A Rapid and Efficient Method to Dissect Pupal Wings of Drosophila Suitable for Immunodetections or PCR Assays
Posted by Ernesto on Tuesday, 3 June 2025
Autores:
Carmen Bolatto 1 , Cristina Parada 2 , Victoria Colmenares 2
Revista (o libro):
J Vis Exp
Año:
2017
Mes-dia:
1230
issue, vol, paginas, etc:
(130):55854
doi:
10.3791/55854
PMID:
29364201
Abstract:
Wing development in Drosophila melanogaster is an ideal model for studying morphogenesis at tissue level. These appendages develop from a group of cells named wing imaginal discs formed during embryonic development. In the larval stages the imaginal discs grow, increasing its number of cells and forming monolayered epithelial structures. Inside the pupal case, the imaginal discs bud out and fold into bilayers along a line that becomes the future margin of the wing. During this process, the longitudinal primodia veins originate vein cells on the prospective dorsal and ventral surfaces of the wing. During the pupal stage the stripes of vein cells of each surface communicate in order to generate tight tubes; at the same time, the cross-veins begin their formation. With the help of appropriate molecular markers, it is possible to identify the major elements composing the wing during its development. For this reason, the ability to accurately detect transcripts or proteins in this structure is critical for studying their abundance and localization related to the development process of the wing. The procedure described here focuses on manipulating pupal wings, providing detailed instructions on how to dissect the wing during the pupal stage. The dissection of pupal tissue is more difficult to perform than their counterparts in third instar larvae. This is why this approach was developed, to obtain rapid and efficient high quality samples. Details of how to immunostain and mount these wing samples, to allow the visualization of proteins or cell components, are provided in the protocol. With little expertise it is possible to collect 8-10 high quality pupal wings in a short amount of time.
Afiliaciones:
1 Departamento de Histología y Embriología, Facultad de Medicina, Laboratorio de Biología del Desarrollo; cbolatto@fmed.edu.uy.
2 Departamento de Histología y Embriología, Facultad de Medicina, Laboratorio de Biología del Desarrollo.
Enlace pubmed:
https://pubmed.ncbi.nlm.nih.gov/29364201/
Enlace full text:
https://doi.org//10.3791/55854
Cita:
Bolatto C, Parada C, Colmenares V. A Rapid and Efficient Method to Dissect Pupal Wings of Drosophila Suitable for Immunodetections or PCR Assays. J Vis Exp. 2017 Dec 30;(130):55854. doi: 10.3791/55854. PMID: 29364201; PMCID: PMC5908410.