ant006  (Alomone Labs)

Journal: bioRxiv

Article Title: Prelimbic proBDNF facilitates memory destabilization by regulation of neuronal function in juveniles

doi: 10.1101/2021.12.30.474526

Figure Lengend Snippet: The increase proBDNF alters synaptic currents, promotes LFS-induced synaptic depression and strengthens the theta phase-gamma amplitude coupling during the PR-LTM test. (A) Schematic describing the timeline for morphological analysis (Top-left). Illustration of the region of interest in prelimbic images (Top-middle), and dendritic segment analysis for spine quantification (Top-right). Red circles indicated the mushroom type spine, yellow circles indicated thin type spine and blue circles indicated stubby type spine. Sample images were projected at minimal intensity and inverted, background was then subtracted, followed by brightness/contrast adjustment. Scale bars, 5 μm. Quantification of spine density (Bottom-left) and the proportion of spine (Bottom-right). No statistical difference in spine density was found between juvenile and adult groups. However, a significant higher proportion of thin type spine but a lower mushroom type spine was observed in juveniles compared with adults. (B) Schematic describing the timeline for EPSCs recordings (Top-left). Representative continuous traces (Top-middle) and average waveform (Top-right) of the pharmacologically isolated NMDA EPSCs in the prelimbic neurons of adult, juvenile and juvenile+anti groups. No change in the amplitude of EPSCs (Bottom-left) was found but the frequency (Bottom-middle) and decay time (Bottom-right) were significantly increased in juvenile group. The enhanced frequency and decay time of NMDA currents in juvenile group were inhibited after infusions of anti-proBDNF antibody. (* P

Article Snippet: Needles were inserted into bilateral cannulae and then cleavage-resistant proBDNF (2 ng/ml; Cat#B257 Alomone Labs), anti-proBDNF antibody (10 μg/μL; Cat#ANT-006, Alomone Labs), TAT-Pep5 (4 ng/μL; Cat#506181, EMD Millipore), K252a (25 μg/μL; Cat#82497; Sigma-Aldrich), 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP; 32 ng/μL; Cat#01773, Tocris Bioscience), NVP-AAM077 (0.8 ng/μL; Cat#P1999, Sigma-Aldrich), Ro25-6981 (2.0 ng/μL; Cat#1594, Tocris Bioscience), mature BDNF (1.5 μg/mL; Cat#B250; Alomone Labs) or artificial CSF (ACSF, Cat#3525, Tocris Bioscience) into prelimbic area (at a rate of 0.5 μL/min/side for 2 min) was infused immediately or one day following memory retrieval.

Techniques: Isolation

Journal: bioRxiv

Article Title: Prelimbic proBDNF facilitates memory destabilization by regulation of neuronal function in juveniles

doi: 10.1101/2021.12.30.474526

Figure Lengend Snippet: The higher prelimbic proBDNF expression during the juvenile period facilitates retrieval-dependent memory destabilization. Quantification of the proBDNF (A) and its receptor p75 NTR (B) levels in prelimbic cortex of juvenile and adult rats. Representative immunoblots the expression of proBDNF and p75 NTR (Top). A significant increase in the proBDNF levels was detected in juvenile group, as well the p75 NTR levels (Bottom). (* P

Article Snippet: Needles were inserted into bilateral cannulae and then cleavage-resistant proBDNF (2 ng/ml; Cat#B257 Alomone Labs), anti-proBDNF antibody (10 μg/μL; Cat#ANT-006, Alomone Labs), TAT-Pep5 (4 ng/μL; Cat#506181, EMD Millipore), K252a (25 μg/μL; Cat#82497; Sigma-Aldrich), 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP; 32 ng/μL; Cat#01773, Tocris Bioscience), NVP-AAM077 (0.8 ng/μL; Cat#P1999, Sigma-Aldrich), Ro25-6981 (2.0 ng/μL; Cat#1594, Tocris Bioscience), mature BDNF (1.5 μg/mL; Cat#B250; Alomone Labs) or artificial CSF (ACSF, Cat#3525, Tocris Bioscience) into prelimbic area (at a rate of 0.5 μL/min/side for 2 min) was infused immediately or one day following memory retrieval.

Techniques: Expressing, Western Blot

Journal: bioRxiv

Article Title: Prelimbic proBDNF facilitates memory destabilization by regulation of neuronal function in juveniles

doi: 10.1101/2021.12.30.474526

Figure Lengend Snippet: Up-regulation of proBDNF-p75NTR signaling mediated by NMDA-GluN2B contributes to enhance the modulation of existing fear memory traces in juvenile rats. (A) Schematic describing the behavioral timeline for the retrieval-dependent memory destabilization experiment using rats conditioned with four tones (Top). Immediately following the memory retrieval, the rats infused with TAT-Pep5, K252a or vehicle into the prelimbic cortex 15 min prior to the mBDNF, proBDNF or vehicle infusion. Two days later, PR-LTM was assessed by exposed the rats to the novel context. Similar, no significant difference in the percentage freezing during the memory retrieval but the percentage freezing level during the PR-LTM test was significant lower in juvenile group than adult group (Bottom). No obvious effect of mBDNF on freeze behavior was found. Infusions of p75 NTR blocker TAT-Pep5 could significantly enhance the percentage of freeze behavior. Meanwhile, infusions of TAT-Pep5, but not K252a, markedly blocked the effects of proBDNF treatment. (* P

Article Snippet: Needles were inserted into bilateral cannulae and then cleavage-resistant proBDNF (2 ng/ml; Cat#B257 Alomone Labs), anti-proBDNF antibody (10 μg/μL; Cat#ANT-006, Alomone Labs), TAT-Pep5 (4 ng/μL; Cat#506181, EMD Millipore), K252a (25 μg/μL; Cat#82497; Sigma-Aldrich), 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP; 32 ng/μL; Cat#01773, Tocris Bioscience), NVP-AAM077 (0.8 ng/μL; Cat#P1999, Sigma-Aldrich), Ro25-6981 (2.0 ng/μL; Cat#1594, Tocris Bioscience), mature BDNF (1.5 μg/mL; Cat#B250; Alomone Labs) or artificial CSF (ACSF, Cat#3525, Tocris Bioscience) into prelimbic area (at a rate of 0.5 μL/min/side for 2 min) was infused immediately or one day following memory retrieval.

Techniques: