bluuebunny/crossref_metadata_2025 huggingface dataset:
Two distinct endothelial lineages in ontogeny, one of them related to hemopoiesis
Authors: Pardanaud Luc , Luton Dominique , Prigent Magali , Bourcheix Laurent-Marie , Catala Martin , Dieterlen-Lièvre Françoise
Published: April 2021

We have shown previously by means of quail/chick transplantations that external and visceral organs, i.e., somatopleural and splanchnopleural derivatives, acquire their endothelial network through different mechanisms, namely immigration (termed angiogenesis) versus in situ emergence of precursors (or vasculogenesis).

We have traced the distribution of QH1-positive cells in chick hosts after replacement of the last somites by quail somites (orthotopic grafts) or lateral plate mesoderm (heterotopic grafts). The results lead to the conclusion that the embryo becomes vascularized by endothelial precursors from two distinct regions, splanchnopleural mesoderm and paraxial mesoderm. The territories respectively vascularized are complementary, precursors from the paraxial mesoderm occupy the body wall and kidney, i.e., they settle along with the other paraxial mesoderm derivatives and colonize the somatopleure.

The precursors from the two origins have distinct recognition and potentialities properties: endothelial precursors of paraxial origin are barred from vascularizing visceral organs and from integrating into the floor of the aorta, and are never associated with hemopoiesis; splanchnopleural mesoderm grafted in the place of somites, gives off endothelial cells to body wall and kidney but also visceral organs. It gives rise to hemopoietic precursors in addition to endothelial cells.
The chinless mutation and neural crest cell interactions in zebrafish jaw development
Authors: Schilling Thomas F. , Walker Charline , Kimmel Charles B.
Published: April 2021

During vertebrate development, neural crest cells are thought to pattern many aspects of head organization, including the segmented skeleton and musculature of the jaw and gills. Here we describe mutations at the gene chinless, chn, that disrupt the skeletal fates of neural crest cells in the head of the zebrafish and their interactions with muscle precursors. chn mutants lack neural-crest-derived cartilage and mesoderm-derived muscles in all seven pharyngeal arches. Fate mapping and gene expression studies demonstrate the presence of both undifferentiated cartilage and muscle precursors in mutants. However, chn blocks differentiation directly in neural crest, and not in mesoderm, as revealed by mosaic analyses. Neural crest cells taken from wild-type donor embryos can form cartilage when transplanted into chn mutant hosts and rescue some of the patterning defects of mutant pharyngeal arches. In these cases, cartilage only forms if neural crest is transplanted at least one hour before its migration, suggesting that interactions occur transiently in early jaw precursors. In contrast, transplanted cells in paraxial mesoderm behave according to the host genotype; mutant cells form jaw muscles in a wild-type environment. These results suggest that chn is required for the development of pharyngeal cartilages from cranial neural crest cells and subsequent crest signals that pattern mesodermally derived myocytes.
Effects of GGF/neuregulins on neuronal survival and neurite outgrowth correlate with erbB2/neu expression in developing rat retina
Authors: Bermingham-McDonogh Olivia , McCabe Kathryn L. , Reh Thomas A.
Published: April 2021

We have examined the expression of neuregulin and its putative receptors, erbB2/neu, erbB3 and erbB4/tyro2 during retinal development, and tested several potential functions of this class of molecules in dissociated rat retinal cell cultures. At least one form of neuregulin is expressed in the retina, from the earliest stages of retinal development examined; in addition, all three of the known receptors are expressed by retinal neurons in a developmentally regulated manner. When added to cultures of embryonic or neonatal rat retinal cells, neuregulin (rhGGF2) promotes survival and neurite extension from retinal neurons in a dose-dependent manner. These results indicate that in addition to their well described effects on glia, the neuregulins also have direct effects on central nervous system neurons.
Notochord signals control the transcriptional cascade of myogenic bHLH genes in somites of quail embryos
Authors: Pownall Mary Elizabeth , Strunk Karen E. , Emerson Charles P.
Published: April 2021

