Simulation of gene pyramiding in Drosophila melanogaster

Gene pyramiding has been successfully practiced in plant breeding for developing new breeds or lines in which favorable genes from several different lines were integrated.But it has not been used in animal breeding,and some theoretical investigation and simulation analysis with respect to its strategies,feasibility and efficiency are needed before it can be implemented in animals.In this study,we used four different pure fines of Drosophila melanogaster,each of which is homozygous at a specific mutant gene with a visible effect on phenotype,to simulate the gene pyramiding process and analyze the duration and population size required in different pyramiding strategies.We finally got the ideal individuals,which are homozygous at the four target genes simultaneously.This study demonstrates that gene pyramiding is feasible in animal breeding and the interaction between genes may affect the final results.     

黑腹果蝇成虫生长过程的脂质组分析

果蝇是一种重要的模式生物,为遗传学研究作出了巨大贡献。近几十年来,在发育生物学、行为生物学、神经生物学等领域也都发挥了十分关键的作用。脂质是构成生命的4大类基本物质之一,但关于果蝇脂质组特征研究并不多,尤其是黑腹果蝇在生长发育过程中的脂质组含量变化尚未见报导。为更全面了解果蝇生长发育的特性,本研究应用脂质组学方法,系统地表征了黑腹果蝇成虫生长过程中的337种脂质分子的变化。结果显示,磷脂总量从152.61 ± 4.92 nmol/mg 降低至112.3 ± 3.87 nmol/mg,其中溶血磷脂酰胆碱含量从0.70 ± 0.03 nmol/mg 升高至0.93 ± 0.11 nmol/mg。而中性脂甘油三酯 (TAG) 和甘油二酯 (DAG) 变化最剧烈,伴随果蝇生长,从356.12 ± 34.05 nmol/mg下降至86.99 ± 13.07 nmol/mg。同时,分析了与细胞膜稳定性有关的膜脂碳链长度、双键数目(DBI)和磷脂酰胆碱(PC)/磷脂酰乙醇胺(PE)比值的变化。结果显示,果蝇成虫在生长阶段早中期（8～15 d）细胞膜最稳定,通过增加膜脂双键数目（10%～20%）和升高PC/PE比值（14%）,增强膜稳定性。黑腹果蝇在成虫生长的不同阶段,其脂质分子组成特征不同,我们所发现的黑腹果蝇成虫生长过程中脂质分子含量变化,一方面为在脂质水平研究黑腹果蝇生长调控奠定了重要基础,另一方面也为黑腹果蝇作为研究模型时控制其正常健康生长提供了参考标准。     

Simulation of gene pyramiding in Drosophila melanogaster

Gene pyramiding has been successfully practiced in plant breeding for developing new breeds or lines in which favorable genes from several different lines were integrated.But it has not been used in animal breeding,and some theoretical investigation and simulation analysis with respect to its strategies,feasibility and efficiency are needed before it can be implemented in animals.In this study,we used four different pure lines of Drosophila melanogaster,each of which is homozygous at a specific mutant gene ...     

黑腹果蝇的性别控制

性别的形成包括两个过程,即性别决定和性别分化。果蝇的性别控制研究包括性别决定、性别分化、性别鉴定、性别诱导和性别控制5个方面。性别决定是在两种不同发育途径之间的选择,它提供了一个研究基因调控的模式系统。果蝇的性别决定问题已经研究得相当详细［1］。性别分化是使胚胎向着雌性或雄性发育的过程,决定了性别表型。果蝇的性别分化也取得了不少研究成果。近年来,许多重要的性别调控基因已被克隆和鉴定。随着果蝇基因组全序列测定的完成,果蝇的性别控制研究将会更为深入而完善。本文对与黑腹果蝇性别决定和性别分化相关的一些问题进行综述。     

Cytological location and further characterization of the Third chromosome resistance gene in Drosophila melanogaster

