The Role of Intracellular pH in Fertilization of Sand Dollar Eggs Analyzed by Microinjection Method

The change in intracellular pH (pH_i) upon fertilization and the effects of changing the pH_i by microinjection of pH buffers were investigated in the eggs of the sand dollar, Clypeaster japonicus. The pH_i was determined by the tint of a pH indicator, phenol red, microinjected into eggs. The pH_i ranged from 6.5 to 6.75 in unfertilized eggs and it rose by 0.4 to 0.5 unit within 3 min upon fertilization. The elevated pH_i ranging from 7.0 to 7.25 was maintained at least until the first cleavage. As reported in eggs of other species of sea urchin (1–4), development of fertilized eggs which had been transferred to Na-free sea water immediately after insemination was arrested and the pH_i did not rise remaining at the level of unfertilized eggs. Development was initiated in eggs arrested in Na-free sea water when the pH_i was elevated up to the level of fertilized eggs, i.e. 7.0 to 7.25, by microinjecting 1 M HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid)-KOH buffer at pH 8.0. By microinjection of pH 7.5 buffer, some eggs started development though none of them underwent cleavage. By microinjection of pH 7.0 or pH 6.5 buffer, development was not initiated. The initiation of development depended on the pH value of microinjected pH buffer, and in consequence, on the final pH_i. The elongation of microvilli which had been arrested in eggs in Na-free sea water was also induced by microinjection of pH 8.0 or 7.5 buffer.     

Melanocytes Fail to Survive in Hair Bulbs of the Shiba Goat (Capra hircus) With the Dominant Black-Eyed White Phenotype

Dominant black-eyed white phenotypes are one of the most commonly observed traits in domestic animals. Their genetic control mechanisms, however, have not been elucidated. As the first step to approach the problem, we examined histologically the patterns of the distribution of pigment cells in Shiba goats (two each of day-73-postcoitum and day-112-postcoitum fetuses, and a 15-week-old kid) with the dominant black-eyed white phenotype. Melanocytes were present and fully pigmented in the choroid and the sclera of eyes, as well as in dorsal skin epidermis of the fetuses and of the kid. Melanocytes were also found in approximately 6% of the hair bulbs in the fetal dorsal skin, while the rest (94%) lacked them. Hair follicles of the kid did not harbor melanocytes except for some in the early anagen stage. The results suggest that the survival of melanocytes was inhibited specifically in the hair follicles of the Shiba goat with the dominant black-eyed white phenotype and that the ostensibly similar phenotypes in the Shiba goat and in the SI or W mutants of the mouse, where melanocytes die en route to the hair bulbs, are regulated by different mechanisms.     

Group composition and contributions to breeding by Rufous Vangas Schetba rufa in Madagascar

The social system of the Rufous Vanga Schetba rufa was studied in a deciduous dry forest in Ampijoroa, western Madagascar. The species lived in groups of two to four individuals. Groups typically contained one adult female, one or two adult males and sometimes a (presumed) immature. All group members appeared to defend the territory. The presumed immatures had spotted throats and were smaller than adults in body size. They helped feed young and guard the nest but did not incubate or brood.     

Cytokinin Activity of Benzoylaminodeazapurines, Pentanoylaminodeazapurines and their Corresponding Purine Analogs in Five Bioassays

Cytokinin activities of 6-benzoylamino-1, 6-benzoylamino-3-, 6-pentanoylamino-1- and 6-pentanoylamino-3-deazapurines and their corresponding purine analogs, 6-benzoylaminopurine and 6-pentanoylaminopurine, were examined using five bioassay systems, tobacco callus growth, bud formation on tobacco callus, lettuce seed germination, fresh weight increase of radish cotyledons and retardation of chlorophyll degradation in radish cotyledons. 6-Benzoylamino- and 6-pentanoylamino-1-deazapurines showed stronger cytokinin activity than their corresponding purine analogs in all bioassays used. In tobacco callus growth, 6-benzoylamino-1-deazapurine was nearly as active as zeatin, one of the most active adenylate cytokinins. On the other hand, 6-benzoylamino- and 6-pentanoylamino-3-deazapurines were as active as or less active than corresponding purine analogs.     

