scholarly journals Estudios funcionales comparados de la evolución de la segmentación en insectos

2014 ◽  
Author(s):  
◽  
Natalia Esponda Behrens
Keyword(s):  
Drosophila Melanogaster ◽  
Rhodnius Prolixus

La presente tesis comprende una revisión del desarrollo de Rhodnius prolixus (Stähl, 1859); anotación de genes del desarrollo temprano y estudios de expresión y funcionales de dos de esos genes. El objetivo general ha sido aportar datos que contribuyan a establecer redes génicas como blanco de la evolución de la forma en los insectos. Los objetivos específicos fueron: Identificar y anotar en el genoma de R.prolixus genes ortólogos a los genes HOX de Drosophila melanogaster (Meigen, 1830); caracterizar el cluster HOX y determinar la función de genes HOX mediante genómica funcional. Se identificaron 70 genes, la mayoría de ellos correspondientes al grupo de TF con homeobox. Se analizaron, curaron y anotaron 26 secuencias; incluyendo a los ocho HOX canónicos. Se logró demostrar que los genes HOX de R.prolixus están agrupados en un cluster y se plantean cinco agrupamientos probables. Los ensayos funcionales se realizaron usando un gen HOX ‒scr‒ y un activador HOX ‒caudal‒ involucrado en el establecimiento del eje anteroposterior. Para ello, se pusieron a punto las técnicas de hibridación in situ de embriones completos ‒WMISH‒ y de ARNi parental. La expresión de scr mostró un patrón acorde a lo esperado en relación a las observaciones hechas en otros insectos. La ARNi mostró variantes en comparación con especies relacionadas, pero se ajusta muy bien a lo esperado. La expresión de caudal muestra las siguientes similitudes con respecto a otras especies estudiadas: (1) actúa tempranamente como gen de efecto materno, (2) se expresa en la región posterior del huevo en estado de blastodermo, (3) los efectos de la ARNi son semejantes a los encontrados en otros insectos de banda germinal corta. Sin embargo, en estados de banda germinal, los resultados de expresión difieren respecto a otras observaciones, esto puede estar en relación con mecanismos de sañalización aún no descriptos para este gen.

Análisis genómico y molecular de la embriogénesis de Rhodnius prolixus (Stähl, 1859) (Hemíptera, Reduviidae): implicancias morfológico-evolutivas en insectos

2012 ◽  
Author(s):  
◽  
Lucía Elena Pagola
Keyword(s):  
Drosophila Melanogaster ◽  
Rhodnius Prolixus

Todos los animales que poseen simetría bilateral se encuentran definidos por dos ejes de simetría ortogonales, el eje anteroposterior (A-P) que corre de la boca al ano y un eje perpendicular a este, el eje dorsoventral (D-V). A pesar de la gran variedad de modos de desarrollo embrionario y formas finales encontradas en los animales, las redes regulatorias y factores de transcripción que dan origen a estos ejes se encuentran muy conservados. De aquí surge una pregunta central, cómo estas redes regulatorias tan conservadas crean tanta diversidad morfológica, se adaptan a nuevos ambientes y de qué manera las novedades evolutivas se incorporan en un sistema de patronamiento ya establecido. El eje DV es un buen sistema de estudio ya que se conoce en detalle en Drosophila melanogaster pero no en otros insectos. Los insectos además presentan una gran variedad de especies y modos de desarrollo embrionario lo que nos permite estudiar de qué manera las redes regulatorias se adaptan a novedades evolutivas y la existencia de varias técnicas que permiten testear el funcionamiento de los genes y sus interacciones. En este contexto hemos utilizado a Rhodnius prolixus como modelo para el estudio del establecimiento del eje DV en un embrión de banda germinal intermedia donde al final del desarrollo el embrión posee la misma forma que el adulto. Hemos estudiado en detalle el desarrollo embrionario de R. prolixus para una mejor comprensión de los patrones de expresión de los genes estudiados y su función. Además, se han buscado y anotado varios genes envueltos en la formación del eje DV: toll, dorsal, decapentaplegic, zerknült, twist y zelda. Mostraremos el patrón de expresión y los fenotipos resultado de ARNi parental de toll, dpp y dorsal, los cuales representan puntos clave en la regulación de la cascada D-V.

