CYTOLOGICAL RESPONSES OF BROWN FAT TISSUE IN COLD-EXPOSED RATS

In young adult laboratory rats exposed to cold (6°C) the brown adipose tissue undergoes time-dependent increases in cellularity, vascular supply, and total mass. These changes are largely complete after 16 days in the cold and concurrent generally with the development of a thermoregulatory state not greatly dependent upon shivering. Histologically the brown fat changes from a tissue having both unilocular and multilocular fat cell types to one having almost exclusively the latter. During the first 6 to 12 hours in cold, the multilocular cells lose their lipid vacuoles and decrease in size, but these features are restored to normal by 24 hours. Cell proliferation, as estimated by the DNA synthetic index method (using tritiated thymidine autoradiography), appears in the reticuloendothelial cells of the brown fat at 1 day of cold exposure, becomes maximal at 4 days, and returns to the control level by 16 days. In animals injected with tritiated thymidine on the 3rd day of cold exposure and then maintained for 1 or more additional days in the cold, autoradiographs indicate that new brown fat (multilocular) cells arise by cytogenesis from reticuloendothelial progenitor cells and not by proliferation of existing brown fat cells. Throughout this and subsequent periods, cells of the epididymal white adipose tissue slowly decrease in size. Because a thermogenic role in cold acclimation has been established for the brown fat, the reported changes are regarded as adaptive responses to a cold environment.     

CORRELATED BIOCHEMICAL AND ULTRASTRUCTURAL CHANGES IN NITROGEN-STARVED EUGLENA GRACILIS1

Growth of Euglena gracilis Z Pringsheim under photoheterotrophic conditions in a nitrogen-deprived medium resulted in progressive loss of chloroplastic material until total bleaching of the cells occurred. Biochemical analysis and ultrastructural observation of the first stages of the starvation process demonstrated an early lag phase (from 0 to 9 h) in which cells increased in size, followed by a period of cell division, apparently supported by the mobilization of some chloroplastic proteins such as the photosynthetic CO₂-fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase. The degradation of the enzyme started after 9 h of starvation and was preceded by a transient concentration of this protein in pyrenoidal structures. Protein nitrogen and photosynthetic pigments as well as number of chloroplasts per cell decreased during proliferation through mere distribution among daughter cells. However, after 24 h, when cell division had almost ceased, there was a slow but steady decline of photosynthetic pigments. This was paralleled by observable ultrastructural changes including progressive loss of chloroplast structure and accumulation of paramylon granules and lipid globules in the cytoplasm. These findings reinforce the role of chloroplastic materials as a nitrogen source during starvation of E. gracilis in a carbon-rich medium. The excess of ribulose-1,5-bisphosphate carboxylase/oxygenase acts as a first reservoir that, once exhausted, is superseded by the generalized disassembly of the photosynthetic structures, if the adverse environment persists more than 24 h.     

METABOLIC AND ULTRASTRUCTURAL CHANGES INDUCED IN ADIPOSE TISSUE BY INSULIN

The addition in vitro of insulin to rat adipose tissue (epididymal) produces marked metabolic changes which may be followed by measurement of the net gas exchange of the tissue. Using this method to monitor the metabolic action of insulin, concomitant observations with the electron microscope on the tissue have been made. These reveal that pronounced morphological changes are induced by insulin. The plasma membranes of the adipose cells become invaginated at many sites to form minute finger-like indentations. Numerous tiny, membrane-bounded vesicles are also present and arranged in relationship to the plasma membrane in such a way as to suggest that their formation occurred when a recessed fold was pinched off. Deeper in the cytoplasm, especially in specimens that had been incubated a longer time, numerous large, smooth, membrane-limited vesicles are seen. Finally, in these incubated specimens the cytoplasmic matrix has lost much of its granular nature, small lipid droplets are frequently found in the cytoplasm and suggestive changes have occurred in mitochondria. In control specimens, incubated without insulin for identical periods of time, indentations and vesicles in the plasma membrane are sparse at best and no vesicles or membrane-bound spaces appear deeper in the cytoplasm. The metabolic and morphologic changes induced by insulin seem to be interdependent events. Both changes appear to be initiated rapidly and concomitantly in the tissue. Both processes are initiated by insulin at concentrations considered to be physiological, 0.004 µg. (100 µunits) per ml. Insulin treated with alkali fails to initiate either process. It is concluded that insulin initiates pinocytosis in rat adipose tissue and the possible significance of this process in the mode of action of insulin is discussed.     

