Timing of oral morphogenesis and its relation to commitment to division in Paramecium tetraurelia

The interval between commitment to division and fission in synchronous cell samples is a constant fraction of the cell cycle (0.2) in cell cycles up to 6.5 h in duration. In longer cell cycles this interval has a fixed duration of about 80 min. The point of commitment to division is associated with the six-rowed analage stage of oral primordium development (stage V). At this stage cells carrying the cc1 mutation are not blocked by transfer to restrictive conditions but rather proceed to division. Stage V is also the stabilization point for oral anlagen. When shifted to restrictive conditions prior to this stage, development is arrested and resorption of analgen is initiated. The cc1 mutation also blocks contractile vacuole duplication and migration under restrictive conditions. The cc1 gene function is required continuously prior to the transition point. The timing of morphogenetic stages in asynchronous cells is roughly similar to that in synchronous cells. There are, however, significant differences in timing as estimated by the two experimental procedures.     

IL-15 treatment during acute simian immunodeficiency virus (SIV) infection increases viral set point and accelerates disease progression despite the induction of stronger SIV-specific CD8+ T cell responses

In this study, we examined the effect of in vivo treatment of acutely SIV-infected Mamu-A*01+ rhesus macaques with IL-15. IL-15 treatment during acute infection increased viral set point by 3 logs and accelerated the development of simian AIDS in two of six animals with one developing early minimal lesion SIV meningoencephalitis. Although IL-15 induced a 2- to 3-fold increase in SIV-specific CD8+ T cell and NK cell numbers at peak viremia and reduced lymph node (LN) SIV-infected cells, this had no impact on peak viremia and did not lower viral set point. At viral set point, however, activated SIV-specific CD8+ T cells and NK cells were reduced in the blood of IL-15-treated animals and LN SIV-infected cells were increased. Week 30 LN from IL-15-treated animals had significantly increased Gag-specific CD8+ T cell numbers, whereas total cell, lymphocyte, and CD4+ T cell numbers were reduced. IL-15 treatment significantly reduced anti-SIV Ab concentrations at week 3 and viral set point. IL-15 increased Ki-67+CD4+ T cells at week 1 of treatment and reduced blood CCR5+ and CD45RA-CD62L- CD4+ T cells. The frequency of day 7 Ki-67+CD4+ T cells strongly correlated with viral set point. These findings suggest that CD4+ T cell activation during acute infection determines subsequent viral set point and IL-15 treatment by increasing such activation elevates viral set point. Finally, IL-15-treated acutely SIV-infected primates may serve as a useful model to investigate the poorly understood mechanisms that control viral set point and disease progression in HIV infection.     

Timing of oral morphogenesis and its relation to commitment to division in Paramecium tetraurelia

The interval between commitment to division and fission in synchronous cell samples is a constant fraction of the cell cycle (0.2) in cell cycles up to 6.5 h in duration. In longer cell cycles this interval has a fixed duration of about 80 min. The point of commitment to division is associated with the six-rowed anlage stage of oral primordium development (stage V). At this stage cells carrying the cc1 mutation are not blocked by transfer to restrictive conditions but rather proceed to division. Stage V is also the stabilization point for oral anlagen. When shifted to restrictive conditions prior to this stage, development is arrested and resorption of anlagen is initiated. The cc1 mutation also blocks contractile vacuole duplication and migration under restrictive conditions. The cc1 gene function is required continuously prior to the transition point. The timing of morphogenetic stages in asynchronous cells is roughly similar to that in synchronous cells. There are, however, significant differences in timing as estimated by the two experimental procedures.     

Cell-cycle checkpoint for transition from cell division to differentiation

In general, growth and differentiation are mutually exclusive, but they are cooperatively regulated during the course of development. Thus, the process of a cell's transition from growth to differentiation is of general importance for the development of organisms, and terminally differentiated cells such as nerve cells never divide. Meanwhile, the growth rate speeds up when cells turn malignant. The cellular slime mold Dictyostelium discoideum grows and multiplies as long as nutrients are supplied, and its differentiation is triggered by starvation. A critical checkpoint (growth/differentiation transition or GDT point), from which cells start differentiating in response to starvation, has been precisely specified in the cell cycle of D. discoideum Ax-2 cells. Accordingly, integration of GDT point-specific events with starvation-induced events is needed to understand the mechanism regulating GDTs. A variety of intercellular and intracellular signals are involved positively or negatively in the initiation of differentiation, making a series of cross-talks. As was expected from the presence of the GDT point, the cell's positioning in cell masses and subsequent cell-type choices occur depending on the cell's phase in the cell cycle at the onset of starvation. Since novel and multiple functions of mitochondria in various respects of development including the initiation of differentiation have been directly realized in Dictyostelium cells, they are also reviewed in this article.     

