Identification of cytoplasmically transferred mitochondrial DNA in female germlines of Drosophila and its propagation in the progeny

Summary The composition of mitochondrial DNA (mtDNA) was analyzed in single female flies that developed from fertilized Drosophila melanogaster eggs, into which germ plasm of D. simulans had been introduced. HpaII cleavage patterns showed that all 12 individual female flies examined had developed from eggs in which 37%–71% of the total mtDNA was D. simulans mtDNA (Ds mtDNA) and the rest was D. melanogaster mtDNA (Dm mtDNA). The stability of this heteroplasmic state in these isofemale lines was monitored for seven generations at both individual and population levels. Results showed that the heteroplasmy of Dm and Ds mtDNAs was stably transmitted for at least three generations at the population level, but showed stochastic segregation at the individual level. After 4–6 generations, all individuals lost Ds mtDNA. The mechanisms of preferential loss of Ds mtDNA and of transmission of heteroplasmic mtDNA to descendants are discussed.     

Specific expression of the Drosophila midline-jumper retro-transposon in embryonic CNS midline cells

Here we describe of a novel Drosophila LTR-type retrotransposon that is expressed in the embryonic CNS midline glia and in the embryonic germ cells. The element is related to the gypsy and burdock retrotransposons and was termed midline-jumper. In addition to cDNA clones generated from internal retrotransposon sequences, we have identified one cDNA clone that appears to reflect a transposition event, indicating that the midline-jumper retrotransposon is not only transcribed but also able to transpose during Drosophila development.     

Functional interactions between the gene tetanic and the Shaker gene complex of Drosophila

Different phenotypes associated with the tetanic (tta) mutation such as appendage contraction, maternal effect and low viability and fertility are enhanced by one extra dose of the Shaker gene complex (ShC). The tta mutation is lethal with two extra doses of ShC. In addition, tta embryos have a defective nervous system. In this paper, I analyse the interaction between tta and ShC to gain insight into their relationship. Aneuploid analysis suggests that the lethality is due to an interaction of the tta mutation with the maternal effect (ME) region of this gene complex. Mutations in the ME region of ShC partially suppress this interaction. Trans-heterozygous combinations of MEI[l(1)305] and MEIII [l(1)459] mutations causes dominant lethality in a tta background. Trans-heterozygous combinations of an MEII [l(1)1359] mutation with the cited MEI and MEIII mutations are lethal in a tta background. Double mutant combinations and gene dosage experiments, suggest that tta also interacts with the viable (V) region of ShC. These specific genetic interactions indicate that tta and the ME and V regions of ShC are functionally related. These results, together with the previous electrophysiological, molecular and biochemical studies on these mutants suggest an interaction at the protein level. Thus, in the case of the V region, the tta gene product may modulate the activity of the K⁺ channels encoded in this region. Furthermore, the extreme dosage sensitivity of the interaction between tta and ShC suggests a stoichiometric requirement for the different gene products involved, which might be physically associated and form heteromultimers.     

Probing the Drosophila retinal determination gene network in Tribolium (II): The Pax6 genes eyeless and twin of eyeless

The Pax6 genes eyeless (ey) and twin of eyeless (toy) are upstream regulators in the retinal determination gene network (RDGN), which instructs the formation of the adult eye primordium in Drosophila. Most animals possess a singleton Pax6 ortholog, but the dependence of eye development on Pax6 is widely conserved. A rare exception is given by the larval eyes of Drosophila, which develop independently of ey and toy. To obtain insight into the origin of differential larval and adult eye regulation, we studied the function of toy and ey in the red flour beetle Tribolium castaneum. We find that single and combinatorial knockdown of toy and ey affect larval eye development strongly but adult eye development only mildly in this primitive hemimetabolous species. Compound eye-loss, however, was provoked when ey and toy were RNAi-silenced in combination with the early retinal gene dachshund (dac). We propose that these data reflect a role of Pax6 during regional specification in the developing head and that the subsequent maintenance and growth of the adult eye primordium is regulated partly by redundant and partly by specific functions of toy, ey and dac in Tribolium. The results from embryonic knockdown and comparative protein sequence analysis lead us further to conclude that Tribolium represents an ancestral state of redundant control by ey and toy.     

