title: Molecular changes associated with hippocampal long-lasting depression induced by the serine protease subtilisin-A.
abstract: The serine protease subtilisin-A (SubA) induces a form of long-term depression (LTD) of synaptic transmission in the rat hippocampus, and molecular changes associated with SubA-induced LTD (SubA-LTD) were explored by using recordings of evoked postsynaptic potentials and immunoblotting. SubA-LTD was prevented by a selective inhibitor of SubA proteolysis, but the same inhibitor did not affect LTD induced by electrical stimulation or activation of metabotropic glutamate receptors. SubA-LTD was reduced by the protein kinase inhibitors genistein and lavendustin A, although not by inhibitors of p38 mitogen-activated protein kinase, glycogen synthase kinase-3, or protein phosphatases. It was also reduced by (RS)-α-methyl-4-carboxyphenylglycine, a broad-spectrum antagonist at metabotropic glutamate receptors. Inhibition of the Rho kinase enzyme Rho-associated coiled-coil kinase reduced SubA-LTD, although inhibitors of the RhoGTPase-activating enzymes farnesyl transferase and geranylgeranyl transferase did not. In addition, a late phase of SubA-LTD was dependent on new protein synthesis. There was a small, non-significant difference in SubA-LTD between wild-type and RhoB(-/-)  mice. Marked decreases were seen in the levels of Unc-5H3, a protein that is intimately involved in the development and plasticity of glutamatergic synapses. Smaller changes were noted, at higher concentrations of SubA, in Unc-5H1, vesicle-associated membrane protein-1 (synaptobrevin), and actin, with no changes in the levels of synaptophysin, synaptotagmin, RhoA, or RhoB. None of these changes was associated with LTD induced electrically or by the metabotropic glutamate receptor agonist (RS)-3,5-dihydroxyphenylglycine. These results indicate that SubA induces molecular changes that overlap with other forms of LTD, but that the overall molecular profile of SubA-LTD is quite different.
authors: Institute of Neuroscience and Psychology, College of Medical, Veterinary and Life Sciences, West Medical Building, University of Glasgow, Glasgow UK.
authors: Caroline M
authors: CM
authors: Forrest
authors: Jonas I
authors: JI
authors: Addae
authors: Sahana
authors: S
authors: Murthy
authors: L Gail
authors: LG
authors: Darlington
authors: Brian J
authors: BJ
authors: Morris
authors: Trevor W
authors: TW
authors: Stone
mesh_terms: D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D001565:Benzoates / Q000494:pharmacology; D004558:Electric Stimulation; D005071:Evoked Potentials / Q000502:physiology; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D005998:Glycine / Q000031:analogs & derivatives / Q000494:pharmacology; D006624:Hippocampus / Q000187:drug effects* / Q000502:physiology*; D036881:Long-Term Synaptic Depression / Q000187:drug effects*; D008297:Male; D051379:Mice; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D018094:Receptors, Metabotropic Glutamate / Q000378:metabolism; D012118:Resorcinols / Q000494:pharmacology; D013381:Subtilisins / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D020743:rhoB GTP-Binding Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D001565:Benzoates; D018690:Excitatory Amino Acid Agonists; D018691:Excitatory Amino Acid Antagonists; D011500:Protein Synthesis Inhibitors; D018094:Receptors, Metabotropic Glutamate; D012118:Resorcinols; C079694:alpha-methyl-4-carboxyphenylglycine; C079215:3,5-dihydroxyphenylglycine; D000841:Anisomycin; D010749:Phosphoprotein Phosphatases; D013381:Subtilisins; D020743:rhoB GTP-Binding Protein; D005998:Glycine