Examination of DNA sequences undergoing chromatin conformation changes at a variegating breakpoint in Drosophila melanogaster

Position effect variegation in Drosophila melanogaster is associated with the inability of certain genes to be correctly expressed in a proportion of cells, giving a mosaic phenotype. The lack of expression is thought to be due to alterations in the gene's chromatin structure due to its proximity to a region of heterochromatin. Because of the difficulties involved, there is little biochemical data to support the intuitively appealing model of heterochromatin spreading used to explain this phenomenon.Differences in restriction fragment length were used to distinguish DNA regions from either normal (non-position affected) or rearranged (position affected) chromosomes so as to examine possible changes in gene copy number and the effects of endogenous nucleases. DNA sequences at the breakpoint of In (1)w ^m4, which variegates for the white gene, were assayed under conditions where the chromatin conformation was altered using second site modifier mutations (Su(var) or En(var)). No change in the DNA sequerice copy number was observed at either chromosome breakpoint, relative to wild type, when either suppressor or enhancer mutations were present. Therefore copy number change, through differential polyploidization or somatic gene loss, is not affected by Su(var) or En(var) induced changes in the chromatin conformation.Initial experiments showed a gross difference in the sensitivity of DNA to endogenous nucleases that appeared associated with Su(var) and En(var) mutations. En(var) mutation bearing samples appeared delayed in the digestion, relative to Su(var). This differential sensitivity seemed to be genome-wide as there was no detectable difference between either breakpoint of In(1)w ^m4 or the sequences on the homologous w ^- chromosome. However, after isogenizing the genetic background, the previously noted difference between the Su(var) and En(var) mutations was eliminated. In studies dealing with nuclease digestion of chromatin, the isogenization of genetic background is essential before meaningful comparisons can be made.     

20-OH-ecdysone swells nuclear volume by alkalinization in salivary glands of Drosophila melanogaster

Ecdysteroids play an important role in the larval moulting process of insects. Ecdysone-induced stimulation causes specific puffs in polytene chromosomes of salivary gland cells resulting in nuclear swelling. During this process, changes of intracellular ion composition are thought to act as an early regulatory mechanism of gene activation. By use of video-imaging analysis and electrophysiological techniques, we examined ecdysone-induced nuclear swelling in Drosophila salivary glands in situ and its dependence on pH and calcium. Isolated glands of the third larval stage were superfused with a solution mimicking the haemolymph. Addition of 5×10^–6 mol/l 20-OH-ecdysone led, after a lag period of 50 min, to a sustained Ca²⁺-dependent increase of nuclear volume by 23.0±2.3%. Amiloride, a blocker of plasma membrane Na⁺/H⁺ exchange, prevented 20-OH-ecdysone-induced nuclear swelling. Decreasing pH in the superfusate from 7.15 to 6.8 led to nuclear shrinkage by 16.9±3.9%. Measurments of pH in salivary gland cells with ion-sensitive microelectrodes disclosed an alkalinization of 0.23±0.05 pH units after stimulation with 20-OH-ecdysone. We postulate that 20-OH-ecdysone activates the amilorde-sensitive plasma membrane Na⁺/H⁺ exchanger. This leads to intracellular alkalinization and concomitant decondensation of the nuclear chromatin visible as nuclear swelling. Thus, cell alkalinization could be a potentially important stimulatory mechanism in mediating ecdysteroid-induced activation of the cell nucleus.     

Pre-mating isolation is determined by larval rearing substrates in cactophilicDrosophila mojavensis. II. Effects of larval substrates on time to copulation,mate choice and mating propensity

Summary It has been hypothesized that reproductive character displacement has evolved in mainland Sonora, Mexico populations of cactophilicD. mojavensis due to the presence of a sympatric sibling speciesD. arizonae. In laboratory tests using ancestral Baja California populations and derived, sympatric mainland populations, asymmetrical sexual isolation has been observed among populations ofD. mojavensis where mainland females discriminate against Baja males. Effects of different pre-adult rearing environments on adult mating behaviour were assessed by comparing fermenting cactus tissues like those used in nature for breeding with laboratory media because previous studies have employed synthetic growth media for fly growth and development. Significant behavioural isolation was evident in all cases when larvae were reared on laboratory food, but was non-significant when flies were reared on fermenting cactus, except for the cactus used by most mainland populations, consistent with previous studies. Time to copulation of Baja females was greater than mainland females over all substrates, but male time to copulation did not differ between populations. Time to copulation for both sexes was significantly greater when flies were reared on laboratory food with one exception. The degree of behavioural isolation was weakly correlated with time to copulation across food types (Spearman rank correlation = 0.58,p = 0.099). Therefore, use of laboratory media in this and previous studies exaggerated adult pre-mating isolation and time to copulation in comparison to natural breeding substrates. These experiments suggest that a change in host substrates by saprophagous insects (where chemical differences exist between hosts) may have subtle effects on mating behaviour in a manner which promotes low levels of sexual isolation as a by-product of their utilization of a particular substrate during larval development. ForD. mojavensis, these results suggest that over evolutionary time, radiation into a new environment (from Baja to the mainland) allowed utilization of new host plants that may have incidentally promoted the sexual isolation patterns that have been observed within this species.See Etges (1992) for the first paper in this series.     