Functional relationships between genes of the Shaker gene complex of Drosophila

Different mutations belonging to the HLI and HLII complementation groups of the haplolethal (HL) region of the Shaker complex (ShC) are described. The HLI complementation group includes viable (hdp), recessive lethals [l(1)1614], semidominant lethals [l(1)8384] and dominant lethals [l(1)5051,l(1)9916, l(1)13193], lack-of-function alleles that affect nervous system, cuticle and muscle development. The HLI complementation group encodes troponin I. HLII lack-of-function mutations [l(1)174 and l(l)4058] affect nervous system development. The semidominant lethal HLI mutation 1(1)8384 shows differential complementation with other mutations in the ME and HL regions of ShC. Thus, heterozygous combinations of l(1)8384 with ME mutations l(1)162 and l(1)387 are poorly viable. The same phenomenon is observed for heterozygotes of l(1)8384 with HL mutations l(1)1199, l(1)2288 and l(1)3014. These specific interactions indicate the existence of functional relationships among the genetic elements of ShC. The implications for the understanding of the functional organization of ShC are discussed.     

Genotoxicity is modulated by ascorbic acid: Studies using the wing spot test in Drosophila

The ability of ascorbic acid (Vitamin C) to modulate the genotoxic action of several mutagens was investigated in the wing spot test of Drosophila melanogaster. In this assay, 3-day-old transheterozygous larvae for the multiple wing hairs (mwh, 3-0.3) and flare (flr, 3-38.8) genes were treated with three reference mutagenic compounds, namely cobalt chloride (CoCl₂), 4-nitroquinoline 1-oxide (4-NQO) and potassium dichromate (K₂Cr₂O₇). The results obtained show that the three reference mutagens tested were clearly genotoxic in the Drosophila wing somatic mutation and recombination test (SMART). None of the three concentrations tested of ascorbic acid (25, 75 and 250 mM) induced significant increases in the frequency of the mutant clones recorded. When co-treatment experiments with ascorbic acid were carried out, different results were found. Thus, ascorbic acid was effective in reducing the genotoxicity of K₂Cr₂O₇ virtually to the control level; on the contrary, it did not show any antigenotoxic effect on the genotoxicity of 4-NQO. Finally, co-treatments with CoCl₂ and ascorbic acid show a significant increase in the frequency of mutant clones over the values obtained with CoCl₂ alone.     

Molybdoenzymes in Drosophila. IV. Further characterization of the cinnamon phenotype

Summary Mutations at the cin gene display drastically lowered levels of the molybdoenzymes, xanthine dehydrogenase (XDH) and aldehyde oxidase (AO), and lack pyridoxal oxidase (PO) and sulfite oxidase (SO) activities. Certain mutations at cin also display varying degrees of female sterility, which is maternally affected. Here we characterize five new cin alleles with respect to the molybdoenzyme activities as well as the molybdenum cofactor, commonly required for molybdoenzyme activity. In complementing cin heterozygotes we find that, in addition to the previously reported unusually high levels of XDH and AO activities, there are unusually elevated levels of SO activity, as well as complementation for PO activity. The levels of immunologically crossreacting material in such heterozygotes indicate that the elevated levels of molybdoenzyme activities cannot be due to increases in the number of enzyme molecules. Measurements of the level of molybdenum cofactor activity normally present in XDH, AO, PO, and SO point to the possibility that a larger fraction of the enzyme molecules are active in these heterozygotes. The possible role of SO with respect to cinnamon's female sterility is also discussed.     

果蝇肠道共生菌对宿主作用的研究进展

肠道共生菌是动物体内的重要组成部分,在宿主的生长发育和健康等方面发挥着重要作用,近年来已成为国内外的研究热点.果蝇作为研究肠道微生物菌群功能的优秀模型,在肠道共生菌与宿主关系研究方面已取得许多重要进展.在本文中,我们首先对果蝇肠道微生物的组成和特征作了总结,然后对果蝇肠道共生菌在其生长发育、营养与代谢、行为反应、寿命以...     

Postembryonic Hematopoiesis in Drosophila

We have investigated the blood cell types present in Drosophila at postembryonic stages and have analysed their modifications during development and under immune conditions. The anterior lobes of the larval hematopoietic organ or lymph gland contain numerous active secretory cells, plasmatocytes, few crystal cells, and a number of undifferentiated prohemocytes. The posterior lobes contain essentially prohemocytes. The blood cell population in larval hemolymph differs and consists mainly of plasmatocytes which are phagocytes, and of a low percentage of crystal cells which reportedly play a role in humoral melanisation. We show that the cells in the lymph gland can differentiate into a given blood cell lineage when solicited. Under normal nonimmune conditions, we observe a massive differentiation into active macrophages at the onset of metamorphosis in all lobes. Simultaneously, circulating plasmatocytes modify their adhesion and phagocytic properties to become pupal macrophages. All phagocytic cells participate in metamorphosis by ingesting doomed larval tissues. The most dramatic effect on larval hematopoiesis was observed following infestation by a parasitoid wasp. Cells within all lymph gland lobes, including prohemocytes from posterior lobes, massively differentiate into a new cell type specifically devoted to encapsulation, the lamellocyte.     

