A Yolk-granule Component Acts as an Adhesive Material for Dissociated Gastrula Cells of the Newt, Cynops pyrrhogaster

Yolk granules were collected from full-grown ovarian oocytes of the newt, Cynops pyrrhogaster , and dissolved in 3% acetic acid or 8 M urea solution. Culture dishes were then coated with either of the yolk-granule solutions. The yolk-coated surfaces acted as adhesive substrata for dissociated gastrula cells, which showed active locomotion when spread on the surfaces. Divalent cation was required for cell adhesion and spreading on the yolk-coated surface. Trypsin and glycosidase digestions of dissociated cells or the yolk-coated surfaces inhibited cell adhesion and spreading. Addition of concanavalin A to the culture medium inhibited cell spreading on the yolk-coated surfaces, while high concentration (50 mM) of the saccharides, D-galactose, D-lactose and D-mannose, had a slightly inhibitory effect on cell spreading.
Two yolk components (30-kD and 108-kD proteins) were isolated from yolk granules and applied to the culture dish surfaces. Surfaces coated with the 30-kD protein showed strong adhesiveness for dissociated gastrula cells.     

Effect of Actinomycin D and Cycloheximide on Gonadotropin-Induced 17α, 20β-Dihydroxy-4-pregnen-3-one Production by Intact Ovarian Follicles and Granulosa Cells of the Amago Salmon,Oncorhynchus rhodurus*

The effect of actinomycin D and cycloheximide on gonadotropin (partially purified chum salmon gonadotropin, SGA)-induced 17α, 20β-dihydroxy-4-pregnen-3-one (17α, 20β-diOHprog, a maturation-inducing steroid in amago salmon) production was examined in intact ovarian follicles and granulosa cells of postvitellogenic amago salmon, Oncorhynchus rhodurus. Both actinomycin D and cycloheximide blocked gonadotropin-induced 17α, 20β-diOHprog production by intact follicles. In contrast, gonadotropin-induced 17α-hydroxyprogesterone production by intact follicles was not abolished by actinomycin D, but was abolished by cycloheximide, suggesting that postvitellogenic amago salmon ovarian follicles already contain the RNAs necessary for the synthesis of 17α-hydroxyprogesterone. In isolated granulosa cells, chum salmon gonadotropin was able to stimulate 17α, 20β-diOHprog production only when a precursor, 17α-hydroxyprogesterone was provided in the incubation medium, indicating that gonadotropin acts directly on granulosa cells to enhance the activity of 20β-hydroxysteroid dehyrogenase (20β-HSD). Total inhibition of 20β-HSD enhancement in granulosa cells, judged by 17α, 20β-diOHprog production, was achieved when actinomycin D was added between 1 hr before the start of incubation with 17α-hydroxyprogesterone and gonadotropin to 6 hr after. With cycloheximide total inhibition was observed when added in the period of 1 hr before to 9 hr after the start of the incubation. These results suggest that chum salmon gonadotropin acts on granulosa cells to enhance the de novo synthesis of 20β-HSD by a mechanism involving RNA synthesis.     

Accumulation and Distribution of Extracellular Matrix as Revealed by Scanning Electron Microscopy in Freeze-Dried Newt Embryos Before and During Gastrulation

A scanning electron-microscopic study was carried out on the extracellular matrices (ECMs) in freeze-dried newt embryos from the cleavage to the gastrula stage. The results revealed the appearance, accumulation and distribution of two types of ECMs, a fibrillar ECM in the blastocoel and an amorphous ECM on the inner surface of the blastocoelic wall (BW). The fibrillar ECM first appeared in the blastocoel at the cleavage stage and increased notably in quantity at the blastula and gastrula stages. On the other hand, the amorphous ECM was initially detected on the inner surface of the BW at the beginning of gastrulation and it increased in quantity during gastrulation. With the progress of archenteron invagination, the amorphous ECM was found to be deposited in the space between the BW and migrating cells.     

Citrulline and DRIP-1 Protein (ArgE Homologue) in Drought Tolerance of Wild Watermelon

Drought-affected plants experience more than just desiccationof their organs due to water deficit. Plants transpire 1000times more molecules of water than of CO₂ fixed by photosynthesisin full sunlight. One effect of transpiration is to cool theleaves. Accordingly, drought brings about such multi-stressesas high temperatures, excess photoradiation and other factorsthat affect plant viability. Wild watermelon serves as a suitablemodel system to study drought responses of C₃ plants, sincethis plant survives drought by maintaining its water contentwithout any wilting of leaves or desiccation even under severedrought conditions. Under drought conditions in the presenceof strong light, wild watermelon accumulates high concentrationsof citrulline, glutamate and arginine in its leaves. The accumulationof citrulline and arginine may be related to the induction ofDRIP-1, a homologue of ArgE in Escherichia coli, where it functionsto incorporate the carbon skeleton of glutamate into the ureacycle. Immunogold electron microscopy reveals the enzyme tobe confined exclusively to the cytosol. DRIP-1 is also inducedby treating wild watermelon with 150 mM NaCl, but is not inducedfollowing treatment with 100 µM abscisic acid. The salttreatment causes the accumulation of -aminobutyrate, glutamineand alanine, in addition to a smaller amount of citrulline.Citrulline may function as a potent hydroxyl radical scavenger.     

