Stein, D. S. | bioRxiv Science

bioRxiv2023

Embryogenesis in myrmicine ants combines features of short and long germ-band modes of development

Ants exhibit complex social organization, morphologically distinct castes with division of labor, and the exploitation of diverse ecological niches. The extent to which these features have influenced embryonic development relative to other insects remains unclear. Insect embryogenesis has been classified into one of three modes: long, short, and intermediate germ-band. In long germ-band development, exemplified by the fruit fly Drosophila melanogaster, segments along the entire anterior-posterior axis of the embryonic primordium are established almost simultaneously, prior to gastrulation, with the initial embryonic primordium surrounding almost the entire volume of the egg. In short and intermediate germ-band modes, the embryonic primordium occupies a smaller proportion of the egg surface, with anterior segments initially specified, and remaining segments being added sequentially from a posterior growth zone. Here, we show a novel pattern of development in three myrmicine ants, the fungus-gardening ants Atta texana and Mycocepurus smithii, and the red imported fire ant Solenopsis invicta. Early in embryogenesis, they exhibit features of short germ-band development, while later in development they exhibit a newly-characterized progressive pattern of segmentation that has been associated with some long germ-band-developing insects. Moreover, despite similarities in the size of ant and Drosophila eggs, the duration of embryogenesis in the three ant species is 10 to 20-fold longer than in Drosophila and is also significantly longer than in the honeybee Apis mellifera and the jewel wasp Nasonia vitripennis. In addition, the embryos produced by A. texana foundress queens develop to first instar larvae 25% faster than embryos produced by mature queens. We discuss these results in the context of the eusocial lifestyle of ants.

developmental biology↗

bioRxiv2019

Isolation of secreted proteins from Drosophila ovaries and embryos through in vivo BirA-mediated biotinylation

The extraordinarily strong non-covalent interaction between biotin and avidin (kD = 10-14-10-16) has permitted this interaction to be used in a wide variety of experimental contexts. The Biotin Acceptor Peptide (BAP), a 15 amino acid motif that can be biotinylated by the E. coli BirA protein, has been fused to proteins of interest, making them substrates for in vivo biotinylation. Here we report on the construction and characterization of a modified BirA bearing signals for secretion and endoplasmic reticulum (ER) retention, for use in experimental contexts requiring biotinylation of secreted proteins. When expressed in the Drosophila female germline or ovarian follicle cells under Gal4-mediated transcriptional control, the modified BirA protein could be detected and shown to be enzymatically active in ovaries and progeny embryos. Surprisingly, however, it was not efficiently retained in the ER, and instead appeared to be secreted. To determine whether this secreted protein, now designated secBirA, could biotinylate secreted proteins, we generated BAP-tagged versions of two secreted Drosophila proteins, Torsolike (Tsl) and Gastrulation Defective (GD), which are normally expressed maternally and participate in embryonic pattern formation. Both Tsl-BAP and GD-BAP were shown to exhibit normal patterning activity. Co-expression of Tsl-BAP together with secBirA in ovarian follicle cells resulted in its biotinylation, which permitted its isolation from both ovaries and progeny embryos using Avidin-coupled affinity matrix. In contrast, co-expression with secBirA in the female germline did not result in detectable biotinylation of GD-BAP, possibly because the C-terminal location of the BAP tag made it inaccessible to BirA in vivo. Our results indicate that secBirA directs biotinylation of proteins bound for secretion in vivo, providing access to powerful experimental approaches for secreted proteins of interest. However, efficient biotinylation of target proteins may vary depending upon the location of the BAP tag or other structural features of the protein.

molecular biology↗