Analysis of the Actin–Myosin II System in Fish Epidermal Keratocytes: Mechanism of Cell Body Translocation

While the protrusive event of cell locomotion is thought to be driven by actin polymerization, the mechanism of forward translocation of the cell body is unclear. To elucidate the mechanism of cell body translocation, we analyzed the supramolecular organization of the actin–myosin II system and the dynamics of myosin II in fish epidermal keratocytes. In lamellipodia, long actin filaments formed dense networks with numerous free ends in a brushlike manner near the leading edge. Shorter actin filaments often formed T junctions with longer filaments in the brushlike area, suggesting that new filaments could be nucleated at sides of preexisting filaments or linked to them immediately after nucleation. The polarity of actin filaments was almost uniform, with barbed ends forward throughout most of the lamellipodia but mixed in arc-shaped filament bundles at the lamellipodial/cell body boundary. Myosin II formed discrete clusters of bipolar minifilaments in lamellipodia that increased in size and density towards the cell body boundary and colocalized with actin in boundary bundles. Time-lapse observation demonstrated that myosin clusters appeared in the lamellipodia and remained stationary with respect to the substratum in locomoting cells, but they exhibited retrograde flow in cells tethered in epithelioid colonies. Consequently, both in locomoting and stationary cells, myosin clusters approached the cell body boundary, where they became compressed and aligned, resulting in the formation of boundary bundles. In locomoting cells, the compression was associated with forward displacement of myosin features. These data are not consistent with either sarcomeric or polarized transport mechanisms of cell body translocation. We propose that the forward translocation of the cell body and retrograde flow in the lamellipodia are both driven by contraction of an actin–myosin network in the lamellipodial/cell body transition zone.     

鱼类体内催乳素的渗透调节作用

该文介绍鱼催乳素的结构、分布、催乳素受体、催乳素渗透调节对象方面的研究进展,并对今后的研究方向及其在海鱼淡水养殖方面的应用潜力进行展望。     

The Mechanism of Heterogeneous Beta-Lactam Resistance in MRSA: Key Role of the Stringent Stress Response

All methicillin resistant S. aureus (MRSA) strains carry an acquired genetic determinant – mecA or mecC - which encode for a low affinity penicillin binding protein –PBP2A or PBP2A′ – that can continue the catalysis of peptidoglycan transpeptidation in the presence of high concentrations of beta-lactam antibiotics which would inhibit the native PBPs normally involved with the synthesis of staphylococcal cell wall peptidoglycan. In contrast to this common genetic and biochemical mechanism carried by all MRSA strains, the level of beta-lactam antibiotic resistance shows a very wide strain to strain variation, the mechanism of which has remained poorly understood. The overwhelming majority of MRSA strains produce a unique – heterogeneous – phenotype in which the great majority of the bacteria exhibit very poor resistance often close to the MIC value of susceptible S. aureus strains. However, cultures of such heterogeneously resistant MRSA strains also contain subpopulations of bacteria with extremely high beta-lactam MIC values and the resistance level and frequency of the highly resistant cells in such strain is a characteristic of the particular MRSA clone. In the study described in this communication, we used a variety of experimental models to understand the mechanism of heterogeneous beta-lactam resistance. Methicillin-susceptible S. aureus (MSSA) that received the mecA determinant in the laboratory either on a plasmid or in the form of a chromosomal SCCmec cassette, generated heterogeneously resistant cultures and the highly resistant subpopulations that emerged in these models had increased levels of PBP2A and were composed of bacteria in which the stringent stress response was induced. Each of the major heterogeneously resistant clones of MRSA clinical isolates could be converted to express high level and homogeneous resistance if the growth medium contained an inducer of the stringent stress response.     

食欲素神经肽在应激过程中的作用

食欲素因其在调节能量代谢、睡眠和唤醒等生理功能中的作用而备受关注.近年来研究逐渐发现,食欲素参与应激和奖赏过程的调节,特别是其在药物成瘾过程中的作用是目前的研究热点.主要介绍食欲素系统与应激相关系统之间的神经联系,阐述了其在应激相关的生理、神经内分泌与行为反应中的作用.并进一步介绍了食欲素系统在应激诱发药物成瘾复吸过程中的作用.食欲素对应激反应的调控作用具有相对特异性,受应激的种类、其他应激相关神经递质系统及食欲素神经元的投射通路等多种因素影响.     

