The phylogenetic position of toadfishes (order Batrachoidiformes) in the higher ray-finned fish as inferred from partitioned Bayesian analysis of 102 whole mitochondrial genome sequences

In a previous study based on 100 whole mitochondrial genome (mitogenome) sequences, we sought to provide a new perspective on the ordinal relationships of higher ray-finned fish (Actinopterygii). The study left unexplored the phylogenetic position of toadfishes (order Batrachoidiformes), as data were unavailable owing to technical difficulties. In the present study, we successfully determined mitogenomic sequences for two toadfish species ( Batrachomoeus trispinosus and Porichthys myriaster ) and found that the difficulties resulted from unusual gene arrangements and associated repetitive non-coding sequences. Unambiguously aligned, concatenated mitogenomic sequences (13 461 bp) from 102 higher actinopterygians (excluding the ND6 gene and control region) were divided into five partitions (1st, 2nd and 3rd codon positions of the protein-coding genes, tRNA genes and rRNA genes) and partitioned Bayesian analyses were conducted. The resultant phylogenies strongly suggest that the toadfishes are not members of relatively primitive higher actinopterygians (Paracanthopterygii), but belong to a crown group of actinopterygians (Percomorpha), as was demonstrated for ophidiiform eels (Ophidiiformes) and anglerfishes (Lophiiformes) in the previous study. We propose revised limits of major unranked categories for higher actinopterygians and a new name (Berycomorpha) for a clade comprising two reciprocally paraphyletic orders (Beryciformes and Stephanoberyciformes) based on the present mitogenomic phylogenies.  © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 85 , 289–306.     

Isolation of polymorphic microsatellite loci in Hemerocallis fulva and Hemerocallis citrina (Hemerocallidaceae)

Twenty microsatellite loci were isolated from a hybrid of two daylilies, Hemerocallis fulva and Hemerocallis citrina. We characterized individuals from two H. fulva populations and two H. citrina populations in Japan and observed three to 20 alleles per locus in H. fulva and one to 19 alleles per locus in H. citrina. Mean observed heterozygosity within populations ranged from 0.35 to 0.85 in H. fulva and from 0 to 0.95 in H. citrina. In about a half of the loci, the observed heterozygosity did not deviate from Hardy–Weinberg equilibrium. These loci are proved useful in studying gene flow and qualitative trait loci mapping using the two species.     

Effects of Succinyl Concanavalin A and Tunicamycin on F9 Embryonal Carcinoma Cells: Inhibition of Cell Spreading

Succinyl concanavalin A at a concentration of 50 μg/ml inhibited spreading of F9 embryonal carcinoma cells, while at this concentration it inhibited cell growth only partially. Thus, in the presence of succinyl concanavalin A, F9 cells grew as rounded cell aggregates that sometimes became detached from the substratum. Tunicamycin at a concentration of 1 μg/ml had a similar effect on F9 cells. These results suggest that a mannosyl glycoprotein(s) is involved in cell-substratum interaction of the cells. Furthermore, tunicamycin partially inhibited the biosynthesis of embryoglycan, the large glycoprotein-bound carbohydrates of early embryonic cells.     

ANALYSIS OF CELL PROLIFERATION DURING EARLY EMBRYOGENESIS

Cell proliferation was examined during early embryogenesis of the newt ( Triturus pyrrhogaster ) by various methods. After the two-cell stage, at 23°C, the blastomere (cell) number per whole embryo increased logarithmically until the mid-blastula stage (for about 19 hr) and the rate of increase slowed down in and after the late blastula stage. On the other hand, the synchronous cleavage of the blastomeres at the animal pole continued for 18 hr until the twelfth cleavage (mid-blastula) and the transition from synchronous to asynchronous division occurred abruptly at and after the thirteenth cell division (late blastula). The study also showed that the presumptive neuro-ectoderm consisted mainly of cells of the fifteenth generation (G-15) at the onset of gastrulation (pigment stage).
The present study suggested that the number of ectodermal cells of the early gastrula (stage 12a) nearly doubled during gastrulation at the presumptive neuro-ectoderm. This means that most of the ectodermal cells are in G-16 at the end of gastrulation. On the other hand, both mitotic activity and the rate of cell increase gradually diminished during gastrulation in the ectoderms of both the presumptive neural and epidermal regions, and there are evidently significant differences in both activities between the neuro-ectoderm and the epidermal ectoderm after stage 13b: the epidermal ectoderm showed greater decrease in the rate of both mitotic activity and cell proliferation than the neuro-ectoderm.
These facts suggested that, whether the ectodermal cells will differentiate into neural cells or epidermal cells is determined during G-15 or G-16 in normal primary induction.     

