Nck adaptor proteins link nephrin to the actin cytoskeleton of kidney podocytes

The glomerular filtration barrier in the kidney is formed in part by a specialized intercellular junction known as the slit diaphragm, which connects adjacent actin-based foot processes of kidney epithelial cells (podocytes). Mutations affecting a number of slit diaphragm proteins, including nephrin (encoded by NPHS1), lead to renal disease owing to disruption of the filtration barrier and rearrangement of the actin cytoskeleton, although the molecular basis for this is unclear. Here we show that nephrin selectively binds the Src homology 2 (SH2)/SH3 domain-containing Nck adaptor proteins, which in turn control the podocyte cytoskeleton in vivo. The cytoplasmic tail of nephrin has multiple YDxV sites that form preferred binding motifs for the Nck SH2 domain once phosphorylated by Src-family kinases. We show that this Nck-nephrin interaction is required for nephrin-dependent actin reorganization. Selective deletion of Nck from podocytes of transgenic mice results in defects in the formation of foot processes and in congenital nephrotic syndrome. Together, these findings identify a physiological signalling pathway in which nephrin is linked through phosphotyrosine-based interactions to Nck adaptors, and thus to the underlying actin cytoskeleton in podocytes. Simple and widely expressed SH2/SH3 adaptor proteins can therefore direct the formation of a specialized cellular morphology in vivo.     

Pinch-3蛋白对细胞形态及牵引力的影响

用基因敲除的方法比较了正常肾小球足细胞和Pinch-3蛋白缺失细胞的形态差异和牵引力的差异,发现当Pinch-3蛋白缺失后,细胞表面出现了许多孔隙,同时细胞投影面积平均增加40%;细胞的牵引力则平均减小40%左右,这说明当肾小球足细胞的Pinch-3基因的表达受到抑制以后,细胞膜上产生的孔隙使细胞膜变得松散,进而使得表面积增大.细胞牵引力减小40%,说明细胞膜上的孔隙对细胞牵引力的形成具有较强的影响.     

青岛文昌鱼硫氧还蛋白基因的克隆及同源性分析

通过对文昌鱼神经胚cDNA文库进行测序 ,获得含文昌鱼硫氧还蛋白基因全部读框的cDNA序列 ,并演绎出对应的氨基酸序列 ,对其可编码蛋白进行了分析预测 ,还与多种脊椎动物和无脊椎动物中同种蛋白的同源性进行比较分析 .发现该蛋白具有典型的硫氧还蛋白活性部位 ,与脊椎动物硫氧还蛋白的同源性大于无脊椎动物 ,说明作为脊椎动物和无脊椎动物之间典型的过渡类型 ,文昌鱼在进化上更接近于脊椎动物 .     

Double cones as a basis for a new type of polarization vision in vertebrates

Many invertebrates and vertebrates are sensitive to the polarization of light. The biophysical basis of invertebrate polarization sensitivity is an intrinsic dichroism, the alignment of chromophores along the photoreceptor microvilli. But such dichroism to axially propagating light is not present in vertebrate photoreceptors, whose chromophores are free to rotate in the plane of the outer-segment disc membranes, and a biophysical mechanism responsible for vertebrate polarization sensitivity has not been established. We hypothesize that the roughly elliptical cross-sectioned double-cone inner segment acts as a birefringent, polarization-sensitive dielectric waveguide, and that the double cone mosaic generates a 'polarization contrast' neural image. Here we confirm three predictions derived from these hypotheses: (1) 90 degrees periodicity for polarization sensitivity; (2) polarization sensitivity maxima corresponding to the absolute orientation of the axes of the double-cone inner-segment cross-sections; and (3) action spectrum for polarization sensitivity corresponding to the absorption spectrum of the double cones. We also present evidence for a polarization-opponent neural encoding in vertebrates.     

Cyclic GMP is involved in the excitation of invertebrate photoreceptors

The hyperpolarizing receptor potential in vertebrate rod photoreceptors appears to be mediated by the second messenger, cyclic GMP. Injection of cGMP into rods or application of cGMP to inside-out membrane patches activates a conductance resembling that produced by light. Light produces a rapid reduction of cGMP in living rods, leading to closure of sodium channels and membrane hyperpolarization. In most invertebrate photoreceptors the response to light is depolarizing. We have investigated whether cGMP is involved in controlling the increase in sodium conductance that underlies this depolarization. We show here that injection of cGMP into Limulus photoreceptors produces a depolarization that mimics the receptor potential. We also show that the cGMP concentration of the squid retina increases rapidly during exposure to light. These results support the hypothesis that cGMP mediates the light-induced depolarization in invertebrate photoreceptors and suggests that vertebrate and invertebrate phototransduction may be more similar than previously thought.     

