Formation of domain structure of erythrocyte membrane in Wistar rat fed with CeCl3 per os

To explore the possibility of absorption of lanthanides via digestive duct and their effects on the membrane structure and permeability of erythrocytes,the fine structure of erythrocyte membrane from Wistar rats,fed for 70 days of daily administration per os with 20 mg CeCl3/kg weight,was imaged by means of atomic force microscopy and FT-IR deconvolution spectra.The results show that,although the erythrocytes maintain the intact shape,the change of secondary structure,aggregation and crosslinking of the protein particles of membrane surface and the enlarged lipid regions lead to the domain structure formation.This structure might be responsible for the increasing permeability of erythrocyte membrane.     

怀柔水库鲫鱼种群繁殖生态学特征

鲫鱼(Carassius auratus)是一种经济价值较高的鱼类。1981～1990年根据怀柔水库的1000余尾标本,讨论了该水域中鲫鱼种群的性比、成熟年龄和体形大小以及与产卵量之间关系。依据体长、体重和年龄建立了绝对产卵量的回归方程,讨论了种群的繁殖力,每年4～5月为鲫鱼产卵时期,应予以保护。     

Formation of domain structure of erythrocyte membrane in Wistar rat fed with CeCl3 per os

To explore the possibility of absorption of lanthanides via digestive duct and their effects on the membrane structure and permeability of erythrocytes, the fine structure of erythrocyte membrane from Wistar rats, fed for 70 days of daily administrationper os with 20 mg CeCl₃/kg weight, was imaged by means of atomic force microscopy and FT-IR deconvolution spectra. The results show that, although the erythrocytes maintain the intact shape, the change of secondary structure, aggregation and crosslinking of the protein particles of membrane surface and the enlarged lipid regions lead to the domain structure formation. This structure might be responsible for the increasing permeability of erythrocyte membrane.     

Partial deficiency of erythrocyte spectrin in hereditary spherocytosis

Hereditary spherocytosis (HS) is a common, clinically heterogeneous haemolytic anaemia in which the primary erythrocyte defect is believed to be some abnormality in the spectrin-actin membrane skeleton, leading to loss of surface membrane. Recessively inherited spectrin deficiency with extreme erythrocyte fragility and spherocytosis has been identified in certain mutant mice and two severely anaemic humans. Although suspected, deficiency of spectrin has not been demonstrated in less severe forms of human HS. We not report the quantitation of erythrocytes spectrin by radioimmunoassay. We found that normal erythrocytes contained 240,000 copies of spectrin heterodimer, whereas erythrocytes from 14 patients with a variety of types of HS were all partially deficient in spectrin (range 74,000-200,000 copies), the magnitude of the deficiency correlating with the severity of the disease. Spectrin deficiency of varying degrees is common in HS and probably represents the principal structural defect leading to loss of surface membrane.     

异育淇鲫及其双亲的过氧化物酶和α-淀粉酶同工酶的比较研究

采用聚丙烯酰胺垂直平板电泳,对异育淇鲫及其母本淇鲫和父本兴国红鲤的6种组织(心脏、肝脏、肾脏、脑、晶状体和骨骼肌)中的过氧化物酶和α淀粉酶同工酶进行比较研究.结果显示:异育淇鲫同工酶的电泳图谱与母本淇鲫相同,与父本兴国红鲤显著不同.因而认为异育淇鲫是雌核发育的产物,父本遗传物质对子代无影响.     

