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1.
SYNOPSIS Cilia detached from mating reactive cells of Paramecium caudatum were fractionated for the purpose of identifying the structural component bearing mating substances. Purified axoneme fractions had no mating reactivity. The membrane fraction obtained by dialyzing against a solution of Tris-EDTA (0.1 m m EDTA, 1 m m Tris-HCI, pH 7.6) and 0.6 m KCI, and then by centrifuging over 40% (w/v) sucrose was strongly reactive. No mating reactivity was detected in the soluble fractions containing axonemal and matrix proteins. The results indicate that the mating substances in active from are localized only on the ciliary membranes.  相似文献   

2.
    
《Current biology : CB》2019,29(17):2942-2947.e2
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3.
    
The phosphoarginine shuttle system effectively regenerates ATP in the cilia of Paramecium caudatum. To estimate the effective concentration of ATP‐regenerating enzymes, we attempted to reconstitute certain ATP‐regenerating systems within the cilia of intact cortical sheets using exogenous enzymes and high‐energy substances. The addition of phosphoenolpyruvate, which is one of the substrates in glycolysis, did not increase the ciliary beat frequency, whereas phosphocreatine together with exogenous creatine kinase, effectively increased the ciliary beat frequency. In the presence of 0.6 mg/ml creatine kinase and 0.4 mM phosphocreatine, the ciliary beat frequency was comparable to that produced by the addition of phosphoarginine. This result indicates that the reconstituted phosphocreatine shuttle system can work as an artificial ATP‐regenerating system for ciliary movements. The effective concentration of creatine kinase in the reconstituted phosphocreatine shuttle system was estimated to be about 7.4 μM based on the molecular mass of creatine kinase (MW 81,000). Therefore, the effective concentration of arginine kinase in the cilia of live Paramecium is approximately 10 μM. This estimated concentration of intraciliary arginine kinase is sufficient to maintain a high ATP concentration throughout the cilia of P. caudatum.  相似文献   

4.
    
Little is known about the influence of magnetic fields on growth of primitive eukaryotes such as the ciliate Paramecium. The latter are known to exhibit interesting characteristics such as electrotaxis, gravitaxis, and membrane excitability not commonly encountered in higher organisms. This preliminary study reports the effects of static magnetic fields on growth of Paramecium caudatum. The microorganisms were either permanently or 24 h on-and-off exposed to North and South polarity magnetic fields of average field gradient 4.3 T/m, for a period of 96 h. The growth rate and lag phase of all exposed populations were not significantly different from control ones exposed to normal geomagnetic field (P > .05). However, a significant negative shift in t(max) (time taken for maximum growth) of 10.5%-12.2% and a significant decrease (P < .05) in population size of 10.2%-15.1% during the 96 h of experimental conditions were recorded for exposed populations compared to control. Our results suggest that magnetic fields, irrespective of polarity and exposure period reduce Paramecium growth by triggering early senescence of the population. The mechanisms underlying the small changes in population growth are unknown at this level, but various hypotheses have been suggested, including disorganization of swimming patterns resulting from (i) changes in cell membrane electric potential due to high speed movement through a gradient magnetic field and (ii) thermodynamic effect of anisotropic magnetic energies on cell membrane components affecting functioning of calcium channels. Altered swimming movements could in turn affect highly orchestrated processes such as conjugation, essential for survival of the organisms during development of adverse environmental conditions as thought to occur in the closed culture system used in this study.  相似文献   

5.
SYNOPSIS. Electron-dense deposits indicating possible Ca-binding sites were found at the ciliary base of Paramecium caudatum fixed in a glutaraldehyde solution containing 5 mM CaCl2. The deposits appeared mainly at the inner surface of the ciliary membrane above the "ciliary necklace" region, although they could also occur in the space between the outer and the central microtubules. In some cases a ring of exactly 9 deposits was found in a ciliary cross section of a cilium.  相似文献   

