三个品种豚鼠血液蛋白多态性的比较分析

目的比较分析白毛黑眼（WHBE）豚鼠和DHP豚鼠、花色豚鼠三个品种豚鼠在13个血液蛋白位点上的多态性。方法采用垂直板浓度和pH均不连续的聚丙烯酰胺凝胶电泳法对WHBE豚鼠、DHP豚鼠和花色豚鼠的66只个体的后白蛋白（Po）、前转铁蛋白1（Prt1）、前转铁蛋白2（Prt2）、转铁蛋白1（Tf1）、转铁蛋白2（Tf2）、后转铁蛋白（Ptf）、慢α球蛋白（Sag）、红细胞酯酶（Es）、血清酯酶1（Est1）、血清酯酶3（Est3）、血红蛋白α（Hbα）、血红蛋白β（Hbβ）和白蛋白（Alb）共13个蛋白位点进行了电泳及染色,再利用电泳图谱对各蛋白位点基因频率、平均杂合度和遗传距离进行计算,然后结合聚类分析。结果 Tf1、Tf2、Ptf、Est1和Es在三个豚鼠品种中表现为多态,其中Tf1可作为识别WHBE豚鼠的遗传标记。Po、Prt1、Prt2、Sag、Est3、Hbα、Hbβ和Alb等位点在三个豚鼠品种中的表型一致。Hardy-Weinberg平衡状态分析表明,Es为DHP豚鼠的高度不平衡位点。Ptf为花色豚鼠的高度不平衡位点。在WHBE豚鼠中,Tf1为高度不平衡位点,Est1为不平衡位点。在三个豚鼠品种中,所检测的13个蛋白位点的平均杂合度的排列顺序为：花色豚鼠（0.350 1）〉WHBE豚鼠（0.339 0）〉DHP豚鼠（0.313 5）。聚类分析结果表明,花色豚鼠和WHBE豚鼠的遗传遗传距离最近（0.064 3）,DHP豚鼠与花色豚鼠的遗传距离最远（0.179 2）。结论利用这些蛋白位点可以有效鉴别WHBE豚鼠、DHP豚鼠和花色豚鼠血液蛋白的遗传多态性。     

Activation of Guinea Pig Herpesvirus Antigen in Leukemic Lymphoblasts of Guinea Pig

Herpesvirus (GPHV) antigen is either present in very small amounts, or absent in leukemic lymphoblasts taken directly from strain 2 guinea pigs. However, after maintenance in tissue culture for 72 hr, almost 100% of these lymphoblasts contained GPHV antigen. The expression of GPHV antigen could be demonstrated by indirect immunofluorescent technique as well as by the direct (125)I labeled antibody technique. However, infectious virus or virus capsids could not be detected in these cells either by infectivity tests or electron microscopy.     

阳离子胶体金标记活细胞阴离子位点的双光子荧光成像及其应用

使用阳离子胶体金标记中国仓鼠卵巢细胞(CHO-K1)的阴离子位点,并采用双光子荧光显微成像和荧光寿命成像技术记录活细胞的阴离子场分布.阳离子胶体金是纳米量级金微粒与多聚L-赖氨酸的结合物,金纳米微粒在超短激光脉冲的照射下可以产生高度局域化的光热效应.当飞秒激光脉冲聚焦在细胞膜上标记的金纳米微粒时会产生这种纳米尺度的微光热效应,并在不影响细胞活性的前提下暂时提高细胞膜的通透性.基于这种效应,使用聚焦的飞秒激光脉冲三维扫描照射CHO-K1细胞,将分子质量为10ku的荧光探针大分子异硫氰酸荧光素葡聚糖(fluorescein isothioeyanate-dextran, FITC-D)递送到CHO-K1细胞的内部,并用双光子荧光图像记录其递送的过程.使用流式细胞仪分析不同实验条件下FITC-D的转导率和细胞死亡率的关系.     

Immunogenic Properties of Colloidal Gold

We studied the capacity of colloidal gold for enhancing specific and nonspecific immune response in laboratory animals (rabbits, rats, and mice) immunized with antigens of various nature. The antibody titers obtained with colloidal gold as a carrier were higher as compared to the standard immunization techniques (free antigen or its combination with Freund's adjuvant). Application of colloidal gold also enhanced nonspecific immune responses, such as lysozyme concentration in the blood, activity of the complement system proteins, as well as phagocytic and bactericidal activities. The antibodies were tested by immunodot assay using gold markers. Immunization of the animals with colloidal gold conjugates with haptens or complete antigens (without other adjuvants) was shown to induce the production of highly active antibodies. In addition, the amount of antigen used for animal immunization with colloidal gold was an order of magnitude lower, compared to immunization with complete Freund's adjuvant. This fact can be evidence for adjuvant properties of colloidal gold proper.     

