The planarian <Emphasis Type="Italic">nanos</Emphasis>-like gene Smednos is expressed in germline and eye precursor cells during development and regeneration

Planarians are highly regenerative organisms with the ability to remake all their cell types, including the germ cells. The germ cells have been suggested to arise from totipotent neoblasts through epigenetic mechanisms. Nanos is a zinc-finger protein with a widely conserved role in the maintenance of germ cell identity. In this work, we describe the expression of a planarian nanos-like gene Smednos in two kinds of precursor cells namely, primordial germ cells and eye precursor cells, during both development and regeneration of the planarian Schmidtea mediterranea. In sexual planarians, Smednos is expressed in presumptive male primordial germ cells of embryos from stage 8 of embryogenesis and throughout development of the male gonads and in the female primordial germ cells of the ovary. Thus, upon hatching, juvenile planarians do possess primordial germ cells. In the asexual strain, Smednos is expressed in presumptive male and female primordial germ cells. During regeneration, Smednos expression is maintained in the primordial germ cells, and new clusters of Smednos-positive cells appear in the regenerated tissue. Remarkably, during the final stages of development (stage 8 of embryogenesis) and during regeneration of the planarian eye, Smednos is expressed in cells surrounding the differentiating eye cells, possibly corresponding to eye precursor cells. Our results suggest that similar genetic mechanisms might be used to control the differentiation of precursor cells during development and regeneration in planarians. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Developmental expression of tryptophan hydroxylase gene in <Emphasis Type="Italic">Ciona intestinalis</Emphasis>

To describe the serotonergic system in a tunicate larva, we cloned a gene encoding for tryptophan hydroxylase (TPH), the rate-limiting enzyme in serotonin synthesis, in the ascidian Ciona intestinalis and studied its expression pattern during development. Ci-TPH expression was found from tailbud stage in the precursor cells of the visceral ganglion and in the tail. In the larva, TPH-expressing neurons formed two clusters in the anterior central nervous system at the level of the visceral ganglion. Moreover, we found Ci-TPH expression at the level of the muscle cells of the tail and suggested that this localisation might be at the level of neuro-muscolar junctions. Moreover, we discussed the involvement of serotonin in the control of larval locomotory activity.     

Evolutionary origin of the Otx2 enhancer for its expression in visceral endoderm

In the mouse, the Otx2 gene has been shown to play essential roles in the visceral endoderm during anterior-posterior axis formation and head induction. While these are primary processes in vertebrate embryogenesis, the visceral endoderm is a tissue unique to mammals. Two enhancers (VE and CM) have been previously found to direct Otx2 expression during early embryogenesis. This study demonstrates that in anterior visceral endoderm the CM enhancer does not have an activity by itself, but enhances the activity of the VE enhancer. These two enhancers also cooperate for the activities in anterior mesendoderm and cephalic mesenchyme. Comparative studies suggest that VE enhancer function was most likely established before the divergence of sarcopterygians into Actinistia, Dipnoi and tetrapods, while the nucleotide sequence corresponding to the VE enhancer was already present in the last common ancestor of bony fishes. The CM enhancer sequence and function would have been also established in ancestral sarcopterygians. The VE/CM enhancers and their gene cascades in the ancestral sarcopterygian head organizer would then have been co-opted by amphibian deep endoderm cells and mammalian visceral endoderm cells for the head development.     