Microsurgical, tissue grafting and in situ hybridization techniques have been used to investigate the role of the neural tube and notochord in the control of the myogenic bHLH genes, QmyoD, Qmyf5, Qmyogenin and the cardiac α-actin gene, during somite formation in stage 12 quail embryos. Our results reveal that signals from the axial neural tube/notochord complex control both the activation and the maintenance of expression of QmyoD and Qmyf5 in myotomal progenitor cells during the period immediately following somite formation and prior to myotome differentiation. QmyoD and Qmyf5 expression becomes independent of axial signals during myotome differentiation when somites activate expression of Qmyogenin and α-actin. Ablation studies reveal that the notochord controls QmyoD activation and the initiation of the transcriptional cascade of myogenic bHLH genes as epithelial somites condense from segmental plate mesoderm. The dorsal medial neural tube then contributes to the maintenance of myogenic bHLH gene expression in newly formed somites. Notochord grafts can activate ectopic QmyoD expression during somite formation, establishing that the notochord is a necessary and sufficient source of diffusible signals to initiate QmyoD expression. Myogenic bHLH gene expression is localized to dorsal medial cells of the somite by inhibitory signals produced by the lateral plate and ventral neural tube. Signaling models for the activation and maintenance of myogenic gene expression and the determination of myotomal muscle in somites are discussed.
Altering cell fates in sea urchin embryos by overexpressing SpOtx, an orthodenticle-related protein
Authors: Mao Chai-An , Wikramanayake Athula H. , Gan Lin , Chuang Chin-Kai , Summers Robert G. , Klein William H.
Published: April 2021

While many general features of cell fate specification in the sea urchin embryo are understood, specific factors associated with these events remain unidentified. SpOtx, an orth-odenticle-related protein, has been implicated as a transcriptional activator of the aboral ectoderm-specific Spec2a gene. Here, we present evidence that SpOtx has the potential to alter cell fates. SpOtx was found in the cytoplasm of early cleavage stage embryos and was translocated into nuclei between the 60- and 120-cell stage, coincident with Spec gene activation. Eggs injected with SpOtx mRNA developed into epithelial balls of aboral ectoderm suggesting that SpOtx redirected nonaboral ectoderm cells to an aboral ectoderm fate. At least three distinct domains on SpOtx, the homeobox and regions in the N-terminal and C-terminal halves of the protein, were required for the morphological alterations. These same N-terminal and C-terminal regions were shown to be transactivation domains in a yeast transactivation assay, indicating that the biological effects of overexpressing SpOtx were due to its action as a transcription factor. Our results suggest that SpOtx is involved in aboral ectoderm differentiation by activating aboral ectoderm-specific genes and that modulating its expression can lead to changes in cell fate.
The TGF-β signaling pathway is essential for Drosophila oogenesis
Authors: Twombly Vern , Blackman Ronald K. , Jin Hui , Graff Jonathan M. , Padgett Richard W. , Gelbart William. M.
Published: April 2021

We examine roles of signaling by secreted ligands of the TGF-β family during Drosophila oogenesis. One family member, the DPP ligand encoded by the decapentaplegic (dpp) gene, is required for patterning of anterior eggshell structures. This requirement presumably reflects the expression pattern of dpp in an anterior subset of somatic follicle cells: the centripetally migrating and the nurse cell-associated follicle cells. Similar requirements are also revealed by mutations in the saxophone (sax)-encoded receptor, consistent with the idea that DPP signaling is, at least in part, mediated by the SAX receptor. A loss of germline sax function results in a block in oogenesis associated with egg chamber degeneration and a failure of the transfer of nurse cell contents to the oocyte, indicating that TGF-β signaling is required for these events. Some phenotypes of sax mutations during oogenesis suggest that SAX responds to at least one other TGF-β ligand as well in the posterior follicle cells.
Two independent and polarized processes of cell elongation regulate leaf blade expansion in Arabidopsis thaliana (L.) Heynh.
Authors: Tsuge Tomohiko , Tsukaya Hirokazu , Uchimiya Hirofumi
Published: April 2021