哺乳动物卵子发生过程中细胞凋亡的研究进展

细胞凋亡是发育过程中的基本生命现象,是多细胞生物体一种重要的自我稳定机制．除各种体细胞凋亡外,生殖细胞在其发生过程中也有细胞凋亡现象产生．对许多脊椎动物和无脊椎动物的研究发现,有大量生殖细胞在其发育的不同阶段丢失．对近10年来哺乳动物卵子发生过程中细胞凋亡的研究进行了综述,主要描述了哺乳动物卵子发生过程中的细胞凋亡及其调控以及凋亡的意义．     

青岛文昌鱼NADH脱氢酶亚基I基因片段的克隆

NADH脱氢酶亚基I是细胞电子传递链的主要成员之一，采用简并引物PCR方法获得青岛文昌鱼NADH脱氢酶亚基I基因片段，将基氨基酸序列与其他生物如佛罗里达文昌鱼，斑马鱼，爪蟾等无脊椎和脊椎动物NADH脱氢酶亚基I基因相应片段进行了同源性分析，均显示较高的同源性，研究结果证实青岛文昌鱼作为脊索动物的代表之一，与脊椎动物有着较近的亲缘关系，是从无脊椎动物到脊椎动物的过渡类型。     

汽轮机缝隙抽湿式隔板中的水份运动特性

本文计算并分析了湿蒸汽汽轮机级隔板中大分散度水份的尺寸分布、运动特性及沉积位置。利用平板上的气-水膜两相流试验装置,研究了流动参数对沉积水份流动形式的影响,并测定了壁面上抽湿缝隙的去湿率与抽吸压差、缝隙宽度、汽流切应力及水膜雷诺数等参数的关系.     

A novel active pentapeptide from chicken brain identified by antibodies to FMRFamide

The tetrapeptide Phe-Met-Arg-Phe-NH2 (FMRFamide) and peptides structurally related to it, have been isolated from molluscan ganglia. They have widespread actions on both invertebrate and vertebrate tissues and there is increasing evidence that they are an important group of invertebrate peptide neurotransmitters. It is of interest that the primary amino acid sequence of FMRFamide forms the C-terminal tetrapeptide of an enkephalin-like heptapeptide (Met-enkephalin-ArgPhe) isolated from bovine adrenal medulla and striatum. Antisera to FMRFamide have been shown to react in radioimmunoassay and immunohistochemistry with material in the central nervous system of various vertebrate species, but the identity of this material, and in particular its relationship to the opioid heptapeptide, remains uncertain. We have used antibodies specific for the C-terminus of FMRFamide in radioimmunoassays to monitor purification of the material in chicken brain. We describe here the sequence of one of the peptides obtained. It is a biologically active peptide which does not seem to be related to other known vertebrate neuropeptides.     

青岛文昌鱼核糖体蛋白Amphil11基因的克隆和同源性分析

对青岛文昌鱼18小时神经胚cDNA文库进行测序，获得了5个AmphiL11的EST，经接接得到文昌鱼核糖体蛋白AmphiL11的编码完整的cDNA序列并演绎出AmphiL11的氨基酸序列，通过对AmphiL11蛋白的结构分析及其与人、鼠、鱼等脊柱动物和果蝇、线虫等无脊动物及酵母中同种核糖体蛋白的同源性分析，发现AmphiL11与脊柱动物核糖体L11蛋白的同源性很高，与高等无脊椎动物甚至酵母的同源性也较高，提示AmphiL11具有较高的进化水平，更接近于脊椎动物的核糖体蛋白L11。同时也表明，真核生物核糖体蛋白L11具有高度的保守性。     

Mitoferrin is essential for erythroid iron assimilation

Iron has a fundamental role in many metabolic processes, including electron transport, deoxyribonucleotide synthesis, oxygen transport and many essential redox reactions involving haemoproteins and Fe-S cluster proteins. Defective iron homeostasis results in either iron deficiency or iron overload. Precise regulation of iron transport in mitochondria is essential for haem biosynthesis, haemoglobin production and Fe-S cluster protein assembly during red cell development. Here we describe a zebrafish mutant, frascati (frs), that shows profound hypochromic anaemia and erythroid maturation arrest owing to defects in mitochondrial iron uptake. Through positional cloning, we show that the gene mutated in the frs mutant is a member of the vertebrate mitochondrial solute carrier family (SLC25) that we call mitoferrin (mfrn). mfrn is highly expressed in fetal and adult haematopoietic tissues of zebrafish and mouse. Erythroblasts generated from murine embryonic stem cells null for Mfrn (also known as Slc25a37) show maturation arrest with severely impaired incorporation of 55Fe into haem. Disruption of the yeast mfrn orthologues, MRS3 and MRS4, causes defects in iron metabolism and mitochondrial Fe-S cluster biogenesis. Murine Mfrn rescues the defects in frs zebrafish, and zebrafish mfrn complements the yeast mutant, indicating that the function of the gene may be highly conserved. Our data show that mfrn functions as the principal mitochondrial iron importer essential for haem biosynthesis in vertebrate erythroblasts.     

