Bidirectional Control of Postsynaptic Density-95 (PSD-95) Clustering by Huntingtin

Huntington disease is associated with early alterations in corticostriatal synaptic function that precede cell death, and it is postulated that ameliorating such changes may delay clinical onset and/or prevent neurodegeneration. Although many of these synaptic alterations are thought to be attributable to a toxic gain of function of the mutant huntingtin protein, the role that nonpathogenic huntingtin (HTT) plays in synaptic function is relatively unexplored. Here, we compare the immunocytochemical localization of a major postsynaptic scaffolding protein, PSD-95, in striatal neurons from WT mice and mice overexpressing HTT with 18 glutamine repeats (YAC18, nonpathogenic). We found that HTT overexpression resulted in a palmitoylation- and BDNF-dependent increase in PSD-95 clustering at synaptic sites in striatal spiny projection neurons (SPNs) co-cultured with cortical neurons. Surprisingly, the latter effect was mediated presynaptically, as HTT overexpression in cortical neurons alone was sufficient to increase PSD-95 clustering in the postsynaptic SPNs. In contrast, antisense oligonucleotide knockdown of HTT in WT co-cultures resulted in a significant reduction of PSD-95 clustering in SPNs. Notably, despite these bidirectional changes in PSD-95 clustering, we did not observe an alteration in basal electrophysiological measures of AMPA and NMDA receptors. Thus, unlike in previous studies in the hippocampus, enhanced or decreased PSD-95 clustering alone was insufficient to drive AMPA or NMDA receptors into or out of SPN synapses. In all, our results demonstrate that nonpathogenic HTT can indeed influence synaptic protein localization and uncover a novel role of HTT in PSD-95 distribution.     

Microbial Culture Collection (MCC) and International Depositary Authority (IDA) at National Centre for Cell Science,Pune

Culture collections are valuable resources for the sustainable use of microbial diversity and its conservation. Advances in biotechnology have further increased their importance and some of these have been recognized as International Depositary Authority (IDA) for the deposition of patent cultures. Microbial Culture Collection at National Centre for Cell Science was established by the Department of Biotechnology, Government of India is country’s newest culture collection with largest holdings. It is recognized as an IDA under the Budapest Treaty and Designated National Repository under the Biodiversity Act 2002. This article describes its various service related activities.     

响应面法优化海洋镰刀腐皮菌产出HMG-CoA还原酶抑制剂的培养条件

【目的】采用响应面法对海洋微生物镰刀腐皮菌FG319发酵产生MFS(Metabolite of Fusarium solani FG319)培养条件进行优化。【方法】在单因素试验结果的基础上,依据Box-Behnken中心组合原则设计诱导物添加量、培养时间、培养温度的3因素3水平响应面实验,以MFS产出量为响应值优化镰刀腐皮菌FG319的培养条件。【结果】镰刀腐皮菌FG319最优培养条件为诱导物添加量0.6%、培养时间7 d、培养温度22°C,在此培养条件下,MFS最高产量达到20.11 mg/L,是优化前的4倍,与理论预测的相对误差为0.64%,实测值与响应面预测值拟合良好。镰刀腐皮菌FG319代谢产物MFS在HMG-CoA还原酶和NADPH构筑的分子评价反应体系,当MFS添加到100 mg/L时,具有类似洛伐他汀抑制HMG-CoA还原酶的最大抑制率。【结论】响应面试验设计对镰刀腐皮菌FG319培养条件的优化是有效的,其次生代谢产物MFS体外抑制HMG-CoA还原酶的效果也是明显的。     

L-天冬酰胺酶工程菌株培养条件及稳定性

L-天冬酰胺酶工程菌株的酶活和表达水平受菌体生物量和诱导时间的影响。在生物量Ａ６００03×１０左右,热诱导4h酶活力和表达水平可达到较高水平。葡萄糖对酶的生成有阻遏作用,当葡萄糖浓度大于025%时,对工程菌酶的合成造成阻遏。确定了工程菌培养的培养基、pH值、接种量等因素。重组质粒pASN在\%E.coli\% JM105,TG1和AS1357等宿主菌中具有很好的稳定性,工程菌培养50代以上重组质粒保留90%以上,在LB和M\|3培养基中也较稳定。     