RNA干扰技术在果蝇中的应用

RNA干扰是双链RNA特异诱导的转录后期基因沉默.该技术随着不断完善而越来越被广泛地运用于果蝇的功能基因组研究上,双链RNA已经成为果蝇中功能基因的一个十分有效的抑制子,势必使RNA干扰技术成为研究果蝇体内基因功能的强有力的反向遗传学研究技术.     

A gain-of-function screen identifies wdb and lkb1 as lifespan-extending genes in Drosophila

The insulin/insulin-like growth factor (IGF) and the target of rapamycin (TOR) signaling pathways are known to regulate lifespan in diverse organisms. However, only a limited number of genes involved in these pathways have been examined regarding their effects on lifespan. Through a gain-of-function screen in Drosophila, we found that overexpression of the wdb gene encoding a regulatory subunit of PP2A, and overexpression of the lkb1 gene encoding a serine/threonine kinase, reduced organ size and extended lifespan. Overexpression of wdb also reduced the level of phosphorylated AKT, while overexpression of lkb1 increased the level of phosphorylated AMPK and decreased the level of phosphorylated S6K. Taken together, our results suggest that wdb- and lkb1-dependent lifespan extension is mediated by downregulation of S6K, a downstream component of the insulin/IGF and TOR signaling pathways.     

Expression and cellular localization of the Toucan protein during Drosophila oogenesis

The toucan (toc) gene is required in the germline for somatic cell patterning during Drosophila oogenesis. To better understand the function of toc, we performed a detailed analysis of the distribution of the Toucan protein during oogenesis. Toc expression is restricted to the germline cells and shows a dynamic distribution pattern throughout follicle development. Mislocalization of the Toc protein in mutant follicles in which the microtubule network is altered indicates that microtubules play a role in Toc localization during oogenesis.     

Regulation of Krüppel expression in the anlage of the Malpighian tubules in the Drosophila embryo

The expression of most Drosophila segmentation genes is not limited to the early blastoderm stage, when the segmental anlagen are determined. Rather, these genes are often expressed in a variety of organs and tissues at later stages of development. In contrast to the early expression, little is known about the regulatory interactions that govern the later expression patterns. Among other tissues, the central gap gene Krüppel is expressed and required in the anlage of the Malpighian tubules at the posterior terminus of the embryo. We have studied the interaction of Krüppel with other terminal genes. The gap genes tailles and huckebein, which repress Krüppel in the central segmentation domain, activate Krüppel expression in the posterior Malpighian tubule domain. The opposite effect on the posterior Krüppel expression is achieved by the interposition of another factor, the homeotic gene fork head, which is not involved in the control of the central domain. In addition, Krüppel activates different genes in the Malpighian tubules than in the central domain. Thus, both the regulation and the function of Krüppel in the Malpighian tubules differ strikingly from its role in segmentation.     

Metabolic pools differ among ecologically diverse Drosophila species

Studies of the genetic mechanisms underlying metabolic storage have focused on a few model organisms. Although very fruitful, these studies have not allowed for the examination of mechanisms across a phylogenetic spectrum. The exploration of natural patterns of metabolic pool size variation across species will help us to better understand the genetics of metabolic adaptation.We examined the metabolic pools size (triglyceride, glycogen and protein) at two ages in 12 Drosophila species with distinctly different ecologies for which complete genome sequences (for 11 of the 12 species) are known. Overall, there were significant differences across species for all three pools, while age and sex appear to affect some metabolic pools more than others. After correcting for the phylogenetic relatedness of the species used, we observed no association between triglyceride and glycogen content. Although within species these two pools sometimes are correlated, at a larger phylogenetic scale control of triglyceride and glycogen contents may have been shaped independently by natural selection.     

