sq-23377 has been researched along with leupeptin* in 3 studies
3 other study(ies) available for sq-23377 and leupeptin
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Activity-dependent cleavage of the K-Cl cotransporter KCC2 mediated by calcium-activated protease calpain.
The K-Cl cotransporter KCC2 plays a crucial role in neuronal chloride regulation. In mature central neurons, KCC2 is responsible for the low intracellular Cl(-) concentration ([Cl(-)](i)) that forms the basis for hyperpolarizing GABA(A) receptor-mediated responses. Fast changes in KCC2 function and expression have been observed under various physiological and pathophysiological conditions. Here, we show that the application of protein synthesis inhibitors cycloheximide and emetine to acute rat hippocampal slices have no effect on total KCC2 protein level and K-Cl cotransporter function. Furthermore, blocking constitutive lysosomal degradation with leupeptin did not induce significant changes in KCC2 protein levels. These findings indicate a low basal turnover rate of the total KCC2 protein pool. In the presence of the glutamate receptor agonist NMDA, the total KCC2 protein level decreased to about 30% within 4 h, and this effect was blocked by calpeptin and MDL-28170, inhibitors of the calcium-activated protease calpain. Interictal-like activity induced by incubation of hippocampal slices in an Mg(2+)-free solution led to a fast reduction in KCC2-mediated Cl(-) transport efficacy in CA1 pyramidal neurons, which was paralleled by a decrease in both total and plasmalemmal KCC2 protein. These effects were blocked by the calpain inhibitor MDL-28170. Taken together, these findings show that calpain activation leads to cleavage of KCC2, thereby modulating GABAergic signaling. Topics: Action Potentials; Analysis of Variance; Animals; Animals, Newborn; Calcium; Calcium Ionophores; Calpain; Cycloheximide; Cysteine Proteinase Inhibitors; Dipeptides; Dizocilpine Maleate; Dose-Response Relationship, Drug; Emetine; Excitatory Amino Acid Antagonists; Gene Expression Regulation; Hippocampus; In Vitro Techniques; Ionomycin; K Cl- Cotransporters; Leupeptins; Magnesium; Male; Membrane Potentials; N-Methylaspartate; Patch-Clamp Techniques; Protein Synthesis Inhibitors; Pyramidal Cells; Rats; Rats, Wistar; Statistics, Nonparametric; Symporters; Valine | 2012 |
Acceleration of Ca2+ ionophore-induced arachidonic acid liberation by thrombin without the proteolytic action toward the receptor in human platelets.
We investigated the regulation of arachidonic acid liberation catalyzed by group-IV cytosolic phospholipase A2 (cPLA2) in human platelets upon stimulation with thrombin through interaction with protease-activated receptor-1 (PAR-1) or glycoprotein Ib. Leupeptin, a protease inhibitor, completely inhibited thrombin-induced arachidonic acid liberation and Ca2+ mobilization, with inhibition of its protease activity. However, preincubation with thrombin in the presence of leupeptin potentiated Ca2+ ionophore-induced arachidonic acid liberation. The preincubation did not affect the intracellular Ca2+ level or cPLA2 activity in response to ionomycin. Human leukocyte elastase, which cleaves glycoprotein Ib, did not inhibit the enhancement of arachidonic acid liberation by thrombin in the presence of leupeptin. However, the effect of thrombin with leupeptin was abolished by a peptide corresponding to residues 54-65 of hirudin (hirudin peptide), which impairs the binding of thrombin to PAR-1. Furthermore, Phe-Pro-Arg chloromethyl ketone (PPACK)-thrombin, which binds to platelets but has no protease activity, also enhanced Ca2+ ionophore-induced arachidonic acid liberation. In contrast, trypsin with leupeptin did not mimic the effect of thrombin with leupeptin, and furthermore trypsin-induced arachidonic acid liberation was insensitive to hirudin peptide. On the basis of the present results, we suggest that thrombin may accelerate cPLA2-catalyzed arachidonic acid liberation through non-proteolytic action toward PAR-1 but not toward glycoprotein Ib in co-operation with the proteolytic action leading to Ca2+ mobilization. Topics: Amino Acid Chloromethyl Ketones; Antithrombins; Arachidonic Acid; Blood Platelets; Calcium; Enzyme Activation; Hirudins; Humans; Ionomycin; Ionophores; Leukocyte Elastase; Leupeptins; Peptide Fragments; Phospholipases A; Phospholipases A2; Platelet Glycoprotein GPIb-IX Complex; Receptor, PAR-1; Receptors, Thrombin; Thrombin; Trypsin | 1999 |
Regulated expression of the tyrosine hydroxylase gene by membrane depolarization. Identification of the responsive element and possible second messengers.
Prolonged depolarization has been used as a model of adaptive changes in the expression of various proteins, such as ion channels and neurotransmitter biosynthetic enzymes, in response to increased trans-synaptic activity in the nervous system. In depolarized PC12 cells, tyrosine hydroxylase (TH) mRNA levels increased severalfold (Kilbourne, E. J., and Sabban, E. L. (1990) Mol. Brain Res. 8, 121-127). In this study, membrane depolarization caused an increase in the expression of the reporter gene chloramphenicol acetyltransferase (CAT), under transcriptional control of the 5' region of the rat TH gene. These results indicate that membrane depolarization leads to increased transcription of the TH gene. Protein kinase C inhibitors had no effect on the induction of TH mRNA by depolarization, as well as the increase in formation of CAT under control of the upstream region of the TH gene. The depolarization responsive element in the TH gene was mapped to the region containing the cAMP responsive element. This region of the TH gene also increased CAT activity in response to the calcium ionophore, ionomycin. Interestingly, combined treatment with cAMP analogs and membrane depolarization had a greater effect than either alone on TH mRNA levels, as well as on CAT activity in PC12 cells transfected with the plasmid containing the cAMP responsive element. Topics: 8-Bromo Cyclic Adenosine Monophosphate; Animals; Calcium; Cell Line; Chloramphenicol O-Acetyltransferase; Drug Interactions; Enzyme Activation; Gene Expression Regulation, Enzymologic; Ionomycin; Leupeptins; Membrane Potentials; PC12 Cells; Plasmids; Protein Kinase C; Regulatory Sequences, Nucleic Acid; RNA, Messenger; Second Messenger Systems; Sphingosine; Transcription, Genetic; Tyrosine 3-Monooxygenase; Veratridine | 1992 |