WATER PERMEABILITY OF THE CELL WALL IN NITELLA

1. A method has been developed to measure the hydraulic conductivityof the wall of the internodal cell of Nitella flexilis.
2. Therate of water penetration through the cell wall varies linearlywith the hydrostatic pressure difference between the two sidesof the wall, showing that water permeability of the cell wallremains independent of the pressure difference applied.
3. Waterpermeability of the cell wall is inversely proportionalto itsthickness It is 30µµmin^–3{dot}atm^–3when the thickness of the wall is 10 µ.
4. Water permeabilityof the cell wall is the same for inward andoutward water flow.The polar water permeability of the entiremembrane system (walland protoplasmic part) of the living celldemonstrated by KAMIYAand TAZAWA (1) is, therefore, due tothe living protoplasmicpart.
5. The ratio of the inward to outward permeability constantsofthe protoplasmic layer alone is higher than that of the entiremembrane system composed of protoplasmic layer and cell wall.

¹ Dedicated to Prof. H. TAMIYA on the occasion of his 60th birthday.The present work was supported in part by a Grant-in-Aid forFundamental Scientific Research from the Ministry of Education. ² Present address: Sh?in Women's College, Kobe. (Received July 21, 1962; )     

Inductive Capacities for the Dorsal Mesoderm of the Dorsal Marginal Zone and Pharyngeal Endoderm in the Very Early Gastrula of the Newt,and Presumptive Pharyngeal Endoderm as an Initiator of the Organization Center*

The organization center of Cynops pyrrhogaster was divided into Parts 1, 2 and 3 of equal size (0.3×0.4 mm²) with presumptive fates as pharyngeal, pharyngeal+prechordal+trunk notochord, and trunk-tail notochord, respectively. Movements and changes in size and shape of each part were followed through gastrulation. Differentiation tendencies of each part were examined under three conditions: I, isolated; II, sandwiched with presumptive ectoderm; 111, sandwiched with presumptive ectoderm after preculture in isolation for various times. In I, Parts 2 and 3 differentiated into dorsal mesoderm. In II, each part induced dorsal mesoderm and neural tissues, the frequency being highest in Part 2 and lowest in Part 3. In III, Parts 1 and 2 realized their presumptive fates, through changes in inductive capacities from trunk-tail to head. This change progressed rapidly in Part 1, and slowly in Part 2. Part 3 required induction by neighbouring Part 2 to realize its presumptive fate. Changes of inductive capacity of Parts 1 and 2 respectively, were chronologically similar in normal development and in preculture experiments. Lastly, the primary presumptive pharyngeal zone at blastula was proposed to act as an initiator of the organization center, its programmed information being transmitted to Part 2, and then to Part 3.     

A Phylogenetic Lineage of Closely Related Trypanosomes (Trypanosomatidae,Kinetoplastida) of Anurans and Sand Flies (Psychodidae,Diptera) Sharing the Same Ecotopes in Brazilian Amazonia1

ABSTRACT. Analysis of the phylogenetic relationships among trypanosomes from vertebrates and invertebrates disclosed a new lineage of trypanosomes circulating among anurans and sand flies that share the same ecotopes in Brazilian Amazonia. This assemblage of closely related trypanosomes was determined by comparing whole SSU rDNA sequences of anuran trypanosomes from the Brazilian biomes of Amazonia, the Pantanal, and the Atlantic Forest and from Europe, North America, and Africa, and from trypanosomes of sand flies from Amazonia. Phylogenetic trees based on maximum likelihood and parsimony corroborated the positioning of all new anuran trypanosomes in the aquatic clade but did not support the monophyly of anuran trypanosomes. However, all analyses always supported four major clades (An01‐04) of anuran trypanosomes. Clade An04 is composed of trypanosomes from exotic anurans. Isolates in clades An01 and An02 were from Brazilian frogs and toads captured in the three biomes studied, Amazonia, the Pantanal and the Atlantic Forest. Clade An01 contains mostly isolates from Hylidae whereas clade An02 comprises mostly isolates from Bufonidae; and clade An03 contains trypanosomes from sand flies and anurans of Bufonidae, Leptodactylidae, and Leiuperidae exclusively from Amazonia. To our knowledge, this is the first study describing morphological and growth features, and molecular phylogenetic affiliation of trypanosomes from anurans and phlebotomines, incriminating these flies as invertebrate hosts and probably also as important vectors of Amazonian terrestrial anuran trypanosomes.     

