Cellular Basis of Neuralization of Induced Neurectoderm in Amphibian Embryogenesis: Changes of Cell Shape, Cell Size, and Cytodifferentiation of the Neurectoderm after Neural Induction

Cellular alterations of the neurectoderm after primary embryonic induction were examined by measuring several indices of shape, volume, and cytodifferentiation of the neurectodermal cells of Cynops embryos during gastrulation and early neurulation.
Results showed that cellular alterations occurs just after the 18 hr embryo stage (stage 13b). The thickness of the neurectoderm layer decreases like that of the epidermal ectoderm during early and middle gastrulation. After the 18 hr embryo stage, however, the neurectoderm thickens, mainly due to formation of columnar cells. Measurement of cell volume indicates that the neurectoderm of the early and middle gastrulae consists of a cell population of heterogeneous size. The heterogeneity diminishes sharply after the 18 hr embryo stage and the neural plate of the 36 hr embryo (stage 18) consists of cells of homogeneous size.
Stages before the 12 hr embryo (stage 12b) and after the 18 hr embryo (stage 13b) also showed differed in cell adhesion to the culture flask and in cytodifferentiating potency. Single cells dissociated from the neurectoderm of 18 hr embryos could adhere to the substratum and differentiate into both nerve-like cells and pigment cells. Both capacities increase during further development.
These results are discussed in relation to the neuralizing determination of neurectoderm after primary embryonic induction.     

Species Specificity of the Insect Prothoracicotropic Hormone (PITH): the Presence of Bombyx-and Samia-Specific PTTHs in the Brain of Bombyx mori

Crude extracts of Bombyx mori brains can provoke adult development when injected into brain-removed dormant pupae of Bombyx mori and Samia Cynthia ricini. From this fact the prothoracicotropic hormone (PTTH) of Bombyx has long been thought to be species-nonspecifically active on Samia. Chemical fractionation of Bombyx brain or head extracts by fractional precipitation with acetone, Sephadex G-50 gel-filtration, and DEAE-Sepharose CL-6B chromatography, however, separated the fractions which activated Bombyx brainless pupae from those which activated Samia. Those results reveal the existence of two species-specific PTTHs.     

Effects of Thioureas, Inhibitors of Melanogenesis, on Lens Transdifferentiation of Cultured Chicken Embryonic Retinal Pigment Cells

The effects of phenylthiourea (PTU) and its analogues on chick embryonic pigmented epithelial cells (PECs) in culture were studied to elucidate the correlation between inhibition of melanogenesis of PECs and enhancement of their transdifferentiation into lens cells.
Both 0.25–0.5 mM PTU and 0.1 mM alpha-naphthylthiourea (ANTU) effectively inhibited melanogenesis of PECs and stimulated their transdifferentiation into lens cells at the same time. Thiourea (TU) also inhibited melanogenesis at a much higher concentration (4 mM), but did not stimulate the lens transdifferentiation at all. Methylthiourea (MTU), on the other hand, did not inhibit melanogenesis, but stimulated the lens transdifferentiation. Testicular hyaluronidase effectively amplified the above-mentioned stimulating effects of thioureas without their altering optimum concentrations, although this enzyme itself never enhanced the lens transdifferentiation of PECs but suppressed their melanogenesis at a concentration of 100 U/ml medium, onward.
These results suggest that the suppression of melanogenesis of PECs by PTU or its analogues does not directly correlate with their transdifferentiation into lens cells. The possible mode of thiourea actions on the lens transdifferentiation of PECs cultured in vitro is discussed.     

Quantitative Analysis of Protein Synthesis Altered by Estrogen in Cultured Xenopus Liver Parenchymal Cell

Using the primary culture system of male Xenopus laevis hepatocytes consisting of more than 95% parenchymal cells, the effect of estradiol-17 β (10⁻⁶M) on protein synthesis was quantitatively analyzed by ³H-leucine incorporation kinetics and the estimation of specific radioactivity of newly synthesized secretory protein. The cells in a well defined culture revealed high plating efficiency and very low DNA synthetic activity. The cultured cells could synthesize several secretory proteins containing serum albumin. The pattern of secreted proteins in SDS-polyacrylamide gel electrophoresis (SDS-PAGE) did not alter with culture time but secretion rate of protein increased for 7 days, starting on the third day following inoculation. Estradiol added to the culture media extensively induced the synthesis of yolk precursor protein vitellogenin which accounted for 40–50% of the overall secretory protein synthesis and 20–30% of the total protein synthesis on day 7 of estradiol treatment. Ultimately, the total protein synthesis and secretory protein synthesis were stimulated 1.2–1.3 fold and 2.0–2.2 fold, respectively, over those of the control cells cultured in the absence of estradiol. These results indicated that the stimulation of protein synthesis was largely due to vitellogenin production. Such an estradiol-dependent stimulation of protein synthesis was also detected in the low molecular weight protein(s). On the other hand, albumin synthesis was evidently reduced by estradiol. Thus, estradiol had two different effects on protein synthesis.
The results obtained in this study will be discussed in relation to the findings o in vivo experiments.     

