Developmental studies with the 14-3-2 antigen and the neuron specific enolase (NSE) associated activities—I

We have compared the rodent developmental pattern of the 14-3-2 antigen estimated by a microcomplement fixation technique with that of the cerebral enolases. Chromatographic separation of enolase isozymes on microcolumns demonstrates that the embryonic neuron specific enolase is firstly and mostly represented by the αγ isozyme. The most important increase in 14-3-2 antigen and γγ enolase occurs between post-natal days 7th and 15th. By post-natal day 30, adult levels have been reached. An interesting observation is—during embryonic development—the decrease in the specific activity of the cerebral enolase isozyme αα. This could be explained by the replacement—in neuroblasts—of αα enolase by neuron specific enolase. A comparison between 14-3-2 antigen and neuron specific enolase (γγ) purified by completely different methods is presented. The 14-3-2 antigen exhibits an enolase specific activity comparable to that of purified enzyme and has the same electrophoretic mobility. Antibodies raised against either antigen have an identical specificity. Pre and post-natal developmental pattern in rodent brains are similar for both proteins. Thus neuron specific 14-3-2 antigen is identical to neuron specific enolase.Thus we have precisely described the ontogenic transition between the three cerebral enolase isozymes at the tissue level. This study is completed by the analysis of these transitions at the neuronal cell level, using homogenous cell lines (Part II of this paper).     

Origin and development of the germ line in sea stars

This review summarizes and integrates our current understanding of how sea stars make gametes. Although little is known of the mechanism of germ line formation in these animals, recent results point to specific cells and to cohorts of molecules in the embryos and larvae that may lay the ground work for future research efforts. A coelomic outpocketing forms in the posterior of the gut in larvae, referred to as the posterior enterocoel (PE), that when removed, significantly reduces the number of germ cell later in larval growth. This same PE structure also selectively accumulates several germ‐line associated factors—vasa, nanos, piwi—and excludes factors involved in somatic cell fate. Since its formation is relatively late in development, these germ cells may form by inductive mechanisms. When integrated into the morphological observations of germ cells and gonad development in larvae, juveniles, and adults, the field of germ line determination appears to have a good model system to study inductive germ line determination to complement the recent work on the molecular mechanisms in mice. We hope this review will also guide investigators interested in germ line determination and regulation of the germ line into how these animals can help in this research field. The review is not intended to be comprehensive—sea star reproduction has been studied for over 100 years and many reviews are comprehensive in their coverage of, for example, seasonal growth of the gonads in response to light, nutrient, and temperature. Rather the intent of this review is to help the reader focus on new experimental results attached to the historical underpinnings of how the germ cell functions in sea stars with particular emphasis to clarify the important areas of priority for future research. genesis 52:367–377, 2014. © 2014 Wiley Periodicals, Inc.     

Cell genetic engineering: Transmission genetics of plants

Achievements of cell and genetic engineering that led to formation of a new genetics chapter— transmission genetics—have been described. Results have been analyzed and new opportunities in the field of transgenomic somatic hybrids and cybrid obtaining, production of transgenic plants with agronomic pharmaceutical application, development of transplastomic plants, and accumulation of recombinant proteins by using the transient expression of foreign genes in plants have been shown.     

Disorder-to-order conformational transitions in protein structure and its relationship to disease

Function in proteins largely depends on the acquisition of specific structures through folding at physiological time scales. Under both equilibrium and non-equilibrium states, proteins develop partially structured molecules that being intermediates in the process, usually resemble the structure of the fully folded protein. These intermediates, known as molten globules, present the faculty of adopting a large variety of conformations mainly supported by changes in their side chains. Taking into account that the mechanism to obtain a fully packed structure is considered more difficult energetically than forming partially “disordered” folding intermediates, evolution might have conferred upon an important number of proteins the capability to first partially fold and—depending on the presence of specific partner ligands—switch on disorder-to-order transitions to adopt a highly ordered well-folded state and reach the lowest energy conformation possible. Disorder in this context can represent segments of proteins or complete proteins that might exist in the native state. Moreover, because this type of disorder-to-order transition in proteins has been found to be reversible, it has been frequently associated with important signaling events in the cell. Due to the central role of this phenomenon in cell biology, protein misfolding and aberrant disorder-to-order transitions have been at present associated with an important number of diseases.     

