High throughput techniques to reveal the molecular physiology and evolution of digestion in spiders.
Spiders are known for their predatory efficiency and for their high capacity of digesting relatively large prey. They do this by combining both extracorporeal and intracellular digestion. Whereas many high throughput ("-omics") techniques focus on biomolecules in spider venom, so far this approach has not yet been applied to investigate the protein composition of spider midgut diverticula (MD) and digestive fluid (DF).



We here report on our investigations of both MD and DF of the spider Nephilingis (Nephilengys) cruentata through the use of next generation sequencing and shotgun proteomics. This shows that the DF is composed of a variety of hydrolases including peptidases, carbohydrases, lipases and nuclease, as well as of toxins and regulatory proteins. We detect 25 astacins in the DF. Phylogenetic analysis of the corresponding transcript(s) in Arachnida suggests that astacins have acquired an unprecedented role for extracorporeal digestion in Araneae, with different orthologs used by each family. The results of a comparative study of spiders in distinct physiological conditions allow us to propose some digestion mechanisms in this interesting animal taxon.



All the high throughput data allowed the demonstration that DF is a secretion originating from the MD. We identified enzymes involved in the extracellular and intracellular phases of digestion. Besides that, data analyses show a large gene duplication event in Araneae digestive process evolution, mainly of astacin genes. We were also able to identify proteins expressed and translated in the digestive system, which until now had been exclusively associated to venom glands.
Laboratory of Biochemistry and Biophysics, Instituto Butantan, São Paulo, 05503-000, Brazil.|Biotechnology Program, University of São Paulo, São Paulo, Brazil.
Felipe J
FJ
Fuzita
Laboratory of Analytical Biotechnology and Innovative Peptide Biology, Delft University of Technology, Delft, The Netherlands.
Martijn W H
MW
Pinkse
Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, Brazil.
José S L
JS
Patane
Laboratory of Analytical Biotechnology and Innovative Peptide Biology, Delft University of Technology, Delft, The Netherlands.|Department of Biology, University of Antwerp, Antwerp, Belgium.
Peter D E M
PD
Verhaert
Laboratory of Biochemistry and Biophysics, Instituto Butantan, São Paulo, 05503-000, Brazil. adriana.lopes@butantan.gov.br.
Adriana R
AR
Lopes
D000818:Animals; D060829:Arthropod Proteins / Q000235:genetics / Q000378:metabolism; D004063:Digestion; D004064:Digestive System / Q000378:metabolism; D019143:Evolution, Molecular; D020440:Gene Duplication; D005786:Gene Expression Regulation; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D040901:Proteomics / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*; D013112:Spiders / Q000235:genetics / Q000502:physiology*
D060829:Arthropod Proteins; D008666:Metalloendopeptidases; C071288:astacin
Arachnida; Astacin; Digestion; Enzyme; High throughput (-omics) techniques; Nephilingis (Nephilengys cruentata); Spider