Crumbs, the Drosophila homologue of human CRB1/RP12, is essential for photoreceptor morphogenesis

The apical transmembrane protein Crumbs is a central regulator of epithelial apical-basal polarity in Drosophila. Loss-of-function mutations in the human homologue of Crumbs, CRB1 (RP12), cause recessive retinal dystrophies, including retinitis pigmentosa. Here we show that Crumbs and CRB1 localize to corresponding subdomains of the photoreceptor apical plasma membrane: the stalk of the Drosophila photoreceptor and the inner segment of mammalian photoreceptors. These subdomains support the morphogenesis and orientation of the photosensitive membrane organelles: rhabdomeres and outer segments, respectively. Drosophila Crumbs is required to maintain zonula adherens integrity during the rapid apical membrane expansion that builds the rhabdomere. Crumbs also regulates stalk development by stabilizing the membrane-associated spectrin cytoskeleton, a function mechanistically distinct from its role in epithelial apical-basal polarity. We propose that Crumbs is a central component of a molecular scaffold that controls zonula adherens assembly and defines the stalk as an apical membrane subdomain. Defects in such scaffolds may contribute to human CRB1-related retinal dystrophies.     

花背蟾蜍角膜的发育和诱导

以花背蟾蜍的胚胎和变态期蝌蚪为材料,采用光镜、电镜、免疫荧光组织化学、放射自显曩、电镜细胞化学、皮肤片移植及类坏死处理等技术,研究：（１）在胚胎发育及变态期间角膜的发育过程;（２）细胞外基质的合成变化及其对角膜发育和诱导的作用;（３）角膜诱导期间的超微结构变化及类坏死对角膜诱导的影响;（４）在角膜下沉发育和诱导过程中的组织化学和视网膜感光细胞特异视蛋白的出现及二者间的可能关系。还提出角膜诱导的可能     

Photoreceptor-specific degeneration caused by tunicamycin

The antibiotic tunicamycin inhibits the biosynthesis of N-acetylglucosaminylpyrophosphoryl polyisoprenol, a key intermediate in the formation of the asparagine-linked oligosaccharides of glycoproteins. The effects of tunicamycin have been studied in various biological systems, primarily with the aim of elucidating the role of the carbohydrate moieties in the cellular function of glycoproteins. Rhodopsin, the visual pigment of retinal rod photoreceptor cells, is a membrane glycoprotein which consists of a single polypeptide chain (opsin) to which a chromophoric prosthetic group (II-cis-retinaldehyde) and two asparagine-linked oligosaccharide chains are covalently attached. The glycosylation of opsin can be blocked with tunicamycin in vitro in conditions where polypeptide synthesis is only slightly decreased. We have reported that tunicamycin can disrupt the normal assembly of rod outer segment membranes in vitro without significantly inhibiting the biosynthesis or intracellular transport of opsin. Here we report that intraocular injection of tunicamycin produces a photoreceptor-specific degeneration characterized by progressive shortening of rod outer segment, decreased membrane assembly, and eventual photoreceptor cell death.     

Transcript localization of four opsin genes in the three visual organs of Drosophila; RH2 is ocellus specific

Drosophila and other Dipteran flies have three different kinds of visual organs; in the adult a pair of compound eyes and three dorsal ocelli; and in the larva a pair of internal photoreceptor organs. They develop in distinct ways, yet have certain features in common. All three organs use retinal-derived chromophores, coupled to distinct opsins, to provide a diversity of spectral sensitivities. Four opsin genes have been identified thus far in Drosophila; Rh1, Rh2, Rh3 and Rh4 (refs 6-11). We have used in situ hybridization to study the messenger RNAs expressed by these four opsin genes in all three visual organs. Rh1, Rh3 and Rh4 are already known to be expressed in different subsets of cells in the compound eye. We found that, in contrast, opsin Rh2 is the predominant opsin expressed in the ocelli. Opsin Rh1 is known to be expressed in the larval photoreceptor. We found that Rh3 and Rh4 are as well, but not Rh2. The ocellar-specific gene expression of Rh2 is of particular interest for its possible bearing on the function of the ocellus.     

