Serotonin(1A)-receptor-dependent signaling proteins in mouse hippocampus.
The serotonin(1A) receptor (5-HT(1A) R) knock-out mouse (KO) is a widely used animal model for anxiety and cognitive function and regulation of signaling cascades by this receptor has been reported. We aimed to determine individual representatives of signaling cascades in order to screen 5-HT(1A) R-dependent signaling proteins (SPs). Hippocampal proteins from wild type and 5-HT(1A) R KO mice were extracted, run on two-dimensional gel electrophoresis, proteins were identified by MALDI and nano-ESI-LC-MS/MS and SPs were quantified by specific software. Nucleoside diphosphate kinase A (NDK A, synonym: nm23), Dual specificity mitogen-activated protein kinase kinase 1 (MAPKK1, synonym: MEK), Serine/threonine-protein phosphatase PP1-gamma catalytic subunit (PP-1G), Septin-5, were reduced in the KO mice. Novel phosphorylation sites at T386 on MAPKK1 and at S225 and Y265 on Septin-5 were observed. MAPKK1 and PP-1G are known 5-HT(1A) R-dependent signaling compounds and are in agreement with receptor knock-out and septin-5 is involved in serotonin transport, although regulation by 5-HT(1A) R has not been reported. 5-HT(1A) R - dependent levels for NDK A have not been demonstrated so far and we herewith propose a role for NDK A in 5-HT(1A) R signaling. Reduced SP levels along with findings of two novel phosphorylation sites may be relevant for interpretation of previous and the design of future studies on this receptor system.
Li
Lin
L
Department of Pediatrics, Medical University of Vienna, Währinger Gürtel 18-20, A-1090 Vienna, Austria.
Whittle
Nigel
N
Klug
Stefanie
S
Chen
Wei-Qiang
WQ
Singewald
Nicolas
N
Toth
Miklos
M
Lubec
Gert
G
D000595:Amino Acid Sequence; D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D030541:Databases, Genetic; D015180:Electrophoresis, Gel, Two-Dimensional; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D006624:Hippocampus / Q000378:metabolism*; D048369:MAP Kinase Kinase 1 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D054778:NM23 Nucleoside Diphosphate Kinases / Q000378:metabolism*; D010766:Phosphorylation; D054645:Protein Phosphatase 1 / Q000378:metabolism*; D044282:Receptor, Serotonin, 5-HT1A / Q000235:genetics / Q000378:metabolism*; D058112:Septins; D015398:Signal Transduction; D012984:Software; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003598:Cytoskeletal Proteins; D054778:NM23 Nucleoside Diphosphate Kinases; D044282:Receptor, Serotonin, 5-HT1A; D048369:MAP Kinase Kinase 1; C517939:Nme1 protein, mouse; D054645:Protein Phosphatase 1; D019204:GTP-Binding Proteins; C491612:Sept5 protein, mouse; D058112:Septins