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Glycogen Synthase Kinase-3 is involved in glycogen metabolism control and embryogenesis of Rhodnius prolixus

Published online by Cambridge University Press:  24 August 2016

FLÁVIA B. MURY
Affiliation:
LIBHM-NUPEM, Universidade Federal do Rio de Janeiro, Macaé, RJ Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de Janeiro, RJ
MAGDA D. LUGON
Affiliation:
LQFPP and UEA-RJ, Universidade Estadual do Norte Fluminense Darci Ribeiro, Campos dos Goytacazes, RJ
RODRIGO NUNES DA FONSECA
Affiliation:
LIBHM-NUPEM, Universidade Federal do Rio de Janeiro, Macaé, RJ Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de Janeiro, RJ
JOSE R. SILVA
Affiliation:
LIBHM-NUPEM, Universidade Federal do Rio de Janeiro, Macaé, RJ Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de Janeiro, RJ
MATEUS BERNI
Affiliation:
Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ
HELENA M. ARAUJO
Affiliation:
Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ
MARCIO RIBEIRO FONTENELE
Affiliation:
Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ
LEONARDO ARAUJO DE ABREU
Affiliation:
LIBHM-NUPEM, Universidade Federal do Rio de Janeiro, Macaé, RJ Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de Janeiro, RJ
MARÍLVIA DANSA
Affiliation:
Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de Janeiro, RJ LQFPP and UEA-RJ, Universidade Estadual do Norte Fluminense Darci Ribeiro, Campos dos Goytacazes, RJ
GLÓRIA BRAZ
Affiliation:
Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de Janeiro, RJ Departamento de Bioquímica, Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ
HATISABURO MASUDA
Affiliation:
Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de Janeiro, RJ Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ
CARLOS LOGULLO*
Affiliation:
Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM), Rio de Janeiro, RJ LQFPP and UEA-RJ, Universidade Estadual do Norte Fluminense Darci Ribeiro, Campos dos Goytacazes, RJ
*
*Corresponding author: Universidade Estadual do Norte Fluminense Darci Ribeiro, CBB/LQFPP/UEA-RJ, Av. Alberto Lamego, 2000. Zip code: 28013-620. Campos dos Goytacazes, RJ. E-mail: [email protected]

Summary

Rhodnius prolixus is a blood-feeding insect that transmits Trypanosoma cruzi and Trypanosoma rangeli to vertebrate hosts. Rhodnius prolixus is also a classical model in insect physiology, and the recent availability of R. prolixus genome has opened new avenues on triatomine research. Glycogen synthase kinase 3 (GSK-3) is classically described as a key enzyme involved in glycogen metabolism, also acting as a downstream component of the Wnt pathway during embryogenesis. GSK-3 has been shown to be highly conserved among several organisms, mainly in the catalytic domain region. Meanwhile, the role of GSK-3 during R. prolixus embryogenesis or glycogen metabolism has not been investigated. Here we show that chemical inhibition of GSK-3 by alsterpaullone, an ATP-competitive inhibitor of GSK3, does not affect adult survival rate, though it alters oviposition and egg hatching. Specific GSK-3 gene silencing by dsRNA injection in adult females showed a similar phenotype. Furthermore, bright field and 4’−6-diamidino-2-phenylindole (DAPI) staining analysis revealed that ovaries and eggs from dsGSK-3 injected females exhibited specific morphological defects. We also demonstrate that glycogen content was inversely related to activity and transcription levels of GSK-3 during embryogenesis. Lastly, after GSK-3 knockdown, we observed changes in the expression of the Wingless (Wnt) downstream target β-catenin as well as in members of other pathways such as the receptor Notch. Taken together, our results show that GSK-3 regulation is essential for R. prolixus oogenesis and embryogenesis.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2016 

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