ABSTRACT
ADNP syndrome, involving the ADNP transcription factor of the SWI/SNF chromatin-remodeling complex, is characterized by developmental delay, intellectual disability, and autism spectrum disorders (ASD). Although Adnp-haploinsufficient (Adnp-HT) mice display various phenotypic deficits, whether these mice display abnormal synaptic functions remain poorly understood. Here, we report synaptic plasticity deficits associated with cognitive inflexibility and CaMKIIα hyperactivity in Adnp-HT mice. These mice show impaired and inflexible contextual learning and memory, additional to social deficits, long after the juvenile-stage decrease of ADNP protein levels to ~10% of the newborn level. The adult Adnp-HT hippocampus shows hyperphosphorylated CaMKIIα and its substrates, including SynGAP1, and excessive long-term potentiation that is normalized by CaMKIIα inhibition. Therefore, Adnp haploinsufficiency in mice leads to cognitive inflexibility involving CaMKIIα hyperphosphorylation and excessive LTP in adults long after its marked expressional decrease in juveniles.
Subject(s)
Autistic Disorder , Intellectual Disability , Mice , Animals , Nerve Tissue Proteins/metabolism , Neuronal Plasticity/genetics , Long-Term Potentiation/genetics , Autistic Disorder/metabolism , Cognition , Homeodomain Proteins/metabolismABSTRACT
In the present study, we showed that hydrophilic graphene can serve as an ideal imaging plate for biological specimens. Graphene being a single-atom-thick semi-metal with low secondary electron emission, array tomography analysis of serial sections of biological specimens on a graphene substrate showed excellent image quality with improvedz-axis resolution, without including any conductive surface coatings. However, the hydrophobic nature of graphene makes the placement of biological specimens difficult; graphene functionalized with polydimethylsiloxane oligomer was fabricated using a simple soft lithography technique and then processed with oxygen plasma to provide hydrophilic graphene with minimal damage to graphene. High-quality scanning electron microscopy images of biological specimens free from charging effects or distortion were obtained, and the optical transparency of graphene enabled fluorescence imaging of the specimen; high-resolution correlated electron and light microscopy analysis of the specimen became possible with the hydrophilic graphene plate.
Subject(s)
Graphite , Dimethylpolysiloxanes , Microscopy, Electron, Scanning , Optical Imaging , OxygenABSTRACT
The molecular basis of Z-DNA recognition and stabilization is mostly discovered via X-ray crystallography. The sequences composed with alteration of purine and pyrimidine are known to adopt Z-DNA conformation. Due to the energy penalty for forming Z-DNA, the small molecular stabilizer or Z-DNA-specific binding protein is required for DNA to adopt Z conformation prior to crystallizing Z-DNA. Here we described the methods ranging from preparation of DNA and Z-alpha protein to crystallization of Z-DNA in detail.
Subject(s)
DNA, Z-Form , Crystallization , Models, Molecular , Base Sequence , DNA/chemistry , Nucleic Acid Conformation , Crystallography, X-RayABSTRACT
The left-handed Z-DNA is surrounded by right-handed canonical B-DNA, and thus the junction between B- and Z-DNA has been occurred during temporal Z-DNA formation in the genome. The base extrusion structure of the BZ junction may help detect Z-DNA formation in DNAs. Here we describe the BZ junction structural detection by using 2-aminopurine (2AP) fluorescent probe. BZ junction formation can be measured in solution by this method.
Subject(s)
DNA, B-Form , DNA, Z-Form , DNA/genetics , 2-Aminopurine/chemistry , DNA Replication , Nucleic Acid ConformationABSTRACT
In the present study, we used the electrochemical transparency of graphene to show that the direct intercalation of alkali-metal cations is not a prerequisite for the redox reaction of Prussian blue (PB). PB thin films passivated with monolayer graphene still underwent electrochemical redox reactions in the presence of alkali-metal ions (K+ or Na+) despite the inability of the cations to penetrate the graphene and be incorporated into the PB. Graphene passivation not only preserved the electrochemical activity of the PB but also substantially enhanced the stability of the PB. As a proof of concept, we showed that a transparent graphene electrode covering PB can be used as an excellent hydrogen peroxide transducer, thereby demonstrating the possibility of realizing an electrochemical sensor capable of long-term measurements.
ABSTRACT
Visual arrestin (Arr1) terminates rhodopsin signaling by blocking its interaction with transducin. To do this, Arr1 translocates from the inner to the outer segment of photoreceptors upon light stimulation. Mounting evidence indicates that inositol phosphates (InsPs) affect Arr1 activity, but the Arr1-InsP molecular interaction remains poorly defined. We report the structure of bovine Arr1 in a ligand-free state featuring a near-complete model of the previously unresolved C-tail, which plays a crucial role in regulating Arr1 activity. InsPs bind to the N-domain basic patch thus displacing the C-tail, suggesting that they prime Arr1 for interaction with rhodopsin and help direct Arr1 translocation. These structures exhibit intact polar cores, suggesting that C-tail removal by InsP binding is insufficient to activate Arr1. These results show how Arr1 activity can be controlled by endogenous InsPs in molecular detail.