Visualization of the mitochondria ofToxoplasma gondii-infected mouse fibroblasts by the cationic permeant fluorescent dye rhodamine 123

Summary The mitochondria of living mouse fibroblasts infected withToxoplasma gondii were monitored with the cationic permeant fluorescent dye rhodamine 123. Fluorescence microscopy revealed that host cell mitochondria accumulated at the cytoplasmic surface of parasitophorous vacuoles and increased the dye uptake in the periparasitophorous vacuole asT. gondii multiplied.The author wishes gratefully to acknowledge the work of Miss Mayumi Kato and Mr. Isao Kimata in preparing the photographs. The publication cost was supported by a grant of the Ohyama Health Foundation.     

Experimenteller Beitrag zur Frage der Mitochondrienfunktion

Summary In the phase microscope it can be seen that cloudy swelling is the consequence of an enlargement of the mitochondria, whereby the membrane of the mitochondria is detached from the body of the mitochondrion. An increased water intake seems to be the reason of this phenomenon.The intraperitoneal injection of hen egg albumen is followed after several hours by an accumulation of hyaline droplets in the protoplasma of the renal tubule cells. The PM reveals that this foreign protein is stored within the mitochondria, which are converted into brilliant granules. The same is true for the accumulation of vital dyes in kidney tubule cells, but in this case there is a combination of the dye with protein deriving from a slight lesion of the cells.     

刚地弓形虫RH株感染对BALB／c小鼠情绪行为的影响

目的研究刚地弓形虫RH株感染对BALB／c小鼠学习记忆行为的影响及可能机制。方法将72只周龄、大小相近的雄性BALB／c小鼠采用随机数字表分为生理盐水对照组与不同数量（3×10^3／ml、3×10^4／ml、3×10^5／m1）弓形虫RH株感染组,每组18只。于感染第5周从每组各随机抽取6只分别进行高架十字迷宫实验、旷场实验及强迫游泳实验,观察各组小鼠在实验中情绪行为变化。并记录各项指标进行统计分析。结果在高架迷宫试验与旷场实验中,3×10^3／ml弓形虫感染组小鼠与生理盐水对照组小鼠比较,各项行为学指标无明显差异。在3×10^4／ml、3×10^5／ml弓形虫感染组小鼠与对照组小鼠比较出现明显降低（P〈0．05）．以3×10^5／ml感染组最为明显,其小鼠运动活力（0E＋cE）、进入开放臂次数比例（OE％）、开放臂停留时间比例（OT％）小鼠爬行总格数、中央格在总格数中比例分别为11．08±2．12、28．73±0．59％、25．62±2．33％、32．30±17．26、2．42±0．65％。在强迫游泳试验中,各弓形虫感染组小鼠游泳的静止时间均高于对照组小鼠（P〈0．05）,3×10^5／ml感染组静止时间最长,达226．6±1．9S。结论刚地弓形虫RH株感染可引起小鼠情绪行为改变,具有焦虑抑郁倾向。     

刚地弓形虫培养上清抑制THP-1细胞增殖及诱导凋亡的研究

摘要：目的提取弓形虫体外细胞共培养上清,并研究上清对人急性单核细胞白血病细胞THP-1增殖及凋亡的影响。方法收集对数生长期的THP-1细胞以5X10^7／ml细胞浓度接种于不同培养瓶中,对照组加入含10％胎牛血清的RPMll640,实验组加入相同体积不同数量（2×10^7／ml、4X10^7／ml、8×10^7／m1）弓形虫速殖子培养上清,采用四甲基氮噻唑蓝（MTY）法检测吸光度（A490值）并计算THP-1细胞增殖抑制率;倒置显微镜下观察细胞形态变化;Annexin-V-FITC／PI染色细胞后上流式细胞仪检测各个时间点细胞凋亡率变化,以Western印迹方法分析凋亡相关蛋白Bax、Bcl-2的表达或活性。结果MTY法检测结果弓形虫培养上清呈时间剂量依赖性抑制THP-1细胞株增殖,倒置显微镜下观察处理组细胞有发泡现象和凋亡小体出现。流式细胞仪检测弓形虫感染后的THP-1细胞凋亡率较对照组有升高趋势（P〈0．05）,呈量效依赖性,Westernblot检测刚地弓形虫培养上清作用于THP-l细胞48h后实验组的Bax、Bcl-2蛋白表达较对照组的比值分别有明显的升高与降低（P〈0．05）。结论刚地弓形虫速殖子培养上清对体外培养THP-l细胞增殖有明显的抑制作用,并可诱导THP-1细胞凋亡。     

The interaction of organic cations with the mitochondrial membrane.

Weak and strong organic bases behave in an opposite manner in respect to several mitochondrial functions. The former induce a catalytic exchange with K+ in valinomycon-treated, respiratory-inhibited mitochondria, and act as uncouplers in respiring mitochondria. The latter induce a stoicheometric exchange with K+ and are actively taken up by respiring mitochondria.     