A LONG-BODIED LIZARD FROM THE LOWER CRETACEOUS OF JAPAN

Abstract:  Platynotan lizards underwent a dramatic Late Cretaceous radiation into marine habitats. Beginning with small-bodied forms, the lineage culminated with the mosasaurs, large predatory lizards with a world-wide distribution in the Santonian–Campanian. Moreover, the marine squamate radiations of the Cenomanian–Turonian are remarkable in having produced a range of long-bodied, reduced-limbed swimmers (dolichosaurs, adriosaurs, coniasaurs and limbed snakes) that seem to have thrived in the shallow coastal environments of the Western Tethys region. Until now, none of these long-bodied aquatic squamates has been recorded prior to the Cenomanian, none has been recovered from a non-marine locality and none is known from Asia. Here we describe a small, gracile, long-bodied mosasauroid lizard from a swampy continental deposit in the Lower Cretaceous of Japan.     

Structural Changes of Yolk Platelets and Related Organelles during Development of the Newt Embryo

Structural changes in yolk platelets and related organelles in the cytoplasm of the presumptive ectodermal region up to the stage of gastrulation were studied by light and electron microscopies using full-grown oocytes, mature eggs descending the oviduct and embryos of the newt, Cynops pyrrhogaster . Yolk platelets with a superficial layer are first observed in mature eggs descending the oviduct. During the cleavage and early morula stages, the superficial layer increases in thickness and the main bodies become more slender. The superficial layer decreases in thickness in the blastula stage, and many yolk platelets lose this layer in the gastrula stage.
The amount of rough-surfaced endoplasmic reticulum (r-ER) increases rapidly in the morula stage, while Golgi complexes gradually increase in number between the cleavage and gastrula stages. In the cleavage and early morula stages, most of the r-ER is closely adherent to yolk platelets and is associated with several mitochondria. Two types of free vesicles, large (0.5–4.0 μm diameter) and small (0.15–0.3 μm diameter), were seen in abundance from the early morula stage to the early gastrula stages.
Changes in the structure of yolk platelets are discussed in relation to changes in other cytoplsmic organelles.     

Scanning Electron Microscopy of the Extracellular Matrix of Amphibian Gastrulae by Freeze-drying

In order to demonstrate the extracellular matrix (ECM) which tends to be missing from specimens prepared by ordinary procedures, a freeze-drying method was applied to gastrulae of the newt ( Cynops pyrrhogaster ) without prefixation and the embryos were examined with a scanning electron microscope (SEM). It was revealed that a prominent amount of fibrillar ECM occupied the blastocoel and amorphous ECM covered the inner surface of the blastocoelic wall (BW) as well as the surface of migrating cells. These results were compared with those obtained from observations of specimens fixed chemically.     

Intraspecific competition as a selective pressure on the choice of oviposition site in a phytophagous insect

Our aim was to clarify the factors providing selective pressures on the choice of oviposition sites in phytophagous insects. To this end, we examined the spatial distribution of eggs of the sawfly leaf miner, Profenusa japonica . The study was conducted over five consecutive years on three subpopulations found in riverbeds in central Japan. Profenusa japonica lays eggs on leaflets of Rosa multiflora , and its larvae mine a single leaflet for the entire larval stage. The egg distribution among leaflets was uniform (regular) at all sites examined in each year. The distribution within leaflets was also uniform, and it appeared that females avoided ovipositing on areas of the leaf that already contained eggs. Few parasitoids were found in eggs or larval stages, despite intensive examination. Larval mortality increased, mainly as a result of exploitative competition, when there were two or three larvae per leaflet. Starvation caused by resource competition for leaflet tissues was an important mortality factor. Therefore, we suggest that P. japonica females avoided ovipositing on areas of the leaflet that already contained eggs to prevent exploitative competition among larvae sharing the same leaflet.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 641–650.     

Morphological Changes of the Inner Surface of the Blastocoelic Wall before and during Gastrulation in the Newt, Cynops pyrrhogaster

Ultrastructural observations were made of morphological changes in the inner surface of the blastocoelic wall (BW) in newt embryos during the period from morula to gastrula. A scanning electron microscopic investigation distinguished three phases of process formation on the inner surface of the BW. First, small blebs and filopodia were found in the morula. These then changed to large blebs in the blastula and early gastrula. Finally, they acquired lamella-like structures (lamellipodia) in the middle and late gastrula. In addition to the changes in the cell surface, cytoplasmic changes such as of the organization of the cortical microfilamentous layer (CML) were found by transmission electron microscopy (TEM). The morphological changes on the inner surface of the BW and those in the cytoplasmic ultrastructures are considered to be closely related, playing some important roles in the mechanisms of morphogenetic cell movements.