Long-term potentiation (LTP) of synaptic strength at Schaffer collateral synapses has largely been attributed to changes in the number and biophysical properties of AMPA receptors (AMPARs). Small-conductance Ca 2+ -activated K + channels (SK2 channels) are functionally coupled with NMDA receptors (NMDARs) in CA1 spines such that their activity modulates the shape of excitatory postsynaptic potentials (EPSPs) and increases the threshold for induction of LTP. Here we show that LTP induction in mouse hippocampus abolishes SK2 channel activity in the potentiated synapses. This effect is due to SK2 channel internalization from the postsynaptic density (PSD) into the spine. Blocking PKA or cell dialysis with a peptide representing the C-terminal domain of SK2 that contains three known PKA phosphorylation sites blocks the internalization of SK2 channels after LTP induction. Thus the increase in AMPARs and the decrease in SK2 channels combine to produce the increased EPSP underlying LTP.Activity-dependent changes in synaptic strength are widely believed to underlie the cellular mechanisms of learning and memory. This view has gained significant support from recent studies showing that learning induces long-lasting changes in synaptic strength 1-3 . At Schaffer collateral-to-CA1 synapses in the hippocampus, stimulation protocols that coordinate presynaptic and postsynaptic activity to induce LTP affect the postsynaptic cell through a process that is dependent upon NMDAR activity and Ca 2+ influx into the stimulated dendritic spine 4-6 . LTP inducing protocols act through PKA and calcium/calmodulin-dependent kinase II (CaMKII) to alter the biophysical properties 7-9 and increase the number of AMPARs in the PSD 10-14 .SK2 channels are activated solely by intracellular Ca 2+ ions, with submicromolar Ca 2+ affinity 15 , and are selectively blocked by the peptide toxin apamin. SK2 channels are expressed throughout the dendritic arbor of CA1 neurons and in dendritic spines 16,17 . Whole-cell current-clamp recordings and Ca 2+ imaging revealed that spine SK2 channels are activated by synaptically driven Ca 2+ influx 16 . The repolarizing effect of SK2 channel activity opposes the depolarizing effect of AMPAR activity, reducing the EPSP, favoring Mg 2+ reblocking of NIH Public Access Author ManuscriptNat Neurosci. Author manuscript; available in PMC 2009 January 5. Published in final edited form as:Nat Neurosci. 2008 February ; 11(2): 170-177. doi:10.1038/nn2041. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptNMDARs and reducing the Ca 2+ transient 16 . SK2 channels are therefore ideally suited to modulate the induction of synaptic plasticity. Indeed, field recordings in area CA1 showed that blocking SK2 channels facilitates the induction of synaptic plasticity. In addition, administration of apamin to mice facilitates hippocampal-dependent memory encoding 18 . In contrast, overexpression of SK2 channels in transgenic mice impairs the induction of synaptic plasticity and severely impairs hippoc...

Vitamin D deficiency is associated with muscle weakness, pain, and atrophy. Serum vitamin D predicts muscle strength and age-related muscle changes. However, precise mechanisms by which vitamin D affects skeletal muscle are unclear. To address this question, this study characterizes the muscle phenotype and gene expression of mice with deletion of vitamin D receptor (VDRKO) or diet-induced vitamin D deficiency. VDRKO and vitamin D-deficient mice had significantly weaker grip strength than their controls. Weakness progressed with age and duration of vitamin D deficiency, respectively. Histological assessment showed that VDRKO mice had muscle fibers that were significantly smaller in size and displayed hyper-nuclearity. Real-time PCR also indicated muscle developmental changes in VDRKO mice with dysregulation of myogenic regulatory factors (MRFs) and increased myostatin in quadriceps muscle (>2-fold). Vitamin D-deficient mice also showed increases in myostatin and the atrophy marker E3-ubiqutin ligase MuRF1. As a potential explanation for grip strength weakness, both groups of mice had down-regulation of genes encoding calcium-handling and sarco-endoplasmic reticulum calcium transport ATPase (Serca) channels. This is the first report of reduced strength, morphological, and gene expression changes in VDRKO and vitamin D-deficient mice where confounding by calcium, magnesium, and phosphate have been excluded by direct testing. Although suggested in earlier in vitro work, this study is the first to report an in vivo association between vitamin D, myostatin, and the regulation of muscle mass. These findings support a direct role for vitamin D in muscle function and corroborate earlier work on the presence of VDR in this tissue.