STIGMA OF NICOTIANA: ULTRASTRUCTURAL AND BIOCHEMICAL STUDIES

The aim of the present study has been to elucidate further the cytology of the glandular stigma of Nicotiana tabacum, the mechanism of the secretory process during stigma development and the biochemical composition of the exudate. The stigma consists of two distinct zones: a glandular zone formed by the papillae and 2–3 layers of cells (basal cells) immediately below them, and a non-glandular region formed by vacuolated cells which are in continuity with the transmitting tissue. The stigmatic exudate is a complex mixture of different chemical compounds such as proteins, saccharides, fatty acids and phenols. The role of stigma secretion is discussed in relation to pollen activation, recognition and pollen tube growth.     

冬小麦低温春化过程中茎尖细胞超微结构的变化与玉米赤霉烯酮的关系

玉米赤霉烯酮(zearalenone, ZEN)是Stob等于1962年首先从玉米赤霉菌培养物中分离出的次生代谢产物,它具有动物雌性激素的作用,也能促进某些真菌形成子实体,被认为是真菌的一种性激素,80年代初发现亦存在于高等植物体内。多年的研究结果表明,ZEN与植物的许多发育过程,包括春化作用、光周期诱导以及授粉受精过程均有密切的关系,可能ZEN参与了小麦成花诱导过程的调控,但其调控机制不明。为进一步研究小麦春化过程中茎尖细胞超微结构的变化与ZEN的相关性,我们以春化过程中的冬小麦幼芽为材料,对茎尖细胞超微结构以及与之相应的ZEN含量变化进行了分析。     

黄鳝性逆转时生殖腺的组织学与超微结构的变化

黄鳝由雌转雄性时,其生殖腺滤泡细胞变得肥大和充满分泌物,卵母细胞的微绒毛、线粒体与放射带退化,卵黄液化。滤泡细胞吞噬卵黄颗粒并消化吸收后,最后形成类胡罗卜素瘤。随着卵巢的退化,精细胞与精巢逐渐形成。     

MORPHOLOGICAL AND BIOCHEMICAL CHANGES IN RAT SYNAPTOSOME FRACTIONS DURING NEONATAL DEVELOPMENT

A biochemical and quantitative morphologic study of presynaptic endings during postnatal development was carried out in subcellular fractions from cerebral cortex of 1, 4, 8, 12, and 18 day old and adult rats. Crude mitochondrial fractions were subfractionated in Ficoll gradients and all resulting fractions were examined in the electron microscope. Presynaptic terminals and other intact processes were counted. Protein content and enzyme activities were assayed in the fractions and in total brain homogenate. In the first and fourth day of life, most of the presynaptic terminals were found in two "light" fractions, between supernatant and 7.5% Ficoll, where they accounted, respectively, for 6 and 22% of all the processes. Progressively with age, more presynaptic terminals were found in the traditional "synaptosomal" fractions between 7.5 and 13% Ficoll. In that region of the gradient, 40, 54, 75, and 89% of the processes were presynaptic endings at 8, 12, and 18 postnatal days and in the adult animal, respectively. A similar shift from the lighter to the heavier fractions was observed in the distribution of choline acetyltransferase and acetylcholinesterase between days 8 and 12. The rate of increase of the specific activity of these two enzymes paralleled that of the percentage of the presynaptic endings after day 8. This study indicates that subcellular fractions can be used to study formation and maturation of synapses during postnatal development.     