6.
目的:采用神经心理测验方法探讨海拔高度及时间对移居者记忆与肢体运动能力的影响及返回平原后的恢复情况。方法:选择即将进入高原的241名健康青年,在其进入高原前、初期、5个月、1年及返回平原时分别采用DDX-200型多功能心理生理能力测试康复仪进行左右手交叉敲击动作频率和数字记忆广度顺背数测验。结果:与进入高原前比较,进入高原5个月至1年,数字记忆广度顺背数测验得分降低,左右手交叉敲击动作频率总次数和正确次数减少,错误次数增多(P〈0.05,或P〈0.01),进入高原初期,5个月及返回平原5个月与进入高原1年相比,数字记忆广度顺背数测验得分增高,左右手交叉敲击动作频率总次数和正确次数增多,错误次数减少(P〈0.05,或P〈0.01)。移居5个月后4700m组各项测试指标均优于移居5000m以上地区组(P〈0.05,或P〈0.01)。结论:移居高原5个月后记忆力及肢体运动能力降低,特别是移居5000m以上地区,而这些能力在返回平原5个月将得到恢复。  相似文献   

7.
The electrical membrane properties and the swimming behaviour of the freshwater ciliate Bursaridium difficile were studied by current clamp recordings and video analysis. The resting membrane potential was –45 ± 6 mV (mean ± SD, n = 80), and the input resistance and membrane capacitance were 109 ± 42 megaohms (MΩ) (n = 63) and 457 ± 150 picofarads (pF) (n = 42), respectively. Based on an estimated surface area of 6.8 × 10-4 cm2, the corresponding specific membrane resistance and capacitance are 7.4 × 104Ω× cm2 and 0.7 μF/cm2. Bursaridium difficile generates spontaneous, all-or-nothing action potentials with a well-defined threshold in normal medium. The spontaneous firing frequency was 0.22 ± 0.06 Hz (n = 80). The maximum rate of rise of the action potentials was less than 1 V/s, and they displayed a prolonged plateau phase (0.5–1 s). The action potentials were abolished in nominal Ca2+-free solution and are thus Ca2+-spikes. The swimming pattern of Bursaridium in homogeneous surroundings is composed of forward swimming periods interrupted by regular, short periods of backward swimming followed by a change in the forward swimming direction. The turning frequency corresponded to the spontaneous firing frequency, and only forward swimming was observed in nominal Ca2+-free solution. The periods of backward swimming activity are thus linked to the spontaneous action potentials.  相似文献   

8.
9.
    
SYNOPSIS. Locomotor behavior in the ciliate protozoa is controlled by the cell membrane through electrophysiological principles already familiar in receptor, nerve, and effector cells of the metazoa. This is illustrated by the avoiding reaction (15). When the membrane of the anterior part of the ciliate receives a mechanical stimulus, as during collision, it permits a local influx of Ca++. This constitutes a receptor current which depolarizes the remaining cell membrane by electrotonic spread. Depolarization causes a secondary transient increase in the calcium conductance of the entire cell membrane, and a general influx of Ca++ occurs. The resulting increase in concentration of intracellular Ca++ activates a reorientation (“reversal”) of the ciliary power stroke, causing the organism to swim backward. Forward locomotion is restored as the resting concentration of intracellular Ca++ in the cell cortex is restored by diffusion, active extrusion, or intracellular sequestering. The control and coordination of locomotion in ciliates depend on several factors in addition to the excitable properties of the membrane. These include the sensitivities of the ciliary apparatus to intracellular concentrations of calcium and other regulating substances, the anatomical distribution of sensory receptor properties of the cell membrane, and the cable properties of the cell which permit electrotonic spread of graded potential signals without need of all-or-none conducted signals.  相似文献   

10.
Normal ciliary ultrastructure is thought to be necessary for effective function. There has been little or no attempt to quantify ultrastructural abnormalities in nasal disease and assess their significance. In this study we measured nasal ciliary function and examined ciliary ultrastructure in nasal brushings from 35 patients with perennial nasal symptoms refractory to treatment. Ultrastructural defects included microtubular abnormalities, compound cilia and ciliary ‘blebs’. the incidence of abnormal cilia was 16.7%, compared with 9% in controls, but there was only a poor correlation between ultrastructural defects and ciliary beat frequency. One patient had primary ciliary dyskinesia (PCD) with a typical clinical history and immotile cilia. However, only secondary ultrastructural abnormalities were seen. We have been unable to show that ciliary ultrastructural defects form the basis of impaired function. In patients with suspected PCD, nasal brushings should be taken for functional and ultrastructural studies; ideally, a further sample should be obtained for examination of possible primary ultrastructural abnormalities.  相似文献   

11.
    