SUMMARY Ca2+-activated SK channels and voltage-gated A-type Kv4 channels shape dendritic excitatory postsynaptic potentials (EPSPs) in hippocampal CA1 pyramidal neurons. Synaptically evoked Ca2+ influx through N-methyl-D-aspartate receptors (NMDARs) activates spine SK channels, reducing EPSPs and the associated spine head Ca2+ transient. However, results using glutamate uncaging implicated Ca2+ influx through SNX-482 (SNX) sensitive Cav2.3 (R-type) Ca2+ channels as the Ca2+ source for SK channel activation. The present findings show that using Schaffer collateral stimulation the effects of SNX and apamin are not mutually exclusive and SNX increases EPSPs independent of SK channel activity. Dialysis with 1,2-bis(o-aminophenoxy)ethane-N’N’N’-tetraacetic acid (BAPTA), application of 4-Aminopyridine (4-AP), expression of a Kv4.2 dominant negative subunit, and dialysis with a KChIPs antibody occluded the SNX-induced increase of EPSPs. The results suggest two distinct Ca2+ signaling pathways within dendritic spines, that links Ca2+ influx through NMDARs to SK channels and Ca2+ influx through R-type Ca2+ channels to Kv4.2-containing channels.

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SK2-containing channels are expressed in the postsynaptic density (PSD) of dendritic spines on mouse CA1 pyramidal neurons, and influence synaptic responses, plasticity, and learning. The SK2 gene encodes two isoforms differing only in the length of the N-terminal domain. SK2-Long (SK2-L) and SK2-Short (SK2-S) are co-expressed in CA1 pyramidal neurons and likely form heteromeric channels. In mice lacking SK2-L (SK2-Sonly mice), SK2-S-containing channels were expressed in the extrasynaptic membrane, but were excluded from the PSD. The SK channel contribution to EPSPs was absent in SK2-Sonly mice, and was restored by SK2-L re-expression. In slices from wild type mice, blocking SK channels increased the amount of long-term potentiation (LTP) induced in area CA1 but was without effect in SK2-Sonly mice. Further, SK2-Sonly mice outperformed wild type mice in the novel object recognition task. These results show that SK2-L directs synaptic SK2-containing channel expression, important for normal synaptic signaling, plasticity, and learning.

Objective Intermediate and small conductance KCa channels IK1 (KCa3.1) and SK3 (KCa2.3) are primary targets of endothelial Ca2+ signals in the arterial vasculature and their ablation results in increased arterial tone and hypertension. Activation of IK1 channels by local Ca2+ transients from internal stores or plasma membrane channels promotes arterial hyperpolarization and vasodilation. Here, we assess arteries from genetically altered IK1 knockout mice (IK1−/−) to determine whether IK1 channels exert a positive feedback influence on endothelial Ca2+ dynamics. Approach and Results Using confocal imaging and custom data analysis software we found that while the occurrence of basal endothelial Ca2+ dynamics was not different between IK1−/− and wild-type (WT) mice (p > 0.05), the frequency of acetylcholine (ACh 2 µM)-stimulated Ca2+ dynamics was greatly depressed in IK1−/− endothelium (515 ± 153 vs. 1860 ± 319 events; p < 0.01). In IK1−/−/SK3T/T mice, ancillary suppression (+Dox) or overexpression (−Dox) of SK3 channels had little additional impact on the occurrence of events under basal or ACh-stimulated conditions. SK3 overexpression did, however, restore the depressed event amplitudes. Removal of extracellular Ca2+ reduced ACh-induced Ca2+ dynamics to the same level in WT and IK1−/− arteries. Blockade of IK1 and SK3 with the combination of charybdotoxin (0.1 µM) and apamin (0.5 µM) or TRPV4 channels with HC-067047 (1 µM) reduced ACh Ca2+ dynamics in WT arteries to the level of IK1−/−/SK3T/T+Dox arteries. These drug effects were not additive. Conclusions IK1, and to some extent SK3 channels, exert a substantial positive feedback influence on endothelial Ca2+ dynamics.

