The silkmoth chorion: Morphogenesis of surface structures and its relation to synthesis of specific proteins

The morphogenesis of four spatially differentiated surface regions of the silkmoth eggshell (chorion) has been documented and correlated with differing patterns of chorion protein synthesis within the corresponding secretory cells. During the first half of choriogenesis the polygonal pattern of ridges which cover the entire chorion appears. Regional differences in the morphology of developing ridges are not accompanied by significant protein differences, and thus presumably reflect differences in secretory cell behavior and shape. During the second half of choriogenesis expanding domes of the chorion located immediately beneath three-cell junctions of the overlying secretory surface become prominent surface features exclusively in the aeropyle crown region. Domes are composed of a thin lamellar skin and an inner buttressing “filler.” Continued filler deposition appears to cause a ripping of the lamellar skin, transforming the dome into a multiple-pronged crown that overflows with filler. Continued synthesis of lamellar chorion components elongates and strengthens the crowns until they can stand alone without the support of filler. In the aeropyle crown region, synthesis of regionally specific proteins begins in the second half of choriogenesis and accelerates until the final stages, in parallel with dome/crown formation. The more numerous proteins which are common to all regions are synthesized at approximately equal rates within all regions, and their synthesis decelerates toward the end of choriogenesis. Fifteen of the proteins (excluding filler) which are found predominantly in the aeropyle crown region may be necessary but not sufficient for crown formation, since they also occur in the stripe region (1); presumably the secretory cell surfaces mold the same components differently in the two regions. Filler appears to play an important scaffolding role in crown formation. A group of eight aeropyle crown region-specific chorion proteins which compose filler have been identified on two-dimensional gels and shown to be restricted to one of five previously described classes of chorion proteins.     

Secretory kinetics in the follicular cells of silkmoths during eggshell formation

Procedures for quantitative autoradiography were used for studying the process of secretion of eggshell (chorion) proteins in the follicular epithelium of silkmoths. The method was based on photometric measurements of the reflectance of vertically illuminated autoradiographic silver grains. Results were analyzed and plotted by computer. Secretory kinetics were also determined by analysis of labeled proteins in physically separated epithelium and chorion. Rapid accumulation of radioactivity into "clumps" visualized by light microscope autoradiography and evidence from preliminary electron microscope autoradiography indicate that, within 2 min from the time of synthesis, labeled chorion proteins move to Golgi regions scattered throughout the cytoplasm. The proteins begin to accumulate in the apical area 10-20 min later and to be discharged from the cell. The time for half-secretion is 20-25 min, and discharge is essentially complete 30-50 min after labeling. At the developmental stages examined, the kinetics of secretion appear to be similar for all proteins. Within the chorion the proteins rapidly assume a characteristic distribution, which varies for different developmental stages. Two relatively slow steps have been identified in secretion, associated with residence in Golgi regions and in the cell apex, respectively. By contrast, translocation of proteins across the cell and deposition of discharged proteins in the chorion are rapid steps.     

Sperm penetration and transformation of sperm entry site in eggs of the freshwater teleost Rhodeus ocellatus ocellatus

Eggs of bony fishes are enveloped by an egg envelope (chorion) in which a micropyle is present near the animal pole. Therefore, sperm penetration into the eggs is limited to the sperm entry site (SES), a region of plasma membrane just beneath the micropyle. In rose bitterling eggs, the SES transforms from a tuft of microvilli into a swollen mass (SM) that continues to plug the micropyle after sperm penetration. The present observations using the rose bitterling Rhodeus ocellatus ocellatus were conducted to examine: 1) whether or not sperm penetration is necessary for formation of the SM and 2) whether or not actin microfilaments are involved in the formation of the SM. Water activation without sperm transformed the SES from a tuft of microvilli into the SM, although it took a longer time for the transformation and the SMs were smaller than in the case of inseminated eggs. The SES presumably has the ability to transform into the SM upon activation of eggs in the present species. Cytochalasin B, which acts on actin microfilaments, did not prevent formation of the SM, irrespective of insemination or activation. The present observations suggest that sperm penetration is not necessary for SM formation and actin microfilaments do not participate in SM formation. © 1996 Wiley-Liss, Inc.     