Gao C  Stern HA 《Proteins》2007,68(1):67-75
We perform a systematic examination of the ability of several different high-resolution, atomic-detail scoring functions to discriminate native conformations of loops in membrane proteins from non-native but physically reasonable, or \"decoy,\" conformations. Decoys constructed from changing a loop conformation while keeping the remainder of the protein fixed are a challenging test of energy function accuracy. Nevertheless, the best of the energy functions we examined recognized the native structure as lowest in energy around half the time, and consistently chose it as a low-energy structure. This suggests that the best of present energy functions, even without a representation of the lipid bilayer, are of sufficient accuracy to give reasonable confidence in predictions of membrane protein structure. We also constructed homology models for each structure, using other known structures in the same protein family as templates. Homology models were constructed using several scoring functions and modeling programs, but with a comparable sampling effort for each procedure. Our results indicate that the quality of sequence alignment is probably the most important factor in model accuracy for sequence identity from 20-40%; one can expect a reasonably accurate model for membrane proteins when sequence identity is greater than 30%, in agreement with previous studies. Most errors are localized in loop regions, which tend to be found outside the lipid bilayer. For the most discriminative energy functions, it appears that errors are most likely due to lack of sufficient sampling, although it should be stressed that present energy functions are still far from perfectly reliable.  相似文献   

12.
    
The use of resealed red blood cell membranes (ghosts) allows the study of the transport of a compound in a nonmetabolizing system with a biological membrane. Transmembrane movements of anandamide (N-arachidonoylethanolamine, arachidonoylethanolamide) have been studied by exchange efflux experiments at 0 degrees C and pH 7.3 with albumin-free and albumin-filled human red blood cell ghosts. The efflux kinetics is biexponential and is analyzed in terms of compartment models. The distribution of anandamide on the membrane inner to outer leaflet pools is determined to be 0.275 +/- 0.023, and the rate constant of unidirectional flux from inside to outside is 0.361 +/- 0.023 s(-1). The rate constant of unidirectional flux from the membrane to BSA in the medium ([BSA]o) increases with the square root of [BSA]o in accordance with the theory of an unstirred layer around ghosts. Anandamide passed through the red blood cell membrane very rapidly, within seconds. At a molar ratio of anandamide to BSA of <1, membrane binding of anandamide increases with increasing temperatures between 0 degrees C and 37 degrees C, and the equilibrium dissociation constants are in the nanomolar range. The nature of membrane binding and the mechanism of membrane translocation are discussed.  相似文献   

13.
    
Ciliary neurotrophic factor (CNTF) was first identified and partially purified from embryonic chick eye tissues. Subsequently, it was shown that CNTF is also present in large amounts in sciatic nerves of adult rats and rabbits, which led to its final purification and cloning. CNTF is not secreted by the classical secretory pathway involving the endoplasmatic reticulum and Golgi complex, but can be detected in high quantities within the cytoplasm of myelinating Schwann cells and astrocytes using immunohistochemistry. CNTF supports survival and / or differentiation of a variety of neuronal cell types including sensory, sympathetic and motoneurons. Also, nonneuroanl cells, such as oligodendrocytes, microglial cells, liver cells, and skeletal muscle cells, respond to exogenously administered CNTF, both in vitro and in vivo. During development, expression of CNTF is very low, if indeed it is expressed at all, and the phenotype of mice lacking endogenous CNTF, suggesting that CNTF after inactivation of the CNTF gene by homologous recombination suggests that CNTF does not play a crucial role for responsive cells during embryonic development. However, motoneurons are lost postnatally in mice lacking endogenous CNTF, suggesting that CNTF acts physiologically on the maintenance of these cells. The ability of exogenous CNTF to protect against motoneuron loss following lesion or in other animal models indicates that CNTF might be useful in the treatment of human motoneuron disorders, provided appropriate means of administration can be found. 1994 John Wiley & Sons, Inc.  相似文献   