中国少数民族体成分的变化

本文对中国少数民族23352例（男10070例,女13282例）的体成分进行了分析,以了解中国少数民族脂肪率、肌肉量的现状,探讨体成分随年龄增长的变化规律。研究发现,男性和女性总体上属于超重水平,还没有达到肥胖水平。与南方族群男性相比,北方阿尔泰语系族群的男性四肢脂肪率高、内脏脂肪等级高、水分率低。南方族群中,藏缅语族群、苗瑶语族群、壮侗语族群的四肢脂肪率、内脏脂肪等级、水分率相对接近;南亚语系族群与这3个南方族群差距较大。与南方族群女性相比,北方阿尔泰语系族群的女性躯干、四肢肌肉量大,骨骼重;南方4个族群女性躯干、四肢肌肉量较小,骨骼较轻,骨量、肌肉量彼此接近。随年龄增长,男性骨量下降,水分率增大;上肢脂肪率减小,躯干脂肪率增大,内脏脂肪等级增大,即脂肪向躯干集中,全身总体脂率增大;下肢肌肉量减少,躯干肌肉量下降,最终导致全身总肌肉量下降。随年龄增长,女性上肢的脂肪率和肌肉量没有明显变化,下肢的脂肪率下降,躯干脂肪率和内脏脂肪等级增大,总体脂率增大;躯干肌肉量下降,总肌肉量下降。男性推定骨量下降的节点是50岁,女性是60岁。男性总肌肉量下降的节点是40岁,女性是50岁。男性、女性身体水分率增加的节点都是60岁,内脏脂肪等级增加的节点都是30岁,总脂肪率下降的节点都是60岁,躯干脂肪率增加的节点都是30岁,躯干肌肉量下降的节点都是40岁。研究还发现,体脂率、内脏脂肪等级与血糖呈显著正相关。研究结果反映了中国少数民族从青年到老年的体成分变化的基本规律。     

Lineage analysis of transplanted individual cells in embryos of Drosophila melanogaster

Summary We describe the results of cell transplantation experiments performed to investigate mesodermal lineages in Drosophila melanogaster, particularly the lineages of the somatic muscles, the visceral muscles and the fat body. Cells to be transplanted were labelled by injecting a mixture of horseradish peroxidase (HRP) and fluorescein-dextran (FITC) in wild-type embryos at the syncytial blastoderm stage. For transplantation cells were removed from the ventral furrow, 8–12 min after the start of gastrulation, and individually transplanted into homotopic or heterotopic locations of unlabelled wild-type hosts of the same age. HRP labelling in the resulting cell clones was demonstrated histochemically in the fully developed embryo; histotypes could be distinguished without ambiguity. Mesodermal cells were already found to be committed to mesodermal fates at the time of transplantation. They developed only into mesodermal derivatives and did not integrate in non-mesodermal organs upon heterotopical transplantation. No evidence was found for commitment to any particular mesodermal organ at the time of transplantation. The majority of somatic muscle clones contributed cells to only one segment. However, clones were not infrequently distributed through two or even three segments. Clones of fat body cells were generally restricted to a small region. However, cells of clones of visceral musculature were widely distributed. With respect to the proliferative abilities of transplanted cells the clones were difficult to interpret due to the syncytial character of the somatic musculature and the fact that the organization of the other organs is poorly understood. Evidence from histological observations of developing normal embryos indicates only three mitoses for mesodermal cells. Clones larger than seven cells were not found when embryos were fixed previous to germ-band shortening; larger clones were found in the fat body and visceral musculature after fixing the embryos at the end of organogenesis. Quantitative considerations suggest that a few mesodermal cells might perform more than three mitoses.     

Beta 3 tubulin expression characterizes the differentiating mesodermal germ layer during Drosophila embryogenesis

During embryogenesis, the beta 3 tubulin gene of Drosophila is transcribed predominantly in the mesoderm. We have raised antibodies specific to the C-terminal domain of the beta 3 tubulin and analysed by immunostaining the distribution of this tubulin isotype during Drosophila embryogenesis. The protein is first detectable in the cephalic mesoderm at maximal germband extension. Shortly afterwards, beta 3 tubulin is expressed in single cells at identical positions of the thoracic and abdominal segments. We suggest that these cells represent muscle pioneer cells of Drosophila. During later embryonic development the somatic musclature, visceral musculature, dorsal vessel and macrophages contain beta 3 tubulin. In dorsalizing mutants dorsal, snail and twist, which do not form a ventral furrow during gastrulation, beta 3 expression is greatly reduced but not completely abolished. Our analysis shows that beta 3 tubulin immunostaining characterizes the differentiation of mesodermal derivatives during embryogenesis.     

