Temperature as a universal resetting cue for mammalian circadian oscillators

Environmental temperature cycles are a universal entraining cue for all circadian systems at the organismal level with the exception of homeothermic vertebrates. We report here that resistance to temperature entrainment is a property of the suprachiasmatic nucleus (SCN) network and is not a cell-autonomous property of mammalian clocks. This differential sensitivity to temperature allows the SCN to drive circadian rhythms in body temperature, which can then act as a universal cue for the entrainment of cell-autonomous oscillators throughout the body. Pharmacological experiments show that network interactions in the SCN are required for temperature resistance and that the heat shock pathway is integral to temperature resetting and temperature compensation in mammalian cells. These results suggest that the evolutionarily ancient temperature resetting response can be used in homeothermic animals to enhance internal circadian synchronization.     

Persisting circadian rhythm of cell division in a photosynthetic mutant of Euglena

A persisting, free-running, circadian rhythm of cell division in a heterotrophically grown mutant of Euglena gracilis var. bacillaris having impaired photosynthesis is obtained upon placing a culture that has been previously synchronized by a 10,14 light-dark cycle into continuous darkness at 19 degrees C (but not at 25 degrees C). A similar persisting rhythm is initiated in exponentially increasing cultures (growing in darkness at 19 degrees C) by a single "switch-up" in irradiance to continuous bright illumination. The results implicate an endogenous biological clock which "gates" the specific event of cell division in the cell developmental cycle.     

Detection of GTP-tubulin conformation in vivo reveals a role for GTP remnants in microtubule rescues

Microtubules display dynamic instability, with alternating phases of growth and shrinkage separated by catastrophe and rescue events. The guanosine triphosphate (GTP) cap at the growing end of microtubules, whose presence is essential to prevent microtubule catastrophes in vitro, has been difficult to observe in vivo. We selected a recombinant antibody that specifically recognizes GTP-bound tubulin in microtubules and found that GTP-tubulin was indeed present at the plus end of growing microtubules. Unexpectedly, GTP-tubulin remnants were also present in older parts of microtubules, which suggests that GTP hydrolysis is sometimes incomplete during polymerization. Observations in living cells suggested that these GTP remnants may be responsible for the rescue events in which microtubules recover from catastrophe.     

表皮生长因子在哺乳动物体外受精中的作用

哺乳动物体外受精(In Vitro Fertilization,简称IVF)研究已有百余年的历史.早在1878年,德国科学家Schenk就开始进行了哺乳动物体外受精的尝试.此后,许多学者为了哺乳动物的体外受精进行了不懈努力.20世纪60年代,IVF技术在世界上得到了迅速发展,而在家畜,到了80年代才真正开展起来.IVF技术对于揭示配子发育、成熟、受精和早期胚胎细胞分化等生命现象的奥秘具有重要的意义.     

Structure of DNMT1-DNA complex reveals a role for autoinhibition in maintenance DNA methylation

Maintenance of genomic methylation patterns is mediated primarily by DNA methyltransferase-1 (DNMT1). We have solved structures of mouse and human DNMT1 composed of CXXC, tandem bromo-adjacent homology (BAH1/2), and methyltransferase domains bound to DNA-containing unmethylated CpG sites. The CXXC specifically binds to unmethylated CpG dinucleotide and positions the CXXC-BAH1 linker between the DNA and the active site of DNMT1, preventing de novo methylation. In addition, a loop projecting from BAH2 interacts with the target recognition domain (TRD) of the methyltransferase, stabilizing the TRD in a retracted position and preventing it from inserting into the DNA major groove. Our studies identify an autoinhibitory mechanism, in which unmethylated CpG dinucleotides are occluded from the active site to ensure that only hemimethylated CpG dinucleotides undergo methylation.     

Transgenic inhibition of synaptic transmission reveals role of CA3 output in hippocampal learning

The hippocampus is an area of the brain involved in learning and memory. It contains parallel excitatory pathways referred to as the trisynaptic pathway (which carries information as follows: entorhinal cortex --> dentate gyrus --> CA3 --> CA1 --> entorhinal cortex) and the monosynaptic pathway (entorhinal cortex --> CA1 --> entorhinal cortex). We developed a generally applicable tetanus toxin-based method for transgenic mice that permits inducible and reversible inhibition of synaptic transmission and applied it to the trisynaptic pathway while preserving transmission in the monosynaptic pathway. We found that synaptic output from CA3 in the trisynaptic pathway is dispensable and the short monosynaptic pathway is sufficient for incremental spatial learning. In contrast, the full trisynaptic pathway containing CA3 is required for rapid one-trial contextual learning, for pattern completion-based memory recall, and for spatial tuning of CA1 cells.     