Putative Cholinergic-Specific Gangliosides in Guinea Pig Forebrain

The nature of the cholinergic-specific antigen Chol-1 recognized by an antiserum raised against Torpedo cholinergic electromotor synaptosomal plasma membranes was investigated in guinea pig forebrain to establish whether it has a gangliosidic nature in guinea pig as in Torpedo. Gangliosides extracted from guinea pig forebrain and extensively purified to eliminate peptide contaminants were effective in inhibiting the selective lysis of the cholinergic subpopulation of cortical synaptosomes induced by the antiserum. Neuraminidase, protease, alkali, and heat treatment did not impair the inhibitory activity of gangliosides. Whereas the antiserum recognized many gangliosides from Torpedo electric organ, the immunostaining of guinea pig forebrain gangliosides separated on TLC showed only two immunopositive bands migrating close to GT1b and GQ. After affinity purification on Torpedo electric organ gangliosides the activity of the antiserum in inducing complement-mediated lysis was increased and it still recognized the two ganglioside bands on TLC. These results strongly suggest the existence of two polysialogangliosides bearing antigenic determinants specific for the cholinergic neurons.     

Opsonization of Staphylococcus aureus by Guinea Pig 7S Gamma-Globulin

Opsonization of Staphylococcus aureus to phagocytosis by guinea pig peritoneal exudate cells (PEC) was examined with various isolated serum globulins. Significant enhancement of in vitro phagocytosis was afforded by partially purified 7S_γ1-immunoglobulin. The rate of killing or the rate of ingestion of ³H-thymidine labeled bacteria by PEC was used as an index of phagocytosis. The results indicate that the immunoglobulin can mediate the adsorption of an allogeneic surface independent of a specific antigenic component.     

不同因素诱导豚鼠肝中金属硫蛋白的研究

通过皮下注射的方法诱导豚鼠产生金属硫蛋白（ＭＴ）,研究了重金属元素（Ｃｄ）、微量元素（Ｃｕ,Ｚｎ）及有机试剂（ＣＣｌ４,在体内可产生自由基）等因素的诱导与豚鼠肝脏中ＭＴ不同亚型的含量及金属结合状态的变化关系．实验结果表明,微量元素及有机试剂的诱导可使豚鼠肝脏中ＭＴ１的产量明显高于ＭＴ２,说明在体内ＭＴ１在参与微量元素的储存及清除自由基功能方面比ＭＴ２强．在重金属元素诱导下体内ＭＴ１对重金属元素的结合量远远大于ＭＴ２．表明ＭＴ１的重金属解毒能力比ＭＴ２强．上述实验结果与对不同亚型ＭＴ生物学功能差异的体外研究结果相吻合．此外,无论采用上述何种因素诱导,所得ＭＴ中均结合有Ｃｕ．对Ｃｕ在ＭＴ形成过程中的作用也进行了初步探讨．     

豚鼠精母细胞联会复合体的电镜观察

以微铺展法制备豚鼠精母细胞联会复合体标本,经硝酸银染色后作电镜观察,建立了SC组型．与有丝分裂染色体组型比较,发现二者有良好的一致性．在粗线期,X,Y轴的配对区很短,配对区的X轴和Y轴没有明显变细．未发现银染SC具有着丝粒,并对可能的原因作了分析讨论．     

豚鼠精母细胞联会复合体的电镜观察

以微铺展法制备豚鼠精母细胞联会复合体标本,经硝酸银染色后作电镜观察,建立了SC组型。与有丝分裂染色体组型比较,发现二者有良好的一致性。在粗线期,X,Y轴的配对区很短,配对区的X轴和Y轴没有明显变细。未发现银染SC具有着丝粒,并对可能的原因做了分析讨论。Synaptonemal complexes (SC) in Guinea pig (Cavia porcellus) spermatocytes prepared with micro-spreading technique and silver staningwere analyzed by electron microscopy.The meiotic SC karyotype was constructed from 7 cells and compared with the mitotic chromosome karyotype. There is a good agreement between them. At pachytene, there is only a very a very short pairing region in which X-and Y-axis are not stinctly thin.The kinetochore thin. The kinetochore was not found on each SC in our experiments and probable reason has been discussed.     