The Structure of the Plumule of Nelumbo Nucifera Gaertn. and the Nature of its Scale

The structure of the plumule of Nelumbo nucifera Gaertn. and its feature covered with scale are seldom seen in dicotyledon. The fact that the plumule possesses scale is even more uncommon. This particular phenomenon is investigated by observing the differentiation of the plumule apex and the development of the leaf organs. After the seed is formed, the embryo has two young leaves and a terminal bud covered with scale. In the bud it has already differentiated the 3rd and the 4th leaf primordium and a shoot apex, the differentiation of which is very complex. So the structure of the plumule passes through 4 plastochrons altogether. It is made clear through observation and analysis that, before the 4th leaf primordium is formed, the transforma- tions of the shoot apex of the embryo in each plastochron are fundamentally alike. After the 4th leaf primordium is developed, the shoot apex becomes complex and there appear 3 different active cell regions which become the bases of vegetative bud of the seeding apex. The development of these 3 active cell regions will be stated in “The Structure of the Vegetative Bud of Nelumbo nucifera Gaertn. and the Nature of its Scales.” The apices of the plumule are almost slightly domed in structure. As a rule, their width is from 95 to 107 μ. Their height is from 17 to 20 μ during one plastochron. Before the 3rd leaf initiation, the anatomical structure of apices is examined and the fol- lowing zones may be delimited: zone of tunica initials, zone of corpus initials, peripheral zone, and zone of rib meristems. It is frequently observed that the cell of corpus in subapical peripheral zone develops periclinal division, which is the initial cell of leaf primordium; Procambium will appear before the stage of the appearance of leaf buttress. The apex of the plumule is in an apical position, but when the seedling is formed, as the developing leaves are alternate, the directions of the shoot apex are changed, simultaneously the base part of the leaf encloses the axis, and the adaxial meristem also differentiates the scale which encloses the terminal bud, thus placing the bud in axillary of the leaf and forming a zigzag phenomenon of the axis of the seedling. Above the basal adaxial side of the leaf primordium develops the scale of the plumule with meristem periclinal division of closely attached protoderm as its base. So the scale of the plumule of Nelumbo nucifera Gaertn. and the axillary stipule are of the same origin. To sum up, the scale of the embryo of Nelumbo nucifera Gaertn. is differentiated from the adaxial meristem of the basal part of the leaf primordium, and is the derivative part of the leaf. It has the same function as the coleoptile of the monocotyledon. Whether they are homologous organs or not is still to be investigated.     

Sphingomyelin metabolism is involved in the differentiation of MDCK cells induced by environmental hypertonicity

Sphingolipids (SLs) are relevant lipid components of eukaryotic cells. Besides regulating various cellular processes, SLs provide the structural framework for plasma membrane organization. Particularly, SM is associated with detergent-resistant microdomains. We have previously shown that the adherens junction (AJ) complex, the relevant cell-cell adhesion structure involved in cell differentiation and tissue organization, is located in an SM-rich membrane lipid domain. We have also demonstrated that under hypertonic conditions, Madin-Darby canine kidney (MDCK) cells acquire a differentiated phenotype with changes in SL metabolism. For these reasons, we decided to evaluate whether SM metabolism is involved in the acquisition of the differentiated phenotype of MDCK cells. We found that SM synthesis mediated by SM synthase 1 is involved in hypertonicity-induced formation of mature AJs, necessary for correct epithelial cell differentiation. Inhibition of SM synthesis impaired the acquisition of mature AJs, evoking a disintegration-like process reflected by the dissipation of E-cadherin and β- and α-catenins from the AJ complex. As a consequence, MDCK cells did not develop the hypertonicity-induced differentiated epithelial cell phenotype.     

The fine structure of conidial development in the genus Torula. I. T. herbarum (Pers.) Link ex S. F. Gray and T. herbarum f. quaternella Sacc.

Conidiogenesis in Torula herbarum and T. herbarum f. quaternella was observed by scanning and transmission electron microscopy. Conidia of the former were shown to be made up of three equally sized cells capped by a distinctive, and easily recognizable, conidiogenous cell. Conidiogenous cells also arose terminally on erect hyphae and on prostrate hyphae. The single-layered conidial cell walls were differentiated into an inner hyaline zone and an outer electron-dense zone formed by the deposition of melanin. Conidiogenous cells lacked melanin at the apex and, before conidiation, the lateral walls were strengthened by a further deposition of melanin. The apex bulged outwards and was modified into a new multicelled conidium bearing another apical conidiogenous cell. Continued development of new conidia resulted in an acropetal chain which became disarticulated after cytolysis within the conidiogenous cell. The relative distinctions between holoblastic and enteroblastic development are discussed and it is concluded that the conidia should be referred to as blastoconidia.     