Distribution of Intramembrane Particles and Its Changes during the Acrosome Reaction in Spermatozoa of the Japanese Abalone

The distribution of intramembrane particles in the plasma and acrosomal membranes of sperm of the Japanese abalone, Haliotis discus , and its changes during the acrosome reaction were studied by the freeze-fracture replica technique. The P face of the plasma membrane covering the acrosome has sparse membrane particles except in the apical region, which includes the trigger and 'truncated cone' regions. Large particles with an average diameter of 10 nm are located in this apical region. The E face of the plasma membrane has only a few particles. On the outer acrosomal membrane, many particles are randomly distributed throughout the P face, but only a small number of particles are found on the E face. Numerous particles on the P face of the inner acrosomal membrane show a regular arrangement as a dense lattice or with a concentric circular pattern. The initial change in the acrosome reaction is clearance of membrane particles from both the P and E faces of the plasma and outer acrosomal membranes around the apical region, where fusion of the two membranes occurs. As the acrosomal process elongates, the dense arrangement of particles on the inner acrosomal membrane changes via a loose lattice arrangement to a patchy distribution with particle-free areas. Then the arrangement is further disorganized becoming a sparse, random distribution.     

Translation of RNA from Eggs During Post-Diapause Development of Bombyx mori

Eggs of Bombyx mori are aroused from diapause by long-term chilling and develop when transferred to 25°C. During the first 20 hr of post-diapause development, the polysome content and the presumed rate of protein synthesis increase about 3-fold, while the ribosome content and the total RNA content increase only 1.1-fold. In this study, total RNAs were extracted from chilled eggs (termed 0 hr of development), and post-diapause eggs at 10 and 20 hr of development. The RNAs were purified further by high pressure liquid chromatography to remove RNA-like oligonucleotides. On translation in a protein-synthesizing system derived from wheat germ with a subsaturating amount of RNA, no difference was found in the relative amounts of translatable mRNA activity at 10 and 20 hr of development from that at 0 hr. Moreover, the translation products of the different RNA preparations in a rabbit reticulocyte lysate system appeared very similar when separated by gel electrophoresis and located by fluorography. These facts suggest that protein synthesis in early post-diapause development is controlled at a translational level.     

Activities of Enzymes Relating to Starch Synthesis and Endogenous Levels of Growth Regulators in Potato Stolon Tips During Tuberization

Changes in contents of starch and protein, and activities of enzymes involved in starch synthesis were studied during tubcrization of stolon tips of Solanum tuberosum L. cv. Irish Cobbler. Starch content and activities of phosphorylase and granule-bound starch synthctase based on fresh weight increased rapidly in the early phase (stage I, the stolon tips just before swelling; stage 2, the swelling tips; stage 3, young tubers of 0.2–0.5 cm diameter), and they all remained nearly unchanged in the later phase (stage 3 to stage 6, young tubers of 3.5 cm diameter). The content of soluble protein based on fresh weight remained unchanged. Activities of soluble starch snythetase and ADP-glucose pyrophosphorylase were not detected at stage 1 and 2, but increased at later stages. Endogenous levels of auxin, cytokinin and gibberellin were assayed for the materials at the corresponding developmental stages. Auxin content was high at stages 1 and 2, and lowered at later stages. Cytokinin content increased abruptly at stage 6. Gibberellin content was low at all stages. The internal conditions for starch deposition and tuberization in potato were discussed in regard to regulation of enzyme activities by growth regulators.     

Female dimorphism in Japanese diving beetle Dytiscus marginalis czerskii (Coleoptera: Dytiscidae) evidenced by mitochondrial gene sequence analysis

Three species of the Japanese diving beetle Dytiscus have been identified: D. dauricus Gebler, 1832; D. marginalis czerskii Zeitzev, 1953; and D. sharpi. At present, the latter consists of the subspecies D. sharpi sharpi Wehncke, 1875 and D. sharpi validus Régimbart, 1899 based on the comparative data of mitochondrial DNA cytochrome‐c oxidase I (COI) sequences. Many Dytiscus species have smooth and grooved elytra, which are female dimorphic traits. For many years it has been thought that Japanese D. marginalis czerskii has a single morph, that is, only grooved females, although there were some collecting reports of smooth females occurring at the foot of Mt. Chokaisan in Yamagata and Akita Prefectures. However, the population of smooth females (smooth population) has not yet been identified by DNA markers. To understand the species status of the smooth population, we sequenced 769 bp of COI of a male derived from a smooth mother insect and compared it with the sequence from a known grooved female. The sequences of 769 bp of the COI gene in the smooth population were identical to that in the grooved female, indicating that Japanese D. marginalis czerskii has female dimorphic traits.     

A New Entodiniomorphid Ciliate,Troglocorys cava n. g., n. sp., from the Wild Eastern Chimpanzee (Pan troglodytes schweinfurthii) from Uganda

ABSTRACT. Troglocorys cava n. g., n. sp. is described from the feces of wild eastern chimpanzee, Pan troglodytes schweinfurthii, in Uganda. This new species has a spherical body with a frontal lobe, a long vestibulum, a cytoproct located at the posterior dorsal side of the body, an ovoid macronucleus, a contractile vacuole near the cytoproct, and a large concavity on the left surface of the body. Buccal ciliature is non‐retractable and consists of three ciliary zones: an adoral zone surrounding the vestibular opening, a dorso‐adoral zone extending transversely at the basis of the frontal lobe, and a vestibular zone longitudinally extending in a gently spiral curve to line the surface of the vestibulum. Two non‐retractable somatic ciliary zones comprise arches over the body surface: a short dorsal ciliary arch extending transversely at the basis of the frontal lobe and a wide C‐shaped left ciliary arch in the left concavity. Because of the presence of three ciliary zones in the non‐retractable buccal ciliature, the present genus might be a member of the family Blepharocorythidae, but the large left concavity and the C‐shaped left ciliary arch are unique, such structures have never been described from other blepharocorythids.