HIGH-INTENSITY EXERCISE TRAINING INDUCES MORPHOLOGICAL AND BIOCHEMICAL CHANGES IN SKELETAL MUSCLES

In the present study we investigated the effect of two different exercise protocols on fibre composition and metabolism of two specific muscles of mice: the quadriceps and the gastrocnemius. Mice were run daily on a motorized treadmill, at a velocity corresponding to 60% or 90% of the maximal running velocity. Blood lactate and body weight were measured during exercise training. We found that at the end of training the body weight significantly increased in high-intensity exercise mice compared to the control group (P=0.0268), whereas it decreased in low-intensity exercise mice compared to controls (P=0.30). In contrast, the food intake was greater in both trained mice compared to controls (P < 0.0001 and P < 0.0001 for low-intensity and high-intensity exercise mice, respectively). These effects were accompanied by a progressive reduction in blood lactate levels at the end of training in both the exercised mice compared with controls (P=0.03 and P < 0.0001 for low-intensity and high-intensity exercise mice, respectively); in particular, blood lactate levels after high-intensity exercise were significantly lower than those measured in low-intensity exercise mice (P=0.0044). Immunoblotting analysis demonstrated that high-intensity exercise training produced a significant increase in the expression of mitochondrial enzymes contained within gastrocnemius and quadriceps muscles. These changes were associated with an increase in the amount of slow fibres in both these muscles of high-intensity exercise mice, as revealed by the counts of slow fibres stained with specific antibodies (P < 0.0001 for the gastrocnemius; P=0.0002 for the quadriceps). Our results demonstrate that high-intensity exercise, in addition to metabolic changes consisting of a decrease in blood lactate and body weight, induces an increase in the mitochondrial enzymes and slow fibres in different skeletal muscles of mice, which indicates an exercise-induced increase in the aerobic metabolism.     

采后大蒜鳞茎的生理生化变化及其贮藏技术

采后大蒜鳞茎的生理生化变化及其贮藏技术刘淑娴,李月标,陈芳,张东林,蒋跃明（中国科学院华南植物研究所,广州５１０６５０）摘要大蒜鳞茎在室温下贮藏２个月后,胚芽开始生长．随着胚芽生长,呼吸速率、蛋白质和维生素Ｃ含量逐渐增高;而可溶性糖和干物质含量下降。...     

ULTRASTRUCTURAL TRANSFORMATION IN MITOCHONDRIA ISOLATED FROM KIDNEYS OF NORMAL AND LEAD-INTOXICATED RATS

Mitochondria isolated from kidneys of lead-intoxicated rats have been shown to have decreased oxidative and phosphorylative abilities. The purpose of this study was to determine whether these abnormal mitochondria would undergo ultrastructural transformation during controlled respiration in the absence of phosphate acceptor (State IV), as previously demonstrated for normal liver mitochondria. It was first shown that normal rat kidney mitochondria transforms from a condensed ultrastructural conformation to an orthodox conformation after 5 min of State IV respiration with pyruvate-malate substrate. Reversal to a condensed conformation follows stimulation of respiration with adenosine diphosphate (ADP). A large portion of kidney mitochondria from lead-poisoned rats do not change from condensed to orthodox conformation during State IV respiration. Other mitochondria do transform to the orthodox form but they rapidly degenerate. State IV respiration decreases as these few orthodox mitochondria disintegrate. The conclusion is that those mitochondria that do not undergo change in ultrastructure have impairment of electron transport, and that those that do become orthodox have increased membrane lability and undergo degeneration.     

青年猫和老年猫视神经超微结构观察比较

目的探讨青年猫和老年猫视神经年龄相关的形态学变化及可能造成的生理影响。方法取4只青年猫(2-3岁,2-2.5kg)和4只老年猫(10-13岁,2.5-3.5kg)颅内相对应部分视神经,制作横向半薄切片和超薄切片,半薄切片用甲苯胺蓝硼砂溶液染色,光镜观察、测量视神经的直径(不含外层神经膜);超薄切片标本用醋酸和柠檬酸铅染色,电镜观察、计数视神经纤维密度、测量视神经纤维外径D(含髓鞘)和内径d(不含髓鞘),按一定分级范围算出各种直径的纤维及各种d/D比值的纤维所占百分比,分别画出直方图,对实验结果进行统计学分析并绘制纤维直径谱。结果与青年猫相比,老年猫视神经直径显著增大(P0.05);纤维数量显著下降(P0.05)。纤维直径谱分析结果显示,青、老年猫纤维直径分布范围相似,但老年猫纤维的峰直径及纤维平均直径比青年猫的显著减小(P0.05),老年猫视神经纤维的d/D比值亦明显降低。另外,老年猫视神经中部分轴突肿胀,髓鞘疏松、结构紊乱,板层脱离、空泡化,有的轴索髓鞘溶解。结论在衰老过程中,老年猫视神经纤维丢失,纤维直径减小,d/D比值下降,以及纤维髓鞘的松散解体,这些变化均可能导致视神经纤维对视觉信息的传导速度减慢,是老年个体视觉分析速度下降的重要原因。     