地球同步轨道上物体内的张力分析

处于地球轨道上的物体 ,同时受到引力和离心力的作用。通过张力分析发现 ,一定长度的线状物体有可能悬空在太空 ,形成太空缆绳。其长度满足公式 :L=3- R32 + R6 2 - 82 7( R32R1+ R212 ) 3+3- R32 - 82 7( R32R1+ R212 ) 3- R1其中 :R1为地球半径 ,R2 为地球同步轨道半径。利用太空缆绳 ,可提供一个由地面到太空的廉价运输及信息通道。     

Drosophila Crumbs is a positional cue in photoreceptor adherens junctions and rhabdomeres

Drosophila Crumbs (Crb) is required for apical-basal polarity and is an apical determinant in embryonic epithelia. Here, we describe properties of Crb that control the position and integrity of the photoreceptor adherens junction and photosensitive organ, or rhabdomere. In contrast to normal photoreceptor adherens junctions and rhabdomeres, which span the depth of the retina, adherens junctions and rhabdomeres of Crb-deficient photoreceptors initially accumulate at the top of the retina and fail to maintain their integrity as they stretch to the retinal floor. We show that Crb controls localization of the adherens junction through its intracellular domain containing a putative binding site for a protein 4.1 superfamily protein (FERM). Although loss of Crb or overexpression of the FERM binding domain causes mislocalization of adherens junctions, they do not result in a significant loss of photoreceptor polarity. Mutations in CRB1, a human homologue of crb, are associated with photoreceptor degeneration in retinitis pigmentosa 12 (RP12) and Leber congenital amaurosis (LCA). The intracellular domain of CRB1 behaves similarly to its Drosophila counterpart when overexpressed in the fly eye. Our studies may provide clues for mechanisms of photoreceptor degeneration in RP12 and LCA.     

鳞翅目昆虫精巢融合的研究

昆虫精巢是产生精子并为精子发育提供营养和保护的重要器官．昆虫雄虫在幼虫期通常有左右对称的２个精巢．但是,部分昆虫如农业害虫斜纹夜蛾在从幼虫向蛹变态发育的过程中,２个精巢发生融合,合二为一,在蛹期最终完全融合成单个精巢;有些昆虫如经济昆虫家蚕的２个精巢并不融合,在成虫期仍保持分离．精巢融合显然是昆虫长期自然演化过程中形成的一种繁殖方式,有利于种群的繁殖和扩大．本文结合课题组的研究,对鳞翅目昆虫精巢的类型和结构、精子产生的过程、精巢融合的现象及其生物学意义以及在农业害虫防治上的潜在应用意义进行了简述．     

椰子心叶和成熟叶片对椰心叶甲生育指标的影响

研究发现椰一but-甲[Brontispa longissima（Gestro）]各项生育指标与椰子[Cocos nucifera（Linnaeus）]叶片的生长阶段关系密切．统计表明：从幼虫到成虫整个阶段的存活率心叶较成熟叶片高,分别为81％和77％,椰心叶甲完成一个世代在心叶上需要58．54d,在成熟叶片上需要52．92d．叶片生长阶段对虫质量和蛹质量的影响表现为取食心叶较重,且大部分龄期的幼虫质量和蛹质量的差异均显著．椰子心叶和成熟叶片上的成虫的性比分别为n（雌）：n（雄）=1．27：1和n（雌）：n（雄）=1．17：1,平均产卵量为143．3粒和161．93粒,成虫寿命分别为189．14d和172．36d,椰子心叶和成熟叶上的实验种群趋势指数分别为56．55和56．45．     

Homing behaviour of axons in the embryonic vertebrate brain

In embryonic nervous systems, growing axons must often travel long distances through diverse extracellular terrains to reach their postsynaptic partners. In most embryos, axons grow to their appropriate targets along particular tracts or nerves, as though they were following guidance cues confined to specific pathways. For example, in all vertebrates, axons from the retina invariably grow to the tectum along the well-defined optic tract. Yet, transplant experiments demonstrate that retinal axons make tectal projections even though they enter the brain at locations which are distinctly off the optic tract. Only recently has it become possible to label discreet growing projections in the embryonic vertebrate brain. Thus, it is not yet known whether displaced retinal axons grow directly towards the tectum or find it accidently, through random extension. To resolve this question, pioneering axons from normal and transplanted eyes in embryonic Xenopus were labelled using a short-survival horseradish peroxidase (HRP) method, and their orientation during growth was quantitatively assessed. The finding that the ectopic fibres head towards their distant targets implies that guidance cues are not restricted to specific pathways but are distributed throughout the embryonic brain. The significance of this result is discussed with respect to the ontogeny and evolution of the visual pathway.     

