Tinman is a direct activator of <i>midline</i> in the <i>drosophila</i> dorsal vessel

Ryu, Jae-Ryeon; Najand, Nima; Brook, William J.

doi:10.1002/dvdy.22495

Cited by 16 publications

(14 citation statements)

References 46 publications

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“…Although bPAC is activated by blue light illumination, there is low-level residual cyclase activity in the dark (Schröder-Lang et al, 2007;Weissenberger et al, 2010;Ryu et al, 2011;Stierl et al, 2011). When expressed in the principal cells we observed a 1.7-fold increase in cellular cAMP levels attributable to bPAC when tissues were maintained in the dark, and PDE activity was inhibited by 3-isobutyl-1-methylxanthine (IBMX; data not shown).…”

Section: Cyclic Amp Exerts a Bimodal Control On Fluid Secretionmentioning

confidence: 67%

Separate roles of PKA and EPAC in renal function unraveled by the optogenetic control of cAMP levels in vivo

Efetova

Petereit²,

Rosiewicz³

et al. 2012

Journal of Cell Science

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SummaryCyclic AMP (cAMP) is a ubiquitous second messenger that regulates a variety of essential processes in diverse cell types, functioning via cAMP-dependent effectors such as protein kinase A (PKA) and/or exchange proteins directly activated by cAMP (EPAC). In an intact tissue it is difficult to separate the contribution of each cAMP effector in a particular cell type using genetic or pharmacological approaches alone. We, therefore, utilized optogenetics to overcome the difficulties associated with examining a multicellular tissue. The transgenic photoactive adenylyl cyclase bPAC can be activated to rapidly and reversibly generate cAMP pulses in a cell-type-specific manner. This optogenetic approach to cAMP manipulation was validated in vivo using GAL4-driven UAS-bPAC in a simple epithelium, the Drosophila renal (Malpighian) tubules. As bPAC was expressed under the control of cell-type-specific promoters, each cAMP signal could be directed to either the stellate or principal cells, the two major cell types of the Drosophila renal tubule. By combining the bPAC transgene with genetic and pharmacological manipulation of either PKA or EPAC it was possible to investigate the functional impact of PKA and EPAC independently of each other. The results of this investigation suggest that both PKA and EPAC are involved in cAMP sensing, but are engaged in very different downstream physiological functions in each cell type: PKA is necessary for basal secretion in principal cells only, and for stimulated fluid secretion in stellate cells only. By contrast, EPAC is important in stimulated fluid secretion in both cell types. We propose that such optogenetic control of cellular cAMP levels can be applied to other systems, for example the heart or the central nervous system, to investigate the physiological impact of cAMP-dependent signaling pathways with unprecedented precision.

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Section: Cyclic Amp Exerts a Bimodal Control On Fluid Secretionmentioning

confidence: 67%

Separate roles of PKA and EPAC in renal function unraveled by the optogenetic control of cAMP levels in vivo

Efetova

Petereit²,

Rosiewicz³

et al. 2012

Journal of Cell Science

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“…GST pull-down assay GST pull-down assay was performed as described previously (Ryu et al, 2011). We purified GST, GST-Mid NT , and GST-Mid NT Á F41E following the manufacturer's protocol (GE Healthcare).…”

Section: Co-immunoprecipitationmentioning

confidence: 99%

“…Electrophoretic mobility shift assays (EMSA) were performed as previously described (Ryu et al, 2011). Double-stranded 32 P-endlabeled (10,000 cpm) oligonucleotides and purified GST-Mid NT (50 ng) were mixed in binding buffer containing 15 mM Hepes (pH 7.6), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 12% glycerol, 0.25 mg/ml poly dI:dC, 0.25 mg/ml BSA, and incubated for 15 min on ice.…”

Section: Electrophoretic Mobility Shift Assay For Putative Mid Bindinmentioning

confidence: 99%

The Tbx20 homolog Midline represses wingless in conjunction with Groucho during the maintenance of segment polarity

Formaz‐Preston

Ryu

Svendsen

et al. 2012

Developmental Biology

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The regulation of the segment polarity gene wingless is essential for the correct patterning of the Drosophila ectoderm. We have previously shown that the asymmetric activation of wingless downstream of Hedghog-signaling depends on the T-box transcription factors, midline and H15. Hedgehog activates wingless anterior to the Hedgehog domain. midline/H15 are responsible in part for repressing wingless in cells posterior to the Hedgehog expressing cells. Here, we show that Midline binds the Groucho co-repressor directly via the engrailed homology-1 domain and requires an intact engrailed-homology-1 domain to repress wingless. In contrast, the regulation of Serrate, a second target of midline repression, is not dependent on the engrailed-homology-1 domain. Furthermore, we identify a midline responsive region of the wingless cis-regulatory region and show that Midline binds to sequences within this region. Mutating these sequences in transgenic reporter constructs results in ectopic reporter expression in the midline-expression domain, consistent with wingless being a direct target of Midline repression.

