A Transmembrane Single-Polypeptide-Chain (sc) Linker to Connect the Two G-Protein–Coupled Receptors in Tandem and the Design for an In Vivo Analysis of Their Allosteric Receptor- Receptor Interactions

A transmembrane (TM) single-polypeptide-chain (sc) linker can connect two G-protein–coupled receptors (GPCRs) in tandem. The priority of a gene-fusion strategy for any two class A GPCRs has been demonstrated. In the striatal function, dopamine (DA) plays a critical role. In the striatum, how the GPCR for adenosine, subtype A2A (A2AR), contributes to the DA neurotransmission in the “volume transmission”/dual-transmission model has been studied extensively. In addition to the fusion receptor, i.e., the prototype scA2AR/D2R complex (the GPCR for DA, subtype D2), several types were created and tested experimentally. To further elucidate this in vivo, we designed a new molecular tool, namely, the supermolecule scA2AR/D2R. Here, no experiments on its expression were done. However, the TM linker to connect the nonobligate dimer as the transient class A GPCR nanocluster that has not been identified at the cell surface membrane deserves discussion through scA2AR/D2R. Supramolecular designs, are experimentally testable and will be used to confirm in vivo the functions of the two GPCRs interactive in such a low specific signal to the nonspecific noise (S/N) ratio in the neurotransmission in the brain. The sc also has, at last, become straightforward in the field of GPCRs, similar to in the field of antibody

Regulation of Synaptotagmin Gene Expression during Ascidian Embryogenesis

AbstractThe ascidian embryo, a model for the primitive mode of chordate development, rapidly forms a dorsal nervous system which consists of a small number of neurons. Here, we have characterized the transcriptional regulation of an ascidian synaptotagmin (syt) gene to explore the molecular mechanisms underlying development of synaptic transmission. In situ hybridization showed that syt is expressed in all neurons described in previous studies and transiently in the embryonic epidermis. Neuronal expression of syt requires induction from the vegetal side of the embryo, whereas epidermal expression occurs autonomously in isolated ectodermal blastomeres. Introduction of green fluorescent protein reporter gene constructs into the ascidian embryos indicates that a genomic fragment of the 3.4-kb 5′ upstream region contains promoter elements of syt gene. Deletion analysis of the promoter suggests that syt expression in neurons and in the embryonic epidermis depends on distinct cis-regulatory regions

H-ZSM-5ゼオライト上でのオレフィンの水素化

H-ZSM-5ゼオライトのオレフィン水素化能を調べるため, 1段目の反応管にZn-Cr系メタノール合成触媒, 2段目の反応管にH-ZSM-5 ゼオライトを充てんした2段反応装置を用いて, メタノールを経由する合成ガスからの炭化水素合成を検討した。1段目の反応圧を40kg/cm2とし, 2段目の反応圧を5～45kg/cm2と変化させたときの結果をFig. 1に示す。2段目の反応圧が20kg/cm2以上では, エタン, プロパンは生成したが, エチレン, プロピレンは全く生成しなかった。このことは, これらパラフィンがH-ZSM-5ゼオライト上で水素化あるいは水素移行反応により生成したことを示唆している。この違いを明らかにするためC2H4-H2, C2H4-Heの2種類の混合ガスをH-ZSM-5ゼオライトに導入した (Fig. 2)。C2H4-H2系では, エタン収率は反応温度と共に著しく増大し, 545°Cでは導入したエチレンの95%がエタンとして検出された。一方, 水素移行反応によるパラフィン生成の可能性を調べるため行ったC2H4-He系では, 550°Cで導入したエチレンの10%がエタンとして検出された。したがって, C2H4-H2系で生成したエタンの大部分はエチレンの水素化により生成したことになる。 また, Fig. 3には, 不純物の影響を調べるため, Feを導入したZSM-5型ゼオライトを用いて合成ガスの転化反応を行った結果を示す。Fe(II) より調製したZSM-5型ゼオライトの生成物はオレフィン指向であった。Fe(II) とAlより調製し, プロトン化した酸点をもつH-ZSM-5型ゼオライトの生成物はパラフィン指向であった。 以上の結果より, H-ZSM-5ゼオライトはオレフィン水素化能を有していると結論した。Conversion of synthesis gas to hydrocarbons was carried out in a two-stage system consisting of a methanol synthesis reactor followed by a hydrocarbon-forming reactor containg H-ZSM-5 zeolite. It was found that the H-ZSM-5 zeolite catalyst was active in olefin hydrogenation

Zfp238 Regulates the Thermogenic Program in Cooperation with Foxo1

Summary: Obesity has become an explicit public health concern because of its relevance to metabolic syndrome. Evidence points to the significance of beige adipocytes in regulating energy expenditure. Here, using yeast two-hybrid screening, we show that Zfp238 is a Foxo1 co-repressor and that adipose-tissue-specific ablation of Zfp238 (Adipo-Zfp238KO) in mice leads to obesity, decreased energy expenditure, and insulin resistance under normal chow diet. Adipo-Zfp238KO inhibits induction of Ucp1 expression in subcutaneous adipose tissue upon cold exposure or CL316243, but not in brown adipose tissue. Furthermore, knockdown of Zfp238 in 3T3-L1 cells decreases Ucp1 expression in response to cool incubation or forskolin significantly compared with control cells. In contrast, overexpression of Zfp238 in 3T3-L1 cells significantly increases Ucp1 expression in response to forskolin. Finally, double knockdown of both Zfp238 and Foxo1 normalizes Ucp1 induction. These data suggest that Zfp238 in adipose tissue regulates the thermogenic program in cooperation with Foxo1. : Molecular Interaction; Molecular Mechanism of Behavior; Diabetology; Specialized Functions of Cells Subject Areas: Molecular Interaction, Molecular Mechanism of Behavior, Diabetology, Specialized Functions of Cell