MICROINJECTION OF COLCHICINE INTO SEA URCHIN EGGS

Inhibition of cleavage by colchicine was examined by microinjecting colchicine solution into one of the blastomeres of a sea urchin egg at the two-cell stage. Cleavage was inhibited if the microinjection was made before a critical point prior to the cleavage, whereas cleavage occurred in spite of the destruction of the mitotic apparatus if the microinjection was made after the critical point. The critical point was 10 min before the mid-stage of the cleavage in Clypeaster japonicus and 8 min before the mid-stage in Temnopleurus toreumaticus at 20 ± 1°C, corresponding to the beginning of anaphase. The threshold for the cleavage inhibition of colchicine was estimated to be 3 × 10⁻⁵ M to 3 × 10⁻⁶ M in final concentration in the cell.     

Development of microsatellite markers for the Manila clam Ruditapes philippinarum

We isolated nine polymorphic microsatellites from the Manila clam Ruditapes philippinarum. These loci provide one class of highly variable genetic marker as the number of alleles ranged from six to 22, and the observed and expected heterozygosity ranged from 0.136 to 0.909 and from 0.553 to 0.954, respectively. We consider that these loci are potentially useful for detailing the genetic structure and gene flow among R. philippinarum populations.     

Assessment of diet choice by the yellow fever mosquito Aedes aegypti

To investigate feeding‐related decisions in Aedes aegypti (L.), adults are presented with simple diets of paired gustatory stimuli conveying information concerning energy content, nutrient richness, osmotic balance and food toxicity in a two‐diet matrix assay. Assessment of mosquito gut contents indicates that both sexes accept single sugar diets in a dose‐dependent manner. When presented with a choice between two different yet equimolar sugar solutions, more individuals of both sexes accept the disaccharides, sucrose and trehalose, than the monosacharrides, fructose and glucose. The combination of pyranose and furanose sugars in solution, either physically associated (as in sucrose) or present as monomers (as glucose and fructose), is accepted over solutions containing a single sugar moiety. Using the two‐diet matrix assay, mosquito diet‐choice is also tested between two equimolar sucrose ‘driver’ solutions in which one is presented with various concentrations of another potential feeding cue ‘test’ compound (i.e. each of the 20 naturally‐occurring amino acids, sodium chloride, quinine or caffeine). Diet‐choice between the ‘driver’ sucrose‐only solution and the solution of the ‘driver’ sucrose containing a ‘test’ amino acid is influenced by sex, amino acid concentration and sucrose concentration. There is also an example of synergism between the diet components, leucine and sucrose. Mosquitoes demonstrate a dose‐dependent acceptance of sucrose‐only diets over sodium chloride‐containing sucrose when presented together. Interestingly, the sucrose‐only diet is accepted by more mosquitoes than all concentrations of the saline‐containing sucrose diets except those approximately isotonic to mosquito haemolymph, at which concentration mosquitoes show no clear choice between the diets. More individuals of both sexes accept sucrose‐only diets than the diets of caffeine‐containing sucrose in a dose‐dependent manner. Only females, however, respond to quinine‐containing sucrose diets and modulate this behaviour in relation to the energetic reward: more females imbibed quinine‐containing sucrose at the higher sucrose concentration (1 m ). A systematic characterization of diet selection behaviour of A. aegypti is presented for 27 putative feeding cues potentially involved in nectar/honeydew feeding. This study will be used as a basis from which to investigate further the mosquito's assessment of food quality and ultimately host choice.     

Development of microsatellite markers for the crown‐of‐thorns starfish Acanthaster planci

We isolated nine polymorphic microsatellites from the crown‐of‐thorns starfish, Acanthaster planci. These loci provide one class of highly variable genetic marker as the number of alleles ranged from three to 12 and the observed and expected heterozygosities ranged from 0.130 to 0.783 and from 0.163 to 0.862, respectively. We consider that these loci are potentially useful for detailing the genetic structure and gene flow among A. planci populations.     

Comparative foraging ecology of Madagascar vangids (Vangidae)

The Vangidae, an endemic family in Madagascar, contains 14 species in 11 genera. The foraging behaviour of 13 species of vangids was investigated during the periods August-November 1989 and September-November 1990. These vangids are grouped into six bill types according to their bill shape, using the analogy of gripping tools: (1) forceps. (2) radio pliers, (3) strong pinchers, (4) pliers, (5) standard and (6) flat forceps types. We recognized seven foraging niches: (A) canopy-gleaning, (B) general, (C) ground-snatching, (D) trunk/ branch-probing, (E) leaf and twig/branch-gleaning/probing, (F) leaf and twig/branch-gleaning and (G) trunk-gleaning niches. Two or more coexisting species with the same foraging niche differed in their bill types. Those vangids that exploited similar resources segregated in their bill types, geographic ranges and/or foraging niches, presumably to avoid severe competition. Several vangid species partially occupied the ecological niche that would be filled by woodpeckers elsewhere.