title: Ontogenetic differences in localization of glutamine transporter ApGLNT1 in the pea aphid demonstrate that mechanisms of host/symbiont integration are not similar in the maternal versus embryonic bacteriome.
abstract: BACKGROUND
  Obligate intracellular symbionts of insects are metabolically and developmentally integrated with their hosts. Typically, reproduction fails in many insect nutritional endosymbioses when host insects are cured of their bacterial symbionts, and yet remarkably little is known about the processes that developmentally integrate host and symbiont. Here in the best studied insect obligate intracellular symbiosis, that of the pea aphid, Acyrthosiphon pisum, with the gammaproteobacterium Buchnera aphidicola, we tracked the expression and localization of amino acid transporter ApGLNT1 gene products during asexual embryogenesis. Recently being characterized as a glutamine transporter, ApGLNT1 has been proposed to be a key regulator of amino acid biosynthesis in A. pisum bacteriocytes. To determine when this important mediator of the symbiosis becomes expressed in aphid embryonic bacteriocytes, we applied whole-mount in situ hybridization and fluorescent immunostaining with a specific anti-ApGLNT1 antibody to detect the temporal and spatial expression of ApGLNT1 gene products during asexual embryogenesis.
  
  
  RESULTS
  During embryogenesis, ApGLNT1 mRNA and protein localize to the follicular epithelium that surrounds parthenogenetic viviparous embryos, where we speculate that it functions to supply developing embryos with glutamine from maternal hemolymph. Unexpectedly, in the embryonic bacteriome ApGLNT1 protein does not localize to the membrane of bacteriocytes, a pattern that leads us to conclude that the regulation of amino acid metabolism in the embryonic bacteriome mechanistically differs from that in the maternal bacteriome. Paralleling our earlier report of punctate cytoplasmic localization of ApGLNT1 in maternal bacteriocytes, we find ApGLNT1 protein localizing as cytoplasmic puncta throughout development in association with Buchnera.
  
  
  CONCLUSIONS
  Our work that documents ontogenetic shifts in the localization of ApGLNT1 protein in the host bacteriome demonstrates that maternal and embryonic bacteriomes are not equivalent. Significantly, the persistent punctate cytoplasmic localization of ApGLNT1 in association with Buchnera in embryos prior to bacteriocyte formation and later in both embryonic and maternal bacteriomes suggests that ApGLNT1 plays multiple roles in this symbiosis, roles that include amino acid transport and possibly nutrient sensing.
authors.affiliation: Department of Biology, University of Miami, Coral Gables, FL 33146 USA ; Department of Entomology, College of Bioresources and Agriculture, National Taiwan University, Taipei, Taiwan ; Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Hsiao-Ling
authors.initials: HL
authors.lastname: Lu
authors.affiliation: Department of Biology, University of Miami, Coral Gables, FL 33146 USA.
authors.forename: Daniel R G
authors.initials: DR
authors.lastname: Price
authors.affiliation: Department of Biology, University of Miami, Coral Gables, FL 33146 USA.
authors.forename: Athula
authors.initials: A
authors.lastname: Wikramanayake
authors.affiliation: Department of Entomology, College of Bioresources and Agriculture, National Taiwan University, Taipei, Taiwan ; Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Chun-Che
authors.initials: CC
authors.lastname: Chang
authors.affiliation: Department of Biology, University of Miami, Coral Gables, FL 33146 USA.
authors.forename: Alex C C
authors.initials: AC
authors.lastname: Wilson
keywords: Amino acid transport; Bacteriome; Coevolution; Holobiont; Host/symbiont developmental integration; Symbiosis