Essential role of the mitochondrial apoptosis-inducing factor in programmed cell death

Programmed cell death is a fundamental requirement for embryogenesis, organ metamorphosis and tissue homeostasis. In mammals, release of mitochondrial cytochrome c leads to the cytosolic assembly of the apoptosome-a caspase activation complex involving Apaf1 and caspase-9 that induces hallmarks of apoptosis. There are, however, mitochondrially regulated cell death pathways that are independent of Apaf1/caspase-9. We have previously cloned a molecule associated with programmed cell death called apoptosis-inducing factor (AIF). Like cytochrome c, AIF is localized to mitochondria and released in response to death stimuli. Here we show that genetic inactivation of AIF renders embryonic stem cells resistant to cell death after serum deprivation. Moreover, AIF is essential for programmed cell death during cavitation of embryoid bodies-the very first wave of cell death indispensable for mouse morphogenesis. AIF-dependent cell death displays structural features of apoptosis, and can be genetically uncoupled from Apaf1 and caspase-9 expression. Our data provide genetic evidence for a caspase-independent pathway of programmed cell death that controls early morphogenesis.     

Myosin-dependent junction remodelling controls planar cell intercalation and axis elongation

Shaping a developing organ or embryo relies on the spatial regulation of cell division and shape. However, morphogenesis also occurs through changes in cell-neighbourhood relationships produced by intercalation. Intercalation poses a special problem in epithelia because of the adherens junctions, which maintain the integrity of the tissue. Here we address the mechanism by which an ordered process of cell intercalation directs polarized epithelial morphogenesis during germ-band elongation, the developmental elongation of the Drosophila embryo. Intercalation progresses because junctions are spatially reorganized in the plane of the epithelium following an ordered pattern of disassembly and reassembly. The planar remodelling of junctions is not driven by external forces at the tissue boundaries but depends on local forces at cell boundaries. Myosin II is specifically enriched in disassembling junctions, and its planar polarized localization and activity are required for planar junction remodelling and cell intercalation. This simple cellular mechanism provides a general model for polarized morphogenesis in epithelial organs.     

Self-organizing optic-cup morphogenesis in three-dimensional culture

Balanced organogenesis requires the orchestration of multiple cellular interactions to create the collective cell behaviours that progressively shape developing tissues. It is currently unclear how individual, localized parts are able to coordinate with each other to develop a whole organ shape. Here we report the dynamic, autonomous formation of the optic cup (retinal primordium) structure from a three-dimensional culture of mouse embryonic stem cell aggregates. Embryonic-stem-cell-derived retinal epithelium spontaneously formed hemispherical epithelial vesicles that became patterned along their proximal-distal axis. Whereas the proximal portion differentiated into mechanically rigid pigment epithelium, the flexible distal portion progressively folded inward to form a shape reminiscent of the embryonic optic cup, exhibited interkinetic nuclear migration and generated stratified neural retinal tissue, as seen in vivo. We demonstrate that optic-cup morphogenesis in this simple cell culture depends on an intrinsic self-organizing program involving stepwise and domain-specific regulation of local epithelial properties.     

IkappaB kinase-alpha acts in the epidermis to control skeletal and craniofacial morphogenesis

IkappaB kinase-alpha (IKK-alpha) exhibits protein-kinase-dependent and -independent functions. Its kinase activity is required for lymphoid organogenesis and mammary gland development, whereas a kinase-independent activity is required for epidermal keratinocyte differentiation. In addition to failed epidermal differentiation, IKK-alpha-deficient mice exhibit abnormal skeletal and craniofacial morphogenesis. As similar defects are not exhibited by mice that experience systemic inhibition of NF-kappaB, we postulated that the morphogenetic defects in IKK-alpha-deficient mice are not caused by reduced NF-kappaB activity but instead are due to failed epidermal differentiation that disrupts proper epidermal-mesodermal interactions. We tested this hypothesis by introducing an epidermal-specific Ikka (also known as Chuk) transgene into IKK-alpha-deficient mice. Mice lacking IKK-alpha in all cell types including bone and cartilage, but not in basal epidermal keratinocytes, exhibit normal epidermal differentiation and skeletal morphology. Thus, epidermal differentiation is required for proper morphogenesis of mesodermally derived skeletal elements. One way by which IKK-alpha controls skeletal and craniofacial morphogenesis is by repressing expression of fibroblast growth factor (FGF) family members, such as FGF8, whose expression is specifically elevated in the limb bud ectoderm of IKK-alpha-deficient mice.     

