Applications of fluorescent brighteners in biological techniques

The visual labeling of various microorganisms has been accomplished with specific fluorescent ultraviolet-absorbing compounds that not only have been bound by the cell but have been transferred by it to subsequent growth. The possibility of application to genetic studies is suggested.     

RFLP和RAPD技术及其在植物线虫鉴定中的应用

主要介绍了分子标记技术中RFLP和RAPD技术发展的概况以及在线虫种类鉴定中的应用 ,并对这一技术在该领域的应用前景进行了展望。     

Fertilization events induced by neurotransmitters after injection of mRNA in Xenopus eggs

Fertilization initiates in the egg a dramatic increase in intracellular calcium that opens ion channels and causes exocytosis. To explore the possibility that these events might involve a receptor-mediated pathway, receptors for serotonin or acetylcholine (M1 muscarinic) were expressed in the Xenopus egg; serotonin or acetylcholine then could initiate a series of responses similar to those normally initiated by sperm. Thus, there may be an endogenous receptor in the egg membrane that is activated by sperm, and the serotonin or M1 muscarinic receptor may replace the sperm receptor in this pathway.     

Pathfinding by peripheral pioneer neurons in grasshoppers

Grasshopper neurons accurately project axons across long distances between peripheral structures and the central nervous system. Nerve-trunk pathways followed by these axons are established early in embryogenesis by pioneer neurons. Growth cones from the first pioneers navigate along a chain of cells to the CNS. The placement of these cells may constitute the initial guidance mechanism underlying long-distance pathfinding.     

采用CFDA荧光染色技术监测木薯渣堆肥中的细菌动态

[目的]探讨CFDA荧光染色技术检测高温细菌的效果,阐明木薯酒精渣堆肥过程中的细菌动态,为堆肥进程的调节和评价提供依据.[方法]在30℃和60℃下对高温型和中温型细菌进行CFDA荧光染色,观察染色效果.同时采用CFDA技术和培养法对木薯酒精渣堆肥中的细菌动态和垂直分布进行监测.[结果]60℃下进行CFDA荧光染色能有效识别高温细菌.CFDA技术检测结果显示,木薯渣堆肥前期堆料中细菌以中温型为主,但随堆肥时间延长,数量逐渐减少;高温型细菌数量随堆肥时间延长迅速增加,并在发酵后期成为优势菌群.10~40 cm深度的堆料温度高,细菌以高温型为主;50 cm以下堆料呈酸性,温度降低,细菌以中温型为主.[结论]高温细菌是木薯酒精渣高温堆肥中的重要微生物群体,CFDA荧光染色技术可以检测出高温细菌,可用于检测木薯渣堆肥中的总高温细菌数量.     

杂交水稻品种指纹鉴定研究进展

从蛋白质指纹和DNA指纹两个方面,综述了杂交水稻品种指纹鉴定研究的进展和不足。对SSR标记在品种指纹鉴定上的应用前景,单一特异标记的筛选和实验室标准检测技术的建立作了探讨和展望,以利于加快杂交水稻品种指纹鉴定的发展和应用。     

模拟土壤自然环境鉴定水稻幼苗耐盐性的方法

采用将NaCl琼脂固定液试管插入土壤的方法模拟土壤自然环境条件 ,对水稻幼苗耐盐性进行鉴定。结果表明 :该鉴定方法简便可行 ,对大田环境模拟性强 ,与其他鉴定方法相比较具独特性 ,尤其适合在盐胁迫下对水稻幼苗根部性状的研究     

Corticotropin-releasing factor-producing neurons in the rat activated by interleukin-1

Intraperitoneal administration of human recombinant interleukin-1 (IL-1) to rats can increase blood levels of corticosterone and adrenocorticotropic hormone (ACTH). The route by which IL-1 affects pituitary-adrenal activity is unknown. That the IL-1-induced pituitary-adrenal activation involves an increased secretion of corticotropin-releasing factor (CRF) is indicated by three lines of evidence. First, immunoneutralization of CRF markedly attenuated the IL-1-induced increase of ACTH blood levels. Second, after blockade of fast axonal transport in hypothalamic neurons by colchicine, IL-1 administration decreased the CRF immunostaining in the median eminence, indicating an enhanced release of CRF in response to IL-1. Third, IL-1 did not stimulate ACTH release from primary cultures of anterior pituitary cells. These data further support the notion of the existence of an immunoregulatory feedback circuit between the immune system and the brain.     

