Journal: Neurology Research International

Article Title: Subarachnoid Transplant of the Human Neuronal hNT2.19 Serotonergic Cell Line Attenuates Behavioral Hypersensitivity without Affecting Motor Dysfunction after Severe Contusive Spinal Cord Injury

doi: 10.1155/2011/891605

Figure Lengend Snippet: Neural and human markers with differentiation of the hNT2.19 cell line in vitro. The hNT2.19 cell line was treated for two weeks with retinoic acid and mitotic inhibitors and lifted to substrate-coated 8-well plastic TC slides for differentiation and immunohistochemistry for neuron-specific markers. As soon as 4 days in vitro, a variety of neural markers appeared, which remained strong until at least 6 wks of differentiation: TuJ1 (a), hNSE (b), NFL (c), NFM (d), and NFH (e). For comparison, the negative control hNT2.6 cell line was cultured similarly as the hNT2.19 cells and is here stained for TuJ1 (f). Magnification bar = 20 nm, (a–f).

Article Snippet: For immunohistochemistry of sectioned spinal cord tissues, the polyclonal antibody anti-5HT (ab10385; dilution 1/100 (in vivo)) was purchased from Abcam Inc, Cambridge, MA, and the antihuman TuJ1 antibody (Neuron-specific class III beta-tubulin) was purchased from Neuromics, Edina, MN (MO15013; dilution 1/100 (in vivo).

Techniques: In Vitro, Immunohistochemistry, Comparison, Negative Control, Cell Culture, Staining

Journal: Neurology Research International

Article Title: Subarachnoid Transplant of the Human Neuronal hNT2.19 Serotonergic Cell Line Attenuates Behavioral Hypersensitivity without Affecting Motor Dysfunction after Severe Contusive Spinal Cord Injury

doi: 10.1155/2011/891605

Figure Lengend Snippet: Transplant of hNT2.19 and hNT2.6 cell lines in the severe contusive SCI model: TuJ1 and 5HT immunohistochemistry. Rats were injured with severe contusive SCI followed at two weeks by hNT2.6 (a, b) or hNT2.19 (c, d) cell grafts. Sagittal spinal cord sections were examined at 8 wks after SCI for evidence of surviving lumbar subarachnoid hNT2.6 (a, b) or hNT2.19 (c, d) cell line grafts, utilizing TuJ1 (a, c) or 5HT (b, d) immunohistochemistry. The hNT2.19 and control hNT2.6 (10 6 cells/injection), which had been differentiated for two weeks in vitro, were injected into the subarachnoid space two weeks after the SCI. Cell graft sites were colocalized with 5HT (b, d) and the human-specific marker TUJ1 (neuron-specific class III β -tubulin; (a, c)). There are many surviving hNT2.19 (c) and hNT2.6 (a) grafted cells visible on the pial surface, which stain for TuJ1 (arrows) at the end of the experiment, 56 days after SCI and about 6 weeks after cell transplant. Adjacent sections with the same grafted hNT2.19 (d) and hNT2.6 cells (b) are stained for 5HT, but only the hNT2.19 cells (d) are labeled for 5HT (arrows).

Article Snippet: For immunohistochemistry of sectioned spinal cord tissues, the polyclonal antibody anti-5HT (ab10385; dilution 1/100 (in vivo)) was purchased from Abcam Inc, Cambridge, MA, and the antihuman TuJ1 antibody (Neuron-specific class III beta-tubulin) was purchased from Neuromics, Edina, MN (MO15013; dilution 1/100 (in vivo).

Techniques: Immunohistochemistry, Control, Injection, In Vitro, Marker, Staining, Labeling

Journal: bioRxiv

Article Title: Targeting NGF but not VEGF or BDNF signaling reduces endometriosis-associated pain in mice

doi: 10.1101/2023.12.05.570181

Figure Lengend Snippet: (A) scheme of experimental procedures. (B) representative image from endometriotic lesions stained for NGF and beta-III tubulin. (C) quantification of TrkA + and pNF-κB + TrkA + neurons in dorsal root ganglia (DRG) of sham and endometriosis lesion-bearing mice. Lesions and DRG were dissected at 56 dpi. Results are presented as mean ± SEM of the percentage of positive neurons. n = 6 or 7 mice per group. (*P < 0.05 vs. sham). (D) Representative images of DRG neurons stained for TrkA (red) and p-NF-kB (green) by confocal microscopy.

Article Snippet: Sections were hydrated with PBS, blocked in 5% BSA in PBS 0.5% triton-X 100 (m/v/v) for 1h at RT, following overnight incubation at 4°C with primary antibodies: mouse anti-mouse phosphorylated-NF-κB (pNF-κB, Santa Cruz Biotechnology, Dallas, TX, USA, cat# sc-136548, 1:200 [RRID:AB_10610391]), rabbit anti-mouse TrkA (Invitrogen, Waltham MA, USA, cat. # MA5-32123, 1:100 [RRID:AB_2809414]), rabbit anti-mouse NGF (Abcam, Boston, MA, USA, cat. # AB52918, 1:300 [RRID:AB_881254]), and β-III tubulin (Novus Biologicals, Centennial, CO, USA, cat. # NB120-11314, 1:200 [RRID:AB_792496]).

Techniques: Staining, Confocal Microscopy