For genetic analysis of mechanisms of leaf morphogenesis, we chose Arabidopsis thaliana (L.) Heynh. as a model for leaf development in dicotyledonous plants. Leaves of the angustifolia mutant were the same length as but narrower and thicker than wild-type leaves. The total number of cells in leaf blades of angustifolia plants was the same as in the wild type. At the cellular level in the angustifolia mutant it was found that the cells were smaller in the leaf-width direction and larger in the leaf-thickness direction than in wild type, revealing the function of the ANGUSTIFOLIA gene, which is to control leaf morphology by regulating polarity-specific cell elongation. The existence of similar genes that regulate leaf development in the length direction was, therefore, predicted. Three loci and several alleles associated with short-leaved mutants were newly isolated as rotundifolia mutants. The rotundifolia3 mutant had the same number of cells as the wild type, with reduced cell elongation in the leaf-length direction. The features of the angustifolia rotundifolia3 double mutant indicated that ANGUSTIFOLIA and ROTUNDIFOLIA3 genes act independently. We propose that leaf expansion in Arabidopsis involves at least two independent developmental processes: width development and length development, with the ANGUSTIFOLIA and ROTUNDIFOLIA3 genes playing different polarity-specific roles in cell elongation.
The maize mutant narrow sheath fails to establish leaf margin identity in a meristematic domain
Authors: Scanlon Michael J. , Schneeberger Richard G. , Freeling Michael
Published: April 2021

The maize mutant narrow sheath (ns) displays a leaf shape and plant stature phenotype that suggests the preprimordial deletion of a leaf domain. The ns mutant phenotype is inherited as a recessive, duplicate-factor trait, conditioned upon homozygosity for each of the two unlinked mutations narrow sheath-1 (ns1) and narrow sheath-2 (ns2). Mutant leaves are missing a large domain including the leaf margin, and mutant internodes are shortened on the marginal side of the stem. This domain deletion extends from the internode to beyond the longitudinal mid-length of the blade, and corresponds to an alteration in the organization of a specific region of the shoot apical meristem. The premargin region of mutant founder cells fail to down-regulate expression of Knox genes, markers of nonleaf meristematic identity. Our results indicate that leaf domains may acquire identity in the meristem itself, and that the subdivision of preprimordial developmental fields into differential domains is a common feature of both plant and animal organogenesis.
Complex expression of the zp-50 pou gene in the embryonic zebrafish brain is altered by overexpression of sonic hedgehog
Authors: Hauptmann Giselbert , Gerster Thomas
Published: April 2021

We report the characterization of the zebrafish zp-50 class III POU domain gene. This gene is first activated in the prospective diencephalon after the end of the gastrula period. During somitogenesis, zp-50 is expressed in a very dynamic and complex fashion in all major subdivisions of the central nervous system. After one day of development, zp-50 transcripts are present in the fore- and midbrain in several distinct cell clusters. In the hindbrain, zp-50 expression is found in two types of domains. Correct zp-50 expression in the ventral fore- and midbrain requires genes known to be involved in dorsoventral patterning of the zebrafish CNS. Transcripts of the sonic hedgehog (shh) gene encoding an intercellular signaling molecule are detected in the forming diencephalon shortly prior to the appearance of zp-50 mRNA. Correct expression in this region of both shh, and zp-50, requires a functional cyclops (cyc) locus: shh and zp-50 transcripts are likewise absent from the ventral rostral brain of mutant cyc−/− embryos. Injection of synthetic shh mRNA into fertilized eggs causes ectopic zp-50 expression at more dorsal positions of the embryonic brain. The close spatial and temporal coincidence of expression in the rostral brain, the similar response to the cyc− mutation, and the ectopic zp-50 expression in the injection experiments all suggest that zp-50 may directly respond to the reception of the Shh signal.
Cell fate determination in an annelid equivalence group
Authors: Huang Françoise Z. , Weisblat David A.
Published: April 2021