Mutations in the Third chromosome resistance (Tcr; 3-39.6) gene confer dominant resistance to α-methyl dopa and suggest the gene is involved in catecholamine metabolism. Evidence for involvement in catecholamine metabolism comes from the three phenotypes associated with the mutant Tcr chromosomes dominant resistance, dominant rescue of partially complementing l(2)amd alleles, and recessive lethal phenotypes. Only dominant resistance to αs-methyl dopa, however, was mapped to the Tcr locus. Both recessive lethality and dominant rescue of l(2)amd alleles have now been mapped to the Tcr gene and, through the isolation of a new deletion in the region, we demonstrate these phenotypes are due to a loss of Tcr function. This deletion places the Tcr gene in the 69B4-5 to 69C8-11 region. Additionally, we have tested and verified three predictions of the biochemical model proposed by Bishop, Sherald, and Wright (1989) for the function of the Tcr protein. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic mosaic analysis of decapentaplegic and wingless gene function in the Drosophila leg

 Genetically mosaic flies were constructed which lack a functional decapentaplegic (dpp) or wingless (wg) gene in portions of their leg epidermis, and the leg cuticle was examined for defects. Although dpp has previously been shown to be transcribed both ventrally and dorsally, virtually the only dpp-null clones that affect leg anatomy are those which reside dorsally. Conversely, wg-null clones only cause leg defects when they reside ventrally – a result that was expected, given that wg is only expressed ventrally. Both findings are consistent with models of leg development in which the future tip of the leg is specified by an interaction between dpp and wg at the center of the leg disc. Null clones can cause mirror-image cuticular duplications confined to individual leg segments. Double-ventral, mirror-image patterns are observed with dpp-null clones, and double-dorsal patterns with wg-null clones. Clones that are doubly mutant (null for both dpp and wg) manifest reduced frequencies for both types of duplications. Duplications can include cells from surrounding non-mutant territory. Such nonautonomy implies that both dpp and wg are involved in positional signaling, not merely in the maintenance of cellular identities. However, neither gene product appears to function as a morphogen for the entire leg disc, since the effects of each gene’s null clones are restricted to a discrete part of the circumference. Interestingly, the circumferential domains where dpp and wg are needed are complementary to one another. Received: 25 March 1996 / Accepted: 13 June 1996     

Drosophila melanogaster males respond differently at the behavioural and genome‐wide levels to Drosophila melanogaster and Drosophila simulans females

Drosophila melanogaster are found in sympatry with Drosophila simulans, and matings between the species produce nonfertile hybrid offspring at low frequency. Evolutionary theory predicts that females choose mates, so males should alter their behaviour in response to female cues. We show that D. melanogaster males quickly decrease courtship towards D. simulans females. Courtship levels are reduced within 5 min of exposure to a heterospecific female, and overall courtship is significantly lower than courtship towards conspecific females. To understand changes at the molecular level during mate choice, we performed microarray analysis on D. melanogaster males that courted heterospecific D. simulans females and found nine genes have altered expression compared with controls. In contrast, males that court conspecific females alter expression of at least 35 loci. The changes elicited by conspecific courtship likely modulate nervous system function to reinforce positive conspecific signals and dampen the response to heterospecific signals.     

Differential requirements for the neurogenic gene almondex during Drosophila melanogaster development

During early development, the neurogenic genes of Drosophila melanogaster are involved in the control of cell fates in the neurectoderm; almondex (amx) belongs to this category of genes. We have identified the amx locus and rescued the amx embryonic neurogenic phenotype with a 1.5 kb DNA fragment. Using a small deficiency, we generated a new amx mutant background called amx(m), which is a null allele. Besides the characteristic neurogenic maternal effect caused by loss of amx, amx(m) flies display a new imaginal phenotype resembling loss of function of Notch. We describe amx-induced misregulation of the Notch pathway target E(spl) m7 in embryos and genetic interactions between amx and Notch pathway mutants in adult flies. These data show that wildtype amx acts as a novel positive regulator of the Notch pathway and is required at different levels during development.     

