PMID- 22952960 OWN - NLM STAT- MEDLINE DCOM- 20130207 LR - 20211021 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 7 IP - 8 DP - 2012 TI - Brain-derived neurotrophic factor enhances calcium regulatory mechanisms in human airway smooth muscle. PG - e44343 LID - 10.1371/journal.pone.0044343 [doi] LID - e44343 AB - Neurotrophins (NTs), which play an integral role in neuronal development and function, have been found in non-neuronal tissue (including lung), but their role is still under investigation. Recent reports show that NTs such as brain-derived neurotrophic factor (BDNF) as well as NT receptors are expressed in human airway smooth muscle (ASM). However, their function is still under investigation. We hypothesized that NTs regulate ASM intracellular Ca(2+) ([Ca(2+)](i)) by altered expression of Ca(2+) regulatory proteins. Human ASM cells isolated from lung samples incidental to patient surgery were incubated for 24 h (overnight) in medium (control) or 1 nM BDNF in the presence vs. absence of inhibitors of signaling cascades (MAP kinases; PI3/Akt; NFkappaB). Measurement of [Ca(2+)](i) responses to acetylcholine (ACh) and histamine using the Ca(2+) indicator fluo-4 showed significantly greater responses following BDNF exposure: effects that were blunted by pathway inhibitors. Western analysis of whole cell lysates showed significantly higher expression of CD38, Orai1, STIM1, IP(3) and RyR receptors, and SERCA following BDNF exposure, effects inhibited by inhibitors of the above cascades. The functional significance of BDNF effects were verified by siRNA or pharmacological inhibition of proteins that were altered by this NT. Overall, these data demonstrate that NTs activate signaling pathways in human ASM that lead to enhanced [Ca(2+)](i) responses via increased regulatory protein expression, thus enhancing airway contractility. FAU - Abcejo, Amard J AU - Abcejo AJ AD - Department of Anesthesiology, Mayo Clinic, Rochester, Minnesota, USA. FAU - Sathish, Venkatachalem AU - Sathish V FAU - Smelter, Dan F AU - Smelter DF FAU - Aravamudan, Bharathi AU - Aravamudan B FAU - Thompson, Michael A AU - Thompson MA FAU - Hartman, William R AU - Hartman WR FAU - Pabelick, Christina M AU - Pabelick CM FAU - Prakash, Y S AU - Prakash YS LA - eng GR - HL088029/HL/NHLBI NIH HHS/United States GR - R01 HL090595/HL/NHLBI NIH HHS/United States GR - R01 HL056470/HL/NHLBI NIH HHS/United States GR - R01 HL088029/HL/NHLBI NIH HHS/United States GR - HL090595/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20120829 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (6,7-dihydroxyflavone) RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Flavones) RN - 0 (Receptors, Nerve Growth Factor) RN - SY7Q814VUP (Calcium) SB - IM MH - Brain-Derived Neurotrophic Factor/*pharmacology MH - Calcium/*metabolism MH - Flavones/pharmacology MH - Humans MH - Intracellular Space/drug effects/metabolism MH - Muscle, Smooth/cytology/*metabolism MH - Myocytes, Smooth Muscle/drug effects/metabolism MH - Receptors, Nerve Growth Factor/metabolism MH - Respiratory System/cytology/*metabolism MH - Sarcoplasmic Reticulum/drug effects/metabolism MH - Signal Transduction/drug effects PMC - PMC3430656 COIS- Competing Interests: The authors have declared that no competing interests exist. EDAT- 2012/09/07 06:00 MHDA- 2013/02/08 06:00 PMCR- 2012/08/29 CRDT- 2012/09/07 06:00 PHST- 2012/01/10 00:00 [received] PHST- 2012/08/02 00:00 [accepted] PHST- 2012/09/07 06:00 [entrez] PHST- 2012/09/07 06:00 [pubmed] PHST- 2013/02/08 06:00 [medline] PHST- 2012/08/29 00:00 [pmc-release] AID - PONE-D-12-02629 [pii] AID - 10.1371/journal.pone.0044343 [doi] PST - ppublish SO - PLoS One. 2012;7(8):e44343. doi: 10.1371/journal.pone.0044343. Epub 2012 Aug 29.