Transmembrane orientation of the fibronectin receptor complex (integrin) demonstrated directly by a combination of immunocytochemical approaches

The avian 140-KD cell adhesion receptor or "integrin," a complex of three glycoproteins with molecular masses averaging 140 KD, interacts with extracellular fibronectin and forms a linkage complex that co-localizes with intracellular actin. To probe the molecular interactions involved in this linkage complex, we used monoclonal antibodies and a combination of immunolocalization approaches to determine whether any component was transmembrane. Immunoadsorption and immunoblotting experiments demonstrated that anti-120-KD Mabs recognized the band 3 component of integrin isolated from chicken embryo fibroblasts (CEF) by JG22E immunoaffinity chromatography, and they co-localize with anti-fibronectin and polyclonal anti-integrin at cell contact sites in double-labeling experiments. Immunofluorescence experiments involved comparisons of double-labeled intact cells or substrate-attached, ventral plasma membrane "rip-off" fragments, using anti-fibronectin and each of the anti-120-KD Mabs. The extracellular faces of living intact cells were strongly labeled by a majority (approximately 70%) of the anti-120-KD Mabs at fibronectin-membrane attachment sites. The remainder (approximately 30%) labeled intact cells weakly or not at all. However, although the anti-120-KD Mab ES186 did not stain living cells, it did demonstrate positive staining above substratum contact sites over entire isolated rip-off membranes. In contrast, Mabs directed against putative extracellular epitopes and anti-fibronectin antibodies did not label these sites at the center of rip-offs unless the membranes were detergent permeabilized. Proteolysis experiments suggested that the ES186 epitope was located at one end of the molecule, since removal of short fragments from integrin band 3 concomitantly removed or destroyed the ES186 epitope, whereas the extracellular epitopes still remained. These experiments directly demonstrate that integrin band 3 is a transmembrane polypeptide with at least one epitope recognized by anti-120-KD Mabs on the cytoplasmic side of the plasma membrane and at least one epitope on the extracellular cell surface.     

Pre-lysosomal divergence of alpha 2-macroglobulin and transferrin: a kinetic study using a monoclonal antibody against a lysosomal membrane glycoprotein (LAMP-1)

We have used electron microscopic immunocytochemistry to compare the distribution of LAMP-1, a marker for lysosomal membranes, with the intracellular localization of alpha 2-macroglobulin (alpha 2-M) and transferrin at various time points after their endocytosis into cultured NIH 3T3 cells. The purposes of this study were (a) to determine how soon endocytic ligands reach lysosomal organelles, (b) to examine whether the intermediate endocytic vesicles gained lysosomal markers gradually or in a precipitous, discrete event, and (c) to examine the relationship, if any, between the pathway of recycling ligands and lysosomes. At early time points (0-5 min) after initiation of endocytosis, most structures containing alpha 2-M labeled with colloidal gold (receptosomes) were not labeled by anti-LAMP-1 detected using ferritin bridge or peroxidase immunocytochemistry. At late time points (greater than or equal to 15 min), the structures containing alpha 2-M (lysosomes) were strongly labeled by anti-LAMP-1. In contrast, transferrin that was directly labeled with ferritin was mostly located in LAMP-1-negative structures at all time points studied. The proportion of alpha 2-M-gold containing vesicles strongly labeled for LAMP-1 roughly paralleled the proportion of alpha 2-M-gold-containing structures positive for cytochemically detectable acid phosphatase. Our data indicate that ligands such as transferrin that are internalized through coated pits and receptosomes, but not delivered to lysosomes, do not traverse a lysosomal organelle compartment as marked by LAMP-1 content. Ligands such as alpha 2-M that are destined for lysosomal delivery reach a LAMP-1-positive organelle compartment only after they traverse LAMP-1-negative, non-lysosomal vesicles previously described as receptosomes.     

Modulation of rat granulocyte traffic by a surface active agent in vitro and bleomycin injury

Pluronic F68 (F68) is a nonionic surfactant which has been reported to inhibit the in vitro adherence and migration of polymorphonuclear leukocytes (PMN) obtained from some species. We demonstrated similar effects on PMN obtained from rats, with diminished adherence to nylon wool and diminished chemotaxis toward zymosan-activated serum. We then examined the in vivo effects of 12-hr F68 infusion on the injury induced by intratracheal bleomycin instillation (ITB) in rats. When sacrificed 24 hr following injury, rats demonstrated neutrophilia, neutrophil-prominent lung lavage cellularity, and increased lung weights. F68 decreased lavage leukocyte counts and lung weight gain in ITB-injured animals. Lung weights of ITB-injured animals correlated (r = 0.81, P less than 0.001) with logarithmic values of lavage PMN. F68 also enhanced neutrophilia and decreased spleen weight gain in injured animals. The acute effects of F68 on circulating leukocyte counts, osmolality, and total complement were also examined. The data demonstrate that F68 can affect PMN traffic both in vitro and in vivo. The data also confirm the prominence of PMN in lavage fluid early in ITB injury, and suggest that an influx of relatively few PMN is associated with lung weight gain in this model.     