Cortical Representation of Species-Specific Vocalizations in Guinea Pig

We investigated the representation of four typical guinea pig vocalizations in the auditory cortex (AI) in anesthetized guinea pigs with the aim to compare cortical data to the data already published for identical calls in subcortical structures - the inferior colliculus (IC) and medial geniculate body (MGB). Like the subcortical neurons also cortical neurons typically responded to many calls with a time-locked response to one or more temporal elements of the calls. The neuronal response patterns in the AI correlated well with the sound temporal envelope of chirp (an isolated short phrase), but correlated less well in the case of chutter and whistle (longer calls) or purr (a call with a fast repetition rate of phrases). Neuronal rate vs. characteristic frequency profiles provided only a coarse representation of the calls’ frequency spectra. A comparison between the activity in the AI and those of subcortical structures showed a different transformation of the neuronal response patterns from the IC to the AI for individual calls: i) while the temporal representation of chirp remained unchanged, the representations of whistle and chutter were transformed at the thalamic level and the response to purr at the cortical level; ii) for the wideband calls (whistle, chirp) the rate representation of the call spectra was preserved in the AI and MGB at the level present in the IC, while in the case of low-frequency calls (chutter, purr), the representation was less precise in the AI and MGB than in the IC; iii) the difference in the response strength to natural and time-reversed whistle was found to be smaller in the AI than in the IC or MGB.     

Activation of K-Cl Cotransport by Mild Warming in Guinea Pig Red Cells

Unidirectional, ouabain-insensitive K⁺ influx rose steeply with warming at temperatures above 37°C in guinea pig erythrocytes incubated in isotonic medium. The only component of ouabain-insensitive K⁺ influx to show the same steep rise was K-Cl cotransport (Q₁₀ of 10 between 37 and 41°C); Na-K-Cl cotransport remained constant or declined and residual K⁺ influx in hypertonic medium with ouabain and bumetanide rose only gradually. Similar results were obtained for unidirectional K⁺ efflux. Thermal activation of K-Cl cotransport-mediated K⁺ influx was fully dependent on the presence of chloride in the medium; none occurred with nitrate replacing chloride. The increase of K⁺ influx through K-Cl cotransport from 37 to 41°C was blocked by calyculin A, a phosphatase inhibitor. The Q₁₀ of K-Cl cotransport fully activated by hydroxylamine and hypotonicity was about 2. The time course of K⁺ entry showed an immediate transition to a higher rate when cells were instantly warmed from 37 to 41°C, but there was a 7-min time lag in returning to a lower rate when cells were cooled from 41 to 37°C. These results indicate that the steepness of the response of K-Cl cotransport to mild warming is due to altered regulation of the transporter. Total unidirectional K⁺ influx was equal to total unidirectional K⁺ efflux at 37–45°C, but K⁺ influx exceeded K⁺ efflux at 41°C when K-Cl cotransport was inhibited by calyculin or prevented by hypertonic incubation. The net loss of K⁺ that results from the thermal activation of isosomotic K-Cl cotransport reported here would offset a tendency for cell swelling that could arise with warming through an imbalance of pump and leak for Na⁺ or for K⁺. Received: 1 November 1997/Revised: 5 March 1998     

Characterization of Bromodeoxyuridine-Induced Endogenous Guinea Pig Virus

An endogenous virus (GPV) was induced after 5-bromodeoxyuridine treatment of cultured guinea pig cells. Compared to Gross murine leukemia virus (G-MuLV) GPV has a reproducibly heterogenous density of about 1.16 to 1.18 g/ml. The virion-associated RNA is slightly larger than that in G-MuLV. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of dissociated GPV resolved five major structural proteins: I (molecular weight 70,000), II (molecular weight 36,000), III (molecular weight 24,000), IV (molecular weight 18,000), and V (molecular weight 16,000) which are similar to but distinct from G-MuLV proteins. Proteins I and II were demonstrated to be glycoproteins by incorporation of [(3)H]glucosamine. GPV and G-MuLV did not have any appreciable genetic homology or any common group-specific antigens when analyzed by immunodiffusion, radioimmunoassay, and indirect immunofluorescence. Morphogenesis of GPV also differed from that of a typical type C oncornavirus and proceeded via two pathways: (i) a majority of virus particles were formed in cytoplasmic vacuoles and were released after cellular disruption; and (ii) a minor population of particles were assembled in the cytoplasmic matrix and then migrated to the plasma membrane where they budded into the extracellular space. To date, GPV has been unable to initiate or maintain a productive replication in any cell line tested.