Respiration in Eggs of the Echiuroid, Urechis unicinctus, Before and After Fertilization

In eggs of the echiuroid Urechis unicinctus the respiration rate, which is not altered by fertilization, is inhibited by rotenone, antimycin A and cyanide. The respiration in echiuroid eggs is probably mediated by the mitochondrial respiratory chain. In fertilized eggs, the respiration was inhibited by oligomycin and stimulated by the uncouplers of oxidative phosphorylation 2,4-dinitrophenol and carbonylcyanide p-trifluoromethoxyphenylhydrazone, whereas respiration in unfertilized eggs was insensitive to these compounds. Insemination increased the respiratory rate in eggs in the presence of uncouplers and reduced it in the presence of oligomycin. These findings suggest that the capacity of electron transport in mitochondira is elevated by fertilization but becomes latent on fertilization-induced coupling of respiration with oxidative phosphorylation. Strong stimulation of the respiration in unfertilized eggs was induced by dichlorophenol indophenol, phenazine methosulfate and tetramethyl p-phenylenediamine, suggesting possible sites at which electron transport is regulated in unfertilized eggs. The resulting stimulation of respiration in unfertilized eggs was insensitive to uncouplers and oligomycin, but became sensitive to them after fertilization simultaneously with considerable decrease in its rate. Fertilization-induced coupling of the respiration seemed to reduce the respiratory rate enhanced artificially by these redox compounds.     

Inhibitory Effect of Calmodulin Antagonists and Calcium Antagonists on Fertilization-Induced Cyanide-Insensitive Respiration in Sea Urchin Eggs

During initial several minutes after fertilization, sea urchin eggs exhibited high rate of respiration which was only slightly inhibited by cyanide. This cyanide-insensitive respiration was inhibited by calcium antagonists, diltiazem and verapamil, and calmodulin antagonists, N-(6-aminohexyl)-5-chloro-1-naphthalensulfonamide hydrochloride (W-7), N-(6-aminohexyl)-1-naphthalenesulfonamide hydrochloride (W-5) and chlorpromazine, which were added within 1 min after insemination. The inhibitory effect of W-7 on cyanide-insensitive respiration was higher than that of W-5. Cyanide-sensitive respiration of fertilized eggs observed after this initial period was not inhibited by these compounds. Ca²⁺ influx in eggs just after fertilization was inhibited by calcium antagonists but was rather enhanced by calmodulin antagonists. Fertilization-induced stimulation of cyanide-insensitive respiration probably results from calmodulin-dependent reactions which are activated by Ca²⁺ influx.     

The relation between calcium and cell wall in growing rice leaf

The percentage of pectic substances in the cell wall of riceleaf decreased with the ageing of the leaf but other componentsin the cell wall changed little during leaf growth. Cell wallcomponents were not affected by a nutritional deficiency ofcalcium. At the beginning of the growth of rice leaf, calciumin the cell wall existed only in the pectic substance fractionboth in the calcium sufficient and deficient leaves. However,in the cell wall of the mature leaf, a considerable amount ofcalcium was found in the lignin fraction. The amount of calciumin this form was larger in calcium sufficient leaves than indeficient leaves. Calcium seems to occur in two forms in the cell wall, combinedwith pectic substances and with ligneous substances. This assumptionwas further supported by calcium distribution in enzymaticallydegradated fractions of the cell wall. Calcium seems to be combined with pectic substances by a strongerchemical bond than with the ligneous substances. ¹Present address: Tohoku Agricultural Experiment Station, Omagari,Akita.     

Effect of Aminopterin and Deoxyribonucleosides on the Cleavage and Embryogenesis of the Sea Urchin, Hemicentrotus pulcherrimus

In the embryos of the sea urchin, Hemicentrotus pulcherrimus , reared with 150 μM aminopterin from the time of fertilization, cessation of the development occurred at the blastula stage, at which the dTTP level became quite low. Another addition of thymidine to the embryo culture containing aminopterin resulted in an elevation of dTTP concentration in the embryos and allowed them to develop normally. Decrease in the dTTP level, resulting from the inhibition of thymidylate synthesis by aminopterin, probably causes a failure of egg cleavage and development. 5-Bromo-2'-deoxyuridine (BUdR) also released the aminopterin-inhibition of egg cleavage and allowed the treated embryos to develop to early gastrulae. Thereafter, the degeneration of archenteron occurred and these embryos became large permanent blastulae. Other deoxyribonucleosides failed to cancel the inhibition by aminopterin of egg cleavage. In the embryos kept with both BUdR and aminopterin, BUdR incorporation into DNA occurred at a similar rate as in thymidine incorporation in the embryos kept with thymidine and aminopterin, and was inhibited by another addition of thymidine. Without aminopterin treatment, BUdR incorporation hardly occurred and the embryos developed normally. BUdR incorporation into DNA in place of thymidine probably occurs in aminopterin-treated embryos, resulting in abnormal development.     