从“湖北光敏感核不育水稻”的未受精子房和花药培养出单倍体植株

采用10种诱导培养基,培养湖北光敏感核不育水稻农垦58品种的未受精子房和花药。共培养未受精子房2790个,获得胚囊愈伤组织17块,最高诱导频率达3.33%,其中2块分化出绿苗。培养花药16740个,获得花药愈伤组织15块,最高诱导频率为0.92%,其中3块分化出苗,2丛白苗,1株绿苗。胚囊植株和花粉植株经根尖染色体检查为单倍体,2n=x=12。实验证明,液体培养、2,4-D0.2-0.5 mg/1、低温预处理对诱导胚囊愈伤组织及花粉愈伤组织的形成具良好效果。     

诱导小麦-天兰偃麦草-黑麦三属杂种花粉植株的研究

以法国六倍体小黑麦为母本,分别与普通小麦(Triticum aestivum)和天兰偃麦草(Elytrigia intermedia of Agropyron glaucum)的杂交后代中的中间类型3号和5号杂交。由此获得的三属杂种F_1性状介于亲本之间,兼有三属亲本类型的特征,呈中间类型。用马铃薯-Ⅱ培养基培养三属杂种F_1的花药,诱导花粉愈伤组织。将所获得的愈伤组织转入190-2培养基进行分化,已成功地诱导出一批三属间杂种花粉植株,并用Giemsa显带技术鉴定花粉植株的染色体组组成。     

D10S20, a previously unmapped RFLP (OS-3), is located on 10q near D10S4

The locus recognized by the probe OS-3 is assigned to chromosome 10 both by Southern blot analysis of a panel of somatic cell hybrid DNAs and by genetic linkage to markers already assigned to chromosome 10. In Caucasians this probe recognizes a three-allele TaqI RFLP as well as two-allele BanII and RsaI RFLPs which are both in strong linkage disequilibrium with each other and with the TaqI RFLP. The D10S20 locus defined by this probe maps 5.5 cM distal to D10S4 on the long arm of chromosome 10. Because this human clone hybridizes with mouse genomic DNA, it will be useful in comparative mapping studies.     

国产姜科植物的染色体计数(5)

<正> 本文继续对7属15种国产姜科植物作染色体计数观察,其中10种是染色体计数的新纪录(表、图)。通过对黄花大苞姜(Caulokaempferia coenobialis)和大苞姜(C.yunnanensis)以及凹唇姜(Bosenbevgia rotunda)和心叶凹唇姜(B.fallax)的染色体观察,确定了大苞姜属(Caulokaempferia)和凹唇姜属(Bosenbergia)的染色体基数分别为12和9。     

一阶导数光谱法测定猫豆中左旋多巴的含量

川一阶导数光谱法测定猫豆中主旋多巴的含量,可以排除无关杂质吸收的干扰,省去复杂的分离提取步骤。试验结果表明:相关系数达0.9999,平均回收率101.5％,重现性较好。     

内蒙古石拐群古鳕类一新属

内蒙古石拐地区石拐群召沟组中的长腹鳍大青山鳕(新属、新种) Daqingshaniscus longiventralis gen. et sp. nov. 是在我国中侏罗世地层中发现的一比较原始的古鳕类.其头骨眶后部分短,鳃盖骨大于下鳃盖骨,背鳍位于腹鳍与臀鳍之间,腹鳍基线长,鳍条都从基部分节、远端分叉,棘鳞仅见于尾上叶,全歪型尾,鳞片呈菱形.大青山鳕既与苏联南哈萨克斯坦 Karatau 地区的 Pteroniscus 很接近,又与我国新疆的维吾尔鳕 Uighuroniscus 及西德北部的 Stadthagen 地区的 Indaginilepis 相似.     

MASTODONT REMAINS FROM THE MIOCENE OF JUNGGAR BASIN IN XINJIANG

<正> The mastodont materials described in the present paper, associated with Amblycastor tunggurensis, Amphicyon sp., Anchitherium cf. aurelianense, Brachypotherium sp., ?Chilotherium sp., Stephanocemas thomsoni, Dicrocerus grangeri, Eotragus sp., Oioceros grangeri and O. noverca, were collected from the Haramagai formation of Junggar Basin in Xinjiang by an IVPP field team in 1982. The geology of the area has already been reported by Tong (1986, 1987). The mastodont fossils found at 5 sites on the north and west banks of the Ulungur river, Junggar Basin are abundant. They comprise 5 species, among which is one new species.The author is greatly indebted to the IVPP field team (Tong, Y., the head of the 1982 Junggar field team) allocating the mastodont materials for me to study.