SPECIFIC PROTEIN SYNTHESIS IN CELLULAR DIFFERENTIATION : Production of Eggshell Proteins by Silkmoth Follicular Cells

Silkmoth follicles, arranged in a precise developmental sequence within the ovariole, yield pure and uniform populations of follicular epithelial cells highly differentiated for synthesis of the proteinaceous eggshell (chorion). These cells can be maintained and labeled efficiently in organ culture; their in vitro (and cell free) protein synthetic activity reflects their activity in vivo. During differentiation the cells undergo dramatic changes in protein synthesis. For 2 days the cells are devoted almost exclusively to production of distinctive chorion proteins of low molecular weight and of unusual amino acid composition. Each protein has its own characteristic developmental kinetics of synthesis. Each is synthesized as a separate polypeptide, apparently on monocistronic messenger RNA (mRNA), and thus reflects the expression of a distinct gene. The rapid changes in this tissue do not result from corresponding changes in translational efficiency. Thus, the peptide chain elongation rate is comparable for chorion and for proteins synthesized at earlier developmental stages (1.3–1.9 amino acids/sec); moreover, the spacing of ribosomes on chorion mRNA (30–37 codons per ribosome) is similar to that encountered in other eukaryotic systems.     

平滑肌22α蛋白在血管稳态和血管重构中的作用

有关血管稳态和重构的分子机制一直是近年来的研究热点,也被视为治疗血管损伤性疾病的突破点.大量研究证实,血管损伤修复及病理性重构过程与血管平滑肌细胞(vascular smooth muscle cells,VSMCs)的表型转化、异常增殖与迁移、细胞衰老关系密切.平滑肌22α(smooth muscle 22α,SM2...     

Analysis of cell movements and fate mapping during early embryogenesis in Drosophila melanogaster

Four spatially differentiated surface regions, called aeropyle crown, flat, stripe, and micropyle, are found on the mature eggshell (chorion). Specializations of the apical surfaces of the secretory follicular epithelial cells are implicated in the formation of regional patterns on the chorion. Some of these specializations are restricted to cells overlying certain regions; others are shared by more than one region. Differences between regions are more apparent on the surface than within the bulk of the chorion. Evidence is presented that distinct cell populations, corresponding to the regions, are present long before the start of choriogenesis. One hundred eighty-six chorion-specific polypeptides have been resolved by two-dimensional gel electrophoresis. Fifteen of these are found entirely or predominantly in the aeropyle crown and stripe regions, while eight others are restricted to the aeropyle crown region. Certain of the spatially restricted components are quite unusual in their amino acid compositions when compared with previously analyzed chorion components. Others are closely related, although clearly distinct.     

Phenotypic changes in the regenerating rabbit bladder muscle. Role of interstitial cells and innervation on smooth muscle cell differentiation

 We have studied the phenotypic changes in regenerating smooth muscle (SM) tissue of detrusor muscle after local application of a necrotizing, freeze–thaw injury to the serosal surface of rabbit bladder. Bromo-deoxyuridine (BrdU) incorporation and immunofluorescence studies were performed on bladder cryosections from day 2 up to day 15 after surgery with monoclonal antibodies specific for some cytoskeletal markers [desmin, vimentin, non-muscle (NM) myosin] and for SM-specific proteins (α-actin, myosin, and SM22). Four days after lesion, some clls incorporated in regenerating SM bundles are BrdU positive, but all display a phenotypic pattern identical to that of the interstitial, highly proliferating cells, i.e., expression of vimentin. By days 7–15 the differentiation profile of regenerating SM returns to that of uninjured SM tissue (appearance of desmin, SM-type α-actin, and SM myosin). A chemical denervation achieved by 6-hydroxydopamine treatment for 2 weeks induces the formation of vimentin/SM α-actin/NM myosin/SM22-containing myofibroblasts in the interstitial, fibroblast-like cells of uninjured bladder. In the bladder wall, alteration of reinnervation during the regenerating SM process produces: (1) in the outer region, the activation of vimentin/SM α-actin/desmin myofibroblasts in the de novo SM cell bundles; and (2) in the inner region of bladder, including the muscularis mucosae, the formation of proliferating, fully differentiated SM cells peripherally to newly formed SM cell bundles. These findings suggest that: (1) the de novo SM tissue formation in the bladder can occur via incorporation of interstitial cells into growing SM bundles; and (2) the alteration of reinnervation during the regenerating process induces a spatial-specific differentiation of interstitial myofibroblasts in SM cells before SM cell bundling. Accepted: 14 May 1997     

Chorion gene amplification in Drosophila: A model for metazoan origins of DNA replication and S-phase control.