Migratory neural crest-like cells form body pigmentation in a urochordate embryo

The neural crest, a source of many different cell types in vertebrate embryos, has not been identified in other chordates. Current opinion therefore holds that neural crest cells were a vertebrate innovation. Here we describe a migratory cell population resembling neural crest cells in the ascidian urochordate Ecteinascidia turbinata. Labelling of embryos and larvae with the vital lipophilic dye DiI enabled us to detect cells that emerge from the neural tube, migrate into the body wall and siphon primordia, and subsequently differentiate as pigment cells. These cells express HNK-1 antigen and Zic gene markers of vertebrate neural crest cells. The results suggest that migratory cells with some of the features of neural crest cells are present in the urochordates. Thus, we propose a hypothesis for neural crest evolution beginning with the release of migratory cells from the CNS to produce body pigmentation in the common ancestor of the urochordates and vertebrates. These cells may have gained additional functions or were joined by other cell types to generate the variety of derivatives typical of the vertebrate neural crest.     

A proteome-wide screen identifies valosin-containing protein as an essential regulator of podocyte endoplasmic reticulum stress

To investigate proteins expressed in the renal tissue of the passive Heymann nephritis (pHN) rat model,we prepared pHN rat models with anti-FxA1 serum and analyzed the proteins differentially expressed in the kidney tissue with label-free liquid chromatography-tandem mass spectrometry.We then analyzed in depth the endoplasmic reticulum stress (ERS)-related protein using an online bioinformatics platform.Forty-one differential proteins and their annotations were obtained.Gene Ontology (GO) function analysis showed that 16 proteins were involved in cellular metabolism and 22 were proteins related to catalytic activity,including protein folding or ATPase.Protein-GO networks indicated that VCP could interact with the ERS marker HSPa5,with both involved in a single pathway.On inhibition of podocyte VCP by RNAi under normal conditions,the HSPa5 expression level did not change,but when the cell was subjected to ERS by tunicamycin,HSPa5 expression significantly increased with RNAi of VCP when compared with the tunicamycin-treated group.Our results showed that ERS plays an important role in podocyte injury of membranous nephropathy and is mediated by an HSPa5-VCP signaling pathway,in which the most predominant proteins are those related to cellular metabolism and catalytic activity.     

细胞挤入狭缝的润滑理论分析

血细胞的入口阻力构成血液循环外周阻力的重要组成部分。本文首次提出一个简化的力学模型,试图探讨细胞挤入二维狭缝的运动特性。本模型的细胞外形根据显微录相设为已知,细胞膜沿其表面滑移。对膜与缝壁间的薄血浆层应用润滑理论,算出压力和剪应力分布,给出了细胞所受阻力。本文得到的细胞运动规律,与显微录相定性一致地模拟了细胞的入口过程。     

Ancient deuterostome origins of vertebrate brain signalling centres

Neuroectodermal signalling centres induce and pattern many novel vertebrate brain structures but are absent, or divergent, in invertebrate chordates. This has led to the idea that signalling-centre genetic programs were first assembled in stem vertebrates and potentially drove morphological innovations of the brain. However, this scenario presumes that extant cephalochordates accurately represent ancestral chordate characters, which has not been tested using close chordate outgroups. Here we report that genetic programs homologous to three vertebrate signalling centres-the anterior neural ridge, zona limitans intrathalamica and isthmic organizer-are present in the hemichordate Saccoglossus kowalevskii. Fgf8/17/18 (a single gene homologous to vertebrate Fgf8, Fgf17 and Fgf18), sfrp1/5, hh and wnt1 are expressed in vertebrate-like arrangements in hemichordate ectoderm, and homologous genetic mechanisms regulate ectodermal patterning in both animals. We propose that these genetic programs were components of an unexpectedly complex, ancient genetic regulatory scaffold for deuterostome body patterning that degenerated in amphioxus and ascidians, but was retained to pattern divergent structures in hemichordates and vertebrates.     

