Sexual selection in Drosophila melanogaster after long-term maintaining on different substrates

Veoma izražene morfološke i ponašajne osobine kod životinja često su rezultat delovanja seksualne selekcije. Kod insekata, ovaj proces za rezultat ima prisustvo intenzivne pigmentacije, neuobičajenih morfoloških struktura i kompleksnih feromona, koji se odlikuju velikom varijabilnošću u zavisnosti od sredinskih faktora. Za voćnu mušicu, D. melanogaster, jedan od najvažnijih faktora sredine je ishrana, koja ima značajan efekat na fiziološke, semiohemijske, morfološke i ponašajne osobine. Cilj ove doktorske disertacije je praćenje efekta višegodišnjeg gajenja pet linija D. melanogaster na različitim supstratima (standardni laboratorijski supstrat i supstrati koji sadrže paradajz, bananu, šargarepu i jabuku) na uspeh u parenju, morfološke i semiohemijske osobine. Rezultati ove doktorske disertacije su pokazali da su mužjaci gajeni na supstratu koji sadrži paradajz imali najveći uspeh u parenju, dok su najmanji uspeh u parenju imali mužjaci gajeni na podlozi koja sadrži jabuku. Razlike u uspehu u parenju, između ostalog, se mogu pripisati i razlikama u sastavu supstrata, ali i razlikama u komponentama adaptivne vrednosti. Značajne razlike između linija su uočene i na semiohemijskom i morfološkom nivou. Seksualna izolacija je utvrđena između linija održavanih na supstratima koji sadrže bananu i šargarepu. Na uspeh u parenju značajno je uticao oblik krila mušica, dok veličina krila nije imala značajan efekat. Takođe, kutikularni ugljovodonici, pre svega seksualni feromoni kakvi su 7-trikozen, 7-pentakozen, cVA, 7,11-nonakozadien i 7,11- heptakozadien, su imali važnu ulogu u odabiru partnera.Very pronounced morphological and behavioural traits in animals are often the result of acting sexual selection. In insects, this process leads to intense pigmentation, unusual morphological structures and complex pheromones, exhibiting great variability depending on the environments. For fruit fly, D. melanogaster, one of the most important environmental factors is food, which has a significant effect on physiological, semiochemical, morphological and behavioural traits. The aim of this doctoral dissertation was to determine the impact of long-term maintaining of five D. melanogaster strains on different nutritive substrates (standard laboratory substrate and substrates which contain tomato, banana, carrot and apple) on mating success, morphological and semiochemical traits. The results of this doctoral dissertation showed that males reared on tomato diet had the highest mating success, while males reared on apple diet had the lowest mating success. Differences in mating success could be related with differences in the substrate composition, as well with differences in the fitness traits. Significant differences among strains were also observed on semiochemical and morphological levels. Sexual isolation has been established between strains reared on banana and apple diets. Mating success was significantly influenced by the wing shape, while the wing size showed no effect. Also, cuticular hydrocarbons, primarily sexual pheromones 7-tricosene, 7-pentacosene, cVA, 7,11-nonacosadiene and 7,11-heptacosadiene, had important role in mate choice

Penile morphology in six populations of Branchipus schaefferi Fischer, 1834 (Crustacea: Branchiopoda) from Serbia

Male genital morphology of six populations of Branchipus schaefferi from different locations in Serbia is described. Both qualitative and quantitative morphological traits were taken into consideration. The majority of males from lowland parts of the country (Pannonian Plane) had a long sickle-curved or arched spine situated on the basal part of the penis. Males sampled in the south-eastern part of the country (the region of Stara Planina Mountains) possessed short, straight or slightly bowed basal spine. A distal penile part appeared as a more stable character than the proximal one. Males from one of the examined populations possessed roundish and spineless penile tip, a feature heretofore not described in B. schaefferi.Acta Zoologica Bulgarica (2017), 69(1): 17-2

Cuticular chemoprofile of the fruit fly Drosophila Subobscura (Diptera, drosophilidae)

