High temperature causes arrest of cell cycle in G2 phase in BmN cells derived from the silkworm, Bombyx mori

Abstract.  The influence of temperature on the insect cell line, BmN, derived from the silkworm, Bombyx mori is investigated. These cells proliferate at an accelerated pace as the temperature increases from 22 to 30 °C, but the growth rate slows at 34 °C, and proliferation stops at 38 °C. At high temperatures, abnormal cellular morphology is observed. Cells treated at 38 °C have cytoplasmic bilateral protrusions and they gradually aggregate and float in the medium. BmN cells without proliferation at 38 °C are viable but have reduced DNA synthesis. At high temperatures, the cell cycle of BmN cells halts at the G₂ phase. After heat treatment of the larvae, an accumulation of larval haemocytes with high DNA content is found, which suggests that the cell cycle arrest at G₂ also occurs in the silkworm at high temperatures.     

Quantitative Analysis of Intercellularly-Transported Photoassimilates in Chara corallina

Using a thin-layer chromatographic technique, we have identifiedthe photoassimilates that are transported intercellularly frombranchlets to internodes in Chara corallina. An internode-branchletcomplex having a primary apex was used in these experiments.After feeding 1 mol m³ NaH¹⁴CO₃ to a branchlet for 10 min, the¹⁴C-labelled photoassimilates (¹⁴C-photoassimilates) found inthe sol endoplasm of the branchlet were composed of sucrose,amino acids, malate, and sugarphosphates. The composition ofthe ¹⁴C-photoassimilates transported from the source branchletto the sink internode in 10 min was the same as that in thesol endoplasm of the source branchlet. From the proportion ofeach ¹⁴C-photoassimilate in both the source branchlet and thesink internode, it was deduced that the main photoassimilatesinvolved in the intercellular transport were sucrose and aminoacids. We found previously that polar transport of photoassimilatesoccurs from a branchlet to an internode with an apex. Determinationof the amount of sucrose, amino acids, glucose-6-phosphate,and malate in both branchlet and internode with or without anapex revealed that there were gradients in the concentrationsof sucrose, serine, and glutamic acid between the sol endoplasmof the two cells. The levels were higher in the branchlet andlower in the internode and the gradients decreased when theapex was detached. Therefore, it is concluded that sucrose andthese amino acids are the compounds involved in the polar intercellulartransport. Key words: Chara corallina, intercellular transport, photoassimilates, ¹⁴C     

Intercellular Transport and Subcellular Distribution of Photoassimilates in Chara corallina

The mechanism of polar transport of ¹⁴C-photoassimilates betweenChara corallina branchlets and internodes was investigated usingan intracellular perfusion procedure. When the apex remained,more ¹⁴C-photoassimilates were transported from branchlet tointernode than in the opposite direction. When the apex wasdetached, this polarity disappeared. To study the polar transportmechanism, we examined the subcellular distribution of ¹⁴C-photoassimilatesin the three main intracellular compartments, the cortical chloroplastlayer, streaming sol endoplasm and vacuole. When the internode or branchlet of an internode-branchiet complexwas exposed to ¹⁴C-bicarbonate for 10 min, 70 to 80% of thetotal fixed carbon was found in the sol endoplasm. In the subsequent3 h chase, the ¹⁴C-photoassimilates in the sol endoplasm ofthe source cell decreased greatly to 20 to 40% of the totallabelled substances. When the apex remained, the branchlet showed higher photosyntheticactivity than the internode, which resulted in a concentrationgradient of 13·5 mol m^–3 in the sol endoplasm frombranchlet to internode after photosynthesis for 10 mm. In the3 h chase period, this gradient decreased to about 1·0mol m^–3. In the other three cases, involving transportfrom branchlet to internode when the apex was detached, or transportfrom internode with or without apex to branchlet, the intercellulargradient of ¹⁴C after 10 mm photosynthesis was about 70 molm^–3 in the sol endoplasm. Thus, when the apex remains,the greater gradient of photoassimilates between the sol endoplasmof branchlet and internode seems to be one of the reasons forpolar intercellular transport. Key words: Chara corallina, polar transport, ¹⁴C photoassimilates     

CELL POPULATION KINETICS OF THE SCLEROBLAST DURING ETHISTERONE-INDUCED ANAL-FIN PROCESS FORMATION IN ADULT FEMALES OF THE MEDAKA, ORYZIAS LATIPES

Proliferation and migration of the scleroblasts and their precursor cells during ethisterone-induced anal-fin process formation in adult females of the medaka, Oryzias latipes , were investigated using a radioautographic technique with ³H-thymidine. In the posterior margin of the joint plate, rapid increases in labeling indexes and scleroblast population began simultaneously in the central portion on the third day of ethisterone treatment, and the population became approximately ten times that in the control fish on the fifth day. However, nomarked increase in scleroblast population occurred in the peripheral portion, although the scleroblasts showed nearly the same labeling index as those in the central portion throughout the experiment. These observations indicate that the scleroblasts and their precursor cells migrated from the peripheral to the central portion along the proxi-distal axis of the joint plate. Some cells in the inner and hind spaces also migrated to the posterior margins and subsequently developed into scleroblasts. It may be concluded, therefore, that the scleroblasts are derived from precursor cells situated not only along the posterior margins but also in the adjacent spaces. These precursor cells proliferate and/or migrate to the central portion of the posterior margins and aggregate to form the scleroblast mass.     

Historical divergence of mechanical isolation agents in the ground beetle Carabus arrowianus as revealed by phylogeographical analyses

In the carabid genus Carabus subgenus Ohomopterus , diverged body size and genital morphology serve as mechanical reproductive barriers. To elucidate the diverging process of body and genital sizes in Carabus arrowianus , which exhibits marked morphological diversity among geographical populations and may represent an early stage of speciation, we analysed a mitochondrial gene sequence for 1051 individuals from 63 populations and male morphology for 359 individuals from 47 populations. Two discrete morphological groups segregated by geographical barriers were distinguished, one of which possessed smaller bodies and shorter genitalia ( S group) than the other ( L group), which exhibited larger bodies and exaggerated genitalia. Genetic divergence between the two groups was significant but not large. Phylogeographical and population genetic analyses indicated that the L group was derived from the S group, and a coalescent simulation revealed that the two groups diverged during the latest middle Pleistocene (0.13 million years ago), with a much larger effective population size in the L group than the S group. Because the body size divergence could not be explained by adaptation to climatic conditions and genital morphology is considered to be subject to sexual selection, we postulated that a population division and colonization in favourable habitats caused by the Pleistocene climatic and geographical change might facilitate natural and sexual selection for enlarged body and genital sizes in the L group.