Structural basis for Duffy recognition by the malaria parasite Duffy-binding-like domain

Molecular processes that govern pathogenic features of erythrocyte invasion and cytoadherence in malaria are reliant on Plasmodium-specific Duffy-binding-like domains (DBLs). These cysteine-rich modules recognize diverse host cell-surface receptors during pathogenesis. DBLs of parasite erythrocyte-binding proteins mediate invasion, and those from the antigenically variant P. falciparum erythrocyte membrane protein 1 (PfEMP1) have been implicated in cytoadherence. The simian and human malarial parasites, P. knowlesi and P. vivax, invade human erythrocytes exclusively through the host DARC receptor (Duffy antigen receptor for chemokines). Here we present the crystal structure of the P. knowlesi DBL domain (Pkalpha-DBL), which binds to DARC during invasion of human erythrocytes. Pkalpha-DBL retains the overall fold observed in DBLs from P. falciparum erythrocyte-binding antigen (EBA)-175 (ref. 4). Mapping the residues that have previously been implicated in binding highlights a fairly flat but exposed site for DARC recognition in subdomain 2 of Pkalpha-DBL; this is in sharp contrast to receptor recognition by EBA-175 (ref. 4). In Pkalpha-DBL, the residues that contact DARC and the clusters of residues under immune pressure map to opposite surfaces of the DBL, and suggest a possible mechanism for immune evasion by P. vivax. Our comparative structural analysis of Pkalpha-DBL and P. falciparum EBA-175 provides a framework for the understanding of malaria parasite DBLs, and may affect the development of new prophylactic and therapeutic strategies.     

Effect on membrane transport in the erythrocytes by band 3 cross-linking

Band 3 and glucose transport protein (GluT1) are two kinds of important proteins in the human erythrocyte membranes. Bis(sulfosuccinimidyl)suberate (BS³), an impermeable cross-linker of band 3, inhibited NO₂ ⁻ transport, showing that anion exchange is affected by the association state of band 3 in the intact erythrocyte membranes. At the same time, the rates of glucose transport of both exit and entry declined. The amount of monomers of band 3 was decreased after treatment of the erythrocytes with BS³, but there was no change in GluT1 according to the SDS-PAGE patterns. This demonstrates that band 3 and GluT1 would be linkaged together in the erythrocyte membranes for the requirement of rapid and cooperative performance of physiological functions of the membrane proteins.     

嗜水气单胞菌HEC毒素苗对鲫鱼的免疫效果的初步研究

用嗜水气单胞菌强毒株BSK－10和TPS－30的HEC毒素苗分别免疫鲫鱼,以不同嗜水气单胞菌菌株攻毒,测定免疫保护率,结果显示：BSK－10HEC毒素苗对同型菌株攻击保护率达90％－100％,对异型菌株攻击的保护无效,为0－20％;TPS－30HEC毒素苗对,同源菌株攻击的保护率达100％,对同型菌的保护率为75．5％－100％,对异型菌株的保护率为44．4％－100％,经统计检验,TPS－30毒素苗对10株攻毒株的保护除1株无效外,其余均效果极显著或显著,对血清型不同的菌株的免疫保护率没有区别。     

Modulation of spectrin-actin assembly by erythrocyte adducin

The spectrin-based membrane skeleton, an assembly of proteins tightly associated with the plasma membrane, determines the shape and mechanical properties of erythrocytes. Spectrin, the most abundant component of this assembly, is an elongated and flexible molecule that, with potentiation by protein 4.1, is cross-linked at its ends by short actin filaments to form a lattice beneath the membrane. These and other proteins stabilize the plasma membrane, organize integral membrane proteins and maintain specialized regions of the cell surface. A membrane-skeleton-associated calmodulin-binding protein of erythrocytes is a major substrate for Ca2+- and phospholipid-dependent protein kinase C (ref. 5), and thus is a target for Ca2+ by two regulatory pathways. Here we demonstrate that this protein, called adducin: (1) binds tightly in vitro to spectrin-actin complexes but with much less affinity either to spectrin or to actin alone; (2) promotes assembly of additional spectrin molecules onto actin filaments; and (3) is inhibited in its ability to induce the binding of additional spectrin molecules to actin by micromolar concentrations of calmodulin and Ca2+. Adducin may be involved in the action of Ca2+ on erythrocyte membrane skeleton and in the assembly of spectrin-actin complexes.     