Macrophages kill capillary cells in G1 phase of the cell cycle during programmed vascular regression

Programmed capillary regression occurs during normal development of the eye and serves as a useful model for assessing the forces that drive vascular involution. Using a combination of S-phase labeling and liposome-mediated macrophage elimination, we show that during regression, macrophages induce apoptosis of both pericytes and endothelial cells in a cell cycle stage-dependent manner. Target cells are signaled to die by macrophages approximately 15 hours after S-phase labeling and this corresponds to a point in mid-G1 phase of the cell cycle. The tight correlation between the restriction point of the cell cycle and the point where the macrophage death signal is received suggests that the mitogen, matrix and cytoskeletal signals essential for cell-cycle progression may be inhibited by macrophages as a means of inducing cell death. Furthermore, these experiments show that cells from two distinct lineages are induced to die as a consequence of macrophage action, and this provides evidence that macrophage-induced cell death may be a general phenomenon during development and homeostasis.     

Simultaneous measurement of passage through the restriction point and MCM loading in single cells

Passage through the Retinoblastoma protein (RB1)-dependent restriction point and the loading of minichromosome maintenance proteins (MCMs) are two crucial events in G1-phase that help maintain genome integrity. Deregulation of these processes can cause uncontrolled proliferation and cancer development. Both events have been extensively characterized individually, but their relative timing and inter-dependence remain less clear. Here, we describe a novel method to simultaneously measure MCM loading and passage through the restriction point. We exploit that the RB1 protein is anchored in G1-phase but is released when hyper-phosphorylated at the restriction point. After extracting cells with salt and detergent before fixation we can simultaneously measure, by flow cytometry, the loading of MCMs onto chromatin and RB1 binding to determine the order of the two events in individual cells. We have used this method to examine the relative timing of the two events in human cells. Whereas in BJ fibroblasts released from G0-phase MCM loading started mainly after the restriction point, in a significant fraction of exponentially growing BJ and U2OS osteosarcoma cells MCMs were loaded in G1-phase with RB1 anchored, demonstrating that MCM loading can also start before the restriction point. These results were supported by measurements in synchronized U2OS cells.     

Induction of meiosis in Saccharomyces cerevisiae at different points in the mitotic cycle

Summary Saccharomyces cells induced to undergo meiosis when in late G ₁ or early S-phase, proceed mitotically until a point between completion of the S-phase and nuclear division. From that point, the cells start meiotic development without intervention of a round of premeiotic DNA replication. Cells induced at any other point in the cell cycle, enter meiosis from G ₁.     

颈卵器植物颈细胞结构与功能的研究进展

颈卵器是苔藓、蕨类、裸子植物的雌性生殖器官,由卵细胞、腹沟细胞和颈细胞(颈沟细胞)构成。其中,颈细胞是这类植物雄配子进入颈卵器并完成受精作用的唯一通道,在颈卵器的发育和受精过程中发挥着重要作用。该文就近年来国内外有关对颈细胞的发生和发育、结构特点和功能等进行较为系统全面的分析和总结,并对未来的研究方向进行展望。     

Stepwise development of thymic microenvironments in vivo is regulated by thymocyte subsets

T-cell development is under the tight control of thymic microenvironments. Conversely, the integrity of thymic microenvironments depends on the physical presence of developing thymocytes, a phenomenon designated as 'thymic crosstalk'. We now show, using three types of immunodeficient mice, i.e. CD3(epsilon) transgenic mice, RAG(null) mice and RAG(null)-bone-marrow-transplanted CD3(epsilon) transgenic mice, that the control point in lymphoid development where triple negative (CD3(-),CD4(-),CD8(-)) thymocytes progress from CD44(+)CD25(-) towards CD44(-)CD25(+), influences the development of epithelial cells, critically inducing the extra, third dimension in the organization of the epithelial cells in the cortex. This tertiary configuration of the thymic epithelium is a typical feature for the thymus, enabling lymphostromal interaction during T-cell development. Crosstalk signals at this control point also induce the formation of thymic nurse cells. Moreover, our data indicate that establishment of a thymic cortex is a prerequisite for the development of the thymic medulla. Thus, differentiating thymocytes regulate the morphogenesis of thymic microenvironments in a stepwise fashion.     