Segmental differentiation processes in embryonic muscle development of the grasshopper

To analyse segmental differentiation processes in muscle development, we studied the embryogenesis of the ventral body wall muscles in thoracic and abdominal segments of the grasshopper Schistocerca gregaria at the identified cell level. We visualized differentiating muscle pioneer and muscle precursor cells by staining with a muscle-specific monoclonal antibody and with rhodamine-coupled phalloidin. Our results show that a similar pattern of serially reiterated early muscle pioneers is initially established in all segments. Subsequently, two major segmental differentiation processes occur. First, segment-specific sets of additional, later differentiating muscle pioneers are generated de novo. Second, segment-specific sets of existing early muscle precursors are eliminated through atrophy and eventual loss. These events have consequences for matching homonomy of muscles and their innervating motoneurons. Taken together, these processes in the embryo, in concert with postembryonic differentiation events, play critical roles in shaping the highly specialized muscular structures of the mature animal.     

Hypaxial muscle migration during primary myogenesis in Xenopus laevis.

In contrast to many vertebrates, the ventral body wall muscles and limb muscles of Xenopus develop at different times. The ventral body wall forms in the tadpole, while limb (appendicular) muscles form during metamorphosis to the adult frog. In organisms that have been examined thus far, a conserved mechanism has been shown to control migratory muscle precursor specification, migration, and differentiation. Here, we show that the process of ventral body wall formation in Xenopus laevis is similar to hypaxial muscle development in chickens and mice. Cells specified for the migratory lineage display an upregulation of pax3 in the ventro-lateral region of the somite. These pax3-positive cells migrate ventrally, away from the somite, and undergo terminal differentiation with the expression of myf-5, followed by myoD. Several other genes are selectively expressed in the migrating muscle precursor population, including neural cell adhesion molecule (NCAM), Xenopus kit related kinase (Xkrk1), and Xenopus SRY box 5 (sox5). We have also found that muscle precursor migration is highly coordinated with the migration of neural crest-derived melanophores. However, by extirpating neural crest at an early stage and allowing embryos to develop, we determined that muscle precursor migration is not dependent on physical or genetic interaction with melanophores.     

Fate of amplified nucleoli in Xenopus laevis embryos

We have followed the fate of two components of extrachromosomal nucleoli, amplified ribosomal DNA (rDNA) and 7.5 kb precursor rRNA, during early embryogenesis of Xenopus laevis. Other workers have shown that the amount of amplified rDNA accumulated during oogenesis remains unchanged through the 16-cell stage of embryogenesis. Here we show that as embryonic cleavage continues, the amount of amplified rDNA decreases until it is no longer detectable in the early gastrula embryo. In contrast, the amount of 7.5 kb precursor rRNA in eggs, early cleavage stage embryos, or blastula stage embryos is the same as in oocyte nuclei. Since no rRNA synthesis occurs during these early stages, we conclude that the precursor rRNA sequences synthesized in the oocyte are neither processed nor degraded during early development. The amplified rDNA is not replicated in the early embryo even though the chromosomal DNA of the embryo replicates every 30 min during the first 7.5 hr of embryogenesis. When amplified rDNA is purified and then injected into cleaving embryos, however, we find that it is replicated. This finding suggests that some factor(s) prevents the endogenous amplified rDNA from responding to the cellular replication signals. We show that methylation of cytosine in the rDNA is not related to the DNA's capacity for replication in this system since amplified (unmethylated) and chromosomal (methylated) rDNA are both replicated when injected into embryos. The methylation pattern of these rDNAs appears to be maintained after replication in the embryo.     

Differential expression of sex-related fat body proteins during the larval–pupal developmental stages of the silkworm (Bombyx mori)

The silkworm fat body is the site of many intermediary metabolic processes, and a source of sustenance for growth throughout the life cycle. Fat body proteins are responsible for storing nutrients, providing energy, and regulating hormones, and they have been identified using proteomic approaches. However, detailed differential expression of sex-related fat body proteins has not previously been evaluated. In the present study, we characterized the differential expression of sex-related fat body proteins, by using 2-dimensional gel electrophoresis (2-DE) followed by mass spectrometry identification and bioinformatics methods. We extracted the fat body proteins from 5-day-old fifth instar larvae (L5), 10-day-old fifth instar larvae (corresponding to the end of spinning [LE]), and 0-day-old pupae (P0) of the multivoltine silkworm variety “Da Zao”. We confirmed the presence of 11 important sex-specific expression proteins and 14 stage-specific expression proteins. We accurately identified 13 of these specific expression proteins, including actin, calponin-like protein, 75 kDa subunit NADH, receptor for activated protein kinase C from Bombyx mori (BmRACK), IMP (inosine monophosphate) cyclohydrolase, tropomyosin 1, β-tubulin, hypothetical protein, antichymotrypsin precursor, and 30 K protein precursor. We showed that BmRACK was differentially expressed between male and female silkworms. We discuss the biological roles of the specific expression proteins during the larval–pupal developmental stages.     