诱导风信子再生花芽不断分化花被片的研究

通过外源激素及外植体年龄的控制,诱导风信子再生花芽不断分化花被片已经获得成功。在２５０ｄ的继代培养中平均每个花芽可分化７０多片花被片,最多的可分化１４０多片。对这种花芽不断分化花被片的形态发生过程以及生长发育特点的观察表明,花芽的第１轮器官与风信子野生型花基本相同,查花被,它由花被筒及其上部的裂片－花被片组成。     

Gari as culture medium for moulds

No significant differences (P<0.05 occurred in the frequency of isolation from soil of four common moulds – Rhizopus stolonifer, Monilia sitophila, Aspergillus niger and Penicillium sp., on malt extract agar (MEA) and gelled gari (carbohydrate gel). Growth of the moulds on slide microcultures of potato dextrose agar (PDA) and gari showed typical diagnostic features of the organisms, which were often clearer on gari. It is concluded that gari, which is much cheaper and far more readily available in Nigeria than MEA or PDA, is an effective alternative to the two standard mycological media.     

藻类批量培养中的比增长率最大值

批量培养谷皮菱形藻、莱茵衣藻、粉核小球藻、斜生栅藻、羊角月牙藻和铜绿微囊藻等六种藻类,在对数生长期,逐日计数而确定比增长率最大值（μｍａｒ）,发现（μｍａｒ）值出现的时间在各种藻类中不同。同时讨论了每种藻类的（μｍａｒ）与营养物质、光强、温度和起始浓度之间的关系。铜绿微囊藻的（μｍａｒ）在一定的总磷（ＴＰ）浓度范围内随ＴＰ浓度呈上升趋势。根据获得增大率最大值的条件,对它们在浮游植物群落中的生态位作     

小鼠胚胎体外培养条件的研究

为了探讨小鼠胚胎在体外发育过程中的最佳方案,我们将从超排昆明小鼠取出的５４３枚受精卵经过不同的培养液（Ｍ１６、Ｍ１６＋ＯＥＣ、Ｇｌｕ－ＦｒｅｅＭ１６和Ｇｌｕ－ＦｒｅｅＭ１６＋ＯＥＣ）及不同的培养微环境处理后,观察胚胎体外发育的过程。结果表明,葡萄糖在胚胎发育早期（２细胞期）有比较明显的发育阻断作用,同种输卵管上皮（ＯＥＣ）共同培养能够有效地抑制这种阻断作用;葡萄糖对胚胎８－１６细胞期及以后阶段的发育具有十分重要的作用,而此时ＯＥＣ的作用则不明显;除了葡萄糖外,在早期可能还有其它一些象磷酸盐、重金属离子的物质也能起胚胎发育阻断作用。同时还发现培养微环境的稳定也是胚胎发育的重要保证条件,石蜡油的封盖能够保持某种特定微环境而支持胚胎的体外发育。     

Potential of ligninolytic enzymatic complex produced by white‐rot fungi from genus Trametes isolated from Bulgarian forest soil

Because of the crucial role of ligninolytic enzymes in a variety of industrial processes, the demand for a new effective producer has been constantly increasing. Furthermore, information on enzyme synthesis by autochthonous fungal strains is very seldom found. Two fungal strains producing ligninolytic enzymes were isolated from Bulgarian forest soil. They were identified as being Trametes trogii and T. hirsuta. These two strains were assessed for their enzyme activities, laccase (Lac), lignin peroxidase (LiP) and Mn‐dependent peroxidase (MnP) in culture filtrate depending on the temperature and the type of nutrient medium. T. trogii was selected as the better producer of ligninolytic enzymes. The production process was further improved by optimizing a number of parameters such as incubation time, type of cultivation, volume ratio of medium/air, inoculum size and the addition of inducers. The maximum activities of enzymes synthesized by T. trogii was detected as 11100 U/L for Lac, 2.5 U/L for LiP and 4.5 U/L for MnP after 14 days of incubation at 25°C under static conditions, volume ratio of medium/air 1:6, and 3 plugs as inoculum. Among the supplements tested, 5% glycerol increased Lac activity to a significant extent. The addition of 1% veratryl alcohol had a positive effect on MnP.