The mechanisms controlling duplication of the metazoan genome are only beginning to be understood. It is still unclear what organization of DNA sequences constitutes a chromosomal origin of DNA replication, and the regulation of origin activity during the cell cycle has not been fully revealed. We review recent results that indicate that chorion gene amplification in follicle cells of the Drosophila ovary is a model for investigating metazoan replication. Evaluation of cis sequence organization and function suggests that chorion loci share attributes with other replicons and provides insights into metazoan origin structure. Moreover, recent results indicate that chorion origins respond to S-phase control, but escape mechanisms that inhibit other origins from firing more than once in a cell cycle. Several identified genes that mediate amplification are critical for the cell cycle control of replication initiation. It is likely that further genetic screens for mutations that disrupt amplification will identify the cadre of proteins associated with origins and the regulatory pathways that control their activity. Furthermore, the recent development of methods to detect amplification in situ has uncovered new aspects of its developmental control. Examining this control will reveal links between developmental pathways and the cell cycle machinery. Visualization of amplifying chorion genes with high resolution also represents an opportunity to evaluate the influence of nuclear and chromosome structure on origin activity. The study of chorion amplification in Drosophila, therefore, provides great potential for the genetic and molecular dissection of metazoan replication.     

Purification and cell-surface marker characterization of quiescent satellite cells from murine skeletal muscle by a novel monoclonal antibody

A novel monoclonal antibody, SM/C-2.6, specific for mouse muscle satellite cells was established. SM/C-2.6 detects mononucleated cells beneath the basal lamina of skeletal muscle, and the cells co-express M-cadherin. Single fiber analyses revealed that M-cadherin+ mononucleated cells attaching to muscle fibers are stained with SM/C-2.6. SM/C-2.6+ cells, which were freshly purified by FACS from mouse skeletal muscle, became MyoD+ in vitro in proliferating medium, and the cells differentiated into desmin+ and nuclear-MyoD+ myofibers in vitro when placed under differentiation conditions. When the sorted cells were injected into mdx mouse muscles, donor cells differentiated into muscle fibers. Flow cytometric analyses of SM/C-2.6+ cells showed that the quiescent satellite cells were c-kit-, Sca-1-, CD34+, and CD45-. More, SM/C-2.6+ cells were barely included in the side population but in the main population of cells in Hoechst dye efflux assay. These results suggest that SM/C-2.6 identifies and enriches quiescent satellite cells from adult mouse muscle, and that the antibody will be useful as a powerful tool for the characterization of cellular and molecular mechanisms of satellite cell activation and proliferation.     

血清饥饿可诱导人血管平滑肌细胞再分化

体外培养的分化型血管平滑肌细胞 (vascularsmoothmusclecells ,VSMC)以特异性标志基因表达、长梭形外观及对兴奋剂刺激产生收缩反应为其表型特征 .以血清饥饿法培养处于超汇合 (overconfluence)状态的人VSMC ,观察其分化型标志基因表达活性及其与细胞形态特征和收缩反应性之间的关系 ,探讨细胞生存环境对VSMC基因表达及表型的影响 .研究显示 ,生长至超汇合的VSMC由含血清培养转为血清饥饿后 ,收缩蛋白如SMα肌动蛋白 (SMα actin)、SM2 2α、h1 calponin、肌球蛋白重链 (MHC)SM1和SM2亚型的表达活性明显上调 ,证实血清饥饿诱导的收缩蛋白基因表达和血清应答因子 (serumresponsefactor ,SRF)与CArG顺式元件结合活性的增强有关 .同时 ,血清饥饿还可激活参与VSMC分化调节的转录调控因子SmLIM、Gax和分化相关蛋白HRG 1基因的转录 .随着血清饥饿培养时间的延长 ,VSMC逐渐形成多层、束状、成极性排列的形式 ,对兴奋剂刺激产生的收缩反应明显增强 .结果表明 ,超汇合状态的去分化型VSMC脱离血清刺激后 ,可以再分化成熟并重新获得收缩能力     

The vitelline envelope,chorion, and micropyle of Fundulus heteroclitus eggs

The architecture and transformation of the vitelline envelope of the developing oocyte into the chorion of the mature egg of Fundulus heteroclitus have been examined by scanning and transmission electron microscopy. The mature vitelline envelope is structurally complex and consists of about nine strata. The envelope is penetrated by pore canals that contain microvilli arising from the oocyte and macrovilli from follicle cells. During the envelope's transformation into the chorion, the pore canals are lost and the envelope becomes more fibrous and compact and its stratified nature less apparent. The micropyle, of pore, through which the sperm gains access to the enclosed egg is located at the bottom of a small funnel-shaped depression in the envelope. Internally, the micropyle opens on the apex of a cone-like elevation of the chorion. During the development of the envelope, structured chorionic fibrils, the components of which are presumed to be synthesized by the follicle cells, become attached to its surface. These chorionic fibrils are though to aid in the attachment of the egg to the substratum and perhaps to help prevent water loss during low tides when the egg may be exposed.     

