Mapeamento físico de genes expressos de resposta imune e sensorial de Anopheles (Nyssorhynchus) Darlingi, vetor da malária neotropical

Abstract

Malaria in Brazil occurs especially in the Amazon (99% of cases), where environmental conditions are favorable for the proliferation of the etiologic and their mosquito vectors agents, whose main transmitter is the Anopheles darlingi. Environmental factors and human activities are some of the main aspects that contribute to the adaptive and evolutionary process that anopheles, which can be reflected in chromosomal rearrangements. We carried out the physical chromosome mapping by fluorescence in situ hybridization (FISH) of inume response genes in the genome of A. darlingi, GNBP (Gran-negative binding protein), Toll-interacting protein (Toll), defensin, Chymotrypsin-like serine protease (AdChyL), 28S ribosomal protein S7- mitochondrial precursor (RpS7), gambicin anti-microbial peptide (gambicin) and also the gene sensory response: OBP (odorant binding protein) to assist in more precise assembly of its structural genomics. These genes have been mapped in single chromosomal regions, with the exception of RpS7, which mapped in more than one location. Other genes have been mapped in inversion regions: GNBP (inversion 2 Rd); AdChyL in the region 38D (inversion 3Lb) and RpS7 in 22C region (inversion 2Lb). The OBP gene mapped the X chromosome from five locations individuals, where only A. darlingi of São Gabriel da Cachoeira/AM (SGC/AM) OBP gene marked the region 3A compared to four locations Coari/AM, Manaus/AM, Barra do Bugres/MT and Macapá/AP, where the probe mapped the region 4A. Two new investments were described for A. darlingi and one on chromosome X (Xb) in the population of SGC/AM and another in the arm 2L (2LC), population Bugres/MT. Geographical distances and ecoregions (geographical barriers) are environmental factors that can favor the appearance of the two new inversions described in this work. In the phylogenetic analysis of GNBP genes Toll, defensin, RpS7 and gambicin, we obtained three clades for GNBP and most of the analyzed sequence was homologous to the subfamily B, including GNBP Anopheles gambiae (87%), suggesting that GNBP A . darlingi belongs to the subfamily B. Phylogenetic trees for RpS7 genes, gambicin, defensin and Toll showed a high degree of conservation between these genes in A. gambiae and Anopheles arabiensis. Immune response genes of A. darlingi and Anopheles albimanus are phylogenetically close but not always a reliable support. This may suggest some level of evolutionary conservation of the genes of the immune response of both species. The genes mapped in situ were considered cytogenetic useful markers for chromosomal variability studies and developments in A. darlingi as they are conserved genes, and assist in the improvement of sequence analyzes not finalized the assembly and annotation of the genome of A. darlingi.