The interaction of organic cations with the mitochondrial membrane

Summary Weak and strong organic bases behave in an opposite manner in respect to several mitochondrial functions. The former induce a catalytic exchange with K⁺ in valinomycon-treated, respiratory-inhibited mitochondria, and act as uncouplers in respiring mitochondria. The latter induce a stoicheometric exchange with K⁺ and are actively taken up by respiring mitochondria.     

Suppression and enhancement of humoral immune response to Toxoplasma gondii by passive antibody

Anti-toxoplasma antibodies administered passively to mice may lead to suppression of enhancement (reported for the first time with Protozoan parasites) of subsequent antibody response when these animals are later infected with Toxoplasma gondii. The outcome is dependent on infecting strain of Toxoplasma and the antigen-antibody ratio.     

Respiratory control in liver mitochondria of rats hosting the walker 256 carcinoma tumor.

The decrease in the respiratory control ratio of mitochondria is a first order process when these organelles are incubated in isotonic sucrose. Furthermore, the initial respiratory control ratios and the rates of loss in respiratory control in liver mitochondria from rats hosting the Walker 256 carcinoma are not significantly different form the same properties of mitochondria from untreated animals.     

Schwann cell mitochondria as key regulators in the development and maintenance of peripheral nerve axons

Formation of myelin sheaths by Schwann cells (SCs) enables rapid and efficient transmission of action potentials in peripheral axons, and disruption of myelination results in disorders that involve decreased sensory and motor functions. Given that construction of SC myelin requires high levels of lipid and protein synthesis, mitochondria, which are pivotal in cellular metabolism, may be potential regulators of the formation and maintenance of SC myelin. Supporting this notion, abnormal mitochondria are found in SCs of neuropathic peripheral nerves in both human patients and the relevant animal models. However, evidence for the importance of SC mitochondria in myelination has been limited, until recently. Several studies have recently used genetic approaches that allow SC-specific ablation of mitochondrial metabolic activity in living animals to show the critical roles of SC mitochondria in the development and maintenance of peripheral nerve axons. Here, we review current knowledge about the involvement of SC mitochondria in the formation and dysfunction of myelinated axons in the peripheral nervous system.     

Abnormal mitochondria in retinoblastoma.

In examination of six retinoblastoma tumor specimens, bizzare mitochondria were often found. Some are irregular forms with focal expansion and constrictions. Occasionally, a portion of the mitochondria forms rings. Branching mitochondria are also seen. Other striking features of the mitochondria from tumor cells are the alternation of cristae. Dense bodies are also occasionally observed within the mitochondria. Morphological modifications of the mitochondria may be as results of pathological conditions of the tumor cells.     

Abnormal mitochondria in retinoblastoma

Summary In examination of six retinoblastoma tumor specimens, bizzare mitochondria were often found. Some are irregular forms with focal expansion and constrictions. Occasionally, a portion of the mitochondria forms rings. Branching mitochondria are also seen. Other striking features of the mitochondria from tumor cells are the alteration of cristae. Dense bodies are also occasionally observed within the mitochondria. Morphological modifications of the mitochondria may be as results of pathological conditions of the tumor cells.     

The effects of 4 detergents on the respiration of mitochondria isolated from rat liver]

The effects of these four detergents (two non-ionic: Titron X100 and Tween 80, two ionic: sodium cholate and sodium dedeoxycholate) upon the respiratory intensities of mitochondria and upon the ADP/0 and respiratory control ratios were observed. At low concentrations and in the absence of exogenous ADP, non ionic as well as ionic detergents provoked a threefold (or fourfold) increase of the respiratory intensities of mitochondria. At higher concentrations, the four detergents were inhibitory for mitochondrial oxidations in the order: Triton X 100 greater than DOC greater than cholate greater than Tween 80. At increasing doses, the four detergents progressively decreased the phosphorylating capacities of mitochondria.     

Mitochondrial association of alpha-synuclein causes oxidative stress

α-Synuclein is a neuron-specific protein that contributes to the pathology of Parkinson’s disease via mitochondria-related mechanisms. The present study investigated possible interaction of α-synuclein with mitochondria and consequences of such interaction. Using SHSY cells overexpressing α-synuclein A53T mutant or wild-type, as well as isolated rat brain mitochondria, the present study shows that α-synuclein localizes at the mitochondrial membrane. In both SHSY cells and isolated mitochondria, interaction of α-synuclein with mitochondria causes release of cytochrome c, increase of mitochondrial calcium and nitric oxide, and oxidative modification of mitochondrial components. These findings suggest a pivotal role for mitochondria in oxidative stress and apoptosis induced by α-synuclein. Received 27 December 2007; received after revision 7 February 2008; accepted 8 February 2008     

Morphofunctional changes in the mitochondrial subpopulations of conceptus tissues during the placentation process

