SPERMIOGENESIS OF THE TELEOST,ORYZIAS LATIPES,WITH SPECIAL REFERENCE TO THE FORMATION OF FLAGELLAR MEMBRANE *

In the early stage of Oryzias spermiogenesis, an axonemal bud appears at the distal end of a centriole characterized by its electron dense accessories. When the axoneme begins to grow in the cytoplasm, small vesicles come to surround it. These vesicles are similar to those produced by the Golgi apparatus which lies close to the growing axoneme. At this stage, the spermatid cell membranes disappear, causing transformation of the mononuclear spermatids into a multinucleated syncytium. As each axoneme elongates in the syncytium, it is enveloped by a cylindrical array of vesicles which are most likely derived from the Golgi apparatus. Shortly after this stage, the syncytium is again partitioned by cell membranes, restoring the existence of mononuclear spermatids. The arrayed vesicles fuse with each other to form two concentric membranes surrounding the axoneme. The inner membrane becomes the flagellar membrane and the outer one, the membrane of a flagellar sheath. These observations lead to the conclusion that the formation of the flagellar membrane is due to the fusion of vesicles surrounding the axoneme which are derived from the Golgi apparatus. In the course of spermiogenesis, no indication of an acrosomal structure is observed.     

Assessing the effects of a sequestered germline on interdomain lateral gene transfer in Metazoa

A sequestered germline in Metazoa has been argued to be an obstacle to lateral gene transfer (LGT), though few studies have specifically assessed this claim. Here, we test the hypothesis that the origin of a sequestered germline reduced LGT events in Bilateria (i.e., triploblast lineages) as compared to early‐diverging Metazoa (i.e., Ctenophora, Cnidaria, Porifera, and Placozoa). We analyze single‐gene phylogenies generated with over 900 species sampled from among Bacteria, Archaea, and Eukaryota to identify well‐supported interdomain LGTs. We focus on ancient interdomain LGT (i.e., those between prokaryotes and multiple lineages of Metazoa) as systematic errors in single‐gene tree reconstruction create uncertainties for interpreting eukaryote‐to‐eukaryote transfer. The breadth of the sampled Metazoa enables us to estimate the timing of LGTs, and to examine the pattern before versus after the evolution of a sequestered germline. We identified 58 LGTs found only in Metazoa and prokaryotes (i.e., bacteria and/or archaea), and seven genes transferred from prokaryotes into Metazoa plus one other eukaryotic clade. Our analyses indicate that more interdomain transfers occurred before the development of a sequestered germline, consistent with the hypothesis that this feature is an obstacle to LGT.     

Soluble components of the flagellar export apparatus,FliI, FliJ,and FliH,do not deliver flagellin,the major filament protein,from the cytosol to the export gate

Flagella, the locomotion organelles of bacteria, extend from the cytoplasm to the cell exterior. External flagellar proteins are synthesized in the cytoplasm and exported by the flagellar type III secretion system. Soluble components of the flagellar export apparatus, FliI, FliH, and FliJ, have been implicated to carry late export substrates in complex with their cognate chaperones from the cytoplasm to the export gate. The importance of the soluble components in the delivery of the three minor late substrates FlgK, FlgL (hook–filament junction) and FliD (filament-cap) has been convincingly demonstrated, but their role in the transport of the major filament component flagellin (FliC) is still unclear.     

Precambrian geobiology

The appearance of Bacteria sensu lato, Eukaryota, Metaphyta, Metazoa, etc., along with the oxygenization of the atmosphere, are shown to have occurred much earlier than was previously assumed. Paleontological data clearly indicate that the difference between surface temperature of the Earth in the Archaen and now was no more than 30°C.     