Circadian rhythm and light regulate opsin mRNA in rod photoreceptors

Disk membranes in the outer segment of rod photoreceptors are continuously renewed, being assembled at the outer segment base, displaced outward by new disks and eventually shed at the tip. In lower vertebrates, disk assembly occurs with a diurnal rhythm with 2-4% of the outer segment length produced daily. We have discovered that in toad and fish retinas the level of mRNA for opsin, the most abundant protein in rod disks, fluctuates with a daily rhythm and is regulated both by light and by a circadian oscillator. The mRNA level rises before light onset, remains high during the light phase of a diurnal cycle and decreases four to tenfold during the dark phase. In constant darkness, mRNA elevation occurs during subjective daytime. At night, rod opsin mRNA can be elevated by exposure to light.     

Targeted misexpression of a Drosophila opsin gene leads to altered visual function

Drosophila mutants transformed with a chimaeric gene that expresses the ocellar visual pigment in the major class of photoreceptor cells of the retina were used to investigate the properties of this minor pigment. The photoreceptor cells in which this opsin was misexpressed showed new spectral characteristics and physiology.     

Temporal identity in axonal target layer recognition

The segregation of axon and dendrite projections into distinct synaptic layers is a fundamental principle of nervous system organization and the structural basis for information processing in the brain. Layer-specific recognition molecules that allow projecting neurons to stabilize transient contacts and initiate synaptogenesis have been identified. However, most of the neuronal cell-surface molecules critical for layer organization are expressed broadly in the developing nervous system, raising the question of how these so-called permissive adhesion molecules support synaptic specificity. Here we show that the temporal expression dynamics of the zinc-finger protein sequoia is the major determinant of Drosophila photoreceptor connectivity into distinct synaptic layers. Neighbouring R8 and R7 photoreceptors show consecutive peaks of elevated sequoia expression, which correspond to their sequential target-layer innervation. Loss of sequoia in R7 leads to a projection switch into the R8 recipient layer, whereas a prolonged expression in R8 induces a redirection of their axons into the R7 layer. The sequoia-induced axon targeting is mediated through the ubiquitously expressed Cadherin-N cell adhesion molecule. Our data support a model in which recognition specificity during synaptic layer formation is generated through a temporally restricted axonal competence to respond to broadly expressed adhesion molecules. Because developing neurons innervating the same target area often project in a distinct, birth-order-dependent sequence, temporal identity seems to contain crucial information in generating not only cell type diversity during neuronal division but also connection diversity of projecting neurons.     

Substrate phosphoprotein availability regulates eclosion hormone sensitivity in an insect CNS

The final step in the moulting of all insects is ecdysis, the shedding of the cuticle of the previous instar, which is triggered in Lepidoptera by the neurosecretory peptide eclosion hormone. This hormone acts directly on the nervous system to release the appropriate motor patterns for larval, pupal and adult ecdysis, but there are only brief periods near the end of each moult when the nervous system is competent to respond to the hormone. Previous experiments have shown that the action of eclosion hormone on the nervous system at pupal ecdysis in the tobacco hornworm, Manduca sexta, is mediated by the second messenger cyclic GMP. Here we report that the hormone-stimulated increase in cGMP results in the phosphorylation of two proteins, each with an apparent relative molecular mass (Mr) of 54,000. Moreover, the brief periods during which the central nervous system (CNS) is responsive to eclosion hormone seem to result from the transient presence of these substrate proteins within the nervous system. This provides a novel mechanism by which hormonal responsiveness can be regulated.     

