Functional morphology of the muscles in Philodina sp. (Rotifera: Bdelloidea)

Whole-mounts of Philodina sp., a bdelloid rotifer, were stained with fluorescent-labeled phalloidin to visualize the musculature. Several different muscle types were identified including incomplete circular bands, coronal retractors and foot retractors. Based on the position of the larger muscle bands in the body wall, their function during creeping locomotion and tun formation was inferred. Bdelloid creeping begins with the contraction of incomplete circular muscle bands against the hydrostatic pseudocoel, resulting in an anterior elongation of the body. One or more sets of ventral longitudinal muscles then contract bringing the rostrum into contact with the substrate, where it presumably attaches via adhesive glands. Different sets of ventral longitudinal muscles, foot and trunk retractors, function to pull the body forward. These same longitudinal muscle sets are also used in `tun' formation, in which the head and foot are withdrawn into the body. Three sets of longitudinal muscles supply the head region (anterior head segments) and function in withdrawal of the corona and rostrum. Two additional pairs of longitudinal muscles function to retract the anterior trunk segments immediately behind the head, and approximately five sets of longitudinal retractors are involved in the withdrawal of the foot and posterior toes. To achieve a greater understanding of rotifer behavior, it is important to elucidate the structural complexity of body wall muscles in rotifers. The utility of fluorescently-labeled phalloidin for the visualization of these muscles is discussed and placed in the context of rotifer functional morphology.     

Different extent of F-actin bundling in walled cells and in protoplasts ofMougeotia scalaris

Summary F-actin was localized inMougeotia interphase cells by rhodamine phalloidin (RLP) using an extended, formaldehyde-based fixation protocol, which included a minimal concentration of 0.05% (v/v) glutardialdehyde and stabilization of the calcium-binding vesicles by presaturation with neutral red. Staining revealed a low level of RLP-fluorescence throughout the cytoplasm. An enhanced level of RLP-fluorescence was found around the nucleus and in mostly two parallel fringes along each longitudinal chloroplast edge; also close to the chloroplast edge, quite regularly spaced patches of RLP-fluorescence were seen possibly associated with dictyosomes. The diffuse staining indicates lack of F-actin bundles inMougeotia filamentous cells, in contrast toSpirogyra interphase cells orMougeotia protoplasts. The observations upon staining with RLP confirm previous findings by electron microscopy and indicate seemingly single actin filaments throughout the entireMougeotia filamentous cell. Thus, a special F-actin organization is evident here which for the chloroplast movement is in support of the hypothesis of pigment regulated plasmalemma anchorage sites to actin filaments.Abbreviations CaBV calcium-binding vesicle - DIC differential interference contrast - EGTA ethyleneglycol-bis-(-aminoethyl ether) N, N, N, N tetraacetic acid - FA formaldehyde - GA glutardialdehyde - MFSB microfilament stabilizing buffer - PIPES piperazine-N, N-bis(2-ethanesulfonic acid) - RLP rhodamine (labeled) phalloidin Dedicated to the memory of Professor Oswald Kiermayer     

神经生长锥内α-spectrin与F-actin的免疫细胞化学研究

目的 为了探讨α血影蛋白（α-spectrin）和纤维型肌动蛋白（F-actin)在神经生长锥激烈运动与神经生长过程中的作用机制。方法 采用免疫双荧光染色法,以共聚焦激光扫描显微镜观察初代培养脊神经节细胞生长锥的连续水平断面上观察α-spectrin和F-actin的三维分布特点及其相互关系。结果 在水平面上,α-spectrin和F-actin分布在细胞质中,尤以细胞膜下,生长锥的P区伸展方向的前端和丝状伪足内分布最多,且α-spectrin比Factin更接近细胞膜,在纵轴方向,α-spectrin从基质侧到表达游离侧都有分布,而F-actin在生长锥体部C区和表面游离侧分布较少。结论 提示 F-actin共同参与生长锥运动,神经生长外,还参与构筑生长锥内的三维网架及维持生长锥的极性。     

Cytoskeleton organization during the cell cycle in two stramenopile microalgae, Ochromonas danica (Chrysophyceae) and Heterosigma akashiwo (Raphidophyceae), with special reference to F-actin organization and its role in cytokinesis