Flagellar root contraction and nuclear movement during flagellar regeneration in Chlamydomonas reinhardtii

When Chlamydomonas cells are deflagellated by pH shock or mechanical shear the nucleus rapidly moves toward the flagellar basal apparatus at the anterior end of the cell. During flagellar regeneration the nucleus returns to a more central position within the cell. The nucleus is connected to the flagellar apparatus by a system of fibers, the flagellar roots (rhizoplasts), which undergo a dramatic contraction that coincides with anterior nuclear movement. A corresponding extension of the root system, back to its preshock configuration is observed as the nucleus retracts to a central position. Anterior displacement of the nucleus and flagellar root contraction require free calcium in the medium. Nuclear movement and flagellar root contraction and extension are not sensitive to inhibitors of protein synthesis (cycloheximide), or drugs that influence either microtubules (colchicine) or actin-based microfilaments (cytochalasin D). Detergent-extracted cell models contract and extend their flagellar roots and move their nuclei in response to alterations of free calcium levels in the medium. Cycles of nuclear movement in detergent-extracted models require ATP to potentiate the contractile mechanism for subsequent calcium-induced contraction. Flagellar root contraction and nuclear movement in Chlamydomonas may be causally related to signaling of induction of flagellar precursor genes or to the transport of flagellar precursors or their messages to sites of synthesis or assembly near the basal apparatus of the cell.     

Maintenance of motility in mouse sperm permeabilized with streptolysin O

One approach to studying the mechanisms governing sperm motility is to permeabilize sperm and examine the regulation of motility by manipulating the intracellular milieu of the cell. The most common method of sperm permeabilization, detergent treatment, has the disadvantage that the membranes and many proteins are extracted from the cell. To avoid this problem, we have developed a method that uses streptolysin O to create stable pores within the plasma membrane while leaving internal membranes intact. Sperm were permeabilized, preincubated, and then treated with 0.6 U/ml of streptolysin O. Permeabilization was assessed by fluorescent dye technologies and endogenous protein phosphorylation using exogenously added [gamma-32P]ATP. Streptolysin O-induced permeabilization rendered the sperm immotile, and the effect was Ca2+-dependent. When the cells were treated simultaneously with a medium containing ATP, streptolysin O-treated sperm maintained flagellar movement. These results demonstrate that the streptolysin O permeabilization model system is a useful experimental method for studying the mechanisms that regulate sperm motility since it allows the flagellar apparatus to be exposed to various exogenously added molecules.     

Biomechanics and allometric scaling in primate locomotion and morphology

Body size has a dominant influence on locomotor performance and the morphology of the locomotor apparatus. In locomotion under the influence of gravity, body mass acts as weight force and is a mechanical variable. Accordingly, the application of biomechanical principles and methods allows a functional understanding of scaling effects in locomotion. This is demonstrated here using leaping primates as an example. With increasing body size, the decreasing ratio of muscle force available for acceleration during takeoff to the body mass that has to be accelerated dictates both the movement pattern and the proportions of the hindlimbs. In an arm-swinging movement, the long, heavy arms of the large-bodied leapers are effectively used to gain additional momentum. A new perspective on decreasing size identifies the absolutely small acceleration distance and time available for propulsion as factors limiting leaping distance and extensively determining locomotor behavior and body proportions. As the mechanical constraints differ according to body size for a given mode of locomotion, a typological approach to morphology in relation to locomotor category is ruled out. Across locomotor categories, dynamic similarity (sensu Alexander) can be expected if the propulsive mechanisms as well as the selective pressures acting upon locomotion are the same.     

Flagellar apparatus structure of choanocyte in Sycon sp. and its significance for phylogeny of Porifera

The problem of the origin of Metazoa has been one of the most discussed for nearly the last one and a half centuries. Some 20 years ago, morphological approaches were replaced with molecules, but the problem then became more complex. At the same time, morphological data were incomplete, and therefore, this approach can still help in comparison with choanoflagellates and sponges—two sister groups having uniflagellated cells with a collar. The structure of the flagellar apparatus has phylogenetic significance, but sponge choanocytes are poorly studied in this respect, and we still do not know what the ancestral kinetid of Porifera looks like. The kinetid structure of choanocytes in Sycon sp. is investigated here for the first time, and a 3D reconstruction of the kinetid provided. It is composed of a flagellar kinetosome with a nuclear fibrillar root and a basal foot with a few microtubules; the accessory centriole lies orthogonal to and just below the foot of the kinetosome, and a dictyosome is near the centriole. This kinetid is similar to that of the choanocyte of Corticium candelabrum (Homoscleromorpha) and is considered to be the ancestral type for the whole branch Calcarea + Homoscleromorpha.     