UHRF1 plays a role in maintaining DNA methylation in mammalian cells

Epigenetic inheritance in mammals relies in part on robust propagation of DNA methylation patterns throughout development. We show that the protein UHRF1 (ubiquitin-like, containing PHD and RING finger domains 1), also known as NP95 in mouse and ICBP90 in human, is required for maintaining DNA methylation. UHRF1 colocalizes with the maintenance DNA methyltransferase protein DNMT1 throughout S phase. UHRF1 appears to tether DNMT1 to chromatin through its direct interaction with DNMT1. Furthermore UHRF1 contains a methyl DNA binding domain, the SRA (SET and RING associated) domain, that shows strong preferential binding to hemimethylated CG sites, the physiological substrate for DNMT1. These data suggest that UHRF1 may help recruit DNMT1 to hemimethylated DNA to facilitate faithful maintenance of DNA methylation.     

Sponge paleogenomics reveals an ancient role for carbonic anhydrase in skeletogenesis

Sponges (phylum Porifera) were prolific reef-building organisms during the Paleozoic and Mesozoic approximately 542 to 65 million years ago. These ancient animals inherited components of the first multicellular skeletogenic toolkit from the last common ancestor of the Metazoa. Using a paleogenomics approach, including gene- and protein-expression techniques and phylogenetic reconstruction, we show that a molecular component of this toolkit was the precursor to the alpha-carbonic anhydrases (alpha-CAs), a gene family used by extant animals in a variety of fundamental physiological processes. We used the coralline demosponge Astrosclera willeyana, a "living fossil" that has survived from the Mesozoic, to provide insight into the evolution of the ability to biocalcify, and show that the alpha-CA family expanded from a single ancestral gene through several independent gene-duplication events in sponges and eumetazoans.     

The Gs-linked receptor GPR3 maintains meiotic arrest in mammalian oocytes

Mammalian oocytes are held in prophase arrest by an unknown signal from the surrounding somatic cells. Here we show that the orphan Gs-linked receptor GPR3, which is localized in the oocyte, maintains this arrest. Oocytes from Gpr3 knockout mice resume meiosis within antral follicles, independently of an increase in luteinizing hormone, and this phenotype can be reversed by injection of Gpr3 RNA into the oocytes. Thus, the GPR3 receptor is a link in communication between the somatic cells and oocyte of the ovarian follicle and is crucial for the regulation of meiosis.     

Action spectra for phase shifts of a circadian rhythm in Drosophila

Action spectra were determined for lightinduced phase shifts of the circadian rhythm of adult emergence in Drosophila pseudoobscura. The action spectra for advance and delay phase shifts are similar; the most effective wavelengths are in the blue region, with a sharp cutoff above 500 nanometers. This similarity suggests that the same photoreceptive pigment mediates both advance and delay phase shifts.     

In vivo imaging reveals an essential role for neutrophils in leishmaniasis transmitted by sand flies

Infection with the obligate intracellular protozoan Leishmania is thought to be initiated by direct parasitization of macrophages, but the early events following transmission to the skin by vector sand flies have been difficult to examine directly. Using dynamic intravital microscopy and flow cytometry, we observed a rapid and sustained neutrophilic infiltrate at localized sand fly bite sites. Invading neutrophils efficiently captured Leishmania major (L.m.) parasites early after sand fly transmission or needle inoculation, but phagocytosed L.m. remained viable and infected neutrophils efficiently initiated infection. Furthermore, neutrophil depletion reduced, rather than enhanced, the ability of parasites to establish productive infections. Thus, L.m. appears to have evolved to both evade and exploit the innate host response to sand fly bite in order to establish and promote disease.     

Characterization and fine mapping of a semi-rolled leaf mutant srl3 in rice

Moderate leaf rolling can maintain leaf erectness, improve light transmittance in the population, and improve light energy utilization, thereby increasing rice yield. This study used ethyl methanesulfonate (EMS) to treat Yunjing 17 (YJ17) and obtained a semi-rolled leaf mutant that was named semi-rolledleaf3 (srl3). We found that the rolled-leaf phenotype was due to the aberrant development of bulliform cells and the loss of sclerenchymatous cells. In addition, the shoot and root length of srl3 seedlings differed from the wild type. The srl3 mutant had significantly lower plant height and seed-setting rate but notably greater tiller number, panicle length, and primary branch number per panicle than the wild type. Genetic analysis showed that a single recessive nuclear gene defined the srl3 mutant, and it was precisely located in a 144-kb region between two insertion-deletion (InDel) markers, M8 and M19, on chromosome 2. In this region, no leaf-rolling-related genes have been reported previously. Thus, the study indicated that SRL3 is a novel leaf-rolling-related gene, and the results laid the foundation for the cloning and functional analysis of the SRL3 gene.     

