title: Developmental transition to bilaterally symmetric cell divisions is regulated by Pax-mediated transcription in embryos of the leech Helobdella austinensis.
abstract: The leech embryo develops by spiral cleavage, and establishes the symmetry properties of its adult body plan through the bilaterally symmetric divisions of mesodermal proteloblast DM″ and ectodermal proteloblast DNOPQ‴. We here show that transcriptional inhibitors α-amanitin and actinomycin D specifically disrupt the symmetry and orientation of these two proteloblast cell divisions while having no apparent effect on the timing or geometry of other divisions. Transcriptional inhibition had a similar effect on both proteloblasts, i.e. cytokinesis was highly asymmetric and the cleavage plane roughly orthogonal to that seen during normal development. These findings suggest that zygotic gene product(s) are required, either directly or indirectly, for the correct placement of the proteloblast cleavage furrow. The same phenotypes were also observed following in vivo expression of dominant-negative Pax gene constructs. These dominant-negative phenotypes depended on protein/DNA interaction, and could be rescued by coexpression of full length Pax proteins. However, symmetric cleavage of the mesodermal proteloblast was rescued by full length constructs of either Hau-Paxβ1 or Hau-Pax2/5/8, while only Hau-Paxβ1 rescued the symmetry of ectodermal cleavage. We conclude that both proteloblasts need Pax-mediated transcription to adopt their normally symmetric cleavage patterns, but differ in terms of the specific Pax proteins required. The implication of these findings for the evolution of spiral cleavage is discussed.
authors.affiliation: Section of Molecular Cell and Developmental Biology, University of Texas at Austin, Austin, TX 78712, USA. Electronic address: leechman@austin.utexas.edu.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Schmerer
authors.forename: Ryan W
authors.initials: RW
authors.lastname: Null
authors.forename: Marty
authors.initials: M
authors.lastname: Shankland
mesh_terms: D000818:Animals; D001757:Blastomeres / Q000166:cytology / Q000378:metabolism; D002452:Cell Count; D002455:Cell Division; D004247:DNA / Q000378:metabolism; D004475:Ectoderm / Q000166:cytology / Q000196:embryology; D004625:Embryo, Nonmammalian / Q000166:cytology* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D005799:Genes, Dominant; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007865:Leeches / Q000166:cytology* / Q000196:embryology* / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology / Q000196:embryology; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic; D015053:Zygote / Q000166:cytology / Q000378:metabolism
chemical_list: D051761:Paired Box Transcription Factors; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D004247:DNA
keywords: Cytokinesis; Dominant-negative; Leech embryo; Pax gene; Spiral cleavage; Zygotic transcription