Estradiol-concentrating neurons: topography in the hypothalamus by dry-mount autoradiography

Serial section autoradiograms were prepared of different planes of the hypothalamus and diencephalon of immature female, immature male, and ovariectomized mature rats injected with 6,7-(3)H-estradiol-17 beta. Known causes of diffusion and redistribution of the label, such as fixation, embedding, and thawing, were eliminated by the use of an autoradiographic technique based on the dry-mounting of freeze-dried sections. Neurons that concentrate estradiol exist in distinct and definable anatomical areas that are independent of the sex and hormonal state of the animals. Distribution of these neurons follows known terminations of the stria terminalis, which supports the concept of an endocrine amygdaloid-hypothalamic-hypophysial axis.     

Control of larval development by chemosensory neurons in Caenorhabditis elegans

Larval development of the nematode Caenorhabditis elegans is controlled by the activities of four classes of chemosensory neurons. The choice between normal development and development into a specialized larval form called a dauer larva is regulated by competing environmental stimuli: food and a dauer pheromone. When the neuron classes ADF, ASG, ASI, and ASJ are killed, animals develop as dauer larvae regardless of environmental conditions. These neurons might sense food or dauer pheromone, or both, to initiate the specialized differentiation of many cell types that occurs during dauer formation. Entry into and exit from the dauer stage are primarily controlled by different chemosensory neurons. The analysis of mutants defective in dauer formation indicates that the chemosensory neurons are active in the absence of sensory inputs and that dauer pheromone inhibits the ability of these neurons to generate a signal necessary for normal development.     

Release of coordinated behavior in crayfish by single central neurons

By stimulating and recording from the same interneuron at two separate points, we have shown that coordinated output to the postural abdominal muscles of crayfish can be produced by electrical stimulation of a single cell. Several central neurons can individually initiate one type of movement(for example, flexion),each producing a unique abdominal geometry.     

雌激素调控的绿色荧光蛋白在酵母细胞中的表达

采用绿色荧光蛋白(GFP)作为报告基因,用酵母细胞构建环境雌激素(EEH)的评价体系,可敏感、快速、方便地筛选出EEH化合物。通过分子生物学技术,将GFP基因插入到pM P 206/ERE启动子CYC 1的下游,用GFP取代L ac Z基因,并转化于酵母细胞。DNA测序发现ERE-CYC 1-GFP框架正确。对转化子进行PCR鉴定,发现有雌激素受体基因(hER cDNA)和GFP基因特异条带,说明hER cDNA和GFP已重组于酵母细胞。荧光显微镜下发现大量的绿色荧光颗粒,表明GFP基因在酵母细胞中成功表达。酵母细胞经不同浓度雌二醇作用后,与GFP表达量有剂量效应关系。与其他酵母评价体系相比,以GFP为报告基因评价EEH不需要破坏细胞壁,也不需要底物,可在96孔板中完成,这为高通量筛选EEH化合物奠定了基础。     

Visualization of specific angiotensin II binding sites in the brain by fluorescent microscopy

The organum vasculosum of the lamina terminalis has been implicated as the site of receptors mediating central responses of angiotensin II. Up to now, this had been based on indirect evidence, but direct visualization of angiotensin II at its site of action has now been achieved by the use of a biologically active fluorescent angiotensin II agonist. The ventricular surface of the organum vasculosum lamina terminalis showed intense fluorescence, which was virtually eliminated by an excess of unlabeled angiotensin II.     

Onset and progression in inherited ALS determined by motor neurons and microglia

Dominant mutations in superoxide dismutase cause amyotrophic lateral sclerosis (ALS), a progressive paralytic disease characterized by loss of motor neurons. With the use of mice carrying a deletable mutant gene, expression within motor neurons was shown to be a primary determinant of disease onset and of an early phase of disease progression. Diminishing the mutant levels in microglia had little effect on the early disease phase but sharply slowed later disease progression. Onset and progression thus represent distinct disease phases defined by mutant action within different cell types to generate non-cell-autonomous killing of motor neurons; these findings validate therapies, including cell replacement, targeted to the non-neuronal cells.     