F-actin organization during the cell cycle was investigated in two stramenopile microalgae, Ochromonas danica (Chrysophyceae; UTEX LB1298) and Heterosigma akashiwo (Raphidophyceae; NIES-6) using FITC-phalloidin. In the interphase cell of O. danica, F-actin bundles were localized forming a network structure in the cortical region, which converged from the anterior region to the posterior, whereas in the interphase cell of H. akashiwo, F-actin bundles were observed forming a network structure in the cortical region without any polarity. In both O. danica and H. akashiwo, at the initial stage of mitosis the cortical F-actin disappeared, and then during cytokinesis assembly of an actin-based ring-like structure occurred in the cell cortex in the plane of cytokinesis. The ring-like structure initiated from aster-like structures was composed of F-actin in both O. danica and H. akashiwo. Different from animal cells, later stages of cytokinesis of O. danica seemed to be promoted by microtubules, although the early stages of cytokinesis progressed with a constriction of the ring-like structure, whereas cytokinesis of H. akashiwo was apparently completed by constriction of the cell mediated by the F-actin ring, as in animal cells.     

Detection of small differences in actomyosin function using actin labeled with different phalloidin conjugates

This study shows that there is only a negligible difference in actomyosin function in the in vitro motility assay among actin filaments labeled with Rhodamine phalloidin (RhPh), Alexa-488 phalloidin (APh), and biotin-XX phalloidin (BPh). Similar results were obtained at varying ionic strengths (0.02-0.13 M), in the presence of imidazole or 3-[N-morpholino]propanesulfonic acid (MOPS) buffer, and at varying MgATP concentrations (0.1-3 mM). If RhPh- and APh-labeled filaments were studied in a given flow cell, there was minimal variability in sliding velocity between the fluorophores (standard deviation of 3% of the absolute sliding velocity). The variability was considerably smaller than that between flow cells, allowing us to use dual labeling of different actin types and then apply analysis of variance to detect minor functional differences between them. Using this method, we could statistically verify a 4% difference (P<0.001) in sliding velocity (3mM Mg ATP) between cardiac and skeletal muscle actin. Suggested improvements of the method would readily allow the detection of even smaller differences. We discuss implications of the results for nanotechnological applications, understanding actomyosin function, and reducing experimental costs and the use of laboratory animals.     

Despite expression in embryonic visceral mesoderm,H2.0 is not essential for Drosophila visceral muscle morphogenesis

H2.0, a homeobox gene identified by homology to the Sex combs reduced homeobox of Drosophila, is expressed in all the cellular precursors of the visceral musculature. By analogy to the essential function of most other known homeobox genes in determining the fate of cells where they are expressed, we hypothesized that mutation of H2.0 would disrupt gut muscle development. In this paper, we show that a small deletion, which eliminates H2.0, has no detectable effect on normal gut morphogenesis, visceral muscle actin organization, or larval peristalsis.     

中国涡虫一新纪录科肠口涡虫属一新纪录种格氏肠口涡虫(原卵黄目, 柱口科)

原卵黄目(Prolecithophora)柱口科(Cylindrostomidae)肠口涡虫属(Enterostomula)为海栖涡虫,全球记录3种,在中国未见报道。作者在广东省深圳市深圳湾海边(22°31'N,113°56'E)采集到一种海栖涡虫。本文对该种涡虫进行了较为详细的比较研究,鉴定为柱口科肠口涡虫属格氏肠口涡虫(E.graffi de Beauchamp,1913),为中国新纪录科新纪录属一新纪录种。该种涡虫多数个体背部具2条黑色横纹,部分个体背部花纹有明显变化。眼点2对。雌雄同体,精巢、卵巢各1个。受精囊位于卵巢与子宫之间;子宫位于体末端,后接一微小的外阴道。本文发现:①该涡虫的尾部腹面具有一个交配囊及一根雌性生殖管,交配囊是一个由肌肉层组成的袋状囊,同时与子宫、雌性生殖管相连;②雌性生殖管外围有明显的生殖腺包裹;③雌性生殖管孔、阴茎孔与口孔一起通往体外;④此种涡虫喜分泌黏液将微小杂质粘结呈半球状窝,虫体隐藏于窝内。     

斜口涡虫(原卵黄目,斜口科)的生物学特性

摘要：本文报道了采自广东惠州市东江支流(23°09′00. 31\"N,114°22′26.59″E)的中国涡虫一新纪录目原卵黄目(Prolecithophora)斜口涡虫(Plagiostomum sp.)。该虫体长4.51 ～6.00 mm,体被不规则的褐色细网纹,头钝圆,尾圆锥形,体中部圆柱状,眼点4只。该涡虫运动...     