In embryos of glossiphoniid leeches such as Helobdella triserialis and H. robusta, pairs of adjacent ‘o/p’ ectodermal blast cells are known to be developmentally equipotent and yet eventually contribute distinct sets of ‘O’ and ‘P’ progeny to the nervous system and epidermis of the mature leech. It has been thought that the fate-determining interactions in this ‘O-P equivalence group’ take place between the equipotent cells themselves. We show here that such intra-group interactions are neither necessary nor sufficient. Instead, transient contact with cells in another ectodermal lineage is necessary and sufficient to induce o/p blast cells to assume the P fate. In the absence of this contact they assume the O fate.
Oligodendrocyte progenitors are generated throughout the embryonic mouse brain, but differentiate in restricted foci
Authors: Hardy Rebecca J. , Friedrich Victor L.
Published: April 2021

Recent evidence from studies mapping the expression of putative oligodendrocyte progenitor specific mRNAs has suggested that oligodendrocyte progenitors arise during embryogenesis, in specific foci of the neuroectoderm. In order to test this hypothesis, we have assayed different regions of the embryonic central nervous system for their ability to generate oligodendrocytes following transplantation into neonatal cerebrum. To allow identification of donor-derived oligodendrocytes in wild-type host brain, we used the MβP transgenic mouse, which expresses lacZ in oligodendrocytes, as donor tissue. We found that tissue fragments derived from several levels of the anteriorposterior axis of the neural tube at E14.5 and E12.5, chosen to include (hindbrain, cervical and lumbar spinal cord), or exclude (dorsal telencephalon) putative foci of oligodendrocyte progenitors, all produced oligodendrocytes following transplantation. In addition, these same regions taken from E10.5, prior to the appearance of putative oligodendrocyte progenitor markers, also all yielded oligodendrocytes on transplantation. This indicates that precursor cells that can generate oligodendrocytes are widespread throughout the neuroectoderm as early as E10.5.

We have also used the oligodendrocyte lineage-specific glycolipid antibodies O4, R-mAb and O1 to identify those regions of the developing brain that first support the differentiation of oligodendrocytes from their progenitor cells. We found that the first oligodendrocytes arise in prenatal brain at E14.5, in a restricted zone adjacent to the midline of the medulla. These cells are mitotically inactive, differentiated oligodendrocytes and, using light and electron microscopy, we show that they become functional, myelinbearing oligodendrocytes. We have mapped the subsequent appearance of differentiated oligodendrocytes in the prenatal brain and show that they appear in a restricted, tract-specific manner.

Our results suggest that oligodendrocytes are generated from neuroectodermal cells positioned throughout the rostrocaudal axis of the neural tube, rather than at restricted locations of the neuroectoderm. By contrast, the differentiation of such cells into oligodendrocytes does occur in a restricted manner, consistent with local regulation of oligodendrocyte progenitor differentiation.
Mothers against dpp encodes a conserved cytoplasmic protein required in DPP/TGF-β responsive cells
Authors: Newfeld Stuart J. , Chartoff Elena H. , Graff Jonathan M. , Melton Douglas A. , Gelbart William M.
Published: April 2021

The proteins necessary for signal transduction in cells responding to ligands of the TGF-β family are largely unknown. We have previously identified Mad (Mothers against dpp), a gene that interacts with the TGF-β family member encoded by decapentaplegic (dpp) in Drosophila. Assay of Mad’s role in the DPP-dependent events of embryonic midgut development demonstrates that Mad is required for any response of the visceral mesoderm or endoderm to DPP signals from the visceral mesoderm. Replacement of the normal DPP promoter with a heterologous (hsp70) promoter fails to restore DPP-dependent responses in Mad mutant midguts. Experiments utilizing Mad transgenes regulated by tissue-specific promoters show that MAD is required specifically in cells responding to DPP. Immunohistochemical studies localize MAD to the cytoplasm in all tissues examined. Experiments in Xenopus embryos demonstrate that Drosophila MAD can function in the signaling pathway of BMP-4, a vertebrate homolog of dpp. Based on these results, we propose that Mad is a highly conserved and essential element of the DPP signal transduction pathway.
A direct measurement of increased divalent cation influx in fertilised mouse oocytes
Authors: McGuinness Orla M. , Moreton Roger B. , Johnson Martin H. , Berridge Michael J.
Published: April 2021