Climatic adaptations in the Drosophila immigrans species group: seasonal migration and thermal tolerance

Abstract. 1. In central Japan, Drosophila curviceps Okada and Kurokawa was collected in spring and autumn but not in summer at lowlands (alt. 500–1200 m), while it was collected only in summer at highlands (1500–2000 m). Experiments on its thermal tolerance suggested that summer heat at the lowlands and winter low temperatures at the highlands were adverse to this species. It is considered that this species escapes from these extreme temperatures by undergoing seasonal migration between the lowlands and the highlands. This species had no photo-periodic diapause and bred at both lowlands and highlands.
2. D.immigrans Sturtevant was less cold-hardy but more heat-tolerant than D.curviceps. It is considered that this species is unable to overwinter outdoors at least in the study areas (i.e. alt. 500m or higher in central Japan) and its populations in these areas originate with migrants from warmer areas.
3. D.albomicans Duda, a subtropical species, was less cold-hardy but more heat-tolerant than the above two species.
4. Climatic adaptations and distributions of these species are discussed with reference to their thermal tolerance.     

Drosophila acidic ribosomal protein rpA2: Sequence and characterization

A cDNA encoding the Drosophila melanogaster acidic ribosomal protein rpA2 was cloned and sequenced. rpA2 is homologous to the Artemia salina acidic ribosomal protein eL12′. In situ hybridization to salivary gland polytene chromosomes localizes the rpA2 gene to band 21C. It is a single copy gene, with an mRNA of 0.8 kb. Two-dimensional gel electrophoresis of Drosophila ribosomal proteins followed by immuno-blotting showed that the rpA2 protein has an apparent relative mobility in SDS of 17 kD and an isoelectric point less than pH 5.0. Although the Drosophila gene rp21C may be the same as rpA2, the reported sequences differ. Comparisons of the aligned nucleotide sequences coding for the acidic ribosomal proteins rpA1 and rpA2 of Drosophila with those of other eukaryotes support the view of two separate, though closely related, groups of acidic proteins. Comparison with the Artemia homologues suggests that nucleotide identity may have been conserved by some constraint that acts in addition to the requirement for substantial similarity of amino acid sequences. © 1993 Wiley-Liss, Inc.     

基于果蝇口腔感染的肠道训练免疫模型研究（英文）

目的 利用果蝇作为遗传工具从个体和分子层面研究果蝇的训练免疫效应,并为后续深入研究其分子机制提供依据。方法 首先构建无菌果蝇模型,在此基础上构建果蝇成虫及跨发育阶段训练免疫模型,用两种革兰氏阴性菌——胡萝卜软腐欧文氏菌（Erwinia carotovora carotovora 15）及铜绿假单胞菌（Pseudomonas aeruginosa）分别经口腔感染果蝇。在第一次感染完全消退后进行再次感染,然后通过比较果蝇在两个感染阶段的存活率和细菌量来衡量训练免疫的潜在效果。通过实时荧光定量PCR检测相应先天免疫相关基因的表达水平,研究革兰氏阴性菌对免疫缺陷（IMD）通路的诱导作用。结果 果蝇成虫及幼虫初次感染均可提高二次感染后的生存率、细菌清除效率及死亡时能承受的最高细菌负荷;二次感染的果蝇中,IMD通路中免疫反应基因的基础表达比未感染的高,这提供了获得感染抗性的分子基础;果蝇的免疫反应主要发生在中肠,二次免疫比初次免疫的效应更迅速且剧烈;二次免疫的果蝇中,肠道干细胞的数量显著多于初次感染。结论 果蝇肠道中强大的训练免疫可由同源或异源革兰氏阴性菌口腔感染引发,且免疫记忆可在整个发育阶段持...     

Neuronal specificity and its development in the Drosophila wing disc and its derivatives

The imaginal wing disc of flies gives rise to the adult wing blade and dorsal thorax (notum). A great deal has been learned in recent years about the process of neurogenesis in this disc; a number of genes that play crucial roles in the formation of sensory mother cells and in the differentiation of the sensory organs have been identified and their roles defined. Given this extensive background of developmental genetics, it has seemed profitable to summarize what is known about the end-products of neural development, the adult sensory organs. Discussed are their physiological function and role in behavior, the pathways followed by their axons in the CNS, and both genes and epigenetic processes that might play some role in the later stages of neural development and in adult function. The highly individual characteristics of certain of the sensory organs is emphasized, both in the context of their adult roles and as a challenge for future studies in developmental genetics. © 1993 John Wiley & Sons, Inc.     

HETEROCHRONIC DEVELOPMENTAL PLASTICITY IN LARVAL SEA URCHINS AND ITS IMPLICATIONS FOR EVOLUTION OF NONFEEDING LARVAE

Preexisting developmental plasticity in feeding larvae may contribute to the evolutionary transition from development with a feeding larva to nonfeeding larval development. Differences in timing of development of larval and juvenile structures (heterochronic shifts) and differences in the size of the larval body (shifts in allocation) were produced in sea urchin larvae exposed to different amounts of food in the laboratory and in the field. The changes in larval form in response to food appear to be adaptive, with increased allocation of growth to the larval apparatus for catching food when food is scarce and earlier allocation to juvenile structures when food is abundant. This phenotypic plasticity among full siblings is similar in direction to the heterochronic evolutionary changes in species that have greater nutrient reserves within the ova and do not depend on particulate planktonic food. This similarity suggests that developmental plasticity that is adaptive for feeding larvae also contributes to correlated and adaptive evolutionary changes in the transition to nonfeeding larval development. If endogenous food supplies have the same effect on morphogenesis as exogenous food supplies, then changes in genes that act during oogenesis to affect nutrient stores may be sufficient to produce correlated adaptive changes in larval development.