新时期提高农民收入的途径选择

系统分析了新时期下提高我国农民收入的新途径,认为提高农民收入是一个庞大、复杂的系统工程,研究和解决农民增收不能单纯就农业论农业、就农村论农村,而应该从整个国民经济和全球一体化的大背景去考虑.既要从生产（产品）、产业化经营、投资渠道、政策性补贴等多方面千方百计提高农民的收入,又要从减免税费、降低农民市民化费用、机构改革等方面尽可能地减轻农民的负担,要从整治农民增收环境、打击非法收入等方面切实保护农民的合法利益.     

A kinetic framework for a mammalian RNA polymerase in vivo

We have analyzed the kinetics of assembly and elongation of the mammalian RNA polymerase I complex on endogenous ribosomal genes in the nuclei of living cells with the use of in vivo microscopy. We show that components of the RNA polymerase I machinery are brought to ribosomal genes as distinct subunits and that assembly occurs via metastable intermediates. With the use of computational modeling of imaging data, we have determined the in vivo elongation time of the polymerase, and measurements of recruitment and incorporation frequencies show that incorporation of components into the assembling polymerase is inefficient. Our data provide a kinetic and mechanistic framework for the function of a mammalian RNA polymerase in living cells.     

樱桃番茄长季节无土栽培技术

樱桃番茄近年来在全国各地广泛栽培,尤其是无土栽培的主要品种之一。在无土栽培条件下,樱桃番茄可以保持长盛不衰,结果期长达8～9个月。其主要栽培技术如下:1品种选择选择生长势强、抗病力强、耐弱光、品质好的无限生长类型品种,如圣女。2栽培方式可根据实际条件选择各种栽培方     

产氨短杆菌的诱变以及TMTD抗性突变株发酵培养基条件的优化

通过对产氨短杆菌进行微波诱变,筛选TMTD(N,N-四甲基二硫双硫羰胺)抗性突变株以期获得辅酶A的高产菌株,并对其中菌株A3进行了培养基的初步优化.获得的优化培养基配方为:葡萄糖1.5％,蛋白胨1.0％,酵母膏0.5％,硫酸镁1％,磷酸氢二钾1％,磷酸二氢钾1％:接种量为15％.在此优化条件下.菌株的OD600可达512.     

The three-dimensional structure of Asn102 mutant of trypsin: role of Asp102 in serine protease catalysis

The structure of the Asn102 mutant of trypsin was determined in order to distinguish whether the reduced activity of the mutant at neutral pH results from an altered active site conformation or from an inability to stabilize a positive charge on the active site histidine. The active site structure of the Asn102 mutant of trypsin is identical to the native enzyme with respect to the specificity pocket, the oxyanion hole, and the orientation of the nucleophilic serine. The observed decrease in rate results from the loss of nucleophilicity of the active site serine. This decreased nucleophilicity may result from stabilization of a His57 tautomer that is unable to accept the serine hydroxyl proton.     

Adenosine 3',5'-monophosphate: electrophysiological evidence for a role in synaptic transmission

Synaptic potentials and changes in resting membrane potentials of superior cervical ganglia of the rabbit were measured in the presence of adenosine 3',5'-monophosphate and agents that affect its metabolism. Adenosine 3',5'-monophosphate and its mono- and dibutyryl derivatives caused a hyperpolarization of the postganglionic neurons. Theophylline potentiated the slow inhibitory postsynaptic potential that follows synaptic transmission, as well as the hyperpolarization of postganglionic neurons caused by exogenous dopamine. Conversely, prostaglandin E(1) inhibited both the slow inhibitory postsynaptic potential and the dopamine-induced hyperpolarization. We hypothesize that the slow inhibitory postsynaptic potential as well as the dopamine-induced hyperpolarization result from increased amounts of adenosine 3'5'-monophosphate in the postganglionic neurons. The dibutyryl derivative of guanosine 3'5'-monophosphate caused a depolarization of the postganglionic neurons, which is consistent with the possibility that guanosine 3'5'-monophosphate mediates synaptic transmission at muscarinic cholinergic synapses.