Twisted gastrulation can function as a BMP antagonist

Bone morphogenetic proteins (BMPs), including the fly homologue Decapentaplegic (DPP), are important regulators of early vertebrate and invertebrate dorsal-ventral development. An evolutionarily conserved BMP regulatory mechanism operates from fly to fish, frog and mouse to control the dorsal-ventral axis determination. Several secreted factors, including the BMP antagonist chordin/Short gastrulation (SOG), modulate the activity of BMPs. In Drosophila, Twisted gastrulation (TSG) is also involved in dorsal-ventral patterning, yet the mechanism of its function is unclear. Here we report the characterization of the vertebrate Tsg homologues. We show that Tsg can block BMP function in Xenopus embryonic explants and inhibits several ventral markers in whole-frog embryos. Tsg binds directly to BMPs and forms a ternary complex with chordin and BMPs. Coexpression of Tsg with chordin leads to a more efficient inhibition of the BMP activity in ectodermal explants. Unlike other known BMP antagonists, however, Tsg also reduces several anterior markers at late developmental stages. Our data suggest that Tsg can function as a BMP inhibitor in Xenopus; furthermore, Tsg may have additional functions during frog embryogenesis.     

Energetic constraints on the diet of terrestrial carnivores

Species in the mammalian order Carnivora exhibit a huge diversity of life histories with body sizes spanning more than three orders of magnitude. Despite this diversity, most terrestrial carnivores can be classified as either feeding on invertebrates and small vertebrates or on large vertebrates. Small carnivores feed predominantly on invertebrates probably because they are a superabundant resource (sometimes 90% of animal biomass); however, intake rates of invertebrate feeders are low, about one tenth of those of vertebrate feeders. Although small carnivores can subsist on this diet because of low absolute energy requirements, invertebrate feeding appears to be unsustainable for larger carnivores. Here we show, by reviewing the most common live prey in carnivore diets, that there is a striking transition from feeding on small prey (less than half of predator mass) to large prey (near predator mass), occurring at predator masses of 21.5-25 kg. We test the hypothesis that this dichotomy is the consequence of mass-related energetic requirements and we determine the predicted maximum mass that an invertebrate diet can sustain. Using a simple energetic model and known invertebrate intake rates, we predict a maximum sustainable mass of 21.5 kg, which matches the point where predators shift from small to large prey.     

Deuterostome phylogeny reveals monophyletic chordates and the new phylum Xenoturbellida

Deuterostomes comprise vertebrates, the related invertebrate chordates (tunicates and cephalochordates) and three other invertebrate taxa: hemichordates, echinoderms and Xenoturbella. The relationships between invertebrate and vertebrate deuterostomes are clearly important for understanding our own distant origins. Recent phylogenetic studies of chordate classes and a sea urchin have indicated that urochordates might be the closest invertebrate sister group of vertebrates, rather than cephalochordates, as traditionally believed. More remarkable is the suggestion that cephalochordates are closer to echinoderms than to vertebrates and urochordates, meaning that chordates are paraphyletic. To study the relationships among all deuterostome groups, we have assembled an alignment of more than 35,000 homologous amino acids, including new data from a hemichordate, starfish and Xenoturbella. We have also sequenced the mitochondrial genome of Xenoturbella. We support the clades Olfactores (urochordates and vertebrates) and Ambulacraria (hemichordates and echinoderms). Analyses using our new data, however, do not support a cephalochordate and echinoderm grouping and we conclude that chordates are monophyletic. Finally, nuclear and mitochondrial data place Xenoturbella as the sister group of the two ambulacrarian phyla. As such, Xenoturbella is shown to be an independent phylum, Xenoturbellida, bringing the number of living deuterostome phyla to four.     

青岛文昌鱼核糖体蛋白AmphiL 37a基因的克隆和同源性分析

对青岛文昌鱼神经胚cDNA进行测序,获得了3个AmphiL 37a的EST,经拼接得到编码文昌鱼核糖体蛋白的AmphiL37a基因全长cDNA序列并演绎出AmphiL37a的氨基酸序列.通过对AmphiL37a蛋白的结构分析及其与多种脊椎动物和无脊椎动物中同种蛋白的同源性进行比较,发现AmphiL37a具有典型的四个半胱氨酸的锌指结构,与人、鼠、鸡的L37a,尤其与果蝇、隐板石鳖等高等无脊椎动物核糖体L37a具有较高的同源性,说明文昌鱼在进化上更接近于高等无脊椎动物;同时说明核糖体蛋白L37a基因在进化上具有较高保守性.     

卵黄蛋白原的性质及热休克蛋白90对其合成代谢的调控

卵黄蛋白原(VTG)广泛存在于卵生脊椎动物和无脊椎动物的卵母细胞中,为卵母细胞的生长和胚胎的发育提供营养。VTG一般在卵巢、肝胰脏和(或)脂肪等组织中合成。VTGcDNA具有较强的保守性,为7 kB左右。VTG具有单体、二聚体等多种形式,其中二聚体有同源二聚体和异源二聚体两种形式。脊椎动物体内,热休克蛋白90(HSP90)通过和雌激素受体结合间接诱导VTG的合成,近年来的研究报道表明HSP90同样对无脊椎动物体内VTG合成也具有诱导作用,但其作用机制有待进一步研究。