METABOLIC AND ULTRASTRUCTURAL CHANGES IN THE FROG OVARIAN FOLLICLE IN RESPONSE TO PITUITARY STIMULATION

Frog ovarian fragments were prevented from ovulating in vitro by the addition of actinomycin D up to 3 hr following pituitary stimulation; but addition of Actinomycin D 6 hr after stimulation was far less effective. Puromycin, on the other hand, effectively inhibited ovulation when added as late as 6 hr after pituitary stimulation. Although actinomycin D reduced uptake of uridine-³H, and puromycin reduced uptake of leucine-³H and lysine-¹⁴ by pituitary-stimulated ovarian tissue minus oocytes (OTMO) in vitro, it was found that pituitary stimulation did not significantly increase uptake of these compounds by OTMO. Radioautographs of ovarian follicles fixed 6 hr after the addition of pituitary extract and uridine-³H in vitro revealed increased RNA synthesis in the peritoneal surface epithelium, compared with unstimulated controls, while the ovarian sac epithelium showed no increase. Gross ultrastructural changes occurred in the peritoneal area of ovarian follicles following pituitary stimulation in vivo, including loss of collagen fibrils, and general disorganization of the connective tissue theca. Changes in the rough endoplasmic reticulum of the peritoneal epithelial cells, while frequently encountered, were less pronounced. None of these changes was observed in the ovarian sac area, or in the interfollicular region. The above data are consistent with the hypothesis that pituitary stimulation of the frog ovary results in increased synthesis of RNA and protein by the peritoneal epithelial cells, and that the protein may be collagenase.     

METHOD FOR COMBINED ULTRASTRUCTURAL AND BIOCHEMICAL ANALYSIS OF NEURAL TISSUE

No significant change was found in the electrolytes and lipids of the brain analyzed after glutaraldehyde fixation by perfusion of laboratory animals; such fixation also satisfactorily preserves neural tissues for electron microscopy. The brains of normal and tumor-bearing C3H mice, Wistar rats, and New Zealand rabbits were studied. Little difference was found in the dry weight and the content of sodium, potassium, total lipid and lipid fractions, and in the sulfate space (S³⁵O₄) between specimens from unperfused and perfused animals, whether normal or tumor-bearing. The results suggest the possibility of using selected regions of the nervous system, dissected after fixation, for chemical study and at the same time characterizing similar regions morphologically with the electron microscope.     

BIOCHEMICAL AND ULTRASTRUCTURAL PROPERTIES OF A MITOCHONDRIAL INNER MEMBRANE FRACTION DEFICIENT IN OUTER MEMBRANE AND MATRIX ACTIVITIES

Treatment of the inner membrane matrix fraction of rat liver mitochondria with the nonionic detergent Lubrol WX solubilized about 70% of the total protein and 90% or more of the following matrix activities: malate dehydrogenase, glutamate dehydrogenase, and isocitrate dehydrogenase (NADP). The Lubrol-insoluble fraction was enriched in cytochromes, phospholipids, and a Mg⁺⁺-stimulated ATPase activity. Less than 2% of the total mitochondrial activity of monoamine oxidase, an outer membrane marker, or adenylate kinase, an intracristal space marker could be detected in this inner membrane fraction. Electron micrographs of negatively stained preparations showed vesicles (≤0.4 µ diameter) literally saturated on the periphery with the 90 A ATPase particles. These inner membrane vesicles, which appeared for the most part to be inverted with respect to the normal inner membrane configuration in intact mitochondria, retained the succinicoxidase portion of the electron-transport chain, an intact phosphorylation site II with a high affinity for ADP, and the capacity to accumulate Ca⁺⁺. A number of biochemical properties characteristic of intact mitochondria and the inner membrane matrix fraction, however, were either absent or markedly deficient in the inner membrane vesicles. These included stimulation of respiration by either ADP or 2,4-dinitrophenol, oligomycin-sensitive ADP-ATP exchange activity, atractyloside sensitivity of adenine nucleotide requiring reactions, and a stimulation of the Mg⁺⁺-ATPase by 2,4-dinitrophenol.     