实验红鲫对137Cs 辐射的氧化应激响应

摘要: 目的探讨实验红鲫对137 Cs 辐射的氧化应激响应和氧化应激生物标记物。方法实验红鲫分别以0 Gy、1. 94 Gy、3. 88 Gy、7. 76 Gy、15. 53 Gy137Cs 辐照,分别于24 h,48 h,72 h,96 h 取出肝脏、性腺,按试剂盒方法检测SOD、GSH-PX 活性,于7 d 和14 d 取出肝脏、肾脏、心脏、脑,按Western blot 方法检测HSP70 含量。结果1. 94 Gy至15. 53 Gy137Cs 辐照能引起实验红鲫氧化应激,SOD、GSH-PX 活性和HSP70 含量发生改变。1. 94 Gy137Cs 辐照诱导实验红鲫肝脏SOD 活性升高, 3. 88 Gy、7. 76 Gy、15. 53 Gy137Cs 辐照诱导实验红鲫肝脏SOD 活性下降,且随辐射剂量升高,其SOD 活性逐渐降低。1. 94 Gy 以上137Cs 辐照诱导实验红鲫性腺SOD 活性下降,并随辐射剂量升高和时间的延长而下降,且下降幅度较大。各辐照处理组实验红鲫肝脏和性腺的GSH-PX 活性变化不大,表现出随辐射剂量升高则活性降低的变化趋势。137Cs 辐照处理后7 d 和14 d,各辐照处理组实验红鲫肝脏、肾脏、心脏和脑的HSP70 表达量均比对照组的高,且随着137Cs 辐照剂量的增加和时间的延长而升高,均存在着一定的“剂量－ 时间－效应”关系。结论实验红鲫性腺对137Cs 辐射的氧化应激响应较敏感,性腺SOD 和肝脏或肾脏的HSP70 可作为氧化应激标志物。     

Cell contact- and shape-dependent regulation of vinculin synthesis in cultured fibroblasts

Recent studies have demonstrated the fundamental role of cell-substrate contacts and changes in cell shape in the regulation of cell growth, motility and differentiation, but the molecular basis for these phenomena is poorly understood. Because of the involvement of cytoskeletal networks in cell morphogenesis and contact formation, it is of interest that the expression of genes encoding several cytoskeletal proteins is markedly affected by changes in cell contacts and configuration. Because most of these phenomena involve changes in the form, extent or topology of cell contacts, we sought to determine whether the expression of components directly involved in the formation of cell-cell or cell-substrate contacts is affected by the respective cellular interactions. A suitable candidate for such analysis is vinculin, a cytoskeletal protein of relative molecular mass (Mr) 130,000 (130K), which is localized in focal contacts and intercellular adherens junctions. The assembly of vinculin into a membrane-bound junctional plaque seems to be one of the earliest cellular responses to contact with exogenous substrates, leading to the subsequent local assembly of the actin-rich microfilament bundles. Here we report on the regulation of vinculin synthesis in response to environmental conditions that affect cell shape and contacts.     

暗纹东方鲀（Takifugu obscurus）性别分化的组织学及芳香化酶基因表达研究

采用组织切片技术和实时定量PCR（qRT-PCR）方法研究了暗纹东方鲀性别分化过程中性腺发育的组织学及芳香化酶基因CYP19A的表达变化.结果显示：孵化后11d的仔鱼切片中可看到原始生殖细胞（PGC）;孵化后17d可看到隆起的性腺原基（GA）;孵化后26d发育成为原始性腺（PG）并从体腔膜上游离出来.卵巢分化时间早于精...     

Pregnenolone stabilizes microtubules and promotes zebrafish embryonic cell movement

Embryonic cell movement is essential for morphogenesis and the establishment of body shapes, but little is known about its mechanism. Here we report that pregnenolone, which is produced from cholesterol by the steroidogenic enzyme Cyp11a1 (cholesterol side-chain cleavage enzyme, P450scc), functions in promoting cell migration during epiboly. Epiboly is a process in which embryonic cells spread from the animal pole to cover the underlying yolk. During epiboly, cyp11a1 is expressed in an extra-embryonic yolk syncytial layer. Reducing cyp11a1 expression in zebrafish using antisense morpholino oligonucleotides did not perturb cell fates, but caused epibolic delay. This epibolic defect was partially rescued by the injection of cyp11a1 RNA or the supplementation of pregnenolone. We show that the epibolic delay is accompanied by a decrease in the level of polymerized microtubules, and that pregnenolone can rescue this microtubule defect. Our results indicate that pregnenolone preserves microtubule abundance and promotes cell movement during epiboly.     

