David Stafford, Victoria E. Prince  Current Biology 

Slides:



Advertisements
Similar presentations
Volume 94, Issue 4, Pages (August 1998)
Advertisements

Single-cell internalization during zebrafish gastrulation
Federica Bertocchini, Claudio D. Stern  Developmental Cell 
Volume 10, Issue 1, Pages (January 2006)
Volume 25, Issue 5, Pages (June 2013)
Karen Lunde, Heinz-Georg Belting, Wolfgang Driever  Current Biology 
Iain Patten, Marysia Placzek  Current Biology 
Volume 35, Issue 2, Pages (July 2002)
Distinct Origins of Adult and Embryonic Blood in Xenopus
Smoothened Mutants Reveal Redundant Roles for Shh and Ihh Signaling Including Regulation of L/R Asymmetry by the Mouse Node  Xiaoyan M. Zhang, Miguel.
Volume 14, Issue 4, Pages (April 2008)
Volume 10, Issue 9, Pages (May 2000)
Vagus Motor Neuron Topographic Map Determined by Parallel Mechanisms of hox5 Expression and Time of Axon Initiation  Gabrielle R. Barsh, Adam J. Isabella,
Reprogramming Hox Expression in the Vertebrate Hindbrain: Influence of Paraxial Mesoderm and Rhombomere Transposition  Nobue Itasaki, James Sharpe, Alastair.
Metabolic Regulation of Cellular Plasticity in the Pancreas
Robert G. Kelly, Nigel A. Brown, Margaret E. Buckingham 
Volume 8, Issue 4, Pages (April 2005)
Volume 21, Issue 4, Pages (February 2011)
Germ cell-less Acts to Repress Transcription during the Establishment of the Drosophila Germ Cell Lineage  Judith L. Leatherman, Lissa Levin, Julie Boero,
Volume 9, Issue 15, Pages S1-840 (August 1999)
Victor Hatini, Stephen DiNardo  Molecular Cell 
Volume 44, Issue 2, Pages e5 (January 2018)
A Crucial Interaction between Embryonic Red Blood Cell Progenitors and Paraxial Mesoderm Revealed in spadetail Embryos  Laurel A. Rohde, Andrew C. Oates,
Helge Amthor, Bodo Christ, Miguel Weil, Ketan Patel  Current Biology 
Volume 1, Issue 1, Pages (July 2001)
BMP Signaling Protects Telencephalic Fate by Repressing Eye Identity and Its Cxcr4- Dependent Morphogenesis  Holger Bielen, Corinne Houart  Developmental.
Neuropeptides: Developmental Signals in Placode Progenitor Formation
Kathleen S. Christine, Frank L. Conlon  Developmental Cell 
Single-Cell Transcript Analysis of Pancreas Development
Intrinsic Differences between the Superficial and Deep Layers of the Xenopus Ectoderm Control Primary Neuronal Differentiation  Andrew D Chalmers, David.
Stem Cells and Diabetes
A molecular pathway leading to endoderm formation in zebrafish
Nadine Peyriéras, Uwe Strähle, Frédéric Rosa  Current Biology 
Stem Cells and Diabetes
Experimental Conversion of Liver to Pancreas
Volume 20, Issue 21, Pages (November 2010)
Early Lineage Segregation between Epiblast and Primitive Endoderm in Mouse Blastocysts through the Grb2-MAPK Pathway  Claire Chazaud, Yojiro Yamanaka,
Sonic hedgehog and vascular endothelial growth factor Act Upstream of the Notch Pathway during Arterial Endothelial Differentiation  Nathan D. Lawson,
The BMP Signaling Gradient Patterns Dorsoventral Tissues in a Temporally Progressive Manner along the Anteroposterior Axis  Jennifer A. Tucker, Keith.
Volume 19, Issue 12, Pages (June 2009)
Developmental Basis of Phallus Reduction during Bird Evolution
Volume 18, Issue 4, Pages (April 2010)
Mark Van Doren, Anne L. Williamson, Ruth Lehmann  Current Biology 
Zebrafish pea3 and erm are general targets of FGF8 signaling
Brian A Hyatt, H.Joseph Yost  Cell 
Xsox17α and -β Mediate Endoderm Formation in Xenopus
Repulsive Interactions Shape the Morphologies and Functional Arrangement of Zebrafish Peripheral Sensory Arbors  Alvaro Sagasti, Matthew R. Guido, David.
Bmp2 Signaling Regulates the Hepatic versus Pancreatic Fate Decision
Volume 12, Issue 11, Pages (June 2002)
Specification of Motor Neuron Identity by the MNR2 Homeodomain Protein
Won-Suk Chung, Didier Y.R. Stainier  Developmental Cell 
Julie E. Cooke, Hilary A. Kemp, Cecilia B. Moens  Current Biology 
Distinct mechanisms regulate slow-muscle development
Åsa Apelqvist, Ulf Ahlgren, Helena Edlund  Current Biology 
Jeffrey D Amack, H.Joseph Yost  Current Biology 
FGF Signaling Controls Somite Boundary Position and Regulates Segmentation Clock Control of Spatiotemporal Hox Gene Activation  Julien Dubrulle, Michael.
Autonomous Modes of Behavior in Primordial Germ Cell Migration
Volume 8, Issue 4, Pages (April 2005)
Pharyngeal arch patterning in the absence of neural crest
Single-cell internalization during zebrafish gastrulation
Islet mass and β-cell proliferation is decreased in Hhip–/– embryos (Hip1 in figure). Islet mass and β-cell proliferation is decreased in Hhip–/– embryos.
Volume 5, Issue 2, Pages (August 2003)
Zebrafish as a Model Organism for the Identification and Characterization of Drugs and Genes Affecting p53 Signaling  Ulrike Langheinrich, Elisabeth Hennen,
Critical role of biklf in erythroid cell differentiation in zebrafish
Lefty-Dependent Inhibition of Nodal- and Wnt-Responsive Organizer Gene Expression Is Essential for Normal Gastrulation  William W. Branford, H.Joseph.
The Anterior-Posterior Axis Emerges Respecting the Morphology of the Mouse Embryo that Changes and Aligns with the Uterus before Gastrulation  Daniel.
Morphogenetic Movements Underlying Eye Field Formation Require Interactions between the FGF and ephrinB1 Signaling Pathways  Kathryn B. Moore, Kathleen.
Volume 12, Issue 23, Pages (December 2002)
Reprogramming Hox Expression in the Vertebrate Hindbrain: Influence of Paraxial Mesoderm and Rhombomere Transposition  Nobue Itasaki, James Sharpe, Alastair.
Volume 19, Issue 19, Pages (October 2009)
Presentation transcript:

Retinoic Acid Signaling Is Required for a Critical Early Step in Zebrafish Pancreatic Development  David Stafford, Victoria E. Prince  Current Biology  Volume 12, Issue 14, Pages 1215-1220 (July 2002) DOI: 10.1016/S0960-9822(02)00929-6

Figure 1 RA Signaling Is Necessary for Pancreatic Development Embryos were processed for one- or two-color whole-mount in situ hybridization (dorsal views, anterior to the left). The left-hand panels show wild-type or mock-treated control embryos. The right-hand panels show RA signaling-deficient embryos (either neckless−/− (nls), identified by morphology, or treated with 10−6 M BMS493). BMS493 stock solution at 10−2 M in 70% ethanol/30% H2O was diluted in embryo medium [32], and treatment commenced at 6 hpf (but equivalent results were obtained when treatment commenced immediately after fertilization). (A) insulin (red) and somatostatin (sst; blue). (B) insulin and sst expression is absent in all BMS493-treated embryos (insulin 585; sst 58) and in the majority of nls embryos (insulin 17/18; sst 5/5). Glucagon expression is also absent after treatment with BMS493 (n = 45; data not shown [DNS]). (C) trypsin (blue; bracket) and insulin (red). (D) trypsin is undetectable after BMS493 treatment (n = 40). (E and F) trypsin is significantly reduced in nls embryos (n = 5). (G) islet1 expression in cranial nerves (bracket) and the pancreas (bar). (H) Pancreatic islet1 expression is lost in BMS493-treated embryos (n = 33) and nls embryos (n = 5; DNS). (I) Pancreatic pdx1 expression (bar). (J) pdx1 is not expressed in BMS493-treated embryos (DNS; n = 60, at stages from 14 to 48 hpf) or nls embryos (n = 23, at 24 hpf). (K) insulin (red) and cepb4 (blue) expression. cepb4 is expressed in the liver (arrowhead), at lower levels in gut endoderm, and in isolated clusters of cells in the anterior of the embryo. (L) cepb4 is not expressed in the liver of BMS493-treated embryos (n = 45). (M) ceruloplasmin (cp; red) and insulin (blue). cp is expressed in the liver (arrowhead). (N) cp is not expressed in BMS493-treated embryos (n = 63). (O) hhex (blue) is expressed in the liver (arrowhead) and thyroid (arrow). (P) liver expression of hhex is radically reduced in nls embryos (arrowhead; n = 5) and is absent from BMS493-treated embryos (n = 35; DNS). The following probes were used: cepb4[18], ceruloplasmin[19], glucagon and somatostatin[12], insulin and pdx1[13], islet1[16], trypsin[14], and hhex[20]. Current Biology 2002 12, 1215-1220DOI: (10.1016/S0960-9822(02)00929-6)