Effects of brefeldin A on the development of the Casparian strip in pea epicotyls

Summary The Casparian strip, a structure that is present in roots, is also present in epicotyls of dark-grown pea seedlings. In a dark-grown epicotyl, the cells in each stage of the development of the Casparian strip have been suggested to be lined up basipetally in the region 3 to 37 mm below the bending point of the hook, in order of the developmental stage. Brefeldin A (BFA), a specific inhibitor of secretory transport, was administrated at 200 M. to dark-grown pea epicotyls for 2 h via a thread passed through the epicotyl 40 mm below the bending point. The basipetal sequence of development of the modification of the cell wall at the Casparian strip, as judged by fluorescence microscopy, stopped 5 h after the start of 2 h treatment with BFA and resumed after 30 h. This basipetal sequence of development did not stop in control seedlings. Electron micrographs of endodermal cells in epicotyls treated with BFA showed striking morphological changes in the Golgi stacks and the ER. Histological examination made 20 h after the start of the experiment revealed that the basipetal sequence of development of the cell wall modification stopped at a point which was present at 25.2 ± 1.6 mm (mean with SD, n=5) from the bending point of the hook at the start while the basipetal sequence of development of the tight adhesion of the plasma membrane to the cell wall at the Casparian strip stopped 0.9 ± 0.5 mm (mean with SD, n=5) below this point. These results indicate the involvement of secretory transport not only in the introduction of the modification of the cell wall but also in the completion of the tight adhesion of the plasma membrane.Abbreviations BFA brefeldin A - PBS phosphate-buffered saline - ER endoplasmic reticulum     

JAM-A is both essential and inhibitory to development of hepatic polarity in WIF-B cells

Junctional adhesion molecule (JAM) is involved in tight junction (TJ) formation in epithelial cells. Three JAMs (A, B, and C) are expressed in rat hepatocytes, but only rat JAM-A is present in polarized WIF-B cells, a rat-human hepatic line. We used knockdown (KD) and overexpression in WIF-B cells to determine the role of JAM-A in the development of hepatic polarity. Expression of rat JAM-A short hairpin RNA resulted in approximately 50% KD of JAM-A and substantial loss of hepatic polarity, as measured by the absence of apical cysts formed by adjacent cells and sealed by TJ belts. When inhibitory RNA-resistant human JAM-A (huWT) was expressed in KD cells, hepatic polarity was restored. In contrast, expression of JAM-A that either lacked its PDZ-binding motif (huDeltaC-term) or harbored a point mutation (T273A) did not complement, indicating that multiple sites within JAM-A's cytoplasmic tail are required for the development of hepatic polarity. Overexpression of huWT in normal WIF-B cells unexpectedly blocked WIF-B maturation to the hepatic phenotype, as did expression of three huJAM-A constructs with single point mutations in putative phosphorylation sites. In contrast, huDeltaC-term was without effect, and the T273A mutant only partially blocked maturation. Our results show that JAM-A is essential for the development of polarity in cultured hepatic cells via its possible phosphorylation and recruitment of relevant PDZ proteins and that hepatic polarity is achieved within a narrow range of JAM-A expression levels. Importantly, formation/maintenance of TJs and the apical domain in hepatic cells are linked, unlike simple epithelia.     

Gap junctions and connexin-mediated communication in the immune system

Gap junctions and connexins are present in the immune system. In haematopoiesis, connexin 43, the most widely distributed gap junction protein, appears to be a key player in the development of progenitor cells and their communication with stromal cells. Connexin 43 is expressed by macrophages, neutrophils and mast cells. Lymphocytes also express connexin 43, and inhibition of gap junction channels in these cells by using highly specific connexin mimetic reagents has profound effects on immunoglobulin secretion and synthesis of cytokines. Lymphocytes and leukocytes also communicate directly in vitro with endothelial cells via gap junctions. Connexins are implicated in inflammatory reactions in a range of tissues. Their involvement in atherosclerotic plaque formation in the vascular system is also a current growth point in research, and could lead to the development of therapeutic interventions.     

干细胞治疗感音神经性耳聋

治疗内耳疾病的主要困难之一是找到耳蜗毛细胞或者螺旋神经元丢失所导致的听力损失的治疗方法。本文讨论使用干细胞替代感觉细胞丢失为目的的几个治疗策略。作者最近在成年内耳中发现了可以分化为毛细胞的干细胞,发现了胚胎干细胞可在体外转化为毛细胞并表达毛细胞标记物。在动物模型中,成年内耳干细胞、神经干细胞和胚胎干细胞来源的前体细胞可分化成为毛细胞和神经细胞。本文将讨论使用干细胞再生损伤毛细胞的不同方法,介绍几种可行的动物模型,并讨论发展基于干细胞的细胞替代疗法治疗内耳损伤中存在的困难。     