保幼激素和蜕皮激素对家蚕翅原基生长分化的影响

翅原基（Wing disc）是昆虫幼虫体内成虫原基的一种,经生长分化后最终形成成虫翅。翅原基是研究昆虫变态发育过程的一个理想系统。为探究发育过程中翅原基的生长变化,本文以家蚕翅原基为材料,通过离体和石蜡切片的方法观察了翅原基从5龄第3天至蛹期0天的形态变化,并通过注射外源蜕皮激素活性物质20E和保幼激素类似物Methoprene探究了昆虫激素对家蚕翅原基生长分化的影响。结果显示,翅原基在幼虫阶段发育缓慢,从5龄第6天起生长分化逐渐加快,且翅原基的形态发生了显著变化,原本附着于翅原基腔口处且呈团状的造血器官逐渐分散至消失,而由气管组成的翅脉逐渐形成。外源激素处理的结果显示,2μg剂量的20E可促进翅原基的生长分化,而2μg 的Methoprene抑制了翅原基的生长分化。上述结果暗示了,蜕皮激素和保幼激素共同调控了翅原基的生长分化,并最终实现了翅原基的变态发育。     

Identification of a distinct synaptic glutamate receptor on horizontal cells in mudpuppy retina

The separation of ON and OFF channels and the development of an antagonistic surround occur at the first synapse in the vertebrate retina. This functional differentiation is mediated by the action of the photoreceptor neurotransmitter on the ON bipolar, OFF bipolar and horizontal cells, respectively. Glutamate mimics the action of the photoreceptor transmitter on all second-order neurones in fish, amphibian and mammalian retinas. The diversity of cellular responses produced by one neurotransmitter raises the possibility of multiple postsynaptic receptor-ionophore complexes. We reported previously that one glutamate analogue, 2-amino-4-phosphonobutyrate, reveals that the ON bipolar synaptic receptor is pharmacologically different from those of other second-order neurones. The results presented here demonstrate that another glutamate analogue, D-O-phosphoserine, selectively antagonizes the synaptic responses of horizontal cells. Taken together, these findings indicate that there are three glutamate-like receptor subtypes in the outer retina and suggest a correlation between receptor subtype and the physiological properties of second-order neurones.     

瓜实蝇和南瓜实蝇的种内竞争

研究了种内竞争对瓜实蝇（Ｂａｃｔｒｏｃｅｒａｃｕｃｕｒｂｉｔａｅ）和南瓜实蝇（Ｂｔａｕ）的生长、发育及存活的影响．对不同竞争强度下蛹的生物量、成虫生物量、幼虫存活率、蛹存活率、蛹的大小和成虫大小进行拟合分析,其模拟方程各不相同,但模拟方程的微分形式可用一通式来表示：ｄｙ／ｄｘ＝ｋ（ｘ－ｘ０）ｘｉｙｊ（ｋ,ｘ０,ｉ,ｊ为常数）种内竞争对昆虫性比的影响不具规律性     

Regulation of the G1-S transition in postembryonic neuronal precursors by axon ingrowth.

In the newly cellularized Drosophila embryo, progress through the cell cycle is regulated at the G2-M transition. We have examined cell-cycle regulation later in Drosophila development, in a group of postembryonic neuronal precursors. The S-phase precursor cells, which generate photoreceptor target neurons (lamina neurons) in the central nervous system, are not present in the absence of photoreceptor innervation. Here we report that axons selectively approach G1-phase precursors. Without axon ingrowth, lamina precursors do not enter their final S phase and by several criteria, arrest in the preceding G1 phase. These findings provide evidence that at this stage in development the control of cell division can occur at the G1-S transition.     

南亚实蝇幼虫人工饲料研究

以组成为麦麸800g、蔗糖300g、啤酒废酵母150g、饲料添加剂6．5g和水1．5L的人工饲料连续饲养南亚实蝇3代，与丝瓜饲养对照，研究人工饲料对南亚实蝇生长发育的影响。结果表明，F1蛹重与对照差异极显著，F2、F3则无显著差异，卵孵化率、幼虫存活率、羽化率和飞出率与对照相比无显著差异，说明该人工饲料能充分满足南亚实蝇生长发育所需。     

家蚕促咽侧体素受体基因克隆及发育表达

文章克隆得到家蚕(Bombyx mori)神经肽促咽侧体素受体基因(Allatotropin receptor,BommoATR),开放阅读框全长为1 254 bp,编码416个氨基酸.Bommo-ATR氨基酸序列的二级结构预测为7次穿膜蛋白,符合GPCR家族成员的典型特征.实时荧光定量PCR结果表明,家蚕脑-咽下神经节复合体中的ATR mRNA在5龄幼虫期表达量最高,其次为蛹后期,蛹前期和成虫阶段表达量最低.其中5龄幼虫第2天表达量最高,1～5 d持续维持较高的表达水平,这可能与保幼激素的合成有关.