Phylogenetic Analyses Based on Small Subunit rRNA and Glycosomal Glyceraldehyde-3-Phosphate Dehydrogenase Genes and Ultrastructural Characterization of Two Snake Trypanosomes: Trypanosoma serpentis n. sp. from Pseudoboa nigra and Trypanosoma cascavelli from Crotalus durissus terrificus

ABSTRACT. We sequenced the small subunit (SSU) rRNA and glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes of two trypanosomes isolated from the Brazilian snakes Pseudoboa nigra and Crotalus durissus terrificus . Trypanosomes were cultured and their morphometrical and ultrastructural features were characterized by light microscopy and scanning and transmission electron microscopy. Phylogenetic trees inferred using independent or combined SSU rRNA and gGAPDH data sets always clustered the snake trypanosomes together in a clade closest to lizard trypanosomes, forming a strongly supported monophyletic assemblage (i.e. lizard–snake clade). The positioning in the phylogenetic trees and the barcoding based on the variable V7–V8 region of the SSU rRNA, which showed high sequence divergences, allowed us to classify the isolates from distinct snake species as separate species. The isolate from P. nigra is described as a new species, Trypanosoma serpentis n. sp., whereas the isolate from C. d. terrificus is redescribed here as Trypanosoma cascavelli .     

Isolation and characterization of microsatellite loci from Quercus mongolica var. crispula

Microsatellites were isolated and characterized for the Japanese oak species, Quercus mongolica var. crispula, distributed in temperate deciduous forests of Japan. Eleven of the 48 primer pairs designed successfully amplified unambiguous and polymorphic single loci among 67 Q. mongolica var. crispula individuals within a plot in southwestern Japan. The observed and expected heterozygosities of the 11 microsatellite markers ranged from 0.522 to 0.896 and from 0.536 to 0.882, respectively. These polymorphic microsatellite markers are useful for estimating pollen‐mediated gene flow in Q. mongolica var. crispula.     

Changes of Chromosome Length and Constitutive Heterochromatin in Association with Cell Division during Early Development of Cynops pyrrhogaster embryo

The length of chromosomes in the presumptive ectoderms of Cynops embryos was measured at nine successive cell divisions from the 6th (cleavage stage) to the 14th (gastrula stage). Up to the 10th cell division (cleavage stage) the chromosome length remained constant. At the 11th cell division the chromosomes began to shorten and continued to shorten at every cell division up to the 14th cell division. The degree of shortening and the mode of variation in length corresponded to the respective developmental stages of cleavage, blastula and gastrula. During those periods when chromosomes became shortened, some fine C-bands of the paracentromeric region found in earlier stages fused together. The chromatin of interphase nuclei also showed considerable changes during chromosome shortening. Besides the size reduction of interphase nuclei, the number of C-band granules in an interphase nucleus decreased in parallel with chromosome shortening and fusion of C-bands in mitotic chromosome.     

Isolation and characterization of polymorphic microsatellite DNA markers in the Omono type of ninespine stickleback,genus Pungitius

We isolated and characterized 25 polymorphic microsatellite loci for the Omono type of ninespine stickleback, genus Pungitius, an endangered species in streams including agricultural canals in Japan. The number of observed alleles per locus ranged from two to 15 within 32 individuals each collected from three populations in the Omono River, and the values of observed and expected heterozygosities ranged from 0.031 to 0.906 and from 0.031 to 0.856, respectively. All loci conformed to the Hardy–Weinberg equilibrium. Genetic linkage disequilibrium analysis between all pairs of the loci showed no significant departure from the null hypothesis between any of the loci.     

Changes in Soybean Agglutinin (SBA) and Peanut Agglutinin (PNA) Binding Pattern in the Epidermis of the Developing Chick Embryo

Lectin binding pattern in the developing chick embryonic epidermis was studied using peroxidase labeling method. The epidermis of the 13-day-old embryo is in an undifferentiated state. Little binding of soybean agglutinin (SBA), specific for N-acetyl-D-galactosamine, and peanut agglutinin (PNA), specific for β-D-galactose, was seen in such epidermal cells. As the epidermis developed toward keratinization, the cell membrane of the differentiating flattened cells was positively stained with SBA and PNA. The positive staining was also seen in the supranuclear region of the cells located between the flattened cells and the basal cells. The basal cells remained unstained in all the stages of development. Similar staining pattern with SBA and PNA was seen in the cultured skin explants during the epidermal differentiation in vitro. These observations show that the SBA- and PNA-reactive glycoconjugates accumulate during the epidermal cell differentiation, suggesting their important roles in the maintenance of the ordered structure of the epidermis.     

PHYSICOCHEMICAL STUDIES OF TASTE RECEPTION VI: Interpretation of Anion Influences on Taste Response

Monolayers of lipids from bovine tongue epithelium were preparedas a model system for the gustatory receptor membrane to clarifythe effects of anions on the taste response. Changes in thesurface potential of the monolayers were measured by use ofthe ionizing-electrode method under the presence of sodium andcalcium salts carrying various species of anions in the aqueousphase. The organic and chaotropic anions showed the suppressiveeffect on the surface potentials as compared with Cl^–or NO₃^-. The influences of anion species on the surface potentialwere similar to those on the taste response recorded from glossopharyngealnerve of the frog. This suggested that the effects of anionson the taste response could be explained in terms of the electricalpotential at the interface between the receptor membrane andstimulating solution. Analysis of data on the surface potentialrevealed that a conformational change of the monolayers causedby non-electrical interaction between the monolayers and anionsis responsible for the suppressive effect.