银鲫受精卵中卵黄粒和线粒体的超微结构变化

电镜观察到雌性银鲫肝脏中正在形成的卵黄物质具有晶形主体结构,银鲫卵的卵黄粒内无晶形主体,第一次卵裂前的受精卵卵黄粒内有两种形式的空泡。一种是一些中空的小空泡;另一种是一个大的空泡,泡内含有线粒体和颗粒状的核糖体,泡的边缘有片层状的类脂物。受精卵中的线粒体内部结构多样,有些含有颗粒状物,有些含有片层状的类脂物。       

桔梗小孢子发生和雄配子体发育过程超微结构变化

对桔梗小孢子发生和雄配子体发育过程超微结构的观察表明,减数分裂过程中发生第一次细胞质改组,表现为：粗线期／双线期,核糖体数量消减,质体和线粒体结构简化;末期Ｉ,核糖体数量恢复,四分体时期,质体和线粒体获得正常结构。单核靠边期到二细胞花粉时期,发生第二次细胞质改组,表现同第一次细胞入组相。两次核糖体消减过程,均涉及到粗面内质网的。在和分前和有丝分裂前期存在核周腔膨大形成核液泡现象。结果表明,核糖体数     

ULTRASTRUCTURAL CHANGES OF CHLOROPLASTS IN ATTACHED AND DETACHED,AGING PRIMARY WHEAT LEAVES

Degradation of chloroplasts is shown in mesophyll cells of primary leaves of wheat. The sequence of ultrastructural changes in chloroplasts of naturally senescing leaves is compared with that of detached, aging leaves. In chloroplasts of naturally senescing leaves, the first indications of aging are the appearance of osmiophilic globuli and reorientation of the thylakoidal system. The membranes of the grana and intergrana lamellae then become distended and later dissociate into distinct vesicles. Concurrent with these membrane changes, osmiophilic globuli increase in size and number, and the stroma breaks down. Finally, the chloroplast envelope ruptures and plastid contents disperse throughout the cell's interior. In chloroplasts of mesophyll cells in detached, aging leaves, initial changes also include appearance of osmiophilic globuli, but later stages of chloroplast degradation are different. The chloroplast envelope ruptures before the lamellae break down. Swelling of grana and intergrana lamellae is not pronounced and, additionally, the thylakoidal system degenerates without forming vesicles or numerous osmiophilic globuli. These differences in the sequence of chloroplast degradation indicate that naturally senescing leaves rather than detached, aging leaves should be used in studies of chloroplast senescence.     

CEREBRAL METABOLIC AND CIRCULATORY CHANGES INDUCED BY HYPOXIA IN STARVED RATS

In order to study cerebral metabolic and circulatory effects of hypoxia under conditions of restricted glucose supply, the arterial P_o2, was reduced to 25–30mm Hg in artificially ventilated and lightly anaesthetized rats that were starved for 24 or 48 h prior to experiments. Arterial glucose concentrations, that were initially around 6μmol g^-1, were significantly reduced after 15min of hypoxia, and decreased to 50^o of control after 30min. In animals studied after 30min of hypoxia (24 h of starvation), cerebral blood flow had increased 4-fold and there was a moderate (25%) rise in cerebral oxygen consumption. During the course of hypoxia, cerebral cortical concentrations of glucose fell to low values. In spite of this, concentrations of pyruvate and lactate rose with time, and the sum of citric acid cycle intermediates (citrate, α-ketoglutarate, fumarate. malate and oxaloacetate) increased. Changes in amino acids were dominated by a fall in aspartate and a rise in alanine concentration. There was a moderate reduction in phosphocreatine and a slight rise in ADP concentration, but concentrations at ATP and AMP were unchanged. The changes observed are similar to those previously obtained in fed animals. It is concluded that even if blood glucose concentrations fall to 3μmol g^-1, and cerebral energy flux is maintained, substrate supply is sufficient to cover the energy requirements of the tissue. Hypoxia was accompanied by increases in the lactate/pyruvate and β-hydroxybutyrate acetoacetate ratios of blood. In the tissue, NADH/NAD⁺ ratios derived from the lactate, malate and β-hydroxybutyrate dehydrogenase systems rose, while that derived from the glutamate dehydrogenase reaction fell. It is concluded that the latter system is not well suited for estimating mitochondrial redox changes in brain tissue.