scholarly journals The Alcohol Dehydrogenase Gene Is Nested in the outspread Locus of Drosophila melanogaster

Genetics ◽  
1996 ◽  
Vol 143 (2) ◽  
pp. 897-911 ◽  
Author(s):  
S McNabb ◽  
S Greig ◽  
T Davis
Keyword(s):  
Drosophila Melanogaster ◽  
Alcohol Dehydrogenase ◽  
Pcr Amplification ◽  
Transcription Unit ◽  
Adjacent Gene ◽  
Dehydrogenase Gene ◽  
Chromosomal Breakpoints ◽  
Splicing Patterns ◽  
Somatic Musculature

Abstract This report describes the structure and expression of the outspread (osp) gene of Drosophila melanogaster. Previous work showed that chromosomal breakpoints associated with mutations of the osp locus map to both sides of the alcohol dehydrogenase gene (Adh), suggesting that Adh and the adjacent gene Adh' are nested in osp. We extended a chromosomal walk and mapped additional osp mutations to define the maximum molecular limit of osp as 119 kb. We identified a 6-kb transcript that hybridizes to osp region DNA and is altered or absent in osp mutants. Accumulation of this RNA peaks during embryonic and pupal periods. The osp cDNAs comprise two distinct classes based on alternative splicing patterns. The 5′ end of the longest cDNA was extended by PCR amplification. When hybridized to the osp walk, the 5′ extension verifies that Adh and Adh' are nested in osp and shows that osp has a transcription unit of ≥74 kb. In situ hybridization shows that osp is expressed both maternally and zygotically. In the ovary, osp is transcribed in nurse cells and localized in the oocyte. In embryos, expression is most abundant in the developing visceral and somatic musculature.

scholarly journals Direct determination of retrotransposon transposition rates in Drosophila melanogaster

1994 ◽  
Vol 63 (2) ◽  
pp. 139-144 ◽  
Author(s):  
Sergey V. Nuzhdin ◽  
Trudy F. C. Mackay
Keyword(s):  
Drosophila Melanogaster ◽  
Transposable Element ◽  
Inbred Line ◽  
Copy Number ◽  
Direct Determination ◽  
Hybridization Analysis ◽  
Spontaneous Mutations ◽  
Insertion Sites

SummaryRates of transposition and excision of the Drosophila melanogaster retrotransposon elements mdg3, 297, Doc, roo and copia were estimated directly, by in situ hybridization analysis of their cytological insertion sites in 31 replicates of a highly inbred line that had accumulated spontaneous mutations for approximately 160generations. Estimated transposition rates of Doc, roo and copia were, respectively, 4·2 × 10−5, 3·1 × 10−3 and 1·3 − 10−3; no transpositions of 297 nor mdg3 were observed. Rates of transposition of copia varied significantly among sublines. Excisions were only observed for roo elements, at a rate of 9·0 × 10−6 per element per generation. Copy number averaged over these element families increased 5·9 %; therefore, in these lines the magnitude of the forces opposing transposable element multiplication were weaker than transposition rates.