Figure 2 Endoderm Formation, Survival, and Differentiation Are Independent of RA Signaling (A–J) Embryos were treated from 6 hpf until fixation and were processed for whole-mount in situ hybridization (A–D; G–J), or they were treated from 9 hpf (E and F) and were processed for in situ end labeling (ISEL) to label cells undergoing apoptotic cell death. (A–D) Expression of general markers of the endoderm is not RA dependent. (A and B) sox17 at 8 hpf, whole-mounts; dorsal is oriented toward the right. (A) Expression is in noninvoluting dorsal forerunner cells (arrowhead) and endodermal precursors, in a characteristic sparse monolayer over the yolk. (B) Expression in BMS493-treated embryos is indistinguishable from controls (n = 45). (C) fkd7 expression at 24 hpf in the gut tube, pharyngeal endoderm (arrowheads), thalamus (bracket), and pancreatic primordium (bar). (D) Expression is maintained in BMS493-treated embryos (n = 15). (E and F) ISEL staining at 11 hpf in whole mounts (dorsal toward reader) showing sparsely distributed apoptotic cells. BMS493-treated embryos do not exhibit consistently different apoptosis patterns relative to controls (n = 65; 11, 13, and 22 hpf; DNS). ISEL staining was performed essentially as previously described [33], using reagents from Intergen. (G–J) Differentiation of other endodermal derivatives is not RA dependent. (G and H) nkx2.3 expression at 24 hpf in bilateral clusters of cells in the pharyngeal endoderm is unaltered by BMS493 treatment (n = 56). (I) insulin (red) and nk2.1a (blue) expression in the forebrain (bracket) and in the developing thyroid (blue arrowhead). (J) insulin expression is lost, and thyroid expression of nk2.1a (blue arrowhead) is present, but it is shifted posteriorly relative to the otic vesicle (green arrowhead) in BMS493-treated embryos (n = 55). The following probes were used: sox17[21], axial[23], fkd7[22], nkx2.3[25], and nk2.1a[26]. Current Biology 2002 12, 1215-1220DOI: (10.1016/S0960-9822(02)00929-6)

Figure 3 Exogenous RA Treatment Results in the Anterior Expansion of the Liver and Pancreas at the Expense of More Anterior Endodermal Derivatives RA (Sigma) stock solution was made in DMSO at 10−3 M, and working dilutions were made in embryo medium. Embryos were treated with RA from 9–10 hpf. The bars indicate the normal location of the pancreas, and the brackets indicate the normal location of the liver. (A) 10−6 M RA treatment causes pdx1 expression to extend anteriorly to the AP level of the eye (e). (B) islet1 is ectopically expressed in bilateral endodermal domains through the anterior extent of the embryo; note the loss of islet1 expression in the cranial nerves (compare to Figure 1G; n = 45). (C) Clusters of ectopic insulin and sst-expressing cells extend anteriorly from the normal location of the pancreas (n = 50). (D–I) Anterior expansion of pancreas and liver cells in response to RA is dose dependent. (D) 10−6 M RA treatment; insulin expression extends to the anterior of the embryo (n = 600). (E) 10−7 M RA treatment; insulin expression does not extend as far anterior (n = 20). (F) 10−6 M RA treatment; hhex liver expression extends anteriorly to the level of the posterior margin of the eye (n = 60). (G) 10−7 M RA; hhex expression extends only to hindbrain levels; also note the absence of the thyroid expression domain (n = 20). (H) trypsin (blue) and insulin (red). Both markers are expressed bilaterally in expanded anterior domains relative to untreated controls (n = 20). (I) 10−7 M RA shifts nkx2.3 pharyngeal endodermal expression anteriorly in comparison to untreated controls (arrowhead indicates posterior limit of nkx2.3 expression in controls; compare to Figure 2G). In embryos treated with 10−6 M RA, nkx2.3 expression cannot be detected (n = 20; DNS). Current Biology 2002 12, 1215-1220DOI: (10.1016/S0960-9822(02)00929-6)

Figure 4 RARs Are Expressed at an Appropriate Time and Place to Transduce RA Signals during Pancreatic Specification In situ hybridizations at 10.5 hpf. The insets show whole mounts with the dorsal side toward the reader and the animal pole toward the top of page (the precise location of the pancreatic primordium at this stage is not known; however, we postulate that converging endodermal cells in the vicinity of the equator of the embryo are most likely to contribute to the pancreas). The arrow indicates the level of the transverse section (main panel). Sectioning was performed as previously described [34]. (A) RARα2a[31] is expressed primarily in the ectoderm. (B) RARα2b (accession number L03399) is expressed in all three germ layers (the arrowhead indicates the endoderm). (C) RARγ[31] is expressed primarily in the endoderm (arrowhead). Current Biology 2002 12, 1215-1220DOI: (10.1016/S0960-9822(02)00929-6)