Biomolecules. 2023 May 11
Pengcheng Chu, Wei Guo, He You, ORCID and Bai Lu
Products used in the paper Details Operation
plasmid construction & AAV vector packaging pAAV-miniCMVEGFP (PackGene, Guangzhou, China, #K104) was used as the backbone to generate the AAV plasmid of e2-BDNF-Myc. The plasmid was transfected into HEK293T cells, and 48 h later, cell lysates were harvested to detect the protein expression using an anti-Myc antibody in western blot experiment. AAV2-miniCMV-e2-BDNF-Myc was packaged by PackGene. Request Quote

Research Field: CNS

Keywords: brain-derived neurotrophic factor; Bdnf promoters; ventromedial hypothalamus; obesity; hyperphagia

Dose: The brain stereotaxic apparatus was assembled with a 5-uL Hamilton syringe plus a 33-gauge needle that inhaled the appropriate amount of adeno-associated virus (AAV) in the dilution of 1 x E12 titer. After anesthetization with Avertin (200 mg/kg), Bdnf-e2-/- mice were settled in apparatus, and then received bilaterally 320ul administration of AAVe2-BDNF-Myc or AAV-GFP into VMH (AP -1.60 mm, DV -5.70 mm, ML 0.30 mm). For DVC rescue experiments, 300 L AAV-e2-BDNF-Myc or AAV-GFP was injected into DVC (AP -7.60 mm, DV -1.80 mm, ML 0.00 mm). AAV-syn-DIO-HM3D(Gq)-mCherry (AAV2/9, OBiO) and AAV-syn-HM3D(Gq)-mCherry (AAV2/9, OBiO) were used to infect Bdnf-ires-Cre and Bdnf-e2-/- mice respectively. AAV-syn-EGFP-T2A-Cre (AAV2/9, OBiO) and AAV-syn-EGFP-P2A-MCS (AAV2/8, OBiO) were used to infect TrkB-flox mice.

Routes of Administration: Stereotactic Injection

Animal or cell line strain: C57/B6J mice


The transcripts for Bdnf (brain-derived neurotrophic factor), driven by different promoters, are expressed in different brain regions to control different body functions. Specific promoter(s) that regulates energy balance remain unclear. We show that disruption of Bdnf promoters I and II but not IV and VI in mice (Bdnf-e1−/−, Bdnf-e2−/−) results in obesity. Whereas Bdnf-e1−/− exhibited impaired thermogenesis, Bdnf-e2−/− showed hyperphagia and reduced satiety before the onset of obesity. The Bdnf-e2 transcripts were primarily expressed in ventromedial hypothalamus (VMH), a nucleus known to regulate satiety. Re-expressing Bdnf-e2 transcript in VMH or chemogenetic activation of VMH neurons rescued the hyperphagia and obesity of Bdnf-e2−/− mice. Deletion of BDNF receptor TrkB in VMH neurons in wildtype mice resulted in hyperphagia and obesity, and infusion of TrkB agonistic antibody into VMH of Bdnf-e2−/− mice alleviated these phenotypes. Thus, Bdnf-e2-transcripts in VMH neurons play a key role in regulating energy intake and satiety through TrkB pathway.

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