Inducible and selective erasure of memories in the mouse brain via chemical-genetic manipulation

Rapid and selective erasures of certain types of memories in the brain would be desirable under certain clinical circumstances. By employing an inducible and reversible chemical-genetic technique, we find that transient alphaCaMKII overexpression at the time of recall impairs the retrieval of both newly formed one-hour object recognition memory and fear memories, as well as 1-month-old fear memories. Systematic analyses suggest that excessive alphaCaMKII activity-induced recall deficits are not caused by disrupting the retrieval access to the stored information but are, rather, due to the active erasure of the stored memories. Further experiments show that the recall-induced erasure of fear memories is highly restricted to the memory being retrieved while leaving other memories intact. Therefore, our study reveals a molecular genetic paradigm through which a given memory, such as new or old fear memory, can be rapidly and specifically erased in a controlled and inducible manner in the brain.     

Roles of autophagy and mTOR signaling in neuronal differentiation of mouse neuroblastoma cells

Cell differentiation is often associated with decreased cell growth, indicating an altered rate of macromolecule synthesis and degradation. In this study, we present evidence that autophagy, a process for bulk degradation of cytoplasm, is activated during retinoic acid-induced neuronal differentiation of neuroblastoma N2a cells. Chemical inhibitors of autophagy, including 3-MA and LY294002, abrogate cell differentiation. RNA interference of autophagy gene beclin 1 markedly delays the process of differentiation. We also find that cell differentiation is accompanied by decreased activity of mTOR, a major controller of cell growth and a negative regulator of autophagy. However, completely inhibiting mTOR by rapamycin decreases neurite outgrowth, cell size and the immunoreactivity for neuronal markers. Our study suggests that an appropriate level of mTOR activity is important in cell differentiation for a balance between macromolecule synthesis and degradation.     

Cytotoxic ent-abietane diterpenes from Gelonium aequoreum

Seventeen ent-abietane diterpenes, including gelomulides K-X (1-14), and three known compounds, were isolated from a dichloromethane-soluble extract of Gelonium aequoreum through bioassay-guided fractionation. Their structures were identified by spectroscopic methods, and stereochemistry was confirmed by X-ray crystallographic analysis, CD spectral data, and Mosher's method. The isolates were evaluated for in vitro cytotoxic activity, and compounds 1 and 3 showed moderate cytotoxicity against lung (A549), breast (MDA-MB-231 and MCF7), and liver (HepG2) cancer cell lines.     

Quantification of N2O fluxes from soil–plant systems may be biased by the applied gas chromatograph methodology

With regard to measuring nitrous oxide (N₂O) emissions from biological sources, there are three most widely adopted methods that use gas chromatograph with an electron capture detector (GC–ECD). They use: (a) nitrogen (N₂) as the carrier gas (DN); (b) ascarite as a carbon dioxide (CO₂) trap with DN (DN-Ascarite); and (c) a mixture gas of argon and methane as the carrier (AM). Additional methods that use either a mixture of argon and methane (or of CO₂ and N₂) as a make-up gas with the carrier nitrogen or soda lime (or ascarite) as a CO₂ trap with the carrier helium have also been adopted in a few studies. To test the hypothesis that the use of DN sometimes considerably biases measurements of N₂O emissions from plants, soils or soil–plant systems, experiments were conducted involving DN, AM and DN-Ascarite. When using DN, a significant relationship appeared between CO₂ concentrations and the apparent N₂O concentrations in air samples. The use of DN led to significantly overestimated N₂O emissions from detached fresh plants in static chamber enclosures. Meanwhile, comparably lower emissions were found when using either the DN-Ascarite or AM methods. When an N₂O flux (from a soil or a soil–plant system), measured by DN in combination with sampling from the enclosure of a static opaque chamber, was greater than 200 μg N m^?2 h^?1, no significant difference was found between DN and DN-Ascarite. When the DN-measured fluxes were within the ranges of <?30, ?30–0, 0–30, 30–100 and 100–200 μg N m^?2 h^?1, significant differences that amounted to ?72, ?22, 5, 38 and 64 μg N m^?2 h^?1, respectively, appeared in comparison to DN-Ascarite. As a result, the DN measurements in rice–wheat and vegetable fields overestimated both annual total N₂O emissions (by 7–62%, p?<?0.05) and direct emission factors for applied nitrogen (by 6–65%). These results suggest the necessity of reassessing the available data determined from DN measurements before they are applied to inventory estimation. Further studies are required to explore appropriate approaches for the necessary reassessment. Our results also imply that the DN method should not be adopted for measuring N₂O emissions from weak sources (e.g., with intensities less than 200 μg N m^?2 h^?1). In addition, we especially do not recommend the use of DN to simultaneously measure N₂O and CO₂ with the same ECD.     