In insects, cuticular hydrocarbon (CHC) profile is involved in many important biological functions and may vary in different conditions. Among fruit fly species, Drosophila subobscura is one of the most frequently used in genetic, ecological and evolutionary research, because of its rich chromosomal polymorphism, specific behavioral repertoires and habitat preferences. In this work, we identified and quantified cuticular chemoprofile of D. subobscura. Using gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS), 25 chemical compounds were found in males and 23 compounds were found in females. Further, ANOVA confirmed significant sexual dimorphism in cuticular chemoprofile amounts. Knowledge of cuticular chemistry could contribute to further research in D. subobscura, starting from behavioral, up to ecological, since this species is recognized as an important model system for the study and monitoring of global climate changes. © 2018, Pakistan Agricultural Scientists Forum. All rights reserved

Cuticular chemoprofile of the fruit fly Drosophila Subobscura (Diptera, drosophilidae)

In insects, cuticular hydrocarbon (CHC) profile is involved in many important biological functions and may vary in different conditions. Among fruit fly species, Drosophila subobscura is one of the most frequently used in genetic, ecological and evolutionary research, because of its rich chromosomal polymorphism, specific behavioral repertoires and habitat preferences. In this work, we identified and quantified cuticular chemoprofile of D. subobscura. Using gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS), 25 chemical compounds were found in males and 23 compounds were found in females. Further, ANOVA confirmed significant sexual dimorphism in cuticular chemoprofile amounts. Knowledge of cuticular chemistry could contribute to further research in D. subobscura, starting from behavioral, up to ecological, since this species is recognized as an important model system for the study and monitoring of global climate changes. © 2018, Pakistan Agricultural Scientists Forum. All rights reserved

Sexual selection in Drosophila melanogaster after long-term maintaining on different substrates

Veoma izražene morfološke i ponašajne osobine kod životinja često su rezultat delovanja seksualne selekcije. Kod insekata, ovaj proces za rezultat ima prisustvo intenzivne pigmentacije, neuobičajenih morfoloških struktura i kompleksnih feromona, koji se odlikuju velikom varijabilnošću u zavisnosti od sredinskih faktora. Za voćnu mušicu, D. melanogaster, jedan od najvažnijih faktora sredine je ishrana, koja ima značajan efekat na fiziološke, semiohemijske, morfološke i ponašajne osobine. Cilj ove doktorske disertacije je praćenje efekta višegodišnjeg gajenja pet linija D. melanogaster na različitim supstratima (standardni laboratorijski supstrat i supstrati koji sadrže paradajz, bananu, šargarepu i jabuku) na uspeh u parenju, morfološke i semiohemijske osobine. Rezultati ove doktorske disertacije su pokazali da su mužjaci gajeni na supstratu koji sadrži paradajz imali najveći uspeh u parenju, dok su najmanji uspeh u parenju imali mužjaci gajeni na podlozi koja sadrži jabuku. Razlike u uspehu u parenju, između ostalog, se mogu pripisati i razlikama u sastavu supstrata, ali i razlikama u komponentama adaptivne vrednosti. Značajne razlike između linija su uočene i na semiohemijskom i morfološkom nivou. Seksualna izolacija je utvrđena između linija održavanih na supstratima koji sadrže bananu i šargarepu. Na uspeh u parenju značajno je uticao oblik krila mušica, dok veličina krila nije imala značajan efekat. Takođe, kutikularni ugljovodonici, pre svega seksualni feromoni kakvi su 7-trikozen, 7-pentakozen, cVA, 7,11-nonakozadien i 7,11- heptakozadien, su imali važnu ulogu u odabiru partnera.Very pronounced morphological and behavioural traits in animals are often the result of acting sexual selection. In insects, this process leads to intense pigmentation, unusual morphological structures and complex pheromones, exhibiting great variability depending on the environments. For fruit fly, D. melanogaster, one of the most important environmental factors is food, which has a significant effect on physiological, semiochemical, morphological and behavioural traits. The aim of this doctoral dissertation was to determine the impact of long-term maintaining of five D. melanogaster strains on different nutritive substrates (standard laboratory substrate and substrates which contain tomato, banana, carrot and apple) on mating success, morphological and semiochemical traits. The results of this doctoral dissertation showed that males reared on tomato diet had the highest mating success, while males reared on apple diet had the lowest mating success. Differences in mating success could be related with differences in the substrate composition, as well with differences in the fitness traits. Significant differences among strains were also observed on semiochemical and morphological levels. Sexual isolation has been established between strains reared on banana and apple diets. Mating success was significantly influenced by the wing shape, while the wing size showed no effect. Also, cuticular hydrocarbons, primarily sexual pheromones 7-tricosene, 7-pentacosene, cVA, 7,11-nonacosadiene and 7,11-heptacosadiene, had important role in mate choice