Characterization of cell clusters in larval hemolymph of the cabbage armyworm Mamestra brassicae and their role in maintenance of hemocyte populations

Maintenance of hemocyte populations is critical for both development and immune responses. In insects, the maintenance of hemocyte populations is regulated by mitotic division of circulating hemocytes and by discharge from hematopoietic organs. We found cell clusters in the hemolymph of Mamestra brassicae larvae that are composed of small, spherical cells. Microscopic observations revealed that the cells in these clusters are similar to immature or precursor cells present in hematopoietic organs. The results of bromodeoxyuridine (BrdU) incorporation experiments demonstrate that these cells are mitotically active. Furthermore, these cells maintain their immature state and proliferate until late in the last larval instar. The results of in vitro experiments showed that most of the cells changed their morphology to one consistent with plasmatocytes or granulocytes, and that the change was promoted by addition of larval hemolymph to the culture medium, in particular when hemolymph was collected at a prepupal stage. Taken together, our results suggested that cells in clusters may be an additional source of hemocytes during larval development.     

Unique intertesticular tissue complex in larvae of Heliothis virescens (F.) (Lepidoptera : Noctuidae)

A unique heart-testis complex has been found in the last instar of the tobacco budworm, Heliothis virescens (F.) (Lepidoptera : Noctuidae). This complex consists of a previously undescribed muscle system, herein designated the cardiotesticular muscle, that attaches the heart to the testes, and a ubiquitous connective tissue sheath that overlays this muscle and surrounding fat body and pericardial cells. These interconnected organs form a sieve-like septum enclosing a sinus that contains clusters of hemocytes and lipid droplets. Lipid droplets appear to be released from the surface of the testes sheath. The cardiotesticular muscle is innervated by neuroendocrine nerves that may be the route by which the brain peptide, testis ecdysiotropin, reaches the testes. The cardiotesticular muscle and products sequestered in the cardiotesticular sinus may be involved in the fusion of the paired testes and maturation of the male reproductive system, because they are present only during this stage of development.     

Tissue-specificity of hemoglobin synthesis: Localization of heme synthesis in the subepidermal fat body of Chironomus thummi (Diptera)

Possible sites of heme synthesis in the fourth instar of Chironomus thummi were investigated by means of autoradiography of specific isotope incorporation. “Body wall” preparations, which include subepidermal and visceral fat body, oenocytes, muscle, epidermis, and cuticle, were cultured for 1 h in a medium containing tritiated-δ-aminolevulinic acid, a specific precursor to heme biosynthesis. Light-microscopic examination of autoradiographs of sections of the body walls indicates that the subepidermal fat body is the major site of incorporation of the precursor into heme. The visceral fat body shows few silver grains. Oenocytes, as well as muscle and epidermis, are characterized by absence of silver deposits. These findings indicate that the subepidermal fat body of Chironomus is the primary site of heme synthesis, and are discussed in relation to specific hemoglobin synthesis.     

布朗族成人的身体成分分析

本文采用生物阻抗分析法,研究了布朗族成人的体成分特点。我们在云南省测量了604例（男性248例,女性356例）布朗族成人19项身体成分指标,运用Excel 2003、Spss 19.0对其各项指标进行统计分析。结果显示,男性全身脂肪分布特征为躯干和下肢的脂肪率都大于上肢脂肪率,女性脂肪率从大到小依次为下肢、躯干、上肢;男、女性双侧下肢脂肪率和肌肉量接近,左上肢肌肉量低、脂肪率高;布朗族男性的身高、体质量、肌肉量、推定骨量、总能量代谢、水分率、内脏脂肪等级均大于女性,而体脂率、BMI小于女性。随着年龄的增长,布朗族成人身体肌肉量、骨量、下肢脂肪率、能量代谢等呈明显下降,而内脏脂肪等级明显增加。与云南汉族比较,布朗族成人的体脂率较低、肌肉较发达。