表达鸡传染性支气管炎病毒JS/95/03株S1蛋白的重组杆状病毒构建

The recombinant baculovirus expressing S1 glycoprotein of nephropathogenic strain JS/95/03 of infectious bronchitis virus was generated by using the Bac-to-Bac baculovirus expression system.The BamH I-Sal Ⅰ fragment containing S1 gene from the recombinant plasmid pMDJS9503S1 was purified and cloned in frame into the baculovirus transposing vector pFASTBAC HTa under the polyhedrin gene promoter.The recombinant transposing plasmid pFASTJS9503S1 was screened and then transformed into Escherichia coli DH10BAC.The resulting recombinant bacmid rBacmidJS9503S1 was transfected into cells of the insect Spodoptera frugiperda(Sf9)and the recombinant baculoviruse rAcJS9503S1 was obtained.The lysates of cells infected with rAcJS9503S1 were analyzed by SDS-PAGE and the expressed product of S1 gene was detected by Western bloting and immunofluorescence assay(IFA).The results showed the recombinant baculovirus was fully capable of expressing S1 glycoprotein of JS/95/03.Maybe owing to the incomplete glycosylation in insect cells,the S1 gene product had a Mr of only 61000.In immunofluorescence test and Western blotting,the expressed product could react with polycolonal antibody against IBV M41 strain,indicating it possessed the antigenic properties specific for native S1 glycoprotein.     

激光诱变产生无核沙田柚的胚胎学研究

采用激光诱变的方法已经成功的培育出无核(少核)沙田柚,我们采用德国产Leica DMLB生物显微镜MPS60照相系统对采摘回的花蕊(经生物技术处理)进行拍照,与未经激光处理的对照组做比较,分析了雌雄配子体发育过程,对无核果作用机理研究提供参考。     

胡萝卜光自养型愈伤组织的诱导培养及其光合特性

经诱导得到胡萝卜光自养型愈伤组织。以叶绿素为单位计算,获得的光自养型愈伤组织的光合活力达到甚至超过了整体植株叶片水平。同时测定愈伤组织光自养过程中光合特性的变化,结果表明其叶绿素含量逐渐上升、暗呼吸速率和Chl a/Chl b比值逐渐下降。并且用电子显微镜观察到愈伤组织中叶绿体结构逐渐发育的过程。     

Partial depletion and repopulation of microglia have different effects in the acute MPTP mouse model of Parkinson’s disease

ObjectivesParkinson''s disease (PD) is a common neurodegenerative disorder characterized by the progressive and selective degeneration of dopaminergic neurons. Microglial activation and neuroinflammation are associated with the pathogenesis of PD. However, the relationship between microglial activation and PD pathology remains to be explored.Materials and MethodsAn acute regimen of MPTP was administered to adult C57BL/6J mice with normal, much reduced or repopulated microglial population. Damages of the dopaminergic system were comprehensively assessed. Inflammation‐related factors were assessed by quantitative PCR and Multiplex immunoassay. Behavioural tests were carried out to evaluate the motor deficits in MPTP‐challenged mice.ResultsThe receptor for colony‐stimulating factor 1 inhibitor PLX3397 could effectively deplete microglia in the nigrostriatal pathway of mice via feeding a PLX3397‐formulated diet for 21 days. Microglial depletion downregulated both pro‐inflammatory and anti‐inflammatory molecule expression at baseline and after MPTP administration. At 1d post‐MPTP injection, dopaminergic neurons showed a significant reduction in PLX3397‐fed mice, but not in control diet (CD)‐fed mice. However, partial microglial depletion in mice exerted little effect on MPTP‐induced dopaminergic injuries compared with CD mice at later time points. Interestingly, microglial repopulation brought about apparent resistance to MPTP intoxication.ConclusionsMicroglia can inhibit PD development at a very early stage; partial microglial depletion has little effect in terms of the whole process of the disease; and microglial replenishment elicits neuroprotection in PD mice.