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“…Interestingly, the role of Mid in selecting muscle fate choice between LT3 and LT4 is analogous to its function in the nervous system, where it mediates neuroblast fate specification (Buescher et al 2006;Gaziova and Bhat 2009). Mid and H15 also promote cell fate specification and morphogenesis in the Drosophila heart (Miskolczi-McCallum et al 2005;Qian et al 2005;, where mid is both a target and a regulator of the Tin transcription factor (Qian et al 2005;Ryu et al 2011;Jin et al 2013).…”

Section: Mid and The T-box Family Of Proteinsmentioning

confidence: 99%

“…Mid and its paralog H15 play partially redundant roles in specification and differentiation of the embryonic ectoderm (Buescher et al 2004) and in sensory organ precursor (SOP) cells in the eye imaginal disc (Das et al 2013). Mid and H15 also promote cardioblast specification and heart morphogenesis (Miskolczi-McCallum et al 2005;Qian et al 2005;, and it has been reported that Mid is both a target of the Tinman (Tin) transcriptional regulator (Qian et al 2005;Ryu et al 2011;Jin et al 2013) and is required for tin expression . Despite these well-characterized roles in the Drosophila embryo, no study of mid in the somatic mesoderm has been described.…”

mentioning

confidence: 99%

Muscle Cell Fate Choice Requires the T-Box Transcription Factor Midline in Drosophila

et al. 2015

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Drosophila Midline (Mid) is an ortholog of vertebrate Tbx20, which plays roles in the developing heart, migrating cranial motor neurons, and endothelial cells. Mid functions in cell-fate specification and differentiation of tissues that include the ectoderm, cardioblasts, neuroblasts, and egg chambers; however, a role in the somatic musculature has not been described. We identified mid in genetic and molecular screens for factors contributing to somatic muscle morphogenesis. Mid is expressed in founder cells (FCs) for several muscle fibers, and functions cooperatively with the T-box protein H15 in lateral oblique muscle 1 and the segment border muscle. Mid is particularly important for the specification and development of the lateral transverse (LT) muscles LT3 and LT4, which arise by asymmetric division of a single muscle progenitor. Mid is expressed in this progenitor and its two sibling FCs, but is maintained only in the LT4 FC. Both muscles were frequently missing in mid mutant embryos, and LT4-associated expression of the transcription factor Krüppel (Kr) was lost. When present, LT4 adopted an LT3-like morphology. Coordinately, mid misexpression caused LT3 to adopt an LT4-like morphology and was associated with ectopic Kr expression. From these data, we concluded that mid functions first in the progenitor to direct development of LT3 and LT4, and later in the FCs to influence whichever of these differentiation profiles is selected. Mid is the first T-box factor shown to influence LT3 and LT4 muscle identity and, along with the T-box protein Optomotor-blind-related-gene 1 (Org-1), is representative of a new class of transcription factors in muscle specification. KEYWORDS midline; Tbx20; founder cell; muscle identity; differentiation T HE body wall muscles of the Drosophila larva are composed of a stereotypic pattern that includes 30 unique muscle fibers (each a single myotube) per abdominal hemisegment (Bate 1990). These muscles form during embryogenesis, during which time they take on unique characteristics such as size, shape, pattern of innervation, and epidermal attachment Baylies et al. 1998;Frasch 1999;De Joussineau et al. 2012). The unique identity of each muscle fiber is conferred through overlapping signaling pathways and cell autonomous transcription factors that specify muscle progenitors Baylies et al. 1998;Frasch 1999;Tixier et al. 2010), which then undergo asymmetric cell division to generate founder cells (FCs) (Bate 1990;Carmena et al. 1998). A single FC initiates the formation of multinucleate syncytia by fusing with fusion competent myoblasts (FCMs) (Rochlin et al. 2010;Abmayr and Pavlath 2012). FCs dictate the number of cell fusion events that take place, leading to a characteristic number of nuclei for each muscle in the pattern . These fusing nuclei adopt the transcriptional profile of the original FC nucleus (Bourgouin et al. 1992;D'Alessio and Frasch 1996;Ruiz-Gomez et al. 1997;Jagla et al. 1998;Keller et al. 1998;Nose et al. 1998;Crozatier and Vincent 1999;Knirr et al. 1999;D...

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Tinman is a direct activator of midline in the drosophila dorsal vessel

Cited by 16 publications

References 46 publications

Separate roles of PKA and EPAC in renal function unraveled by the optogenetic control of cAMP levels in vivo

Separate roles of PKA and EPAC in renal function unraveled by the optogenetic control of cAMP levels in vivo

The Tbx20 homolog Midline represses wingless in conjunction with Groucho during the maintenance of segment polarity

Muscle Cell Fate Choice Requires the T-Box Transcription Factor Midline in Drosophila

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