阿特拉津对泥鳅性腺及性别分化相关基因的影响

以泥鳅为材料,首先确定了阿特拉津对泥鳅的急性毒性.在此基础上采用组织学切片和实时荧光定量PCR(qRT-PCR)的方法研究了阿特拉津长期处理对泥鳅性腺形态和性别分化相关基因表达模式的影响,以期准确评估阿特拉津的环境毒性和对水生生物生殖发育的作用.结果表明:阿特拉津对泥鳅的24h、48h、96h的半数致死浓度(LC50)分别为31.60mg/L、26.82mg/L、18.98mg/L,安全质量浓度(SC)为5.8mg/L.随着阿特拉津浓度的增加和处理时间的延长,泥鳅的死亡率明显的升高.通过组织学研究发现高浓度的阿特拉津(2.68 mg/L)处理泥鳅21d能够抑制泥鳅精巢精子的发生,促进卵巢细胞的发育.同时qRT-PCR结果显示:与对照组相比不同剂量的阿特拉津(2.68mg/L,0.268mg/L,0.0268mg/L)能够显著的诱导细胞色素P450芳香化酶基因(cyp19a),类固醇生成因子(sf1)的表达,对抗缪勒氏管激素(amh)有显著的抑制效应.因此,阿特拉津对泥鳅有明显的外源雌激素效应;通过干扰类固醇激素合成基因的表达影响生殖细胞的生成;也通过某种未知的机制干扰性别分化上游基因的表达,从而影响性腺分化.     

Auxin transport is sufficient to generate a maximum and gradient guiding root growth

The plant growth regulator auxin controls cell identity, cell division and cell expansion. Auxin efflux facilitators (PINs) are associated with auxin maxima in distal regions of both shoots and roots. Here we model diffusion and PIN-facilitated auxin transport in and across cells within a structured root layout. In our model, the stable accumulation of auxin in a distal maximum emerges from the auxin flux pattern. We have experimentally tested model predictions of robustness and self-organization. Our model explains pattern formation and morphogenesis at timescales from seconds to weeks, and can be understood by conceptualizing the root as an 'auxin capacitor'. A robust auxin gradient associated with the maximum, in combination with separable roles of auxin in cell division and cell expansion, is able to explain the formation, maintenance and growth of sharply bounded meristematic and elongation zones. Directional permeability and diffusion can fully account for stable auxin maxima and gradients that can instruct morphogenesis.     

中国林蛙胚胎期性腺发育的组织学观察

采用常规石蜡切片方法,对产于中国东北地区的中国林蛙南移至华南地区后胚胎发育期的性腺发育进行了组织学观察,并首次探讨了中国林蛙的性别分化模式.结果证明变迁后的环境对中国林蛙胚胎期的生殖腺发育没有造成不良结果.此结果不但丰富了发育生物学的内容,也为林蛙的南移提供了依据.     

波纹巴非蛤性腺发育分期的研究

采用活体性腺观察和组织切片法对广西北海自然海区波纹巴非蛤的性腺周年发育进行了初步研究 ,并以波纹巴非蛤滤泡中生殖细胞所处的主要发育阶段作为性腺发育的分期标准 ,将性腺发育过程分为 :休止期、增殖期、生长期、成熟期、排放期和滞留退化期等 6个时期 ,同时揭示了在广西北海自然海区波纹巴非蛤的性腺发育一年只有一个周期 ;此外 ,还发现雄性早熟现象 .     

Asymmetric leaves1 mediates leaf patterning and stem cell function in Arabidopsis

Meristem function in plants requires both the maintenance of stem cells and the specification of founder cells from which lateral organs arise. Lateral organs are patterned along proximodistal, dorsoventral and mediolateral axes. Here we show that the Arabidopsis mutant asymmetric leaves1 (as1) disrupts this process. AS1 encodes a myb domain protein, closely related to PHANTASTICA in Antirrhinum and ROUGH SHEATH2 in maize, both of which negatively regulate knotted-class homeobox genes. AS1 negatively regulates the homeobox genes KNAT1 and KNAT2 and is, in turn, negatively regulated by the meristematic homeobox gene SHOOT MERISTEMLESS. This genetic pathway defines a mechanism for differentiating between stem cells and organ founder cells within the shoot apical meristem and demonstrates that genes expressed in organ primordia interact with meristematic genes to regulate shoot morphogenesis.     

c-Myc regulates mammalian body size by controlling cell number but not cell size.

Overexpression of the proto-oncogene c-myc has been implicated in the genesis of diverse human tumours. c-Myc seems to regulate diverse biological processes, but its role in tumorigenesis and normal physiology remains enigmatic. Here we report the generation of an allelic series of mice in which c-myc expression is incrementally reduced to zero. Fibroblasts from these mice show reduced proliferation and after complete loss of c-Myc function they exit the cell cycle. We show that Myc activity is not needed for cellular growth but does determine the percentage of activated T cells that re-enter the cell cycle. In vivo, reduction of c-Myc levels results in reduced body mass owing to multiorgan hypoplasia, in contrast to Drosophila c-myc mutants, which are smaller as a result of hypotrophy. We find that c-myc substitutes for c-myc in fibroblasts, indicating they have similar biological activities. This suggests there may be fundamental differences in the mechanisms by which mammals and insects control body size. We propose that in mammals c-Myc controls the decision to divide or not to divide and thereby functions as a crucial mediator of signals that determine organ and body size.