The Role of Cell Calcium in the Contraction of Single Cannulated Muscle Fibers

Intracellular injections of the calcium-binding agent, EGTA,into single cannulated fibers of Balanus and Maia were ableto suppress, almost completely, the contractions induced byvarious contractile agents. The amount oE EGTA required in Maia fibers for the suppressionof the contractile response produced by caffeine and high-Ksaline, as has already been reported, was similar to the meanfiber calcium level. In Balanus fibers, however, although theamount of EGTA needed for the suppression of the caffeine-salineresponse was similar to the estimated level of fiber calcium,the amount required in the raised-K-saline experiments was considerablygreater. It has been suggested that membrane depolarizationunder these conditions allows calcium to enter the fiber fromthe external saline, the amount entering being related, at leastin part, to a large effective sarcolemmal surface area and thepresence of binding agent internally. The results of intracellularand plate-electrode stimulation of Belanus fibers also suggestedthat the fiber under certain conditions could utilize externalcalcium ions, while the results of plate-electrode stimulationof Maia fibers could be explained most easily in terms of mobilizationof mainly intracellular calcium for the process of contraction. Efflux of Sr⁸⁹ ions from Balanus fibers under various conditionssuggested that this ion is bound and mobilized internally ina manner similar to calcium. The results are seen not to contradict a chanelled-current theoryfor e-c coupling of the type proposed for crayfish fibers.     

The Mechanism of Dual Responsiveness in Muscle Fibers of the Grasshopper Romalea microptera

Dually innervated Romalea muscle fibers which respond differently to stimulation of their fast and slow axons are excited by intracellularly applied depolarizing stimuli. The responses, though spike-like in appearance, are graded in amplitude depending upon the strength of the stimuli and do not exceed about 30 mv. in height. In other respects, however, these graded responses possess properties that are characteristic of electrically excitable activity: vanishingly brief latency; refractoriness; a post-spike undershoot. They are blocked by hyperpolarizing the fiber membrane; respond repetitively to prolonged depolarization, and are subject to depolarizing inactivation. As graded activity, these responses propagate decrementally. The fast and slow axons of the dually responsive muscle fibers initiate respectively large and small postsynaptic potentials (p.s.p.'s) in the muscle fiber. These responses possess properties that characterize electrically inexcitable depolarizing activity. They are augmented by hyperpolarization and diminished by depolarization. Their latency is independent of the membrane potential. They have no refractory period, thus being capable of summation. The fast p.s.p. evokes a considerable or maximal electrically excitable response. The combination, which resembles a spike, leads to a twitch-like contraction of the muscle fiber. The individual slow p.s.p.'s elicit no or only little electrically excitable responses, and they evoke slower smaller contractile responses. The functional aspects of dual responsiveness and the several aspects of the theoretical importance of the gradedly responsive, electrically excitable component are discussed.     

Self-Organized Shape and Frontal Density of Fish Schools

Models of swarming (based on avoidance, alignment and attraction) produce patterns of behaviour also seen in schools of fish. However, the significance of such similarities has been questioned, because some model assumptions are unrealistic [e.g. speed in most models is constant with random error, the perception is global and the size of the schools that have been studied is small (up to 128 individuals)]. This criticism also applies to our former model, in which we demonstrated the emergence of two patterns of spatial organization, i.e. oblong school form and high frontal density, which are supposed to function as protection against predators. In our new model we respond to this criticism by making the following improvements: individuals have a preferred ‘cruise speed’ from which they can deviate in order to avoid others or to catch up with them. Their range of perception is inversely related to density, with which we take into account that high density limits the perception of others that are further away. Swarm sizes range from 10 to 2000 individuals. The model is three‐dimensional. Further, we show that the two spatial patterns (oblong shape and high frontal density) emerge by self‐organization as a side‐effect of coordination at two speeds (of two or four body lengths per second) for schools of sizes above 20. Our analysis of the model leads to the development of a new set of hypotheses. If empirical data confirm these hypotheses, then in a school of real fish these patterns may arise as a side‐effect of their coordination in the same way as in the model.     