SCANNING ELECTRON MICROSCOPIC STUDY ON THE SURFACE CHANGE OF EGGS OF THE TELEOST, ORYZIAS LATIPES, AT THE TIME OF FERTILIZATION

The surface change of the egg of the teleost, Oryzias latipes , during fertilization was observed with a scanning electron microscope. The microvilli of the outer surface of the unfertilized egg show a slight difference in density between the animal and vegetal pole areas. In the initial step of the breakdown of cortical alveoli (CA), several small holes or gapes are formed at the apical part of the CA membrane, becoming a large aperture from which the alveolar contents are discharged. The formation of microvilli is observed on the inner surface of the exposed cavity left by the CA, starting from the periphery of the aperture and propagating throughout the whole inner surface in accompaniment with the release of the alveolar contents. After the completion of CA breakdown, the CA membrane cannot be distinguished from the original egg plasma membrane.     

CARBOHYDRATE METABOLISM IN THE EGGS OF THE SILKWORM, BOMBYX MORI. I. ABSENCE OF PHOSPHOFRUCTOKINASE IN THE DIAPAUSING EGG

In the diapausing eggs of the silkworm, Bombyx mori , glycogen is rapidly converted to sorbitol and glycerol, and this conversion is reversed at termination of the diapause (C hino , 1958). To elucidate the pathway leading to this polyol formation and its regulatory mechanisms, enzymes concerning carbohydrate metabolism were surveyed in diapausing as well as in developing eggs of the silkworm.
Most of the enzyme activities concerning citric acid cycle are low at the beginning of the embryogenesis and during diapause, but increase at the later stages of the development. Making an exception, reduction rate of malate and fumarate was rather high from the onset of the embryonic development. Several glycolytic enzymes were also studied. Most remarkable fact is that phosphofructokinase activity could not be demonstrated in the diapausing and also in the early stages of the developing eggs. Other enzymes, viz. α-glycerophosphate dehydrogenase, aldolase, glyceraldehyde-3-phosphate dehydrogenase were detected from the beginning of the embryogenesis.
Absence of phosphofructokinase, together with the high activity in glucose-6-phosphate dehydrogenase, suggests that predominant pathway in carbohydrate metabolism in the early stages of embryogenesis and in the diapause period is by way of pentose phosphate pathway. This supposition is confirmed by the experiments using labeled glucose. Incorporation of the label into glycerol of the diapausing eggs was three to four fold when G-6-¹⁴C was injected into pupae as compared with the case of G-1-¹⁴C injection. The above experiments provide evidence supporting the theory that glycogen is converted into sorbitol and glycerol mostly by way of the pentose phosphate pathway in the diapausing eggs.     

Micromere Differentiation in the Sea Urchin Embryo: Two-Dimensional Gel Electrophoretic Analysis of Newly Synthesized Proteins

A method for large-scale culture of isolated blastomeres of sea urchin embryos in spinner flasks was developed. Micromeres and meso-, macromeres isolated from sea urchin embryos at the 16-cell stage were cultured by this method and the patterns of protein synthesis by their descendants were examined by two-dimensional gel electrophoresis of [³⁵S] methionine-labeled proteins. Six distinct proteins with molecular weights of 140–kDa, 105–kDa, 43–kDa, 32–kDa, and 28–kDa (two components) were specifically synthesized by differentiating micromeres. Quantitative analysis of the two-dimensional gel patterns demonstrated that all these proteins, except the 32–kDa protein, appeared at the time of ingression of primary mesenchyme cells (PMC's) in vivo , several hours earlier than the onset of spicule formation. The synthesis of 32–kDa protein was paralleled to active spicule formation and the uptake of Ca²⁺. Cell-free translation products directed by poly (A)⁺ RNAs isolated from descendant cells of micromeres and meso-, macromeres were compared by two-dimensional gel electrophoresis. Several spots specific to the micromere lineage were detected. However, none of them comigrated with the proteins synthesized specifically by the cultured micromeres. The results suggest that the expression of these proteins specific to differentiating micromeres may involve post-translational modification.