A novel, tissue-specific integrin subunit, beta nu, expressed in the midgut of Drosophila melanogaster

Development ◽  
1993 ◽  
Vol 118 (3) ◽  
pp. 845-858 ◽  
Author(s):  
G.H. Yee ◽  
R.O. Hynes
Keyword(s):  
Drosophila Melanogaster ◽  
Matrix Proteins ◽  
Beta Subunits ◽  
Integrin Subunit ◽  
Integrin Receptor ◽  
Tissue Specific ◽  
Surface Receptors ◽  
Polymerase Chain ◽  
Maximal Expression

The integrins are a family of cell surface receptors for extracellular matrix proteins and counter-receptors on other cells. We have used the polymerase chain reaction to identify a novel integrin receptor beta subunit in Drosophila melanogaster. The deduced amino acid sequence of this subunit, which we have termed beta v (beta-neu), indicates that it has several unusual properties. The beta v subunit is roughly 33% identical with each of the previously sequenced vertebrate and Drosophila beta subunits and is lacking four of the 56 cysteine residues characteristic of most members of this protein family. The expression of the beta v gene is strikingly restricted. It is temporally regulated, with maximal expression occurring at 12–15 hours of embryonic development. In situ hybridization analyses and antibody localization on whole-mount embryos reveal that beta v expression is tissue-specific and confined to the developing midgut endoderm and its precursors during embryogenesis. Tissue specificity of expression is maintained through later stages of development as beta v transcripts are found exclusively in the larval midgut. Within this structure, beta v transcripts are especially concentrated in the cells of the midgut imaginal islands which give rise to the adult midgut.

Quantitative in situ hybridization of ribosomal RNA species to polytene chromosomes of Drosophila melanogaster

1977 ◽  
Vol 115 (3) ◽  
pp. 539-563 ◽  
Author(s):  
Paul Szabo ◽  
Robert Elder ◽  
Dale M. Steffensen ◽  
Olke C. Uhlenbeck
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Ribosomal Rna ◽  
Polytene Chromosomes

scholarly journals The distribution of transposable elements within and between chromosomes in a population of Drosophila melanogaster. I. Element frequencies and distribution

1992 ◽  
Vol 60 (2) ◽  
pp. 103-114 ◽  
Author(s):  
Brian Charlesworth ◽  
Angela Lapid ◽  
Darlene Canada
Keyword(s):  
Population Dynamics ◽  
Drosophila Melanogaster ◽  
Linkage Disequilibrium ◽  
Transposable Elements ◽  
Copy Number ◽  
Polytene Chromosomes ◽  
Chromosome Band ◽  
High Frequencies ◽  
Copy Numbers

SummaryData were collected on the distribution of nine families of transposable elements among second and third chromosomes isolated from a natural population of Drosophila melanogaster, by means of in situ hybridization of element probes to polytene chromosomes. It was found that the copy numbers per chromosome in the distal sections of the chromosome arms followed a Poisson distribution. Elements appeared to be distributed randomly along the distal sections of the chromosome arms. There was no evidence for linkage disequilibrium in the distal sections of the chromosomes, but some significant disequilibrium was detected in proximal regions. There were many significant correlations between different element families with respect to the identity of the sites that were occupied in the sample. There were also significant correlations between families with respect to sites at which elements achieved relatively high frequencies. Element frequencies per chromosome band were generally low in the distal sections, but were higher proximally. These results are discussed in the light of models of the population dynamics of transposable elements. It is concluded that they provide strong evidence for the operation of a force or forces opposing transpositional increase in copy number. The data suggest that the rate of transposition perelement per generation is of the order of 10−4, for the elements included in this study.

scholarly journals A fertility region on the Y chromosome of Drosophila melanogaster encodes a dynein microtubule motor

1993 ◽  
Vol 90 (23) ◽  
pp. 11132-11136 ◽  
Author(s):  
J Gepner ◽  
T S Hays
Keyword(s):  
Drosophila Melanogaster ◽  
Y Chromosome ◽  
Heavy Chain ◽  
Male Fertility ◽  
Chromosome Region ◽  
Cytoplasmic Dynein ◽  
Dynein Heavy Chain ◽  
Microtubule Motor ◽  
Hydrolysis Of