Melittin, a Component of Bee Venom, Activates Unfertilized Sea Urchin Eggs

Melittin, which is known to stimulate phospholipase A , in many cells, caused as much elevation of fertilization membranes and increase in respiration of unfertilized eggs of the sea urchins Anthocidaris crassispina and Hemicentrotus pulcherrimus as normal fertilization.
In melittin-activated eggs, amino acid transport was decreased to less than that of unfertilized eggs, nucleoside transport was only slightly, activated, protein synthesis was rather inhibited and neither DNA synthesis nor cleavage was observed. It is concluded that although melittin induces the cortical reaction and activation of respiration in unfertilized eggs, its cytotoxicity prevents any "late changes".     

Regeneration failure of lakeshore plants under an artificially altered water regime

To reveal the effects of artificial alteration of water level regime on the regeneration of lakeshore plants from seeds, we examined the factors causing regeneration failure in Lake Kasumigaura, Japan. A survey of microtopography within and around a remnant fragment of lakeshore vegetation revealed that, over a large range, the habitat is frequently inundated in spring under the current water regime, although it was rarely inundated under past water regimes. Analysis of the patterns of seedling emergence and establishment at microsites at various elevations revealed a significant negative correlation between number of inundation days and abundance or species-richness of seedlings that emerged in the spring. Most seedling deaths occurred when the study site was inundated. We suggest that regeneration failure caused by the artificial raising of the lakes water level is one of the principal mechanisms of the recent vegetational decline in the lake.     

INHIBITION OF RESPIRATION IN SEA URCHIN SPERMATOZOA FOLLOWING INTERACTION WITH FIXED UNFERTILIZED EGGS

The respiration of spermatozoa of the sea urchin, Hemicentrotus pulcherrimus , was found to be sensitive to rotenone, antimycin A, and cyanide. This suggests that sperm respiration results from electron transport which spans the whole mitochondrial respiratory chain. The sperm respiration was inhibited by oligomycin and this inhibition was released by 2, 4-dinitrophenol (DNP). DNP did not stimulate the respiration of spermatozoa in a diluted suspension (2 × 10⁸/ml), where they were swimming vigorously. The ADP level of spermatozoa in the diluted suspension was markedly higher than that in dry sperm. The spermatozoa, which had reacted with unfertilized eggs fixed with glutaraldehyde, were immotile with a quite low respiratory rate. The respiratory rate of the immotile spermatozoa was enhanced by DNP. In the immotile spermatozoa, ADP level was markedly low and the ATP level was as high as that in dry sperm. From these findings, it is concluded that in the swimming spermatozoa respiration coupled with oxidative phosphorylation occurs at the maximum rate. State 3 respiration probably occurs in the swimming spermatozoa. The low respiratory rate of the immotile spermatozoa is assumed to be due to a shortage of ADP and is practically regarded as state 4 respiration.     

Some further studies on metabolism of acetate-l-14C in potassium deficient rice leaves

Acetate-1-¹⁴C was fed to excised leaves of normal and potassiumdeficient rice plants. The rate of respiratory evolution of¹⁴CO₂ was increased by a potassium deficiency. Malonate, glyoxylateor malate supplied enhanced the metabolism of acetate-l-¹⁴C.These results suggest an accelerated turn of the TCA cycle inpotassium deficient leaves. Malate synthetase activity was notrecognized in either normal or deficient leaves. However, condensingenzyme activity was higher in deficient leaves than in normalones. An addition of DNP to the leaves increased ¹⁴CO₂ productionfrom acetate-l-¹⁴C though its effect was smaller in deficientleaves than it was in normal ones. This result may suggest anincrease in the turnover rate of ATP or loose coupling of electrontransfer with oxidative phosphorylation in deficient leaves.Chromatographic separation of cold acidsoluble nucleotides hasshown that the ATP level was lowered by a potassium deficiency,though the ADP level was not affected. ¹Present address: Sericultural Experiment Station, Suginami-Ku,Tokyo     

Growth allocation between height and stem diameter in nonsuppressed reproducing Abies mariesii trees

To understand the growth patterns with respect to competition and leaf‐mass increase in reproducing trees, growth allocation between height and stem diameter was examined for nonsuppressed reproducing Abies mariesii trees in a subalpine forest in northern Honshu, Japan. The growth allocation was analyzed by dividing the relative growth rate of the stem volume into the relative contributions of height and stem‐diameter growth. During a 9‐yr period, height growth and seed‐cone production showed obvious annual variation, while stem‐diameter growth recorded moderate variation. For two of three years of seed‐cone production during the 9‐yr period, trees with larger seed‐cone production were associated with less height growth in the following year of seed‐cone production; however, there was no trend of height growth in the year of seed‐cone production. In the following year of mast seeding, trees with larger stem‐volume growth were associated with less height growth. This trend was also shown for the relationship between the cumulative stem‐volume growth during the 9‐yr period and growth allocation to height, suggesting that trees with a larger biomass increase depress the allocation of photosynthate to competition with a large expenditure for reproduction. In contrast to this, trees with a smaller biomass increase might allocate photosynthate to competition with surrounding trees. The results of this study suggest that an increase in reproductive organs during life history and annual variation in reproduction are closely associated with the growth patterns of the stem in A. mariesii trees.