Ontogeny of Axillary and Accessory Buds in Clerodendrum phlomidis L.

Plastochronic changes in the vegetative shoot apex and originand development of axillary and accessory buds are studied. The flat shoot apex shows structural and dimensional changesin a plastochron. They are described in three phases, the pre-leafinitiation, the leaf initiation, and the post-leaf initiation.The youngest axillary bud meristem is identified near the axilat the second node when the subtending leaf primordium is 200–12µ long. The corpus of the bud meristem has a more activerole in bud development than has the tunica layers. The shellzone associated with a young bud meristem persists until thebud has attained the structural and functional attributes ofthe main shoot apex. It loses its histological identity by producingderivatives which merge with the ground tissue and procambialcells of bud traces. In a developing bud the provascular systemof the bud appears as an arc, a loop, or as a ring in transversesections at different levels. These configurations are composedof anastomosing procambial strands of bud trace and residualmeristem, both being differentiated from developing bud meristem.     

Changes of membrane phospholipid composition of human erythrocytes in hyperlipidemias. I. Increased phosphatidylcholine and reduced sphingomyelin in patients with elevated levels of triacylglycerol-rich lipoproteins.

The composition of red blood cell membrane and plasma phospholipids has been analyzed in patients with hyperlipidemias. In red cells of patients with elevated levels of triacylglycerol-rich lipoproteins, phosphatidylcholine (PC) was raised and sphingomyelin (SM) reduced, resulting in a 20% increase of the membrane PC/SM ratio. In plasma phospholipids of these patients PC and SM levels were also higher and lower, respectively and the plasma PC/SM ratio was elevated by more than 50%. Close positive correlations between plasma and membrane phospholipids were obtained for PC, SM and the PC/SM ratio in normolipidemic and hyperlipidemic donors. Plasmalogen phosphatidylethanolamine (PE), a supposed endogenous protector against lipid oxidation, was reduced by about 20% in red cell membrane lipids in hyperlipidemic patients. Also plasmalogen-PE in plasma tended to be reduced in hyperlipidemic donors. Plasma HDL levels were positively related to the content of plasmalogen PE in the red cell membrane. In conclusion, there are closely related increases in PC/SM ratios in plasma and the red cell membrane in patients with elevated levels of triacylglycerol-rich lipoproteins. It is speculated that decreases in red cell membrane plasmalogen-PE in hyperlipidemic patients could be related to impaired antioxidant protection, possibly as a consequence of reductions in plasma HDL levels.     

Early patterning of the chorion leads to the trilaminar trophoblast cell structure in the placental labyrinth

The labyrinth of the rodent placenta contains villi that are the site of nutrient exchange between mother and fetus. They are covered by three trophoblast cell types that separate the maternal blood sinusoids from fetal capillaries--a single mononuclear cell that is a subtype of trophoblast giant cell (sinusoidal or S-TGC) with endocrine function and two multinucleated syncytiotrophoblast layers, each resulting from cell-cell fusion, that function in nutrient transport. The developmental origins of these cell types have not previously been elucidated. We report here the discovery of cell-layer-restricted genes in the mid-gestation labyrinth (E12.5-14.5) including Ctsq in S-TGCs (also Hand1-positive), Syna in syncytiotrophoblast layer I (SynT-I), and Gcm1, Cebpa and Synb in syncytiotrophoblast layer II (SynT-II). These genes were also expressed in distinct layers in the chorion as early as E8.5, prior to villous formation. Specifically, Hand1 was expressed in apical cells lining maternal blood spaces (Ctsq is not expressed until E12.5), Syna in a layer immediately below, and Gcm1, Cebpa and Synb in basal cells in contact with the allantois. Cebpa and Synb were co-expressed with Gcm1 and were reduced in Gcm1 mutants. By contrast, Hand1 and Syna expression was unaltered in Gcm1 mutants, suggesting that Gcm1-positive cells are not required for the induction of the other chorion layers. These data indicate that the three differentiated trophoblast cell types in the labyrinth arise from distinct and autonomous precursors in the chorion that are patterned before morphogenesis begins.