Reconstruction of the flagellar apparatus and microtubular cytoskeleton inPyramimonas gelidicola (Prasinophyceae,Chlorophyta)

Summary The absolute configuration of the flagellar apparatus inPyramimonas gelidicola McFadden et al. has been determined and shows identity withP. obovata, indicating that they are closely related. Comparison with the flagellar apparatus of quadriflagellate zoospores from the more advancedChlorophyceae suggest thatPyramimonas may be a primitive ancestral form. The microtubular cytoskeleton has been examined in detail and is shown to be unusual in that it does not attach to the flagellar apparatus. Cytoskeletal microtubules are nucleated individually, and this is interpreted as an adaptation to the methods of mitosis and scale deployment. In view of the primitive nature of these processes, it is proposed that this type of cytoskeletal organization may represent a less advanced condition than that of the flagellar root MTOCs (microtubule organizing centers) observed in theChlorophyceae.     

FLAGELLAR MOTION AND FINE STRUCTURE OF THE FLAGELLAR APPARATUS IN CHLAMYDOMONAS

The biflagellate alga Chlamydomonas reinhardi was studied with the light and electron microscopes to determine the behavior of flagella in the living cell and the structure of the basal apparatus of the flagella. During normal forward swimming the flagella beat synchronously in the same plane, as in the human swimmer's breast stroke. The form of beat is like that of cilia. Occasionally cells swim backward with the flagella undulating and trailing the cell. Thus the same flagellar apparatus produces two types of motion. The central pair of fibers of both flagella appear to lie in the same plane, which coincides with the plane of beat. The two basal bodies lie in a V configuration and are joined at the top by a striated fiber and at the bottom by two smaller fibers. From the area between the basal bodies four bands of microtubules, each containing four tubules, radiate in an X-shaped pattern, diverge, and pass under the cell membrane. Details of the complex arrangement of tubules near the basal bodies are described. It seems probable that the connecting fibers and the microtubules play structural roles and thereby maintain the alignment of the flagellar apparatus. The relation of striated fibers and microtubules to cilia and flagella is reviewed, particularly in phytoflagellates and protozoa. Structures observed in the transitional region between the basal body and flagellar shaft are described and their occurrence is reviewed. Details of structure of the flagellar shaft and flagellar tip are described, and the latter is reviewed in detail.     

The coupled evolution of breathing and locomotion as a game of leapfrog

Because the increase in metabolic rate related to locomotor activity places demands on the cardiorespiratory apparatus, it is not surprising that the evolution of breathing and of locomotion are coupled. As the respiratory faculty becomes more refined, increasingly aerobic life strategies can be explored, and this activity is in turn expedited by a higher-performance respiratory apparatus. This apparent leapfrogging of respiratory and locomotor faculties begins in noncraniate chordates and continues in water-breathing and air-breathing vertebrates. Because both locomotor and cardiorespiratory activities are coordinated in the brain, neurological as well as biochemical coupling is evident. In spite of very different breathing mechanisms in various vertebrate groups, the basic respiratory control mechanisms appear to have been conserved, and respiratory-locomotor coupling is evident in all classes of vertebrates. Hypaxial body wall muscles that were strictly locomotor in fish have respiratory function in amniotes, but some locomotor function remains in all groups.     

Gene expression analysis of six GC-rich Gram-negative phytopathogens

Predicted highly expressed (PHX) genes are comparatively analyzed for six GC-rich Gram-negative phytopathogens, i.e., Ralstonia solanacearum, Agrobacterium tumefaciens, Xanthomonas campestris pv. campestris (Xcc), Xanthomonas axonopodis pv. citri (Xac), Pseudomonas syringae pv. tomato, and Xylella fastidiosa. Enzymes involved in energy metabolism, such as ATP synthase, and genes involved in TCA cycle, are PHX in most bacteria except X. fastidiosa, which prefers an anaerobic environment. Most pathogenicity-related factors, including flagellar proteins and some outer membrane proteins, are PHX, except that flagellar proteins are missing in X. fastidiosa which is spread by insects and does not need to move during invasion. Although type III secretion system apparatus are homologous to flagellar proteins, none of them is PHX, which support the viewpoint that the two types of genes have evolved independently. Furthermore, it is revealed that some biosynthesis-related enzymes are highly expressed in certain bacteria. The PHX genes may provide potential drug targets for the design of new bactericide.     

