The Cell Cycle and Cell Population Kinetics in the Programmed Cell Death in the Limb-Buds of Normal and 5-Bromodeoxyuridine- Treated Chick Embryos

The cell cycle and cell population kinetics have been analyzed in the interdigital regions of chick limb-buds during the course of programmed cell death both in normal and the 5-bromodeoxyuridine (BrdU)-treated embryos. Our previous study has shown that a single administration of BrdU at day 6 1/3 inhibited the programmed cell death occurring in normal development of limb-buds.
Pulse- as well as continuous labelings with tritiated thymidine (³H-TdR) were used. The results obtained from the analyses made on both normal and experimental embryos have demonstrated the presence of a particular DNA-synthetic period, around day 6 1/3, closely related to the programmed death occurring on day 7 1/3. In normal embryos, new cell populations, which did not belong to any phases of normal cell cycle, made their appearances in the process of programmed cell death. A possible correlation between programmed cell death and the cell cycle has been discussed in relation to the morphogenesis of limbs in both normal and BrdU-treated embryos.     

Bombyx acid cysteine proteinase

Summary

Three kinds of yolk proteins (vitellin, egg-specific protein and 30 k-proteins) are found in silkmoth eggs and have been well characterized. Essentially these proteins are considered to be amino acid reserves for developing embryos. Since at an early stage of egg development the cysteine proteinase accounts for the majority of the total proteinase activity, it may be involved in the degradation of yolk proteins. The enzyme is stored in the eggs as an inactive pro-form, indicating that the activation of the enzyme might be one of the key steps in yolk protein degradation. To investigate at the molecular level how yolk proteins degradation takes place, we have studied Bombyx acid cysteine proteinase (BCP) during an early period of embryonic development. We summarize how proteinases are regulated and are involved in the degradation of Bombyx yolk proteins during embryogenesis. These will be discussed mainly in light of recent results obtained from eggs of the silkmoth, Bombyx mori.     

Ontogenetic histology of Stegosaurus plates and spikes

Abstract: The dinosaur Stegosaurus is characterized by osteoderms of alternating plates and terminal paired spikes. Previous studies have described the histological features and possible functions of these osteoderms. However, ontogenetic changes are poorly documented. In this study, the ontogenetic changes of the osteoderms are examined using eight different ontogenetic skeletons (a juvenile, a subadult, a young adult, and five old adults based on the cortical histology of their body skeletons). The juvenile plate and subadult spike show thin cortex and thick cancellous bone. The young adult plates have an extensive vascular network, which is also seen in old adults. Old adult spikes are different from old adult plates in having a thick cortex and a large axial channel. The cortical histology, in both plates and spikes, show well‐vascularized bone tissue consisting of dense mineralized fibres in young adult forms. In old adult forms, the bone tissues in the spikes become more compact and are extensively remodelled. This might contribute to the structural reinforcement of the spikes. The plates in old adult forms also show extensive remodelling and lines of arrested growth, but only limited signs of compaction. The timing for acquisition of features seen in old adults is different between plates (an extensive vascular network in the young adult) and spikes (a thick cortex with a large axial channel in old adults). The result suggests that the timing for plate and spike functions is different. The extensive vascular networks seen in large plates suggest their function is for display and/or thermoregulation. The thick cortical bone of spikes of old adults suggests that spikes acquire a weapon function for defence ontogenetically late.     

Microdissection Studies on the Polarity of Unequal Division in Grasshopper Neuroblasts: II. Cell Division in Binucleate Neuroblasts

The grasshopper neuroblast divides unequally to produce two types of cells: a large daughter neuroblast that contains a doughnut-shaped nucleus and repeats unequal division with definite polarity, and a small daughter ganglion cell that has a spherical nucleus with low mitotic activity. Binucleate neuroblasts were induced by preventing cytokinesis in the course of microdissection experiments, and subsequent divisions were traced to analyze the factors that determine the polarity of unequal division.
In binucleate neuroblasts, both daughter chromosome groups developed into neuroblast-type nuclei. Mitosis of the two nuclei proceeded synchronously. Although the axes of the two mitotic apparatuses formed at late prophase were random in direction, they became parallel with the original division axis at metaphase. The two mitotic apparatuses shifted simultaneously toward the ganglion cell side during anaphase, just as in normal neuroblasts, and the binucleate cell divided unequally. These findings showed that the poearity of unequal division is strictly maintained in grasshpper neuroblasts, even when they contain two nuclei.     

Impacts of light and temperature on shoot branching gradient and expression of strigolactone synthesis and signalling genes in rose

Light and temperature are two environmental factors that deeply affect bud outgrowth. However, little is known about their impact on the bud burst gradient along a stem and their interactions with the molecular mechanisms of bud burst control. We investigated this question in two acrotonic rose cultivars. We demonstrated that the darkening of distal buds or exposure to cold (5 °C) prior to transfer to mild temperatures (20 °C) both repress acrotony, allowing the burst of quiescent medial and proximal buds. We sequenced the strigolactone pathway MAX‐homologous genes in rose and studied their expression in buds and internodes along the stem. Only expressions of RwMAX1, RwMAX2 and RwMAX4 were detected. Darkening of the distal part of the shoot triggered a strong increase of RwMAX2 expression in darkened buds and bark‐phloem samples, whereas it suppressed the acropetal gradient of the expression of RwMAX1 observed in stems fully exposed to light. Cold treatment induced an acropetal gradient of expression of RwMAX1 in internodes and of RwMAX2 in buds along the stem. Our results suggest that the bud burst gradient along the stem cannot be explained by a gradient of expression of RwMAX genes but rather by their local level of expression at each individual position.     

The Inhibitory Effect of 5-Bromodeoxyuridine on the Programmed Cell Death in the Chick Limb

The inhibitory effect of 5-bromodeoxyuridine (BrdU) on the programmed cell death in interdigital mesenchyme of a chick leg bud was studied using vital staining and histological methods. A single administration of BrdU at day 6 1/3 specifically inhibited the programmed cell death in the mesenchyme of interdigital areas of a leg bud, resulting in the formation of a web-like structure. No inhibitory effect was observed on general development of limb buds. Hoechst 33258 staining revealed that many cells of interdigits incorporated BrdU into their nuclei. The simultaneous administration of thymidine blocked the BrdU effect.