The Role of Urine in Parent-Offspring Communication in a Cichlid Fish

The fry of the Midas cichlid, Cichlasoma citrinellum, preferred over blank water, a urine solution from either parent or from a nonparental adult of either sex. They failed to show a preference for the urine of a predator, C. managuense, or for the mucus from the mother. It is possible that steroid titers in urine could provide information on sex, and that peptide chains in urine could indicate species. The chemosensory acuity of fry is high, as they were able to detect a “concentration” of pheromone as low as one sibling per 2 1 of water. This high acuity implies that olfaction rather than gustation mediated the chemosensory responses described above.     

The Role of Carrion Supply in the Abundance of Deep-Water Fish off California

Few time series of deep-sea systems exist from which the factors affecting abyssal fish populations can be evaluated. Previous analysis showed an increase in grenadier abundance, in the eastern North Pacific, which lagged epibenthic megafaunal abundance, mostly echinoderms, by 9–20 months. Subsequent diet studies suggested that carrion is the grenadier''s most important food. Our goal was to evaluate if changes in carrion supply might drive the temporal changes in grenadier abundance. We analyzed a unique 17 year time series of abyssal grenadier abundance and size, collected at Station M (4100 m, 220 km offshore of Pt. Conception, California), and reaffirmed the increase in abundance and also showed an increase in mean size resulting in a ∼6 fold change in grenadier biomass. We compared this data with abundance estimates for surface living nekton (pacific hake and jack mackerel) eaten by the grenadiers as carrion. A significant positive correlation between Pacific hake (but not jack mackerel) and grenadiers was found. Hake seasonally migrate to the waters offshore of California to spawn. They are the most abundant nekton species in the region and the target of the largest commercial fishery off the west coast. The correlation to grenadier abundance was strongest when using hake abundance metrics from the area within 100 nmi of Station M. No significant correlation between grenadier abundance and hake biomass for the entire California current region was found. Given the results and grenadier longevity, migration is likely responsible for the results and the location of hake spawning probably is more important than the size of the spawning stock in understanding the dynamics of abyssal grenadier populations. Our results suggest that some abyssal fishes'' population dynamics are controlled by the flux of large particles of carrion. Climate and fishing pressures affecting epipelagic fish stocks could readily modulate deep-sea fish dynamics.     

The Role of Sodium Current in the Radial Spread of Contraction in Frog Muscle Fibers

The membrane potential of isolated muscle fibers was controlled with a two-electrode voltage clamp, and the radial extent of contraction elicited by depolarizing pulses of increasing magnitude was observed microscopically. Depolarizations of the fiber surface only 1–2 mv greater than the contraction threshold produced shortening throughout the entire cross-section of the muscle fiber. The radial spread of contraction was less effective in fibers exposed to tetrodotoxin or to a bathing medium with a greatly reduced sodium concentration. The results provide evidence that depolarization of a muscle fiber produces an increase in sodium conductance in the T tubule membrane and that the resultant sodium current contributes to the spread of depolarization along the T system.     

赤霉素信号在非生物胁迫中的作用及其调控机制研究进展

赤霉素(GA)能促进种子萌发和植物生长发育。近年来的研究表明赤霉素这一激素在植物响应非生物胁迫中同样发挥着重要的作用。植物通过调节GA生物合成、信号转导及其生物活性提高胁迫耐受性。综述了赤霉素在应对常见的几类非生物胁迫中所起的作用,分析了其合成、信号转导通路调控机制以及赤霉素与其他激素在响应非生物胁迫的关系。     

FGF-2 在细胞凋亡中的作用机制

成纤维细胞生长因子2(fibroblast growth factor 2,FGF-2)具有多种细胞生物学功能。FGF-2在肿瘤组织中呈高水平表达状态,且可抑制多种化疗药物的促凋亡作用,从而曾为肿瘤细胞存活的重要刺激因素。但也有研究表明FGF-2可诱导部分细胞的分化和凋亡。鉴于FGF-2在肿瘤的发生发展中发挥的重要作用,FGF-2与细胞凋亡的关系及其相应的调节机制成为有待于深入研究和迫切需要解决的问题。本文主要阐述在细胞凋亡通路中,FGF-2关键分子的作用机制及其最新研究进展。     