Steroid-sensitive single neurons in rat hypothalamus and midbrain: identification by microelectrophoresis

Minute amounts of Na-dexamethasone-21-phosphate administered by microelectrophoresis to the immediate extracellular environment promptly suppressed electrical activity of 15 out of 115 hypothalamic and mesencephalic neurons, the effect being readily reversible. Such neurons marked with fast green were found to lie in circumscribed areas of the periventricular gray of the third ventricle and aqueduct, and may represent a site of action of adrenocortical steroids in the regulation of corticotrophin releasing factor and/or adrenocortico-tropin secretion by negative feedback.     

Induction of nitric oxide-dependent apoptosis in motor neurons by zinc-deficient superoxide dismutase

Mutations in copper, zinc superoxide dismutase (SOD) have been implicated in the selective death of motor neurons in 2 percent of amyotrophic lateral sclerosis (ALS) patients. The loss of zinc from either wild-type or ALS-mutant SODs was sufficient to induce apoptosis in cultured motor neurons. Toxicity required that copper be bound to SOD and depended on endogenous production of nitric oxide. When replete with zinc, neither ALS-mutant nor wild-type copper, zinc SODs were toxic, and both protected motor neurons from trophic factor withdrawal. Thus, zinc-deficient SOD may participate in both sporadic and familial ALS by an oxidative mechanism involving nitric oxide.     

干旱胁迫下木薯AP2转录因子的 荧光定量PCR 分析

为研究木薯的两个AP2基因在干旱胁迫下的表达模式、对木薯品种华南5 号进行干旱胁迫处理、提取 各个时间点离区部位的RNA、以其反转录的cDNA 为模板、应用SYBR Green玉荧光定量PCR 检测AP2转录因子基 因的差异性表达。结果表明、目的基因的熔解曲线都只出现一个峰、说明引物特异性强、且各基因的扩增效率与参 照基因Actin 接近、试验结果可用2-驻驻CT 计算法进行分析。经分析、两个AP2家族成员在干旱胁迫下均诱导表达、其 表达模式不完全相同且存在一定的相关性、这与之前做过的基因芯片杂交结果相符。     

Dopamine-accumulating retinal neurons revealed by in vitro fluorescence display a unique morphology

Dopamine is the principal catecholamine neurotransmitter in the vertebrate retina. The shape of retinal neurons that accumulate dopamine has been demonstrated in an in vitro preparation of cat retina. This was achieved by the discovery that the combined uptake of dopamine and the indoleaminergic transmitter analog 5,7-dihydroxytryptamine leads to an intense, catecholamine-like fluorescence in the cell bodies and processes of presumed dopaminergic amacrine cells in the living retina. This fluorescence served as an in vitro marker for these cells, and their detailed morphology was analyzed after intracellular injection of horseradish peroxidase under direct microscopic control. The horseradish peroxidase-filled cells show an unprecedented neuronal morphology: each cell gives rise to multiple, axon-like processes that arise from, and extend for millimeters beyond, the dendritic tree. The unique structure of this type of amacrine cell suggests a function for dopamine in long-range lateral interactions in the inner plexiform layer.     

Serotonin-containing neurons in brain: depression of firing by monoamine oxidase inhibitors

Monoamine oxidase inhibitors were administered to rats while the activities of single, serotonin-containing neurons of the midbrain raphe nuclei were being monitored with microelectrodes. All the inhibitors tested (pargyline, tranylcypromine, phenelzine, iproniazid) caused depression of raphe unit firing rate. The ability of monoamine oxidase inhibitors to depress raphe units was impaired by prior treatment with p-chlorophenylalanine, an inhibitor of serotonin synthesis.     

露地瓠瓜无公害搭架栽培技术

从地块选择、科学育苗、移栽、田间管理、病虫害防治、适时收获、留种管理等方面总结露地瓠瓜无公害搭架栽培技术。