 were randomly selected in the parenchymal region of IFP in each section. CGRP-positive nerve fibers > 0.03 mm were counted. There were 6 samples at each time point and 2 sections were randomly selected in each sample. Data are represented as mean and SD values. e Differences in innervation density between ipsilateral and contralateral indicated in ( c ) and ( d ) were calculated in each time point (innervation density of MIA subtracted by innervation density of control) and plotted. f , g The percentage of CGRP-positive neurons among the FG-labeled neurons. There were 6 samples in each time point. Four sections were randomly selected from each sample and the mean ± SD values were recorded ( n = 6). Asterisks indicate statistically significant differences. h Differences in innervation density between ipsilateral and contralateral indicated in ( f ) and ( g ) were calculated in each time point (innervation density of MIA subtracted by innervation density of control) and plotted. CGRP, calcitonin gene-related peptide; DRG, dorsal root ganglion; FG, fluorogold; IFP, infrapatellar fat pad; MIA, monoiodo-acetic acid

Article Snippet: Anti-human CGRP polyclonal antibody was purchased from Peninsula Laboratories LLC (San Carlos, CA, USA).

Techniques: Immunohistochemical staining, Injection, Control, Labeling