Serotonergic lesion of median raphe nucleus alters nerve growth factor content and vulnerability of cholinergic septohippocampal neurons in rat

The cannabinergic system is implicated in the upregulation of central NGF protein by psychotropic drugs

“…In homogenization procedure, tissue samples were individually homogenized on ice in 5 to 6 vol of 0.25 mol/L sucrose, and 10 mMol/L HEPES (pH 7.0) containing 10 mMol/L DTT, then, immediately frozen in a dry ice/acetone bath and stored at −80°C until NGF analysis. As previously reported, NGF content in brain is severalfold higher than generally reported and largely associated with sedimentable fractions (Hoener et al 1996), so, the homogenates were centrifuged at 10,000×g for 10 min at 15°C and the remaining pellets were each dissolved in 750 μl NGF homogenization buffer, treated with ultrasound for 3 min, and processed for quantification of NGF as previously described in detail (Hellweg et al 1989;Hellweg et al 2001;Hellweg et al 2002). The measured and recoverycorrected NGF contents are expressed in ng NGF per g protein in the resuspended NGF homogenate quantified by Bradford protein assay (Bio-Rad, Hercules, CA, USA; Bradford 1976).…”

Subchronic treatment with lithium increases nerve growth factor content in distinct brain regions of adult rats

“…The remaining pellets were each dissolved in 750 l NGF-homogenization buffer, treated with ultrasound for 3 min and processed for quantification of endogenous NGF as described in detail elsewhere. [22][23][24] The measured and recovery-corrected NGF contents were expressed in ng NGF per g protein in the resuspended NGF homogenate, which was quantified by Bio-Rad  protein assays. 25 Our results obtained in untreated brain tissues are highly consistent with those previously reported by Hoener and coworkers.…”

Nerve Growth Factor Serum Levels in Patients with Acute Forms of Atopic Dermatitis

“…Chemicals of analytical grade were purchased from Merck (Darmstadt, Germany). Each 140 µl of serum was diluted (1+4) with sample buffer and endogenous NGF was quantified by a highly sensitive and specific two-site enzyme immunoassay as described in detail previously [Hellweg et al, 1989[Hellweg et al, , 2001. Briefly, black 96-well flat bottom immunoplates (Dynatech Laboratories, Inc., Virginia) were coated with 50 µl per well of 1.0 µg/ml monoclonal anti-mouse-β-NGF antibody 27/21 (Chemicon, Hofheim, Germany).…”

“…After 2 h at room temperature (20°C) the plates were washed 3 times with 200 µl per well of washing buffer. Unlike the previous protocol [Hellweg et al, 1989], this assay did not contain gelatin and the samples were incubated in the coated wells (50 µl each) overnight at 4°C [Hellweg et al, 2001]. After additional 3 washes the immobilized antigen was incubated with 0.75 mU per well of monoclonal antibody 27/21 conjugated with β-D-galactosidase (Chemicon, Hofheim, Germany) for 1.5 h at room temperature.…”