Mechanobiology of the relocation of proteins in advecting cells: in vitro experiments,multi-physics modeling,and simulations

Cell motility—a cellular behavior of paramount relevance in embryonic development, immunological response, metastasis, or angiogenesis—demands a mechanical deformation of the cell membrane and influences the surface motion of molecules and their biochemical interactions. In this work, we develop a fully coupled multi-physics model able to capture and predict the protein flow on endothelial advecting plasma membranes. The model has been validated against co-designed in vitro experiments. The complete picture of the receptor dynamics has been understood, and limiting factors have been identified together with the laws that regulate receptor polarization. This computational approach might be insightful in the prediction of endothelial cell behavior in different tumoral environments, circumventing the time-consuming and expensive empirical characterization of each tumor.

     

Cytochemical observations of larval development in Eunapius fragilis (Leidy): Porifera; spongillidae

Freshwater sponges of the family Spongillidae reproduce sexually through formation of a parenchymula larva. The cytochemical characteristics of parenchymula larval metamorphosis — beginning with the blastula and terminating with the motile escape stage — for the spongillid Eunapius fragilis (Leidy) have been defined using both absorption and fluorescent cytochemical methods, particularly those demonstrating protein end-groups. Morphogenesis of the parenchymula larva of E. fragilis involves the interrelated processes of cytodifferentiation and mobilization of reserve materials. Larval development has been categorized into five stages, from blastula (stage I) through the escape stage (stage V). Parenchymula development is characterized by morphogenetic precocity, a fact influencing the rate of mobilization of cytoplasmic reserves, cytodifferentiation, and the fate of individual cell types. With attainment of the stage V parenchymula, the larva is, essentially, a mobile adult sponge exhibiting flagellated chambers, canal systems, a well defined connective tissue stroma, a diverse cell population consisting of specialized elements and a totipotent archeocyte reserve, and a terminal epitheliocyte line. The present study recognizes differences in development within the spongillids as well as within more remote poriferan taxa — emphasizing the need for detiled understanding of particular processes in individual species before proposing major generalizations about development in this ancient but evolutionally specialized group.     

Cell Churches and Stem Cell Marketing in South Korea and the United States

The commercial provision of putative stem cell‐based medical interventions in the absence of conclusive evidence of safety and efficacy has formed the basis of an unregulated industry for more than a decade. Many clinics offering such supposed stem cell treatments include statements about the ‘ethical’ nature of somatic (often colloquially referred to as ‘adult’ stem cells) stem cells, in specific contrast to human embryonic stem cells (hESCs), which have been the subject of intensive political, legal, and religious controversy since their first derivation in 1998 1 . Christian groups—both Roman Catholic and evangelical Protestant—in many countries have explicitly promoted the medical potential and current‐day successes in the clinical application of somatic stem cells, lending indirect support to the activities of businesses marketing stem cells ahead of evidence 2 . In this article, I make a preliminary examination of how the structures and belief systems of certain churches in South Korea and the United States, both of which are home to significant stem cell marketing industries, has complemented other factors, including national biomedical funding initiatives, international economic rivalries, permissive legal structures, which have lent impetus to a problematic and often exploitative sector of biomedical commerce 3 .     

Jamming and arrest of cell motion in biological tissues

Collective cell motility is crucial to many biological processes including morphogenesis, wound healing, and cancer invasion. Recently, the biology and biophysics communities have begun to use the term ‘cell jamming’ to describe the collective arrest of cell motion in tissues. Although this term is widely used, the underlying mechanisms are varied. In this review, we highlight three independent mechanisms that can potentially drive arrest of cell motion — crowding, tension-driven rigidity, and reduction of fluctuations — and propose a framework that connects all three. Because multiple mechanisms may be operating simultaneously, this emphasizes that experiments should strive to identify which mechanism dominates in a given situation. We also discuss how specific cell-scale and molecular-scale biological processes, such as cell–cell and cell-substrate interactions, control aspects of these underlying physical mechanisms.     