Body shape and differences between species

The variation of body shape among prosimians is reviewed. Special emphasis is placed on the selective advantages, that is the mechanical reasons, to which variants of the locomotor apparatus can be traced back. There are differences found in the cheiridia, but at present they cannot be explained in terms of mechanics; there is nearly no knowledge about the mechanical meaning of their diversity. Myological characteristics of taxa can be explained mechanically, but this has not yet been done. Well known are variations of body proportions. These discriminate higher taxa, and are largely coincident with the often-used locomotor categories. In spite of this, there are only few sound arguments about the real biomechanic value of characteristic proportions for a given locomotor mode. What is known on this field, is reviewed. Progress can be made only, if the mechanical conditions, set by postural behavior and locomotion, are understood completely. The subtle distinctions between lower taxonomic units can normally be identified only on the basis of detailed and quantified analyses of movements on one hand, and of biometrics on the other. In the few cases in which such studies have been made, the differences of morphology fit to the mechanical requirements of locomotion which also differ only in quantitative details.     

Large feet are beneficial for eiders Somateria mollissima

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Characterization of two sets of subpolar flagella in Bradyrhizobium japonicum

Bradyrhizobium japonicum is one of the soil bacteria that form nodules on soybean roots. The cell has two sets of flagellar systems, one thick flagellum and a few thin flagella, uniquely growing at subpolar positions. The thick flagellum appears to be semicoiled in morphology, and the thin flagella were in a tight-curly form as observed by dark-field microscopy. Flagellin genes were identified from the amino acid sequence of each flagellin. Flagellar genes for the thick flagellum are scattered into several clusters on the genome, while those genes for the thin flagellum are compactly organized in one cluster. Both types of flagella are powered by proton-driven motors. The swimming propulsion is supplied mainly by the thick flagellum. B. japonicum flagellar systems resemble the polar-lateral flagellar systems of Vibrio species but differ in several aspects.     

Brainstem pathways of the initiation of locomotion

Conclusion This overview of the brainstem pathways of the initiation of locomotion can be summarized as follows. There are locomotor regions (hypothalamic and mesencephalic) whose stimulation leads to the appearance of rhythmic stepping movements. These regions are nonuniform in composition: transient fibers as well as cells are found. The locomotor effects of electrical stimulation of these regions can largely be explained on the basis of the presence of efferent projections of the neurons to the medial reticular formation, as well as the activation of the transient fibers of other brain systems. The ventromedial parts of the reticular formation of the medulla oblongata, including areas of the macrocellular and (to a lesser extent) gigantocellular nuclei, form the final element in the suprasegmental system of locomotion initiation. The inconclusive data of different scientists who have used chemical microinjections into the locomotor regions make it impossible at present to specify precisely the neurochemical mechanisms underlying the initiation of locomotion. NMDA has been found to play an important role. The activation of the reticular formation during the triggering of stepping movements can take place either from the locomotor regions or by means of signals coming from the collaterals of the ascending sensory tracts. The wide spectrum of possible pathways of the initiation of locomotion apparently affords the organism a choice of ways by which to realize this process and is an important factor in its adaptation to its environment.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 23, No. 4, pp. 488–505, July–August, 1991.     

Responses of ring-necked pheasant chicks (Phasianus colchicus) to conspecific calls

The locomotor responses of pheasant chicks to conspecific calls and the vocalizations of the chicks relating to their locomotor responses were measured in a test apparatus permitting approach or retreat movements. The brood-gathering call and the content call elicited strong approach from chicks. The alarm call and the squeak call inhibited locomotion in chicks. The hiss call, brood-caution call, fright call and flock call do not greatly affect the locomotion of subjects. The flock call appears to be primarily a care-soliciting call. Conspecific calls appear to be important to the survival of young pheasant chicks in the wild.