Journal: Frontiers in Cell and Developmental Biology

Article Title: Age Related Response of Neonatal Rat Retinal Ganglion Cells to Reduced TrkB Signaling in vitro and in vivo

doi: 10.3389/fcell.2021.671087

Figure Lengend Snippet: In vitro study: summary of treatment groups and number of culture wells analyzed.

Article Snippet: RGCs were separated into samples which were resuspended either with or without exogenous BDNF (50 ng/ml; Peprotech) and further subdivided into groups that were resuspended with or without TrkB-Fc (TrkB-Fc + , 500 ng/ml; BioSensis PE–1235–20) or with or without a blocking cocktail (Block + , 500 ng/ml) containing 1:1 of antibody to rh BDNF:lgG (BioSensis S-015-500) and antibody to rh NT4:lgG (Biosensis S-059-500).

Techniques: In Vitro, Blocking Assay

Journal:

Article Title: Neurotrophins are key mediators of the myelination program in the peripheral nervous system

doi: 10.1073/pnas.251543398

Figure Lengend Snippet: Western blot analysis of the effects of endogenous and exogenous neurotrophins on the expression of myelin proteins in Schwann cell/neuronal cocultures. Exogenous BDNF (100 ng/ml), TrkB-Fc (1 μg/ml), NT3 (100 ng/ml), and TrkC-Fc (1 μg/ml) were added to cocultures on the day of induction of myelin formation. After 6 days of induction, cocultures were extracted and probed for MAG and P0 proteins. The results are shown as the mean value ± SD.

Article Snippet: BDNF, NT3, TrkB-Fc, and TrkC-Fc (3 μg each; Regeneron Pharmaceuticals) were injected s.c., starting from the caudal portion of the greater trochanter region and running parallel along the sciatic nerve (total volume of 5 μl).

Techniques: Western Blot, Expressing

Journal:

Article Title: Neurotrophins are key mediators of the myelination program in the peripheral nervous system

doi: 10.1073/pnas.251543398

Figure Lengend Snippet: The effects of endogenous and exogenous neurotrophins in the formation of myelin internodes in Schwann cell/neuronal cocultures determined by immunocytochemical analysis of P0. (A) Control cultures without the addition of primary antibody. (B) Control cultures with the addition of the anti-P0 antibody. (C) Addition of exogenous BDNF (100 ng/ml). (D) Addition of TrkB-Fc (1 μg/ml). (E) Addition of exogenous NT3 (100 ng/ml). (F) Addition of TrkC-Fc (1 μg/ml). All factors were added to cocultures on the day of induction of myelin formation. Immunocytochemistry was performed on cocultures after 6 days of induction.

Article Snippet: BDNF, NT3, TrkB-Fc, and TrkC-Fc (3 μg each; Regeneron Pharmaceuticals) were injected s.c., starting from the caudal portion of the greater trochanter region and running parallel along the sciatic nerve (total volume of 5 μl).

Techniques: Immunocytochemistry

Journal:

Article Title: Neurotrophins are key mediators of the myelination program in the peripheral nervous system

doi: 10.1073/pnas.251543398

Figure Lengend Snippet: Western blot analysis of the effect of neurotrophins and neurotrophin scavengers during the development of the sciatic nerve. Newborn mice (P1) were s.c. injected with 3 μg each of BDNF, NT3, TrkB-Fc, or TrkC-Fc as indicated under Materials and Methods. The contralateral leg was injected with vehicle alone as a control for each one of the individually treated mice. Two days later (P3, 2 days treatment), the sciatic nerves were isolated and processed for Western blot analysis. In some instances, the animals were reinjected at this stage and allowed to proceed for 2 more days (P5, 4 days treatment). The sciatic nerves were then processed and analyzed in the same manner. (A) Quantification of MAG protein content after treatment with the different factors for 2 or 4 days. Representative Western blots are shown (Lower). (B) Quantification of P0 protein content and representative Western blots. The results are shown as the mean value ± SD of the percentage of the levels expressed in the contralateral leg (injected with saline vehicle alone).

Article Snippet: BDNF, NT3, TrkB-Fc, and TrkC-Fc (3 μg each; Regeneron Pharmaceuticals) were injected s.c., starting from the caudal portion of the greater trochanter region and running parallel along the sciatic nerve (total volume of 5 μl).

Techniques: Western Blot, Injection, Isolation

Journal:

Article Title: Neurotrophins are key mediators of the myelination program in the peripheral nervous system

doi: 10.1073/pnas.251543398

Figure Lengend Snippet: Effects of TrkB-Fc treatment on the myelin ultrastructure during sciatic nerve development. Newborn mice were injected with 3 μg of TrkB-Fc and their sciatic nerves analyzed as in Fig. ​Fig.5.5. Electron microphotographs from (A) control nerve (saline alone) and (B) TrkB-Fc-treated nerve. (C) Distribution of myelinated axons against ensheathed axons as a percentage of the sum of both. (D) The thickness of the myelin sheath was determined by counting the number of lamellae. Myelinated axons were distributed as a function of the thickness of the myelin sheath. The distribution is shown as the mean value ± SD of the percentage of myelinated axons that falls within the given range in the number of lamellae.

Article Snippet: BDNF, NT3, TrkB-Fc, and TrkC-Fc (3 μg each; Regeneron Pharmaceuticals) were injected s.c., starting from the caudal portion of the greater trochanter region and running parallel along the sciatic nerve (total volume of 5 μl).

Techniques: Injection