Autophagy and Cancer Therapy

Autophagy is a dynamic process of protein degradation which is typically observed during nutrient deprivation. Recently, interest in autophagy has been renewed among oncologists, because different types of cancer cells undergo autophagy after various anticancer therapies. This type of non-apoptotic cell death has been documented mainly by observing morphological changes, e.g., numerous autophagic vacuoles in the cytoplasm of dying cells. Thus, autophagic cell death is considered programmed cell death type II, whereas apoptosis is programmed cell death type I. These two types of cell death are predominantly distinctive, but many studies demonstrate cross-talk between them. Whether autophagy in cancer cells causes death or protects cells is controversial. In multiple studies, autophagy has been inhibited pharmacologically or genetically, resulting in contrasting outcomes—survival or death—depending on the specific context. Interestingly, the regulatory pathways of autophagy share several molecules with the oncogenic pathways activated by tyrosine kinase receptors. Tumor suppressors such as Beclin 1, PTEN, and p53 also play an important role in autophagy induction. Taken together, these accumulating data may lead to development of new cancer therapies that manipulate autophagy.     

花粉发育的研究进展

近年来,随着现代生物学技术及分子生物学技术在花粉生物学研究领域中广泛的应用,从而使花粉发育过程中一些关键问题及其有关分子机理的研究均获得了重要进展。目前已克隆了许多与花粉发育有关的基因,并鉴定了一些花粉发育的突变体。本文针对花粉发育过程中的细胞质改组;胼胝质壁、淀粉粒和细胞器的动态变化;绒毡层的发育;花粉发育特异基因;以及不对称分裂和细胞命运等几个生物学问题的研究进展作了简要综述,最后还提出了一些研究展望。     

花粉发育的研究进展

近年来,随着现代生物学技术及分子生物学技术在花粉生物学研究领域中广泛的应用,从而使花粉发育过程中一些关键问题及其有关分子机理的研究均获得了重要进展。目前已克隆了许多与花粉发育有关的,并鉴定了一些花粉发育的突变体。本文针对花偻发育过程中的细胞质改组;胼胝质壁、淀粉粒和细胞器的动态变化;绒毡层的发育;花粉发育特异基因;以及不对称分裂和细胞命运等几个生物学问题的研究进展作了简要综述,最后还提出了一些研究展望。     

Stem cells in the embryonic cerebral cortex: Their role in histogenesis and patterning

The cytoarchitectural simplicity of the cerebral cortex makes it an attractive system to study central nervous system (CNS) histogenesis—the process whereby diverse cells are generated in the right numbers at the appropriate place and time. Recently, multipotent stem cells have been implicated in this process, as progenitor cells for diverse types of cortical neurons and glia. Continuous analysis of stem cell clone development reveals stereotyped division patterns within their lineage trees, highly reminiscent of neural lineage trees in arthropods and Caenorhabditis elegans. Given that these division patterns play a critical part in generating diverse neural types in invertebrates, we speculate that they play a similar role in the cortex. Because stereotyped lineage trees can be observed from cells growing at clonal density, cell-intrinsic factors are likely to have a key role in stem cell behavior. Cortical stem cells also respond to environmental signals to alter the types of cells they generate, providing the means for feedback regulation on the germinal zone. Evidence is accumulating that cortical stem cells, influenced by intrinsic programs and environmental signals, actually change with development—for example, by reducing the number and types of neurons they produce. Age-related changes in the stem cell population may have a critical role in orchestrating development; whether these cells truly self-renew is a point of discussion. In summary, we propose that cortical stem cells are the focus of regulatory mechanisms central to the development of the cortical cytoarchitecture. © 1998 John Wiley & Sons, Inc. J Neurobiol 36: 162–174, 1998     