On fertilisation of mouse oocytes, the fusing spermatozoon triggers a series of repetitive calcium (Ca2+) spikes. These Ca2+ spikes seem to be necessary for successful progression through the cell cycle and are regulated in a cell-cycledependent manner. The spikes appear to require the linkage of continuous Ca2+ influx to the periodic release of Ca2+ from intracellular stores by a process of Ca2+-induced Ca2+ release. The precise role of Ca2+ influx was explored using the manganese (Mn2+)-quench technique to monitor unidirectional cation influx into single mouse oocytes. There was a marked stimulation of cation influx associated closely with the upsweep of the first and subsequent fertilisation Ca2+ spikes. A smaller but significant increase in the rate of cation influx persisted in the interspike period in fertilised oocytes. Spike-associated entry was not as apparent in oocytes stimulated to spike repetitively by thimerosal or acetylcholine application. Instead, there was a continuous increase in cation influx underlying Ca2+ spiking which commenced with the onset of the first spike. Using the specific microsomal inhibitor thapsigargin and the Ca2+ionophore ionomycin, we found evidence for a capacitative entry mechanism in mouse oocytes. We propose that the persistent influx of Ca2+ observed in response to all stimuli examined is controlled by a capacitative mechanism and sets the frequency of spiking by determining the time taken to refill the internal stores to a point where they are again sensitive enough to initiate the next spike.
Dual function of the region-specific homeotic gene spalt during Drosophila tracheal system development
Authors: Kühnlein Ronald P. , Schuh Reinhard
Published: April 2021

We report that the region-specific homeotic gene spalt affects the Drosophila tracheal system at two different stages of embryonic development. Both lack-of-function and gain-of-function experiments show that blastodermal spalt activity restricts tracheal development to 10 bilaterally positioned pairs of tracheal placodes in the trunk region by repressing placode formation in parasegments 2, 3 and 14. The results suggest that the activity of the zinkfinger type transcription factor encoded by spalt suppresses the molecular pathway that establishes tracheal development. spalt function is also necessary for the directed migration of the dorsal trunk cells, a distinct subset of tracheal cells. This process is a prerequisite for the formation of the dorsal trunk generated by fusion of adjacent tracheal metameres into a common tubular structure. The directed cell migration, in which spalt gene function participates, seems to be independent of branch fusion and general tracheal cell migration processes.
Specification of cell fates at the dorsal margin of the zebrafish gastrula
Authors: Melby Anna E. , Warga Rachel M. , Kimmel Charles B.
Published: April 2021

Using fate mapping techniques, we have analyzed development of cells of the dorsal marginal region in wild-type and mutant zebrafish. We define a domain in the early gastrula that is located just at the margin and centered on the dorsal midline, in which most cells generate clones that develop exclusively as notochord. The borders of the notochord domain are sharp at the level of single cells, and coincide almost exactly with the border of the expression domain of the homeobox gene floating head (flh; zebrafish homologue of Xnot), a gene essential for notochord development. In flh mutants, cells in the notochord domain generate clones of muscle cells. In contrast, notochord domain cells form mesenchyme in embryos mutant for no tail (ntl; zebrafish homologue of Brachyury).

A minority of cells in the notochord domain in wild-type embryos develop as unrestricted mesoderm, invariably located in the tail, suggesting that early gastrula expression of flh does not restrict cellular potential to the notochord fate. The unrestricted tail mesodermal fate is also expressed by the forerunner cells, a cluster of cells located outside the blastoderm, adjacent to the notochord domain. We show that cells can leave the dorsal blastoderm to join the forerunners, suggesting that relocation between fate map domains might respecify notochord domain cells to the tail mesodermal fate. An intermediate fate of the forerunners is to form the epithelial lining of Kupffer’s vesicle, a transient structure of the teleost tailbud. The forerunners appear to generate the entire structure of Kupffer’s vesicle, which also develops in most flh mutants. Although forerunner cells are present in ntl mutants, Kupffer’s vesicle never appears, which is correlated with the later severe disruption of tail development.
Sensitivity of proneural genes to lateral inhibition affects the pattern of primary neurons in Xenopus embryos
Authors: Chitnis Ajay , Kintner Chris
Published: April 2021

