Cell and Developmental Biology Commons^™

Germline Transgenic Methods For Tracking Cells And Testing Gene Function During Regeneration In The Axolotl, Shahryar Khattak, Maritta Schuez, Tobias Richter, Dunja Knapp, Saori Haigo, Tatiana Sandoval-Guzman, Kristyna Hradlikova, Annett Duemmler, Ryan Kerney, Elly Tanaka

Ryan Kerney

The salamander is the only tetrapod that regenerates complex body structures throughout life. Deciphering the underlying molecular processes of regeneration is fundamental for regenerative medicine and developmental biology, but the model organism had limited tools for molecular analysis. We describe a comprehensive set of germline transgenic strains in the laboratory-bred salamander Ambystoma mexicanum (axolotl) that open up the cellular and molecular genetic dissection of regeneration. We demonstrate tissue-dependent con- trol of gene expression in nerve, Schwann cells, oligodendrocytes, muscle, epidermis, and cartilage. Furthermore, we demonstrate the use of tamoxifen-induced Cre/loxP-mediated recombination to indelibly mark different cell types. Finally, we inducibly …

Do Larval Traits Re-Evolve? Evidence From The Embryogenesis Of A Direct-Developing Salamander, Plethodon Cinereus, Ryan Kerney, David Blackburn, Hendrik Muller, James Hanken

Ryan Kerney

Recent molecular phylogenies suggest the surprising reacquisition of posthatching metamorphosis within an otherwise directdeveloping clade of lungless salamanders (family Plethodontidae). Metamorphosis was long regarded as plesiomorphic for plethodontids, yet the genus Desmognathus, which primarily includes metamorphosing species, is now nested within a much larger clade of direct-developing species. The extent to which the putative reacquisition of metamorphosis in Desmognathus represents a true evolutionary reversal is contingent upon the extent to which both larva-specific features and metamorphosis were actually lost during the evolution of direct development. In this study we analyze development of the hyobranchial skeleton, which is dramatically remodeled during …

Intracellular Invasion Of Green Algae In A Salamander Host, Ryan Kerney, Eunsoo Kim, Roger Hangater, Aaron Heiss, Cory Bishop, Brian Hall

Ryan Kerney

The association between embryos of the spotted salamander (Ambystoma maculatum) and green algae (“Oophila amblystomatis” Lamber ex Printz) has been considered an ectosymbiotic mutualism. We show here, however, that this symbiosis is more intimate than previously reported. A combination of imaging and algal 18S rDNA amplification reveals algal invasion of embryonic salamander tissues and cells during development. Algal cells are detectable from embryonic and larval Stages 26–44 through chlorophyll autofluorescence and algal 18S rDNA amplification. Algal cell ultrastructure indicates both degradation and putative encystment during the process of tissue and cellular invasion. Fewer algal cells were detected in later-stage larvae …

Symbioses Between Salamander Embryos And Green Algae, Ryan Kerney

Ryan Kerney

The symbiosis between Ambystoma maculatum (spotted salamander) embryos and green algae was initially described over 120 years ago. Algae populate the egg capsules that surround individual A. maculatum embryos, giving the intracapsular fluid a characteristic green hue. Early work established this symbiosis to be a mutualism, while subsequent studies sought to identify the material benefits of this association to both symbiont and host. These studies have shown that salamander embryos benefit from increased oxygen concentrations provided by their symbiotic algae. The algae, in turn, may benefit from ammonia excreted by the embryos. All of these early studies considered the associ- …

Early Cranial Patterning In The Direct-Developing Frog Eleutherodactylus Coqui Revealed Through Gene Expression, Ryan Kerney, Joshua Gross, James Hanken

Ryan Kerney

Genetic and developmental alterations associated with the evolution of amphibian direct development remain largely unexplored. Specifically, little is known of the underlying expression of skeletal regulatory genes, which may reveal early modifications to cranial ontogeny in direct-developing species. We describe expression patterns of three key skeletal regulators (runx2, sox9, and bmp4) along with the cartilage-dominant collagen 2a1 gene (col2a1) during cranial development in the direct- developing anuran, Eleutherodactylus coqui. Expression patterns of these regulators reveal transient skeletogenic anlagen that correspond to larval cartilages, but which never fully form in E. coqui. Suprarostral anlagen in the frontonasal processes are detected through …

Life History, Sexual Dimorphism And 'Ornamental' Feathers In The Mesozoic Bird Confuciusornis Sanctus., Winfried S. Peters, Dieter Stefan Peters

Winfried S. Peters

Gene Expression Reveals Unique Skeletal Patterning In The Limb Of The Direct-Developing Frog, Eleutherodactylus Coqui, Ryan Kerney, James Hanken

Ryan Kerney

The growing field of skeletal developmental biology provides new molecular markers for the cellular pre- cursors of cartilage and bone. These markers enable resolution of early features of skeletal development that are otherwise undetectable through conventional staining tech- niques. This study investigates mRNA distributions of skeletal regulators runx2 and sox9 along with the cartilage-dominant collagen 2a1 (col2a1) in embryonic limbs of the direct- developing anuran, Eleutherodactylus coqui. To date, distri- butions of these genes in the limb have only been examined in studies of the two primary amniote models, mouse and chicken. In E. coqui, expression of transcription factors runx2 …

Runx2 Is Essential For Larval Hyobranchial Cartilage Formation In Xenopus Laevis, Ryan Kerney, Joshua Gross, James Hanken

Ryan Kerney

The vertebrate transcription factor protein Runx2 is regarded as a “master regulator” of bone formation due to the dramatic loss of the osseous skeleton in the mouse homozygous knockout. However, Runx2 mRNA also is expressed in the pre-hypertrophic cartilaginous skeleton of the mouse and chicken, where its developmental function is largely unknown. Several tiers of Runx2 regulation exist in the mouse, any of which may account for its seeming biological inactivity during early stages of skeletogenesis. Unlike mouse and chicken, zebrafish require Runx2 function in early cartilage differentiation. The present study reveals that the earlier functional role of Runx2 in …

Pollination Of The Crown Imperial Fritillaria Imperialis By Great Tits Parus Major, Winfried S. Peters, Michael Pirl, Gerhard Gottsberger, Dieter Stefan Peters

Winfried S. Peters