Genome scans of variation and adaptive change: extended analysis of a candidate locus close to the phantom gene region in Drosophila melanogaster

Nucleotide variation in populations originating from the recent range expansion of a species should reflect their adaptation to new habitats as well as their demographic history. A survey of nucleotide variation at 109 noncoding X-chromosome fragments in a European population of Drosophila melanogaster allowed identifying some candidates to have been recently affected by positive selection. Adaptive changes leave a spatial differential footprint that can be used to discriminate among candidates by extending their study to neighboring regions. Here, we surveyed variation at an approximately 190-kb region spanning a locus exhibiting a significantly skewed frequency spectrum. A stretch of approximately 12 kb with reduced variation was detected within a continuously sequenced region that included the focal fragment. Moreover, the regions flanking this stretch exhibited an excess of high-frequency derived variants. Application of maximum likelihood ratio and goodness-of-fit tests suggested that the pattern of variation detected at the studied region (at cytological bands 17C-17D) might have been shaped by a recent selective change, most probably at or around the phantom gene that encodes CYP306A1, a cytochrome P450 enzyme in the ecdysteroidogenic pathway.     

No apparent cost of evolved immune response in Drosophila melanogaster

Maintenance and deployment of the immune system are costly and are hence predicted to trade‐off with other resource‐demanding traits, such as reproduction. We subjected this longstanding idea to test using laboratory experimental evolution approach. In the present study, replicate populations of Drosophila melanogaster were subjected to three selection regimes—I (Infection with Pseudomonas entomophila), S (Sham‐infection with MgSO₄), and U (Unhandled Control). After 30 generations of selection flies from the I regime had evolved better survivorship upon infection with P. entomophila compared to flies from U and S regimes. However, contrary to expectations and previous reports, we did not find any evidence of trade‐offs between immunity and other life history related traits, such as longevity, fecundity, egg hatchability, or development time. After 45 generations of selection, the selection was relaxed for a set of populations. Even after 15 generations, the postinfection survivorship of populations under relaxed selection regime did not decline. We speculate that either there is a negligible cost to the evolved immune response or that trade‐offs occur on traits such as reproductive behavior or other immune mechanisms that we have not investigated in this study. Our research suggests that at least under certain conditions, life‐history trade‐offs might play little role in maintaining variation in immunity.     

果蝇衰老的调控机制

衰老是一个复杂的生物学过程,涉及到有害物质的积累导致整体生命功能的下降,生物的生理状况逐渐恶化,最终导致疾病和死亡。黑腹果蝇Drosophila melanogaster作为最重要的遗传学工具之一,近年来常被用于衰老的研究,以阐明衰老的发生与发展机制。本文结合本实验室的研究进展,综述了果蝇寿命调控的生理生化机制,如保幼激素、胰岛素/类胰岛素生长因子、TOR信号网络、腺苷酸活化蛋白激酶信号通路、热量限制和饮食限制、氧化应激、小分子RNA以及鞘脂类代谢都会对果蝇的寿命产生影响。除此之外,基因调控网络研究还能够发现潜在的与长寿相关的基因组区域,将有可能发现更多寿命相关基因。以果蝇为模式生物的研究,对于其他昆虫衰老、存活等种群生物学问题的研究以及天敌、益虫保育和害虫控制,具有十分重要的指导意义。     

Jun N-terminal kinase signaling makes a face

decapentaplegic (dpp), the Drosophila ortholog of BMP 2/4, directs ventral adult head morphogenesis through expression in the peripodial epithelium of the eye-antennal disc. This dpp expressing domain exerts effects both on the peripodial epithelium, and the underlying disc proper epithelium. We have uncovered a role for the Jun N-terminal kinase (JNK) pathway in dpp-mediated ventral head development. JNK activity is required for dpp's action on the disc proper, but in the absence of dpp expression, excessive JNK activity is produced, leading to specific loss of maxillary palps. In this review we outline our hypotheses on how dpp acts by both short range and longer range mechanisms to direct head morphogenesis and speculate on the dual role of JNK signaling in this process. Finally, we describe the regulatory control of dpp expression in the eye-antennal disc, and pose the problem of how the various expression domains of a secreted protein can be targeted to their specific functions.     