“Scent of a fruit fly”: Cuticular chemoprofilesafter mating in differently fedDrosophilamelanogaster(Diptera: Drosophilidae) strains

n the world of complex smells in natural environment,feeding and mating represent two important olfactory‐guided behaviors inDrosophila melanogaster(Diptera:Drosophilidae). Diet affects the chemoprofile compositionof the individuals, which, indirectly, may significantly affecttheir mating success. In this study, chemoprofiles of re-cently mated flies belonging to fourD. melanogasterstrains,which were fed for many generations on different sub-strates (standard cornmeal—S strain; banana—B strain;carrot—C strain; tomato—T strain) were identified andquantified. In total, 67 chemical compounds were identi-fied: 48 compounds were extracted from males maintainedon banana and carrot, and 47 compounds from malesmaintained on cornmeal and tomato substrates, while totalof 60 compounds were identified in females from allstrains. The strains and the sexes significantly differed inqualitative nature of their chemoprofiles after mating. Sig-nificant differences in the relative amount of three majormale pheromones (cis‐vaccenyl acetate—cVA, (Z)‐7‐pentacosene, and (Z)‐7‐tricosene) and in female pheromone(Z,Z)‐7,11‐nonacosadiene among strains were also re-corded. Furthermore, multivariate analysis of variance(MANOVA) pointed to significant differences between irgin and mated individuals of all strains and within bothsexes. Differences in some of the well known sex pher-omones were also identified when comparing their relativeamount before and after mating. The presence of typicalmale pheromones in females, and vice versa may indicatetheir bidirectional transfer during copulation. Our resultsconfirm significant effect of mating status on cuticular hy-drocarbon (CHC) phenotypes in differently fedD. melano-gasterflies

"Does my Diet Affect my Perfume?' Identification and Quantification of Cuticular Compounds in Five Drosophila melanogaster Strains Maintained over 300 Generations on Different Diets

Cuticular hydrocarbons (CHCs) in Drosophila melanogaster represent the basis of chemical communication being involved in many important biological functions. The aim of this study was to characterize chemical composition and variation of cuticular profiles in five D. melanogaster strains. These strains were reared for approximately 300 generations on five diets: standard cornmeal medium and substrates prepared with apple, banana, tomato, and carrot. Differences in quantity and/or quality in CHCs were assumed as a result of activation of different metabolic pathways involved in food digestion and adaptations to the particular diet type. In total, independently of sex and strain, 66 chemical compounds were identified. In females of all strains, 60 compounds were identified, while, in males, 47 compounds were extracted. Certain new chemical compounds for D. melanogaster were found. MANOVA confirmed that CHC amounts significantly depend on sex and substrates, as well as on their interactions. Discriminant analysis revealed that flies belonging to apple' and carrot' strains exhibited the most noticeable differences in CHC repertoires. A non-hydrocarbon pheromone, cis-vaccenyl acetate (cVA) also contributed to the variation in the pheromone bouquet among the strains. Variability detected in CHCs and cVA may be used in the explanation of differences in mating behaviour previously determined in analyzed fly strains

Cyclohexyl Analogues of Ethylenediamine Dipropanoic Acid Induce Caspase-Independent Mitochondrial Apoptosis in Human Leukemic Cells