We have compared the roles of XASH-3 and NeuroD, two basic helix-loop-helix transcription factors, in the formation of primary neurons in early Xenopus embryos. When ectopically expressed in Xenopus embryos, XASH-3 and NeuroD induce ectopic primary neurons in very different spatial patterns. We show that the pattern of primary neurons induced by XASH-3 and NeuroD can be accounted for by a difference in their sensitivity to inhibitory interactions mediated by the neurogenic genes, X-Delta-1 and X-Notch-1. Both NeuroD and XASH-3 promote the expression of the inhibitory ligand, X-Delta-1. However, XASH-3 appears to be sensitive to the inhibitory effects of X-Delta-1 while NeuroD is much less so. Consequently only a subset of cells that ectopically express XASH-3 eventually form neurons, giving a scattered pattern, while the ectopic expression of NeuroD leads to a relatively dense pattern of ectopic neurons. We propose that differences in the sensitivity of XASH-3 and NeuroD to lateral inhibition play an important role during their respective roles in neuronal determination and differentiation.
Xenopus mothers against decapentaplegic is an embryonic ventralizing agent that acts downstream of the BMP-2/4 receptor
Authors: Thomsen Gerald H.
Published: April 2021

Dorsal-ventral patterning in vertebrate embryos is regulated by members of the TGF-β family of growth and differentiation factors. In Xenopus the activins and Vg1 are potent dorsal mesoderm inducers while members of the bone morphogenetic protein (BMP) subclass pattern ventral mesoderm and regulate ectodermal cell fates. Receptors for ligands in the TGF-β superfamily are serinethreonine kinases, but little is known about the components of the signal transduction pathway leading away from these receptors. In Drosophila the decapentaplegic protein (dpp), a homolog of vertebrate BMP-2 and BMP-4, functions in dorsal-ventral axial patterning, and a genetic screen for components involved in signaling by dpp has identified a gene named mothers against decapentaplegic (Mad). Mad encodes a unique, predicted cytoplasmic, protein containing no readily identified functional motifs. This report demonstrates that a gene closely related to Drosophila Mad exists in Xenopus (called XMad) and it exhibits activities consistent with a role in BMP signaling. XMad protein induces ventral mesoderm when overexpressed in isolated animal caps and it ventralizes embryos. Furthermore, XMad rescues phenotypes generated by a signalingdefective, dominant-negative, BMP-2/4 receptor. These results furnish evidence that XMad protein participates in vertebrate embryonic dorsal-ventral patterning by functioning in BMP-2/4 receptor signal transduction.
Reversal of cell fate determination in Caenorhabditis elegans vulval development
Authors: Euling Susan , Ambros Victor
Published: April 2021

In Caenorhabditis elegans, the fates of the multipotent vulval precursor cells (VPCs) are specified by intercellular signals. The VPCs divide in the third larval stage (L3) of the wild type, producing progeny of determined cell types. In lin-28 mutants, vulva development is similar to wild-type vulva development except that it occurs precociously, in the second larval stage (L2). Consequently, when lin-28 hermaphrodites temporarily arrest development at the end of L2 in the dauer larva stage, they have partially developed vulvae consisting of VPC progeny. During post-dauer development, these otherwise determined VPC progeny become reprogrammed back to the multipotent, signalsensitive state of VPCs. Our results indicate that VPC fate determination by intercellular signals is reversible by dauer larva developmental arrest and post-dauer development.
Integrin α6 expression is required for early nervous system development in Xenopus laevis
Authors: Lallier Thomas E. , Whittaker Charles A. , DeSimone Douglas W.
Published: April 2021