葡萄园果蝇的发生及综合防治研究进展

果蝇因种类多、分布广、寄主广、繁殖力强,世代历期短,近年已成为葡萄的重要害虫之一。葡萄果蝇幼虫不仅蛀食果实,导致果实逐渐软化以致变褐色,还能导致微生物的二次侵染,诱发酸腐病,严重影响葡萄的品质和产量。现已发现危害葡萄的果蝇有7种,其中以黑腹果蝇为优势种,其次为斑翅果蝇和叔白颜果蝇。在山东地区,7月初是斑翅果蝇和叔白颜果蝇的发生高峰期,而黑腹果蝇发生高峰期为8-9月上旬。就不同葡萄品种来说,黑腹果蝇成虫对4个葡萄品种巨峰、马奶、无核白和红提的嗜好程度无显著差异,幼虫对红提的嗜好程度显著高于巨峰、马奶和无核白;斑翅果蝇和叔白颜果蝇对于金手指等早熟品种以及果皮较薄的葡萄品种具有更强的趋性。本文依据葡萄园主要果蝇的种类、生物特征、生活习性、发生规律和危害习性,并从农业防治、物理防治、化学防治和生物防治等方面总结分析了其综合防治措施,以期为葡萄园果蝇的可持续控制提供理论和技术支持。     

Electron microscope immunolocation of gap junctions in Drosophila

Gap junction immunolocation was carried out in thin sections of Lowicryl K4M-embedded Drosophila imaginal wing discs using an affinity-purified polycolonal anti-18 kD gap junction protein anitbody and a colloidal gold-conjugated secondary antibody. Colloidal gold labelling was predominantly associated with obliquely-sectioned gap junctions, the ends of junctional profiles and other regions in which the adjacent junctional membranes were separated or distorted. The pattern of staining suggests that the determinant recognized by the antibody is relatively inaccessible, probably with a topological location in the transmembrane or extracellular domain of the membrane-spanning connexin protein.     

果蝇共生菌降低有害真菌的毒性

【背景】目前对于如何解决有害真菌对黑腹果蝇的致死性病理研究较少,对共生菌抑制有害真菌的研究引起普遍关注。【目的】检测黑腹果蝇共生菌对病原性真菌的拮抗作用,揭示共生菌提高果蝇的适合度。【方法】利用PDA培养基分离黑腹果蝇食物中真菌;利用形态和rDNAITS基因序列比对进行真菌的鉴定;通过测量菌落直径、孢子数量以及菌丝分枝数量以评定真菌的生长;利用存活率评估病原真菌的毒性;建立无菌和悉生模型,通过发育历期验证其共生菌与病原性真菌的竞争作用;利用双向选择食物装置检测共生菌抑制病原真菌的效果。【结果】从果蝇食物中分离出的真菌经鉴定为拟茎点霉(Phomopsis),可显著地降低成年果蝇的存活率和延缓果蝇发育。东方醋酸杆菌在体外可明显抑制拟茎点霉的生长,有效地减轻拟茎点霉对果蝇的致死作用,挽救了拟茎点霉导致的果蝇发育延滞,改善了果蝇产卵对拟茎点霉的趋避作用。【结论】拟茎点霉是果蝇的一株条件性病原真菌,而东方醋酸杆菌可以有效地减轻拟茎点霉对果蝇生长发育和存活率的损害,从而提高果蝇适合度。     

RagA和Nprl2在果蝇肠道发育中的调节作用关系

动物胃肠道是食物消化和营养吸收器官,对机体健康至关重要。果蝇与哺乳动物的肠道在细胞组成、遗传调控等方面高度相似,是研究肠道发育的良好模型。体外培养细胞中的研究发现,Nprl2通过作用于Rag GTPase,抑制雷帕霉素靶点复合物1(target of rapamycin complex 1,TORC1)的活性,参与细胞代谢的调节。前期报道nprl2突变果蝇具有前胃增大、消化能力降低等肠道衰老相关表型。但对于Nprl2是否通过Rag GTPase调控肠道发育等方面尚不清楚。为了探究Rag GTPase在Nprl2调控果蝇肠道发育中的作用,本研究利用遗传杂交结合免疫荧光等方法对RagA敲减和nprl2突变果蝇的肠道形态、肠道细胞组成等方面进行研究。发现单独敲减RagA可以引起肠变粗、前胃增大等表型,敲减RagA能挽救nprl2突变体中肠道变细、分泌型细胞减少的表型,但并不能挽救nprl2突变体中前胃增大的表型。以上结果表明,RagA在肠道发育中发挥重要作用,Nprl2通过作用于Rag GTPase调节肠道细胞分化和肠道形态,但Nprl2对前胃发育和肠道的消化功能的调节可能通过不依赖于Rag GTPase的机制实现。