Journal: Frontiers in Physiology

Article Title: Plasticity of TRPV1-Expressing Sensory Neurons Mediating Autonomic Dysreflexia Following Spinal Cord Injury

doi: 10.3389/fphys.2012.00257

Figure Lengend Snippet: High-thoracic (T3) spinal cord injury-induced selective hypertrophy of sensory neurons expressing the capsaicin receptor (TRPV1) and the artemin receptor (GFRα3) in the L4/L5 DRG . (A) Substance P (SP) – positive DRG neurons and their size-frequency distributions. (B) IB4-binding DRG neurons. (C) P2X3-positive DRG neurons. (D) TRPV1-positive DRG neurons. (E) GFRα3-expressing DRG neurons, known to express TRPV1. The overall proportions of immunopositive neurons [insets (A–E) ] did not change for any subpopulation. Ganglia were harvested 3 months after sham-injury (gray) or complete T3 SCI (black). Scale bar = 50 μm. Asterisks indicate P < 0.05, K–S goodness-of-fit test.

Article Snippet: For immunohistochemistry, slides were incubated in 10% normal donkey serum in PBS plus Triton X-100 (0.1%) for 20 min. We used five antibodies to delineate different subsets of nociceptors: these included antibodies raised against TRPV1 (Neuromics, Edina, MN, USA; 1:2,000), the ionotropic ATP purinoceptor P2X 3 (Millipore, Billerica, MA, USA; 1:1,000), Substance P (Neuromics, 1:1,000), the glycoprotein isolectin B4 (IB4; Neuromics; 1:2,000), the glial cell line-derived neurotrophic factor family receptor α 3 (GFRα3; R&D Systems; 1:500).

Techniques: Expressing, Binding Assay

Journal:

Article Title: Loss of brain-derived neurotrophic factor-dependent neural crest-derived sensory neurons in neurotrophin-4 mutant mice

doi:

Figure Lengend Snippet: Anti-TrkB antibody staining (arrows) of neonatal DRG in wt (a and b) and NT-4 mutant (c and d) mice. Dark-field view of TrkC mRNA expression in neurons (arrows) of neonatal DRG in wt (e and f) and NT-4 mutant (g and h) mice. (i and j) Higher, bright-field magnification of g and h. (i and j) Arrows indicate “positive,” TrkC-expressing neurons whereas arrowheads indicate “negative,” TrkC-expressing neurons. Digoxigenin-labeled NT-4-specific antisense and sense probes (k– o) were hybridized to E10 embryo sections of P0, wt, and NT-4 mutant DRG. (k and l) Transverse thoracic section of E10 embryo showing hybridized (Left) and Nissl-stained tissue (Right). Arrows point to region of forming DRG. NT-4 mRNA expression in neonatal wt (m and o) and NT-4 mutant (n) DRG using antisense (m and n) and sense (o) probes. (p) DRG sections from neonatal wt, NT-4, and BDNF mutant mice were immunostained with antibodies for somatostatin (SS), substance P (SP), calcitonin gene-related protein (cGRP), parvalbumin (PA), and SSEA-3. (q) Number of myelinated axons innervating cutaneous tissue (saphenous nerve) and muscle (common peroneal and tibial nerves) in wt and NT-4 mutant mice. (r) Coordination of wt and NT-4 mutant on a rotating rod over 4 consecutive days.

Article Snippet: The remaining sections were immunostained ( 5 ) every 50 μM with antibodies directed against the following proteins: somatostatin (SS; 1:150; Zymed), substance P (SP; 1:150; Zymed), SSEA-3 (1:300; Hybridoma Bank, Iowa City, IA) ( 20 , 21 ), calcitonin gene-related protein (cGRP; 1:50; Biogenesis, Kingston, NH), and parvalbumin (PA; 1:250; Swant, Bellinzona, Switzerland).

Techniques: Staining, Mutagenesis, Expressing, Labeling