BDNF and TrkB Modulation in Neuroscience

All reagents were dissolved in 1 percent DMSO in saline. Vehicle (VEH) refers 1 percent DMSO in saline. Human BDNF (Peprotech, Rocky Hill, NJ, USA) was microinjected at 2 μg/μl (i.e. 1 μg/0.5 μl/side), a dose within the order of magnitude used in the literature (0.24–2.5 μg/0.5 μl/side) (Lu et al. 2004 (link); Graham et al. 2007 (link); Berglind et al. 2009 (link)). The TrkB selective and non-competitive antagonist ANA-12 (SML0209, Sigma-Aldrich, St Louis, MO, USA) (Cazorla et al. 2011 (link)) was microinjected in combination with BDNF or alone at 0.2 pg/μl (i.e. 0.1 pg/0.5 μl/side (Shirayama et al. 2015 (link)). We also tested a higher dose of [ANA-12] = 2 pg/μl (i.e.1 pg/0.5 μl/side), but this dose decreased spontaneous locomotion (data not shown). Human recombinant TrkB/Fc chimera (T8694; Sigma-Aldrich, St Louis, MO, USA), a soluble scavenger form of the TrkB receptor, was microinjected in combination with BDNF or alone at 1.3 μg/μl (i.e. 650 ng/0.5 μl/side) (Revest et al. 2014 (link)).

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Bobadilla A.C., Garcia-Keller C., Chareunsouk V., Hyde J., Camacho D.M., Heinsbroek J.A, & Kalivas P.W. (2018). Accumbens brain-derived neurotrophic factor (BDNF) transmission inhibits cocaine seeking. Addiction biology, 24(5), 860-873.

Publication 2018

Human BDNF BDNF

Chimera Dmso Human Human bdnf Locomotion Saline Scavenger Scavenger receptor Trkb Trkb receptor

Corresponding Organization : Medical University of South Carolina

Other organizations : Western Washington University

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Variable analysis

independent variables

Human BDNF (Peprotech, Rocky Hill, NJ, USA) microinjected at 2 μg/μl (i.e. 1 μg/0.5 μl/side)
ANA-12 (TrkB selective and non-competitive antagonist) microinjected in combination with BDNF or alone at 0.2 pg/μl (i.e. 0.1 pg/0.5 μl/side) and 2 pg/μl (i.e.1 pg/0.5 μl/side)
Human recombinant TrkB/Fc chimera (a soluble scavenger form of the TrkB receptor) microinjected in combination with BDNF or alone at 1.3 μg/μl (i.e. 650 ng/0.5 μl/side)

dependent variables

Not explicitly mentioned

control variables

All reagents were dissolved in 1 percent DMSO in saline. Vehicle (VEH) refers 1 percent DMSO in saline.

controls

Positive control: Vehicle (VEH) refers 1 percent DMSO in saline.
Negative control: Not explicitly mentioned.

Annotations

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