Characterization of Rhipicephalus microplus glycogen synthase kinase 3 Beta protein by inhibition assays
Schuler, Aline DominguesOliveira, Carlos Jorge Logullo deCoutinho, Mariana LonerVaz Junior, Itabajara da Silva
Background: Rhipicephalus microplus is a monogenetic, hematophagous ectoparasite that has a large economic impact due to associated losses in the cattle industry. Glycogen synthase kinase 3 is a highly conserved and ubiquitously expressed protein in several species. It has been identified as GSK-3 isoform in the cattle tick, and is involved in the modulation of glycogen synthase activity, as a regulator of glycogen synthesis with a role in energy metabolism of R. microplus. It is a fundamental kinase for embryo development, and is directly linked with R. microplus reproductive process. Thus, the aim of this study was to investigate the role of GSK-3 in R. microplus physiology by inhibiting its activity.Materials, Methods & Results: In vitro and in vivo assays were carried out to test the effects of immunological and chemical GSK-3 inhibition. Synthetic peptides were designed by an in silico analysis of the protein antigenic index. Rabbit antibodies were raised against the designed synthetic peptides based on R. microplus GSK-3 sequence (anti-SRm0218 and anti-SRm86100). To access if the inhibition of GSK-3 would result in a decrease in fertility and hatching, the purified IgG from rabbit sera was used to feed partially engorged tick females. Results show that the antibodies were not capable of affect egg production. The same antibodies were used in an in vitro...(AU)
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