We investigated the cytotoxicity of recently synthesized (S,S)-ethyleridiamine-N,N'-di-2-(3-cyclohexyl)propanoic acid esters toward human leukemic cell lines and healthy blood mononuclear cells. Cell viability was assessed by acid phosphatase assay, apoptosis, and differentiation were analyzed by flow cytometry and electron microscopy, while intracellular localization of apoptosis-inducing factor (AIF) was determined by immunoblotting. It was demonstrated that methyl, ethyl, and n-propyl esters were toxic to HL-60, REH, MOLT-4, KG-1, JVM-2, and K-562 leukemic cell lines, while the nonesterified parental compound and n-butyl ester were devoid of cytotoxic action. The ethyl ester exhibited the highest cytotoxic activity (IC50 10.7 mu M-45.4 mu M), which was comparable to that of the prototypical anticancer drug cisplatin. The observed cytotoxic effect in HL-60 cells was associated with an increase in superoxide production and mitochondrial membrane depolarization, leading to apoptotic cell death characterized by phosphatidylserine externalization and DNA fragmentation in the absence of autophagic response. DNA fragmentation preceded caspase activation and followed AIF translocation from mitochondria to nucleus, which was indicative of caspase-independent apoptotic cell death. HL-60 cells treated with subtoxic concentration of the compound displayed morphological signs of granulocytic differentiation (nuclear indentations and presence of cytoplasmic primary granules), as well as an increased expression of differentiation markers CD11b and CD15. The cyclohexyl analogues of ethylenediamine dipropanoic acid were also toxic to peripheral blood mononuclear cells of both healthy controls and leukemic patients, the latter being more sensitive. Our data demonstrate that the toxicity of the investigated cyclohexyl compounds against leukemic cell lines is mediated by caspase-independent apoptosis associated with oxidative stress, mitochondrial dysfunction, and AIF translocation.Ministry of Science of the Republic of Serbia [41025, 172035

Cyclohexyl Analogues of Ethylenediamine Dipropanoic Acid Induce Caspase-Independent Mitochondrial Apoptosis in Human Leukemic Cells

We investigated the cytotoxicity of recently synthesized (S,S)-ethyleridiamine-N,N'-di-2-(3-cyclohexyl)propanoic acid esters toward human leukemic cell lines and healthy blood mononuclear cells. Cell viability was assessed by acid phosphatase assay, apoptosis, and differentiation were analyzed by flow cytometry and electron microscopy, while intracellular localization of apoptosis-inducing factor (AIF) was determined by immunoblotting. It was demonstrated that methyl, ethyl, and n-propyl esters were toxic to HL-60, REH, MOLT-4, KG-1, JVM-2, and K-562 leukemic cell lines, while the nonesterified parental compound and n-butyl ester were devoid of cytotoxic action. The ethyl ester exhibited the highest cytotoxic activity (IC50 10.7 mu M-45.4 mu M), which was comparable to that of the prototypical anticancer drug cisplatin. The observed cytotoxic effect in HL-60 cells was associated with an increase in superoxide production and mitochondrial membrane depolarization, leading to apoptotic cell death characterized by phosphatidylserine externalization and DNA fragmentation in the absence of autophagic response. DNA fragmentation preceded caspase activation and followed AIF translocation from mitochondria to nucleus, which was indicative of caspase-independent apoptotic cell death. HL-60 cells treated with subtoxic concentration of the compound displayed morphological signs of granulocytic differentiation (nuclear indentations and presence of cytoplasmic primary granules), as well as an increased expression of differentiation markers CD11b and CD15. The cyclohexyl analogues of ethylenediamine dipropanoic acid were also toxic to peripheral blood mononuclear cells of both healthy controls and leukemic patients, the latter being more sensitive. Our data demonstrate that the toxicity of the investigated cyclohexyl compounds against leukemic cell lines is mediated by caspase-independent apoptosis associated with oxidative stress, mitochondrial dysfunction, and AIF translocation.Ministry of Science of the Republic of Serbia [41025, 172035