Biology subjects

Druzin, M.

Publications and source records attributed to Druzin, M..

2 recordsLinked to original sources

bioRxiv2023

Extracellular Matrix Regulates Neuronal Chloride Concentration via K+-Cl--Cotransporter 2

The neuronal intracellular chloride concentration [Cl-]i is critical for {gamma}-aminobutyric acid type A (GABAA) receptor-mediated transmission. Degradation of the extracellular matrix (ECM) is associated with raised [Cl-]I but neither the mechanisms underlying this effect nor the consequences for GABA- mediated transmission are well understood. Hitherto it has been unclear how to reconcile the effect of the ECM on [Cl-]i with the established role of cation-chloride cotransporters in setting [Cl-]I. In the present work we clarify the role of the ECM in the control of neuronal [Cl-]i. By measuring [Cl-]i in central neurons from male rats we show that the ECM affects basal [Cl-]i as well as the rate of Cl- extrusion after a high load. The mechanism is not via impermeant anions but through regulation of K+-Cl--cotransporter 2 (KCC2). ECM degradation is accompanied by an N-type Ca2+-channel- and calpain-dependent reduction in the amount of KCC2 protein, increased basal [Cl-]i, reduced Cl- extrusion capacity as well as by reduced inhibitory, or even an excitatory, effect of intense GABAA- receptor mediated trans mission. This implies a previously unrecognized pathway for the control of neuronal [Cl-]i and excitability by the ECM. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=144 SRC="FIGDIR/small/527837v1_ufig1.gif" ALT="Figure 1"> View larger version (53K): org.highwire.dtl.DTLVardef@1ee0d30org.highwire.dtl.DTLVardef@1a3eb1aorg.highwire.dtl.DTLVardef@a00b44org.highwire.dtl.DTLVardef@143b43f_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

bioRxiv2023

Heparan sulfates regulate axonal excitability and context generalization through Ca2+/calmodulin-dependent protein kinase II

Our previous studies demonstrated that enzymatic removal of highly sulfated heparan sulfates with heparinase 1 impaired axonal excitability and reduced expression of ankyrin G at the axon initial segments in the CA1 region of the hippocampus ex vivo, impaired context discrimination in vivo, and elevated Ca2+/calmodulin-dependent protein kinase II (CaMKII) activity in vitro. Here, we show that in vivo delivery of heparinase 1 in the CA1 region of the hippocampus elevated autophosphorylation of CaMKII 24 hours after injection in mice. Patch-clamp recording in CA1 neurons revealed no significant heparinase effects on the amplitude or frequency of miniature excitatory and inhibitory postsynaptic currents, while the threshold for action potential generation was increased and fewer spikes were generated in response to current injection. Delivery of heparinase on the next day after contextual fear conditioning induced context overgeneralization 24 hours after injection. Co-administration of heparinase with the CaMKII inhibitor (autocamtide-2-related inhibitory peptide) rescued neuronal excitability and expression of ankyrin G at the axon initial segment. It also restored context discrimination, suggesting a key role of CaMKII in neuronal signaling downstream of heparan sulfate proteoglycans and highlighting a link between impaired CA1 pyramidal cell excitability and context generalization during recall of contextual memories.

neuroscience↗