Munc13 genotype regulates secretory amyloid precursor protein processing via postsynaptic glutamate receptors.
The amyloid precursor protein (APP) can be proteolytically degraded via non-amyloidogenic α-secretase and amyloidogenic β-secretase pathways. Previously, we have identified the presynaptic protein Munc13-1 as a diacylglycerol/phorbolester (DAG/PE) receptor that contributes to secretory, non-amyloidogenic APP processing after PE stimulation. Here, we used organotypic brain slice cultures from wild-type mice and from Munc13-1 knock-out (KO), Munc13-2 KO and Munc13-1/2 double KO (DKO) mice for pharmacological stimulation experiments. First, we demonstrate that neuronal populations and synaptic components important for secretory APP processing develop normally in organotypic brain slice cultures of all genotypes analyzed. Blockade of voltage-gated Na(+) channels by tetrodotoxin reduced the PE-stimulated secretory APP processing, whereas depolarization by high extracellular K(+) concentration evoked APP secretion. Additionally, the PE-stimulated APP secretion from Munc13-1 KO brain slices was significantly lower than that from wild-type brain slices. This effect was not observed in brain slices from Munc13-2 KO mice, which is consistent with the lower abundance and subpopulation-specific distribution of Munc13-2 in presynaptic elements. In Munc13-1/2 DKO brain slices, the deficiency of Munc13-1 dominated the effect of APP processing. The Munc13-1 KO effect on APP processing could be rescued by the stimulation of postsynaptic glutamatergic receptors. This indicates that lack of postsynaptic glutamate receptor stimulation in Munc13-1 KO brain slice cultures but not presynaptic mechanisms account for compromised APP processing. We conclude that organotypic brain slices cultures are a valuable tool for studying APP processing pathways in intact neuronal circuits and that neuronal activity is important for maintenance of the non-amyloidogenic APP processing.
Paul Flechsig Institute for Brain Research, University of Leipzig, 04109 Leipzig, Germany.
Maike
M
Hartlage-Rübsamen
Alexander
A
Waniek
Steffen
S
Roßner
D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D000779:Anesthetics, Local / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000166:cytology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D005838:Genotype; D018698:Glutamic Acid / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins / Q000172:deficiency / Q000378:metabolism*; D007736:Kynurenic Acid / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000172:deficiency / Q000378:metabolism*; D009474:Neurons / Q000648:ultrastructure; D009924:Organ Culture Techniques; D010703:Phorbol Esters / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D017470:Receptors, Glutamate / Q000378:metabolism*; D013569:Synapses / Q000378:metabolism*; D013744:Tetanus Toxin / Q000494:pharmacology; D013779:Tetrodotoxin / Q000494:pharmacology; D013997:Time Factors
D016564:Amyloid beta-Protein Precursor; D000779:Anesthetics, Local; D002121:Calcium Channel Blockers; D018691:Excitatory Amino Acid Antagonists; D047908:Intracellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; D010703:Phorbol Esters; D017470:Receptors, Glutamate; D013744:Tetanus Toxin; C503321:Unc13a protein, mouse; C485549:Unc13b protein, mouse; D018698:Glutamic Acid; D013779:Tetrodotoxin; D011189:Potassium Chloride; D007770:L-Lactate Dehydrogenase; D007736:Kynurenic Acid