Ankyrin binding to (Na+ + K+)ATPase and implications for the organization of membrane domains in polarized cells

The interaction between membrane proteins and cytoplasmic structural proteins is thought to be one mechanism for maintaining the spatial order of proteins within functional domains on the plasma membrane. Such interactions have been characterized extensively in the human erythrocyte, where a dense, cytoplasmic matrix of proteins comprised mainly of spectrin and actin, is attached through a linker protein, ankyrin, to the anion transporter (Band 3). In several nonerythroid cell types, including neurons, exocrine cells and polarized epithelial cells homologues of ankyrin and spectrin (fodrin) are localized in specific membrane domains. Although these results suggest a functional linkage between ankyrin and fodrin and integral membrane proteins in the maintenance of membrane domains in nonerythroid cells, there has been little direct evidence of specific molecular interactions. Using a direct biological and chemical approach, we show here that ankyrin binds to the ubiquitous (Na+ + K+)ATPase, which has an asymmetrical distribution in polarized cells.     

Folding patterns of chromatin in chicken erythrocyte studied by using AFM and TEM

Four folding patterns (～15, ～30-40, ～60 and ～90-110 nm) of the higher_order structure of chromatin in chicken erythrocytes have been observed with an atomic force microscope (AFM) under near natural conditions. Results showed that the chromatin in the interphase nucleus of chicken erythrocyte was not in a disorganized form but represented a higher_order conformation. The thin sections of the chromatin in the nucleus of chicken erythrocyte were examined with a transmission electron microscope (TEM). Compared with TEM, AFM had the advantages that the samples did not need the preparation procedures such as fixing, sectioning and staining, and thus were closer to their native condition.     

重金属离子(Hg~(2+),Cu~(2+),Ag~+)对鲫鱼和鲤鱼呼吸运动机能的影响

本文研究了不同浓度的醋酸酮、醋酸汞和醋酸银溶液对鲫鱼(Carassius auratus)和鲤鱼(Cyprinus Carpio)呼吸运动机能的影响.结果表明,上述三种含重金属离子的溶液.其浓度在0.1,0.3,0.5,1.0mg/l范围内,均能迅速地引起鲫鱼和鲤鱼呼吸运动机能的改变.随着离子浓度的增高,这两种鱼的鳃盖运动频率有些减慢,但清除污物的洗涤运动(Clean-ing movement,又称Cough response)频率则急剧地增加.同一种重金属离子对不同鱼类的呼吸运动机能有不同的影响.Cu~(2+)对鲫鱼的作用大于鲤鱼;相反,Hg~(2+)对鲤鱼的作用大于鲫鱼,而Ag~+对这两种鱼类的作用都特别明显.在这三种重金属离子中,Ag~+对鱼类的呼吸运动机能的影响最大.鲫鱼和鲤鱼对重金属离子具有相当高的敏感性,而且其洗涤运动与重金属离子密切相关,这提示有可能利用这两种淡水养殖鱼类的呼吸运动对重金属离子这种反应特性作为水质重金属污染的生物监测指标.     

Rapid growth and sterility of growth hormone gene transgenic triploid carp

Triploid carp (100%) with 150 (3n=150) chromosomes were obtained by crossing the females of improved tetraploid hybrids (♀, 4n=200) of red crucian carp (♀)×common carp (♂) with the males of diploid yellow river carp (♂, 2n=100). The crosses yielded transgenic triploid carp (positive triploid fish, 44.2% of the progeny) and non-transgenic triploid carp (negative triploid fish). Histological examination of the gonads of 24-month-old positive triploid fish suggested they were sterile and the fish were not able to produce mature gametes during the breeding season. Morphologically, both the positive and negative triploid fish were similar. They had a spindle-shaped, laterally compressed, steel grey body with two pairs of barbells. Most of the quantifiable traits of the triploid carp were intermediate between those of the two parents. The positive and negative triploid fish were raised in the same pond for 2 years. The mean body weight of the positive triploid fish was 2.3 times higher than the negative triploid fish. The weight of the largest positive triploid fish was 2.91 times higher than that of the largest negative triploid fish. Thus, we produced fast-growing transgenic triploid carp that have a reduced ecological risk because of their inability to mate and produce progeny.     

