PMID- 11007900 OWN - NLM STAT- MEDLINE DCOM- 20010222 LR - 20191023 IS - 1529-2401 (Electronic) IS - 0270-6474 (Print) IS - 0270-6474 (Linking) VI - 20 IP - 19 DP - 2000 Oct 1 TI - Activity-dependent release of endogenous brain-derived neurotrophic factor from primary sensory neurons detected by ELISA in situ. PG - 7417-23 AB - To define activity-dependent release of endogenous brain-derived neurotrophic factor (BDNF), we developed an in vitro model using primary sensory neurons and a modified ELISA, termed ELISA in situ. Dissociate cultures of nodose-petrosal ganglion cells from newborn rats were grown in wells precoated with anti-BDNF antibody to capture released BDNF, which was subsequently detected using conventional ELISA. Conventional ELISA alone was unable to detect any increase in BDNF concentration above control values following chronic depolarization with 40 mM KCl for 72 hr. However, ELISA in situ demonstrated a highly significant increase in BDNF release, from 65 pg/ml in control to 228 pg/ml in KCl-treated cultures. The efficacy of the in situ assay appears to be related primarily to rapid capture of released BDNF that prevents BDNF binding to the cultured cells. We therefore used this approach to compare BDNF release from cultures exposed for 30 min to either continuous depolarization with elevated KCl or patterned electrical field stimulation (50 biphasic rectangular pulses of 25 msec, at 20 Hz, every 5 sec). Short-term KCl depolarization was completely ineffective at evoking any detectable release of BDNF, whereas patterned electrical stimulation increased extracellular BDNF levels by 20-fold. In addition, the magnitude of BDNF release was dependent on stimulus pattern, with high-frequency bursts being most effective. These data indicate that the optimal stimulus profile for BDNF release resembles that of other neuroactive peptides. Moreover, our findings demonstrate that BDNF release can encode temporal features of presynaptic neuronal activity. FAU - Balkowiec, A AU - Balkowiec A AD - Department of Neurosciences, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106, USA. FAU - Katz, D M AU - Katz DM LA - eng PT - Journal Article PT - Research Support, U.S. Gov't, P.H.S. PL - United States TA - J Neurosci JT - The Journal of neuroscience : the official journal of the Society for Neuroscience JID - 8102140 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 660YQ98I10 (Potassium Chloride) SB - IM MH - Animals MH - Animals, Newborn MH - Brain-Derived Neurotrophic Factor/*analysis/metabolism MH - Cell Count MH - Cell Survival/drug effects MH - Cells, Cultured MH - Electric Stimulation MH - Enzyme-Linked Immunosorbent Assay/methods MH - Extracellular Space/metabolism MH - Ganglia, Sensory/cytology MH - Neurons, Afferent/cytology/drug effects/*metabolism MH - Potassium Chloride/pharmacology MH - Rats MH - Rats, Sprague-Dawley PMC - PMC6772775 EDAT- 2000/09/29 11:00 MHDA- 2001/03/03 10:01 PMCR- 2001/04/01 CRDT- 2000/09/29 11:00 PHST- 2000/09/29 11:00 [pubmed] PHST- 2001/03/03 10:01 [medline] PHST- 2000/09/29 11:00 [entrez] PHST- 2001/04/01 00:00 [pmc-release] AID - 4550 [pii] AID - 10.1523/JNEUROSCI.20-19-07417.2000 [doi] PST - ppublish SO - J Neurosci. 2000 Oct 1;20(19):7417-23. doi: 10.1523/JNEUROSCI.20-19-07417.2000.