14.
The movements of Sagitta are conditioned by the presence of Ca(2+) in the external medium. When this ion is removed from artificial sea water, animals do not move. They swim again when Ca(2+) is present. Among the problems raised by this observation, we have studied the role of Ca(2+) in the contraction of the primary musculature. Physiological experiments show the central importance of the extracellular Ca(2+) and of its translocation through the membrane during the initiation of the contraction. Cytochemical data correlate these observations. They show that Ca(2+) is localized mainly at the level of the plasma membrane, its invaginations and in the poorly developed SR (less than 2% of cell). Like SR, mitochondria accumulate Ca(2+) but do not seem to participate in the regulation of these Ca movements except in abnormal situations. La(3+) blocks the entry of extracellular Ca(2+) and attaches to the membranes; this fixation is not the same on the plasma membrane and in its invaginations. The contractile apparatus of Sagitta primary musculature show remarkable specializations (Duvert and Savineau, 1986). It is composed of ribbon-shaped myofibrils of regular thickness surrounded by external membranes implicated in the fixation and the translocation of a pool of Ca(2+) necessary for initiating contraction. The poorly developed SR and the mitochondria probably modulate the functioning of the two types of fibres (A and B).  相似文献   

15.
The actin network has been implicated in the intracellular transport and positioning of the melanosomes, organelles that are specialized in the biosynthesis and the storage of melanin. It contributes also to molecular mechanisms that underlie the intracellular membrane dynamics and thereby can control the biogenesis of melanosomes. Two mechanisms for actin‐based movements have been identified: one is dependent on the motors associated to actin namely the myosins; the other is dependent on actin polymerization. This review will focus on to the role of the actin cytoskeleton and myosins in the transport and in the biogenesis of melanosomes. Myosins involved in membrane traffic are largely seen as transporters of organelles or membrane vesicles containing cargos along the actin networks. Yet increasing evidence suggests that some of the myosins contribute to the dynamics of internal membrane by using other mechanisms. The role of the myosins and the different molecular mechanisms by which they contribute or may contribute to the distribution, the movement and the biogenesis of the melanosomes in epidermal melanocytes and retinal pigmented epithelial (RPE) cells will be discussed.  相似文献   

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17.
K. Murata  M. Okamoto  T. Suzaki 《Protoplasma》2000,214(1-2):73-79
Summary Fourier transform and an image filtering technique were used for structural analysis of the pellicular strip inEuglena gracilis. Freeze-fracture images of a two-dimensional crystalline structure in the plasma membrane were taken from elongated and rounded cells. Their lattice constants were precisely determined from Fourier transform patterns, and masked filter images were generated. Although differences in their lattice constants could not be detected, the Fourier transform patterns showed a significant difference between these two stages of cell shape. In elongated cells, the pseudo-crystal lattice was generated in the half periodicity of the minor striation. In contrast, it disappeared when the cells became rounded. The filtered images confirmed the crystallographic disagreement between these two stages in cell shape, probably reflecting morphological changes of integrated plasma membrane particles. Four particles detected in a unit structure showed quite similar shapes across the pseudo-lattice in elongated cells, though a unit structure in rounded cells consisted of four heterogeneous particles with opaque boundaries. A 39 kDa integral plasma membrane protein (IP39) is known as a major component in the plasma membrane ofE. gracilis. The compositions of monomeric and oligomeric forms of IP39 were determined in both elongated and rounded cells by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which showed the possibility that an oligomerization of IP39 proceeds during rounding-up movement of the cell. These morphological and stoichiometrical changes of the integrated plasma membrane proteins may suggest an active involvement in cell shape change.  相似文献   

18.
As an introduction to a symposium of nematode chemoreception, the anatomy of nematode chemosensilla, their distribution on plant parasitic nematodes, and their possible functional roles is briefly reviewed. Comparison of nematode chemosensilla with those of other animals shows their greater resemblance to olfactory primary sense cells of vertebrates. Although the sensory process is obviously derived from a cilium, the absence of many ciliary features is noted. Retention of the ciliary necklace may be important functionally. A simple model is proposed, wherein binding of stimulant molecules to receptors in the membrane of the cilium-derived process results in entry of Na⁺ and Ca⁺⁺ (the latter via the ciliary necklace) to produce a receptor potential that spreads along the dendrite to the cell body where action potentials continue along the short axon to synapses.  相似文献   

19.
    
A pair of multiciliate, saccular structures containing multilayered lamellar bodies is reported in the head region of freshly hatched juveniles of the polyopisthocotylean monogenean Sphyranura sp. from Necturus maculosus. Similar structures have been recorded in only one other monogenean, in the oncomiracidium of Entobdella soleae, and the looser packing of the lamellar bodies in Sphyranura has permitted a more detailed ultrastructural study. A comparison is made with similar structures found in digenean miracidia.  相似文献   

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