陕北淡水涡虫组织形态学初步观察

对陕北常见淡水涡虫的外部形态和石蜡切片进行组织形态学观察,经过绘图、照相、分析、鉴定,结果表明陕北淡水涡虫的常见种主要为日本三角涡虫(Dugesia japonica)。     

Visualization of actin arrays in growing hyphae of the fungusSaprolegnia ferax

Summary We have observed the distribution of filamentous actin in growing hyphae of the oomyceteSaprolegnia ferax. The actin was stained by electroporating intact hyphae in the presence of 4×10^–8 M rhodamine phalloidin. Hyphae quickly recovered from electroporation and showed an apical cap of densely packed actin filaments. The pores created by the electric shock resealed in 8–10min and within 1/2 h hyphae resumed growth and appeared normal. This technique allows us to observe actin arrays during growth and may prove to be a useful tool in determining the complex roles of actin in apical growth.Abbreviations RP rhodamine phalloidin - F-actin filamentous actin     

Structure and maintenance of the epidermis in Friedmaniella sp. (Prolecithophora)

The epidermis of Friedmaniella sp. has been studied using light and electron microscopy. Three main morphological features characterize its cells, namely (1) DNA bodies in the nuclei, (2) an extensive Golgi apparatus with a well-developed system of transport vesicles, (3) clusters of centrioles mainly in the basal cytoplasm and axonemes and rootlets in the middle and apical cell parts. These peculiarities may indicate continuous physiological regeneration within the cell at the level of cell organelles. DNA bodies may prove to be a taxonomic feature distinguishing the Prolecithophora from other turbellarians.     

Temporary pools are not `enemy-free'

Temporary pools are traditionally considered as refuges where the conspicuous anostracans are protected from predation. While this is true for the size-selective predation by fish, there is compelling evidence that invertebrate predation is an important biotic stress regulating temporary pool communities. In rock pools in southeastern Botswana, we studied the impact of some suspected invertebrate predators on populations of the freshwater anostracan Branchipodopsis wolfi by means of observations and manipulative experiments. In a survey of 45 pools, the relationship between B. wolfi natural population sizes and the abundance of suspected predators were never negative for turbellarians and mosquito larvae. When dragonfly larvae, notonectids or tadpoles were present, the anostracan populations were generally non-existent or very small. In enclosure experiments with turbellarians, there was a significant effect of predation within one hour of the start; the average daily predation rate was about 1/4 anostracan per turbellarian. Anostracans from a pool with few turbellarians were slightly less vulnerable than those from a turbellarian-rich pool. Furthermore, there was an indication of males being predated on more than females. With dragonfly larvae and notonectids, the predation effect was marked with all six anostracans in an experiment eaten in less than one day by a single predator (predation rate: about one anostracan every 2 h per predator). In a behavioral study, both sexes of B. wolfi avoided swimming above sediment that held more turbellarians than the open patches; there was no evidence for chemical communication with respect to this behavior.     

Comparisons of Endothelial Cell G- and F-Actin Distribution in Situ and in Vitro

Numerous studies have described the F-actin cytoskeleton; however, little information relevant to C-actin is available. The actin pools of bovine aortic endothelial cells were examined using in situ and in vitro conditions and fluorescent probes for G-(deoxyribonuclease I.0.3 μM) or F-actin (phalloidin, 0.2 μM). Cells in situ displayed a diffuse G-actin distribution, while F-actin was concentrated in the cell periphery and in fine stress fibers that traversed some cells. Cells of subconfluent or just confluent cultures demonstrated intense fluorescence, with many F-actin stress fibers. Postconfluent cultures resembled the condition in situ; peripheral F-actin was prominent, traversing actin stress fibers were greatly reduced and fluorescent intensity was diminished. Postconfluency had little influence on G-actin. with only an enhancement in the intensity of G-actin punctate fluorescence. When post-confluent cultures were incubated with cytochalasin D (15 min; 10^--⁴ M), F-actin networks were disrupted and actin punctate and diffuse fluorescence increased. G-actin fluorescence was not altered by the incubation. Although its unstructured nature may account for the minor changes observed, the stability of the G-actin pool in the presence of notable F-actin modulations suggested that filamentous actin was the key constituent involved in these actin cytoskeletal alterations. A separate finding illustrated that the concomitant use of actin probes with image enhancement and fluorescent microscopy could reveal simultaneously the G- and F-actin pools within the same cell.