内质网应激在乙型脑炎病毒诱导神经元细胞凋亡中的作用及机制研究

内质网应激(Endoplasmic reticulum stress,ERS)的激活与创伤、缺血再灌注等病理刺激引起的神经元凋亡有关,流行性乙型脑炎病毒(JEV)感染能够促进神经元凋亡、激活ERS,但ERS在JEV诱导神经元细胞凋亡中的作用尚不清楚.为了研究ERS在JEV诱导神经元细胞凋亡中的作用及机制,本研究以神经细胞株SH-SY5Y为对象,感染JEV并加用ERS激动剂、ERS抑制剂或转染阴性对照(NC) siRNA、蛋白激酶R样内质网激酶(PERK)siRNA,检测细胞存活率、凋亡率、ERS蛋白PERK、肌醇必需酶-1α(IRE1α)、活化转录因子6(ATF6)及凋亡蛋白C/EBP同源蛋白(CHOP)、含半胱氨酸的天冬氨酸蛋白水解酶12(Caspase-12)、Bcl-2相关X蛋白(Bax)的表达.结果 显示:JEV组SH-SY5Y细胞的凋亡率及PERK、CHOP、Caspase-12、Bax的表达水平高于对照组,存活率低于对照组(P＜0.05),IRE1α、ATF6的表达水平与对照组比较无显著差异(P＞0.05).与JEV组比较,激动剂组SH-SY5Y细胞的凋亡率及PERK、CHOP、Caspase-12、Bax的表达水平显著增加,存活率显著降低(P＜0.05);抑制剂组SH-SY5Y细胞的凋亡率及PERK、CHOP、Caspase-12、Bax的表达水平显著降低,存活率显著增加(P＜0.05);与si-NC+JEV组比较,si-PERK+JEV组SH-SY5Y细胞的凋亡率及PERK、CHOP、Caspase-12、Bax的表达水平P显著降低,存活率显著增加(P＜0.05).以上结果表明ERS的PERK通路激活与JEV诱导神经元凋亡有关.     

The Role of Cytokinins in Plant Under Salt Stress

The plant hormone cytokinins (CKs) were generally considered associated with plant development. More recently, functions of CKs in plant stress defense including salt have been increasingly characterized. Under saline conditions, not only CKs homeostasis but also its signal transduction pathway is disturbed in a plant species-dependent manner. In turn, through manipulating exogenous CKs application or endogenous CKs metabolism or signaling, plant behavior can also be diverse across species. In this review, we systematically summarized this mutual regulation between CKs and salt stress. Considering the senescence-delaying effect of CKs, its roles in mitigating salt-induced senescence and maintaining crop yields are specifically highlighted. We also discussed here how CKs crosstalk with other phytohormones, including ABA and ethylene, to mediate salt response. In sum, this review provides a comprehensive integration of current knowledge on the regulatory role of CKs upon salt stress and puts forward research directions for future studies.

     

The Role of Oxidative Stress in Nervous System Aging

While oxidative stress is implicated in aging, the impact of oxidative stress on aging in the peripheral nervous system is not well understood. To determine a potential mechanism for age-related deficits in the peripheral nervous system, we examined both functional and morphological changes and utilized microarray technology to compare normal aging in wild-type mice to effects in copper/zinc superoxide dismutase-deficient (Sod1^−/−) mice, a mouse model of increased oxidative stress. Sod1^−/− mice exhibit a peripheral neuropathy phenotype with normal sensory nerve function and deficits in motor nerve function. Our data indicate that a decrease in the synthesis of cholesterol, which is vital to myelin formation, correlates with the structural deficits in axons, myelin, and the cell body of motor neurons in the Sod1^+/+ mice at 30 months and the Sod1^−/− mice at 20 months compared with mice at 2 months. Collectively, we have demonstrated that the functional and morphological changes within the peripheral nervous system in our model of increased oxidative stress are manifested earlier and resemble the deficits observed during normal aging.