Secretion of secretory phospholipase A2 into Spodoptera exigua larval midgut lumen and its role in lipid digestion

Insect Mol Biol. 2019 Dec;28(6):773-784. doi: 10.1111/imb.12588. Epub 2019 May 13.

Abstract

In insects, lipid digestion is controversial because insects have no bile salts to solubilize dietary lipids. One hypothesis is that a secretory type of phospholipase A2 (sPLA2 ) provides lysophospholipid (LPL) from dietary phospholipids (PLs). We identified a sPLA2 , Se-sPLA2 , in beet armyworm, Spodoptera exigua, that hydrolyses PLs at sn-2. Our goal was to investigate its role in lipid digestion. Se-sPLA2 was expressed in the entire alimentary canal. Incubating the isolated midgut in a cell culture medium led to secretion of Se-sPLA2 and other proteins. Ex vivo RNA interference (RNAi) of Se-sPLA2 expression in isolated midgut culture led to significantly decreased Se-sPLA2 secretion into the medium. Feeding double-stranded RNA specific to Se-sPLA2 to larvae suppressed sPLA2 activity in gut contents. A recombinant Se-sPLA2 was susceptible to benzylideneacetone (BZA), a specific PLA2 inhibitor. After feeding BZA to larvae, we recorded significant decreases in gut content sPLA2 activity, body growth and total haemolymph lipid contents. RNAi against Se-sPLA2 resulted in reduced digestibility. Addition of a specific LPL, 1-palmitoyl-sn-glycero-3-phosphocholine, to BZA-treated larvae rescued digestibility and larval growth. These results strongly bolster our hypothesis that Se-sPLA2 secreted from the midgut acts in lipid digestion by providing necessary LPL to solubilize dietary neutral lipids.

Keywords: Spodoptera exigua; RNA interference; digestion; lipid; lysophospholipid; sPLA2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Digestion*
  • Insect Proteins / chemistry
  • Insect Proteins / metabolism*
  • Intestines / physiology
  • Larva / enzymology
  • Larva / growth & development
  • Larva / physiology
  • Lipid Metabolism*
  • Phospholipases A2, Secretory / chemistry
  • Phospholipases A2, Secretory / metabolism*
  • Spodoptera / enzymology
  • Spodoptera / growth & development
  • Spodoptera / physiology*

Substances

  • Insect Proteins
  • Phospholipases A2, Secretory