A clone encoding a portion of the highly conserved ATP-binding domain of a dynein heavy-chain polypeptide was mapped to a region of the Drosophila melanogaster Y chromosome. Dyneins are large multisubunit enzymes that utilize the hydrolysis of ATP to move along microtubules. They were first identified as the motors that provide the force for flagellar and ciliary bending. Seven different dynein heavy-chain genes have been identified in D. melanogaster by PCR. In the present study, we demonstrate that one of the dynein genes, Dhc-Yh3, is located in Y chromosome region h3, which is contained within kl-5, a locus required for male fertility. The PCR clone derived from Dhc-Yh3 is 85% identical to the corresponding region of the beta heavy chain of sea urchin flagellar dynein but only 53% identical to a cytoplasmic dynein heavy chain from Drosophila. In situ hybridization to Drosophila testes shows Dhc-Yh3 is expressed in wild-type males but not in males missing the kl-5 region. These results are consistent with the hypothesis that the Y chromosome is needed for male fertility because it contains conventional genes that function during spermiogenesis.

scholarly journals Rates of movement of transposable elements on the second chromosome of Drosophila melanogaster

2000 ◽  
Vol 75 (3) ◽  
pp. 275-284 ◽  
Author(s):  
XULIO MASIDE ◽  
STAVROULA ASSIMACOPOULOS ◽  
BRIAN CHARLESWORTH
Keyword(s):  
Drosophila Melanogaster ◽  
Transposable Elements ◽  
Polytene Chromosomes ◽  
Mutation Accumulation ◽  
Long Terminal Repeats ◽  
Significant Difference ◽  
Transposable Element Copy ◽  
Excision Rate

The rates of movement of 11 families of transposable elements of Drosophila melanogaster were studied by means of in situ hybridization of probes to polytene chromosomes of larvae from a long-term mutation accumulation experiment. Replicate mutation-accumulation lines carrying second chromosomes derived from a single common ancestral chromosome were maintained by backcrosses of single males heterozygous for a balancer chromosome and a wild-type chromosome, and were scored after 116 generations. Twenty-seven transpositions and 1 excision were detected using homozygous viable and fertile second chromosomes, for a total of 235056 potential sources of transposition events and a potential 252880 excision events. The overall transposition rate per element per generation was 1·15×10−4 and the excision rate was 3·95×10−6. The single excision (of a roo element) was due to recombination between the element's long terminal repeats. A survey of the five most active elements among nine homozygous lethal lines revealed no significant difference in the estimates of transposition and excision rates from those from viable lines. The excess of transposition over excision events is in agreement with the results of other in situ hybridization experiments, and supports the conclusion that replicative increase in transposable element copy number is opposed by selection. These conclusions are compared with those from other studies, and with the conclusions from population surveys of element frequencies.

Transformation of Drosophila melanogaster with a suppressor tRNA gene from Schizosaccharomyces pombe

Genome ◽  
1988 ◽  
Vol 30 (2) ◽  
pp. 211-217 ◽  
Author(s):  
Charles M. Molnar ◽  
Tove Reece ◽  
James A. Williams ◽  
John B. Bell
Keyword(s):  
Drosophila Melanogaster ◽  
Salivary Gland ◽  
In Situ Hybridization ◽  
Schizosaccharomyces Pombe ◽  
Southern Hybridization ◽  
Trna Gene ◽  
P Element ◽  
Suppressor Trna ◽  
Good Agreement

P-element mediated transformation was utilized to introduce a suppressor tRNA gene [Formula: see text] from Schizosaccharomyces pombe into Drosophila melanogaster. Thirteen independently transformed lines were characterized as to the number and cytological locations of the transposons. It was ascertained that the suppressor tRNA gene of interest was introduced into each transformed strain. The helper P element used (pπ25.1) allows further transposition to occur, and it was determined that from one to seven copies of the heterologous [Formula: see text] gene per strain were present among the respective transformed strains. The number of transposons per transformed line was established by in situ hybridization to salivary gland chromosomes as well as by Southern hybridization analyses and there was good agreement in the totals determined by these two techniques.Key words: Drosophila, Schizosaccharomyces, tRNA suppressor, transformation, transposon.

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