Progress in measuring biophysical properties of membrane proteins with AFM single-molecule force spectroscopy

Membrane proteins are crucial in cell physiological activities and are the targets for most drugs.Thus,investigating the behaviors of membrane proteins not only provide deeper insights into cell function,but also help disease treatment and drug development.Atomic force microscopy is a unique tool for investigating the structure of membrane proteins.It can both image the morphology of single native membrane proteins with high resolution and,via single-molecule force spectroscopy(SMFS),directly measure their biophysical properties during molecular physiological activities such as ligand binding and protein unfolding.In the context of molecular biomechanics,SMFS has been successfully used to understand the structure and function of membrane proteins,complementing the static three-dimensional structures of proteins obtained by X-ray crystallography.Here,based on the authors’antigen-antibody binding force measurements in clinical tumor cells,the principle and method of SMFS is discussed,the progress in using SMFS to characterize membrane proteins is summarized,and challenges for SMFS are presented.     

Molecular cloning and polymorphism of major histocompatibility complex class I genes from grass carp (Ctenophayngodon idellus)

In order to clarify the molecular sequences,allelic polymorphism and the tertiary structure of grass carp (Ctenophayngodon idellus) MHC class I,and to further study their relationship with disease resistances,grass carp MHC class I gene (Ctid-MHC I) was cloned from a cDNA library and the allelic polymorphism in the population was investigated.The results showed that most of the variations exist in the peptide-binding domain (PBD) and high polymorphism was identified in the Ctid-MHC I allelic genes from 12 individuals.Based on the genetic distance,Ctid-MHC class I can be classified into 6 types (from Ctid-MHC I-UA to Ctid-MHC I-UF) which were subdivided into 9 lineages (from A to I).Comparison of the Ctid-MHC I among animals and humans showed that the key amino acids of the peptide binding sites are conserved.Analysis of the tertiary structure of the PBD between Grass carp and human crystallographic data of HLA-A2,the variation with insertion or deletion was found in eight regions (A～H).The phylogenetic tree of MHC class I indicates the evolution of MHC class I among grass carp,fish,amphibian,birds,higher vertebrates and humans.     

Membrane structure and interactions with protein and DNA in bacteriophage PRD1

Membranes are essential for selectively controlling the passage of molecules in and out of cells and mediating the response of cells to their environment. Biological membranes and their associated proteins present considerable difficulties for structural analysis. Although enveloped viruses have been imaged at about 9 A resolution by cryo-electron microscopy and image reconstruction, no detailed crystallographic structure of a membrane system has been described. The structure of the bacteriophage PRD1 particle, determined by X-ray crystallography at about 4 A resolution, allows the first detailed analysis of a membrane-containing virus. The architecture of the viral capsid and its implications for virus assembly are presented in the accompanying paper. Here we show that the electron density also reveals the icosahedral lipid bilayer, beneath the protein capsid, enveloping the viral DNA. The viral membrane contains about 26,000 lipid molecules asymmetrically distributed between the membrane leaflets. The inner leaflet is composed predominantly of zwitterionic phosphatidylethanolamine molecules, facilitating a very close interaction with the viral DNA, which we estimate to be packaged to a pressure of about 45 atm, factors that are likely to be important during membrane-mediated DNA translocation into the host cell. In contrast, the outer leaflet is enriched in phosphatidylglycerol and cardiolipin, which show a marked lateral segregation within the icosahedral asymmetric unit. In addition, the lipid headgroups show a surprising degree of order.     