P element regulation and X-chromosome subtelomeric heterochromatin in Drosophila melanogaster

In Drosophila melanogaster, crossing males carrying autonomous P elements with females devoid of P copies results in hybrid dysgenesis in the germline of progeny. The reciprocal cross produces non-dysgenic progeny due to a maternally inherited state non-permissive for P transposition. The capacity of a P copy to repress transposition depends on both its structure and its chromosomal location. Naturally occuring regulatory P elements inserted at the telomere of the X chromosome have been genetically isolated in a genomic context devoid of other P elements. One or two copies of autonomous P elements at this site (1A) are sufficient to elicit a strong P repression in the germline. These elements are flanked by Telomeric Associated Sequences, previously identified and described by Karpen and Spradling (1992) as having heterochromatic properties. The regulatory properties of P elements at 1A are strongly impaired by mutations affecting Su(var)205, which encodes Heterochromatin Protein 1, a non-histone heterochromatin protein. The regulatory properties of classical P strains are not sensitive to Su(var)205. Models based on chromatin structure or on nuclear localisation of the telomeres are discussed in order to explain both the strong regulatory properties of P elements at the X chromosome telomere and their sensitivity to Su(var)205. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characterization of a novel Drosophila melanogaster cis‐regulatory module that drives gene expression to the larval tracheal system and adult thoracic musculature

In a previous bioinformatics analysis we identified 10 conserved Drosophila melanogaster sequences that reside upstream from protein coding genes (CGs). Here we characterize one of these genomic regions, which constitutes a Drosophila melanogaster cis‐regulatory module (CRM) that we denominate TT‐CRM. The TT‐CRM is 646 bp long and is located in one of the introns of CG32239 and resides about 3,500 bp upstream of CG13711 and about 620 bp upstream of CG12493. Analysis of 646 bp‐lacZ lines revealed that TT‐CRM drives gene expression not only to the larval, prepupal, and pupal tracheal system but also to the adult dorsal longitudinal muscles. The patterns of mRNA expression of the transgene and of the CGs that lie in the vicinity of TT‐CRM were investigated both in dissected trachea and in adult thoraces. Through RT‐qPCR we observed that in the tracheal system the pattern of expression of 646 bp‐lacZ is similar to the pattern of expression of CG32239 and CG13711, whereas in the thoracic muscles 646 bp‐lacZ expression accompanies the expression of CG12493. Together, these results suggest new functions for two previously characterized D. melanogaster genes and also contribute to the initial characterization of a novel CRM that drives a dynamic pattern of expression throughout development.     

紫外线对黑尾果蝇的生物学效应

野生型黑尾果蝇经紫外线连续处理3代,随着照射时间延长,每代均表现显著的生物学效应。这些生物学效应主要是子代数量减少,羽化延续时间缩短,体重减轻,而且有致略效应和间接的诱变效应。但处理后的子代性别比例变化不大。     

Proteome reference map of Drosophila melanogaster head

Drosophila melanogaster has been used as a genetic model organism to understand the fundamental molecular mechanisms in human biology including memory formation that has been reported involving protein synthesis and/or post-translational modification. In this study, we employed a proteomic platform based on fluorescent 2DE and MALDI-TOF MS to build a standard D. melanogaster head proteome map for proteome-proteome comparison. In order to facilitate the comparison, an interactive database has been constructed for systematically integrating and analyzing the proteomes from different conditions and further implicated to study human diseases related to D. melanogaster model. In summary, the fundamental head proteomic database and bioinformatic analysis will be useful for further elucidating the biological mechanisms such as memory formation and neurodegenerative diseases.