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Treen, N.

Publications and source records attributed to Treen, N..

5 recordsLinked to original sources

Possible function of Hox2 in atrial siphon fusion of the ascidian Ciona

The hallmark of sessile adult ascidians is a vase-like shape with a single oral and atrial siphon. Ciona, however, develops two atrial siphons after metamorphosis, which subsequently fuse into one. The mechanisms underlying this fusion are unknown. This study suggests that Hox2 controls this process. Hox2-knockout animals using Transcription-Activator-Like Effector Nuclease (TALEN) retain two atrial siphons throughout their lives. During normal fusion, epidermal cells between the siphons flatten along the anterior-posterior axis. This cellular flattening does not occur in Hox2-knockout animals, suggesting that the shape change in the epidermal cells produces tension, allowing the atrial siphon openings to converge at the midline for fusion. Hox2-knockout animals lack cupular organs, which are suspected hydrodynamic sensors in the internal epithelium of the fused atrial siphon and on the sperm duct. Among several knockout attempts, atrial siphon fusion was reproduced by only one TALEN pair, suggesting that this phenotype is driven by a mutation having a broader effect than those abolishing protein function. Many ascidians, unlike Ciona, develop a single atrial siphon shortly after metamorphosis. Our findings suggest that a phylogenetically conserved gene, Hox2, establishes this group-specific atrial siphon formation mechanism in Ciona.

developmental biology↗

Cell Numbers Contribute to Cell Fate During Ciona Cardiopharyngeal Mesoderm Specification

The Ciona heart cell lineage can be accurately traced back to a pair of blastomeres, the B7.5 cells, that form at the 64-cell stage. In addition to the adult heart, the B7.5 cells also contribute to two tail muscle cells in the larva, as well as the muscles that form the siphons for pumping water for feeding. Because of the simplicity of this system, we have a good understanding of how the B7.5 derivatives are specified during development. However, we know less about how the Ciona embryo precisely regulates cell numbers, as well as what effects altering cell numbers will have on development. We found that cell numbers in the B7.5 lineage are controlled by a pulse of transcription of the cell cycle inhibitor Cdkn1.b. Cdkn1.b can be repressed by the paralogues Prdm1-r.a and Prdm1-r.b that are exclusively transcribed in the B7.5 cells at the 110-cell stage. We unexpectedly found that precocious arrest of cell division in the B7.5 cell lineage resulted in a reversion to tail muscle fate, even in cells that can migrate. Our work demonstrates an unexpected connection between the control of cell numbers and cell fate in development. Significance StatementThe control of cell numbers is essential to many developmental processes, but it has been relatively understudied. A clear example of this can be seen in heart development. Hearts develop from precursor cells called cardiopharyngeal mesoderm. These cells must be able to migrate and proliferate in a precise way or heart defects will occur. We took advantage of the simplicity of embryonic development in the chordate Ciona (an invertebrate closely related to vertebrates) to investigate the consequences of disrupting cell numbers on heart development. Our finding that an early arrest in cell division results in a failure of heart development has implications for similar processes in vertebrates, including humans.

developmental biology↗

An FGF Timer for Zygotic Genome Activation

Zygotic genome activation has been extensively studied in a variety of systems including flies, frogs, and mammals. However, there is comparatively little known about the precise timings of gene induction during the earliest phases of embryogenesis. Here we employ high-resolution in situ detection methods, along with genetic and experimental manipulations, to study the timing of zygotic activation in the simple model chordate, Ciona intestinalis with minute-scale temporal precision. We found that two Prdm1 homologs in Ciona are the earliest genes that respond to FGF signaling. We present evidence for a FGF timing mechanism that is driven by derepression of the ERF repressor by ERK activity, which works in concert with localized activators such as Foxa.a. Absence of ERF results in derepression of target genes throughout the embryo. A highlight of this timer is the sharp transition in FGF responsiveness between the 8- and 16-cell stages of development. We propose that this timer is an innovation of chordates that is also employed by other vertebrates.

developmental biology↗

ERK signaling dissolves ERF Repression Condensates in Living Embryos

Phase separation underlies the organization of the nucleus, including the biogenesis of nucleoli and the packaging of heterochromatin. Here we explore the regulation of transcription factor condensates involved in gene repression by ERK signaling in gastrulating embryos of a simple proto-vertebrate (Ciona). ERK signaling induces nuclear export of the transcriptional repressor ERF, which has been linked to various human developmental disorders. Using high resolution imaging we show that ERF is localized within discrete nuclear condensates that dissolve upon ERK activation. Interestingly, we observe dynamic pulses of assembly and dissociation during the cell cycle, providing the first visualization of a nuclear phase separation process that is regulated by cell signaling. We discuss the implications of these observations for producing sharp on/off switches in gene activity and suppressing noise in cell-cell signaling events.

developmental biology↗

Regulation of gene expression by repression condensates during development

There is emerging evidence for transcription condensates in the activation of gene expression1-3. However, there is considerably less information regarding transcriptional repression, despite its pervasive importance in regulating gene expression in development and disease. Here, we explore the role of liquid-liquid phase separation (LLPS) in the organization of the Groucho/TLE (Gro) family of transcriptional corepressors, which interact with a variety of sequence-specific repressors such as Hes/Hairy4. Gro-dependent repressors have been implicated in a variety of developmental processes, including segmentation of the Drosophila embryo and somitogenesis in vertebrates. These repressors bind to specific recognition sequences, but instead of interacting with coactivators (e.g., Mediator) they recruit Gro corepressors5. Gro contains a series of WD40 repeats that are thought to mediate oligomerization6. How putative Hes/Gro oligomers repress transcription has been the subject of numerous studies5, 6. Here we show that Hes/Gro complexes form discrete puncta within nuclei of living Ciona embryos. These puncta rapidly dissolve during the onset of mitosis and reappear in the ensuing cell cycle. Modified Hes/Gro complexes that are unable to bind DNA exhibit the properties of viscous liquid droplets, similar to those underlying the biogenesis of P-granules in C. elegans7 and nucleoli in Xenopus oocytes8. These observations provide vivid evidence for LLPS in the control of gene expression and suggest a simple physical exclusion mechanism for transcriptional repression. WD40 repeats have been implicated in a wide variety of cellular processes in addition to transcriptional repression9. We suggest that protein interactions using WD40 motifs might be a common feature of processes reliant on LLPS.

developmental biology↗