黄颡鱼人工繁殖的初步研究

利用面积为2000m^2的鲫鱼池套养培育黄颡鱼亲鱼；采用不同的药物和剂量进行人工催产；利用家鱼孵化池进行流水孵化。实验结果表明：黄颡鱼在性成熟度基本一致的情况下，用LRH—A2 DOM PG(LRH—A2、DOM、PG分别为4ug、0．8mg、2．2mg)的配合药物来进行生产，催产率可达到95％。     

胞二磷胆碱抑制贮存红细胞溶血作用的研究

贮存红细胞溶血的主要原因,现认为与能量代谢障碍和膜结构功能有密切关系.前文曾报导胞二磷胆碱(简称CDP胆碱)能抑制贮存红细胞的溶血,而且其作用主要不是通过能量代谢.本文试从CDP胆碱与红细胞膜磷脂的关系等方面作一些探索. 将去除血浆的红细胞贮存于代血浆中,看到CDP胆碱仍能抑制溶血.经测定血浆磷脂的含量,也没有发现CDP胆碱血和对照之间有明显差别.此结果提示CDP胆碱抑制红细胞溶血的作用与血浆并无联系,排除了通过与血浆进行磷脂交换来更新红细胞磷脂,从而抑制溶血的可能性.此外,还测定了CDP胆碱血和对照的红细胞膜磷脂含量,并用薄板层析法测定了卵磷脂和神经鞘磷脂的百分含量.结果显示,实验组并不比对照有所增高,这也表明CDP胆碱对红细胞膜磷脂含量没有显著增进作用. 除CDP胆碱以外,CMP也具有抑制贮存红细胞溶血的作用,其意义值得作进一步研究.     

胞二磷胆碱抑制贮存红细胞溶血作用的研究

贮存红细胞溶血的主要原因,现认为与能量代谢障碍和膜结构功能有密切关系。前文曾报导胞二磷胆碱(简称CDP胆碱)能抑制贮存红细胞的溶血,而且其作用主要不是通过能量代谢。本文试从CDP胆碱与红细胞膜磷脂的关系等方面作一些探索。将去除血浆的红细胞贮存于代血浆中,看到CDP胆碱仍能抑制溶血。经测定血浆磷脂的含量,也没有发现CDP胆碱血和对照之间有明显差别。此结果提示CDP胆碱抑制红细胞溶血的作用与血浆并无联系,排除了通过与血浆进行磷脂交换来更新红细胞磷脂,从而抑制溶血的可能性。此外,还测定了CDP胆碱血和对照的红细胞膜磷脂含量,并用薄板层析法测定了卵磷脂和神经鞘磷脂的百分含量。结果显示,实验组并不比对照有所增高,这也表明CDP胆碱对红细胞膜磷脂含量没有显著增进作用。除CDP胆碱以外,CMP也具有抑制贮存红细胞溶血的作用,其意义值得作进一步研究。     

Abnormal display of PfEMP-1 on erythrocytes carrying haemoglobin C may protect against malaria

Haemoglobin C, which carries a glutamate-to-lysine mutation in the beta-globin chain, protects West African children against Plasmodium falciparum malaria. Mechanisms of protection are not established for the heterozygous (haemoglobin AC) or homozygous (haemoglobin CC) states. Here we report a marked effect of haemoglobin C on the cell-surface properties of P. falciparum-infected erythrocytes involved in pathogenesis. Relative to parasite-infected normal erythrocytes (haemoglobin AA), parasitized AC and CC erythrocytes show reduced adhesion to endothelial monolayers expressing CD36 and intercellular adhesion molecule-1 (ICAM-1). They also show impaired rosetting interactions with non-parasitized erythrocytes, and reduced agglutination in the presence of pooled sera from malaria-immune adults. Abnormal cell-surface display of the main variable cytoadherence ligand, PfEMP-1 (P. falciparum erythrocyte membrane protein-1), correlates with these findings. The abnormalities in PfEMP-1 display are associated with markers of erythrocyte senescence, and are greater in CC than in AC erythrocytes. Haemoglobin C might protect against malaria by reducing PfEMP-1-mediated adherence of parasitized erythrocytes, thereby mitigating the effects of their sequestration in the microvasculature.