'Troy-bodies': antibodies as vector proteins for T cell epitopes

A major objective in vaccine development is the design of reagents that give a strong, specific T cell response. Targeting of antigens to antigen presenting cells (APC) results in enhanced antigen presentation and T cell activation. In this paper, we describe a novel targeting reagent denoted 'Troy-bodies', namely recombinant antibodies with APC-specificity and with T cell epitopes integrated in their C regions. We have made such antibodies with V regions specific for either IgD or MHC class II, and five different T cell epitopes have been tested. All epitopes could be introduced into loops of C domains without disrupting immunoglobulin (Ig) folding. Four have been tested in T cell activation studies, and all could be released and presented by APC. Furthermore, whether IgD- or MHC-specific, the molecules tested enhanced T cell stimulation compared to non-specific control antibodies in vitro as well as in vivo. Using this technology, specific reagents can be designed that target selected antigenic peptides to an APC of choice. Troy-bodies may therefore be useful for manipulation of immune responses, and in particular for vaccination purposes.     

Genetic control of macrochaetae development in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The Drosophila head and body have a regular species-specific pattern of strictly defined number of external sensory organs—macrochaetae (large bristles). The pattern constancy and relatively simple organization of each bristle organ composed of only four specialized cells makes macrochaetae a convenient model to study the developmental patterns of spatial structures with a fixed number of elements in specific positions as well as the mechanisms of cell differentiation. The experimental data on the major genes and their products controlling three stages of macrochaetae development—the emergence of proneural clusters in the imaginal disc ectoderm, the precursor cell determination in the proneural clusters, and the specialization of cells of the definitive sensory organ—were reviewed. The role of the achaete-scute gene complex, EGFR and Notch signaling, and selector genes in these processes was considered. Analysis of published data allowed us to propose an integrated diagram of the system controlling macrochaetae development in D. melanogaster.     

Mechanisms of axon regeneration: The significance of proteoglycans

BackgroundTherapeutics specific to neural injury have long been anticipated but remain unavailable. Axons in the central nervous system do not readily regenerate after injury, leading to dysfunction of the nervous system. This failure of regeneration is due to both the low intrinsic capacity of axons for regeneration and the various inhibitors emerging upon injury. After many years of concerted efforts, however, these hurdles to axon regeneration have been partially overcome.Scope of reviewThis review summarizes the mechanisms regulating axon regeneration. We highlight proteoglycans, particularly because it has become increasingly clear that these proteins serve as critical regulators for axon regeneration.Major conclusionsStudies on proteoglycans have revealed that glycans not only assist in the modulation of protein functions but also act as main players—e.g., as functional ligands mediating intracellular signaling through specific receptors on the cell surface. By regulating clustering of the receptors, glycans in the proteoglycan moiety, i.e., glycosaminoglycans, promote or inhibit axon regeneration. In addition, proteoglycans are involved in various types of neural plasticity, ranging from synaptic plasticity to experience-dependent plasticity.General significanceAlthough studies on proteins have progressively facilitated our understanding of the nervous system, glycans constitute a new frontier for further research and development in this field. This article is part of a Special Issue entitled Neuro-glycoscience, edited by Kenji Kadomatsu and Hiroshi Kitagawa.     

Renouvellement des protéines et effet spécifique de la kinétine sur des cultures de cellules de Tabac

Determination and fractionation of proteins of tobacco cell suspensions requireing kinetin for cell division. — Cell suspensions either in stationary phase without kinetin in the medium or dividing in the presence of this factor have been compared. It was found a) that the specific rates of protein synthesis and protein degradation were not changed by the addition of kinetin during the early period of growth. A quantitative change ocurred only after the first generation period, b) Soluble proteins of these cells were mapped by polyacrylamide gel electrophoresis. The observed protein patterns were very similar as well as the patterns or radioactivity incorporated into the same proteins during the incubation period of the cells. However, a small number of specific discrepancies appeared in the pattern of cells growing in the presence of kinetin matched to the patterns of stationary cells. At least one specific difference in these patterns could be observed before the first cell division occurred.