Kinetic aspects of follicle development in the rat

The eighth and ninth generations of follicle growth in rats represent a turning point in development. This stage is characterized by establishing a complete regulatory control based on feedback in follicle development. The feedback between follicles and gonadotropin secretion regulates a number of follicles maturing for ovulation. Gonadotropins seem to play a permissive, and not a directive part in regulation of follicle development in the eighth and ninth generations. By this stage of development, possibilities for theca and granule cells become sharply limited. Whereas expression of many maturation features may be hastened or hindered by changes in hormonal status, the result of development cannot be changed. Although only last generations of theca and granule cells exhibit mature functional features, their precursors seem to become committed to a single direction of development at early stages of follicle development. Neither the stage when precursor cells become irreversibly committed to differentiation into granule or theca cells, neither regulatory factors which determine this process have been identified yet. We suppose that precursor cells become committed to thecal tissue compartment when the follicle is at a primordial stage of development. Precursors of all the follicle components may already be assembled into one unit by the beginning of follicle growth. Accumulation of sufficient number of precursor cells around the primordial follicle may serve as a signal for follicle growth initiation. We think that the understanding of follicle postnatal growth and development should be based on understanding of origins, destiny, and possibilities of cells which form ovary and its compartments. First generations of follicle growth seem to be most promising for future research.     

Microvascular development: learning from pancreatic islets

Microvascular development is determined by the interplay between tissue cells and microvascular endothelial cells. Because the pancreatic islet is an organ composed mainly of endothelial and endocrine cells, it represents a good model tissue for studying microvascular development in the context of a tissue. In this review, we will describe the special morphology of islet capillaries and its role in the physiologic function of islets: secretion of insulin in response to blood glucose levels. We will speculate on how islet-secreted VEGF-A generates a permeable endothelium that allows insulin to pass quickly into the blood stream. In addition, we speculate on how endothelial cells might form a capillary lumen within the islets. At the end, we look at the islet microvasculature from a medical point of view, thus describing its critical role during type I diabetes and islet transplantation.     

The mystique of irreversibility in cyanobacterial heterocyst formation: parallels to differentiation and senescence in eukaryotic cells

The formation of cyanobacterial heterocysts is unique in the prokaryotic world: it is the only irreversible collective process. This terminal differentiation resembles senescence and differentiation in the eukaryotic urkingdom. During their cell cycle eukaryotic cells at the restriction point may reversibly proceed from a vegetative phase (G1) into a quiescent state (G0), and then may irreversibly enter the way towards differentiated or senescent cells. In parallel, at commitment point 1 vegetative cells from filamentous cyanobacteria may reversibly form proheterocysts, and then may proceed irreversibly towards mature heterocysts at commitment point 2. While the signals paving the path for differentiation or senescence in eukaryotes are largely unknown, heterocyst development is clearly triggered by nitrogen starvation. The reasons for the irreversibility in both systems are poorly understood. We discuss these questions, especially in the light of recent advances in the molecular biology of cyanobacteria, with emphasis on self-stabilizing autocatalytic cycles.     

Expression pattern for adrenomedullin during pancreatic development in the rat reveals a common precursor with other endocrine cell types

Adrenomedullin is an α-amidated 52-amino acid peptide involved in many physiological actions, among others the regulation of insulin secretion. Using immunohistochemical methods, we found that adrenomedullin immunoreactivity first appears at day 11.5 of embryonic development in the rat, coinciding with the appearance of pancreatic glucagon. The early appearance of adrenomedullin in the developing pancreas may indicate an active involvement in either the morphogenesis of the organ or its endocrine/paracrine/autocrine hormone regulation during intrauterine life. We also investigated the pattern of colocalizations of adrenomedullin with the other pancreatic hormones. At some point during development all the cell types express adrenomedullin, progressively evolving towards the adult pattern where only the pancreatic polypeptide cells contain a strong immunoreactivity for adrenomedullin. At this point the remaining cells of the islet are, in general, weakly stained. This sequential and time-dependent expression of adrenomedullin suggests a tight regulation similar to that observed for other modulatory substances responsible for embryonic morphogenesis.     

Transverse viscoelastic extension in nitella: I. Relationship to growth rate

Transverse viscoelastic extensibility was measured directly in isolated walls of Nitella internode cells. Cell walls extended transversely exhibit a yield point which is approximately twice the yield point in the longitudinal direction. Walls from young, growing cells are four to seven times more extensible longitudinally than transversely, while walls from mature, nongrowing cells are only two times more extensible longitudinally. Although longitudinal extensibility decreases drastically with the decrease in the growth rate, lateral extensibility is constant through development. There is a discrepancy between the lateral growth rate and transverse creep, since the lateral growth rate is not constant. However, the degree of wall anisotropy observed is consistent with the view that the transversely oriented cellulose microfibrils act as a “reinforcing filler” in Nitella cell walls.