We investigated the temporal and spatial expression of SK2 in the developing mouse hippocampus using molecular and biochemical techniques, quantitative immunogold electron microscopy and electrophysiology. The mRNA encoding SK2 was expressed in the developing and adult hippocampus. Western blotting and immunohistochemistry showed that SK2 protein increased with age. This was accompanied by a shift in subcellular localization. Early in development (P5), SK2 was predominantly localized to the endoplasmic reticulum in the pyramidal cell layer. But by P30 SK2 was almost exclusively expressed in the dendrites and spines. The level of SK2 at the postsynaptic density (PSD) also increased during development. In the adult, SK2 expression on the spine plasma membrane showed a proximal-to-distal gradient. Consistent with this redistribution and gradient of SK2, the selective SK channel blocker apamin increased evoked excitatory postsynaptic potentials (EPSPs) only in CA1 pyramidal neurons from mice older than P15. However, the effect of apamin on EPSPs was not different between synapses in proximal or distal stratum radiatum or stratum lacunosum-moleculare in adult. These results show a developmental increase and gradient in SK2-containing channel surface expression that underlie their influence on neurotransmission, and that may contribute to increased memory acquisition during early development.

Patients with nosocomial pneumonia exhibit elevated levels of neurotoxic amyloid and tau proteins in the cerebrospinal fluid (CSF). In vitro studies indicate that pulmonary endothelium infected with clinical isolates of either Pseudomonas aeruginosa, Klebsiella pneumoniae, or Staphylococcus aureus produces and releases cytotoxic amyloid and tau proteins. However, the effects of the pulmonary endothelium‐derived amyloid and tau proteins on brain function have not been elucidated. Here, we show that P. aeruginosa infection elicits accumulation of detergent insoluble tau protein in the mouse brain and inhibits synaptic plasticity. Mice receiving endothelium‐derived amyloid and tau proteins via intracerebroventricular injection exhibit a learning and memory deficit in object recognition, fear conditioning, and Morris water maze studies. We compared endothelial supernatants obtained after the endothelia were infected with P. aeruginosa possessing an intact [P. aeruginosa isolated from patient 103 (PA103) supernatant] or defective [mutant strain of P. aeruginosa lacking a functional type 3 secretion system needle tip complex (ΔPcrV) supernatant] type 3 secretion system. Whereas the PA103 supernatant impaired working memory, the ΔPcrV supernatant had no effect. Immunodepleting amyloid or tau proteins from the PA103 supernatant with the A11 or T22 antibodies, respectively, overtly rescued working memory. Recordings from hippocampal slices treated with endothelial supernatants or CSF from patients with or without nosocomial pneumonia indicated that endothelium‐derived neurotoxins disrupted the postsynaptic synaptic response. Taken together, these results establish a plausible mechanism for the neurologic sequelae consequent to nosocomial bacterial pneumonia.—Balczon, R., Pittet, J.‐F., Wagener, B. M., Moser, S. A., Voth, S., Vorhees, C. V., Williams, M. T., Bridges, J. P., Alvarez, D. F., Koloteva, A., Xu, Y., Zha, X.‐M., Audia, J. P., Stevens, T., Lin, M. T. Infection‐induced endothelial amyloids impair memory. FASEB J. 33, 10300–10314 (2019). http://www.fasebj.org