To establish the role of mitochondrial subpopulations in the mitochondrial maturation process, we studied morphological and functional changes in the mitochondria of different mammalian conceptus tissues during the organogenic and the placentation processes. Mitochondrial subpopulations of three different conceptus tissues, embryo and visceral yolk sac placenta on gestational days 11, 12 and 13 and placenta on days 12 and 13, were examined morphologically by transmission electron microscopy. Cytochrome oxidase activity and protein levels were also measured in each mitochondrial subpopulation. The results indicate two different mitochondrial subpopulation profiles: a homogeneous one, which corresponds to immature mitochondria, and a heterogeneous one, which represents the mature mitochondria. The three tissues studied show different morphologic and metabolic patterns of mitochondrial maturation during the placentation process, rendering them suitable as experimental models to establish the p ossible relationship between mitochondrial maturation and the mitochondrial subpopulations. Received 5 August 2002; received after revision 23 September 2002; accepted 8 October 2002 RID="*" ID="*"Corresponding author.     

The importance of the terminal hemi-acetal group for the ionophoric properties of nigericin

Opening of the terminal hemi-acetal in dihydronigericin drastically reduces the ionophoric properties of nigericin and dehydroxymethylnigericin with 6 intact heterocycles. This is shown by 2 complementary methods, first with a liquid membrane electrode system, secondly by testing their ionophoric activities in rat liver mitochondria.     

Ultrastructural differences in mitochondria of skeletal muscle in the prerigor and rigor states.

Loss of cristae and matrix occur in the mitochondria of skeletal muscles prior to any observable changes in myofibrillar proteins during the development of rigor mortis. Care must be observed because ultrastructural changes in mitochondria in some studies may be attributed to a specific trauma, whereas the changes may be due to the lower pH in postmortem muscle.     

Role of nitric oxide in the functional response to ischemia-reperfusion of heart mitochondria from hyperthyroid rats

We investigated the role of nitric oxide (NO) in the mitochondrial derangement associated with the functional response to ischemia-reperfusion of hyperthyroid rat hearts. Mitochondria were isolated at 3000 g from hearts subjected to ischemia-reperfusion, with or without N-nitro-L-arginine (L-NNA, an NO synthase inhibitor). During reperfusion, hyperthyroid hearts displayed tachycardia and low functional recovery. Their mitochondria exhibited O₂ consumption similar to euthyroid controls, while H₂O₂ production, hydroperoxide, protein-bound carbonyl and nitrotyrosine levels, and susceptibility to swelling were higher. L-NNA blocked the reperfusion tachycardic response and increased inotropic recovery in hyperthyroid hearts. L-NNA decreased mitochondrial H₂O₂ production and oxidative damage, and increased respiration and tolerance to swelling. Such effects were higher in hyperthyroid preparations. These results confirm the role of mitochondria in ischemia-reperfusion damage, and strongly suggest that NO overproduction is involved in the high mitochondrial dysfunction and the low recovery of hyperthyroid hearts from ischemia-reperfusion. L-NNA also decreased protein content and cytochrome oxidase activity of a mitochondrial fraction isolated at 8000 g. This and previous results suggest that the above fraction contains, together with light mitochondria, damaged mitochondria coming from the heaviest fraction, which has the highest cytochrome oxidase activity and capacity to produce H₂O₂. Therefore, we propose that the high mitochondrial susceptibility to swelling, favoring mitochondrial population purification from H₂O₂-overproducing mitochondria, limits hyperthyroid heart oxidative stress.Received 24 March 2004; received after revision 9 June 2004; accepted 5 July 2004     

Functional impact of PTP1B-mediated Src regulation on oxidative phosphorylation in rat brain mitochondria

Given the presence of Src and PTP1B within rat brain mitochondria, we have investigated whether PTP1B regulates Src activity in mitochondria as in the cytosol. Results showed that Src was stimulated by in vitro addition of ATP to mitochondria, and this stimulation was reversed by a membrane-permeable allosteric inhibitor of PTP1B and by a potent selective Src inhibitor. They also indicated a direct action of PTP1B on phosphorylated tyrosine 527 residue of Src, thus implicating a role for PTP1B in the modulation of Src activity in mitochondria. Putative Src and PTP1B substrates were identified by liquid chromatography tandem mass spectrometry and two-dimensional blue native/SDS-PAGE. Both inhibitors inhibited ADP-stimulated respirations concurrently with Src activation and complex IV activation by ATP, while having no effect or increasing the activity of the other complexes. Our analysis emphasizes the regulatory function of Src and its modulation by PTP1B on oxidative phosphorylation in mitochondria.     

Ultrastructural differences in mitochondria of skeletal muscle in the prerigor and rigor states

Summary Loss of cristae and matrix occur in the mitochondria of skeletal muscles prior to any observable changes in myofibrillar proteins during the development of rigor mortis. Care must be observed because ultrastructural changes in mitochondria in some studies may be attributed to a specific trauma, whereas the changes may be due to the lower pH in postmortem muscle.Scientific Journal Series, Paper No. 9633, Minnesota Agricultural Experiment Station.