The integrin α6 subunit pairs with both the β1 and β4 subunits to form a subfamily of laminin receptors. Here we report the cDNA cloning and primary sequence for the Xenopus homologue of the mammalian integrin α6 subunit. We present data demonstrating the spatial and temporal expression of α6 mRNA and protein during early development. Initially, α6 transcripts are expressed in the dorsal ectoderm and future neural plate at the end of gastrulation. Later in development, α6 mRNAs are expressed in a variety of neural derivatives, including the developing sensory placodes (otic and olfactory) and commissural neurons within the neural tube. Integrin α6 is also expressed in the elongating pronephric duct as well as a subset of the rhombencephalic neural crest, which will form the Schwann cells lining several cranial nerves (VII, VIII and X). In vivo expression of an α6 antisense transcript in the animal hemisphere leads to a reduction in α6 protein expression, a loss of adhesion to laminin, and severe defects in normal development. In 35% of cases, reduced levels of α6 expression result in embryos that complete gastrulation normally but arrest at neurulation prior to the formation of the neural plate. In an additional 22% of cases, embryos develop with severe axial defects, including complete loss of head or tail structures. In contrast, overexpression of the α6 subunit by injection of full-length mRNA has no apparent effect on embryonic development. Co-injection of antisense and sense plasmid constructs results in a partial rescue of the antisense-generated phenotypes. These data indicate that the integrin α6 subunit is critical for the early development of the nervous system in amphibians.
Mutations affecting xanthophore pigmentation in the zebrafish, Danio rerio
Authors: Odenthal Jörg , Rossnagel Karin , Haffter Pascal , Kelsh Robert N. , Vogelsang Elisabeth , Brand Michael , van Eeden Fredericus J. M. , Furutani-Seiki Makoto , Granato Michael , Hammerschmidt Matthias , Heisenberg Carl-Philipp , Jiang Yun-Jin , Kane Donald A. , Mullins Mary C. , Nüsslein-Volhard Christiane
Published: April 2021

In a large-scale screen for mutants with defects in embryonic development we identified 17 genes (65 mutants) specifically required for the development of xan-thophores. We provide evidence that these genes are required for three different aspects of xanthophore development. (1) Pigment cell formation and migration (pfeffer and salz); (2) pigment synthesis (edison, yobo, yocca and brie) and (3) pigment translocation (esrom, tilsit and tofu). The number of xanthophore cells that appear in the body is reduced in embryos with mutations in the two genes, salz and pfeffer. In heterozygous and homozygous salz and pfeffer adults, the melanophore stripes are interrupted, indicating that xanthophore cells have an important function in adult melanophore pattern formation. Most other genes affect only larval pigmentation. In embryos mutant for edison, yobo, yocca and brie, differences in pteridine synthesis can be observed under UV light and by thin-layer chromatography. Homozygous mutant females of yobo show a recessive maternal effect. Embryonic development is slowed down and embryos display head and tail truncations. Xanthophores in larvae mutant in the three genes esrom, tilsit and tofu appear less spread out. In addition, these mutants display a defect in retinotectal axon pathfinding. These mutations may affect xanthophore pigment distribution within the cells or xanthophore cell shape. Mutations in seven genes affecting xanthophore pig-mentation remain unclassified.
Zebrafish mutations affecting retinotectal axon pathfinding
Authors: Karlstrom Rolf O. , Trowe Torsten , Klostermann Stefan , Baier Herwig , Brand Michael , Crawford Alexander D. , Grunewald Barbara , Haffter Pascal , Hoffmann Heike , Meyer Stefan U. , Müller Bernhard K. , Richter Sandra , Eeden Fredericus J. M. van , Nüsslein-Volhard Christiane , Bonhoeffer Friedrich
Published: April 2021

We have isolated mutants in the zebrafish Danio rerio that have defects in axonal connectivity between the retina and tectum. 5-day-old fish larvae were screened by labeling retinal ganglion cells with DiI and DiO and observing their axonal projections to and on the tectum.

82 mutations, representing 13 complementation groups and 6 single allele loci, were found that have defects in retinal ganglion cell axon pathfinding to the tectum. These pathfinding genes fall into five classes, based on the location of pathfinding errors between eye and tectum. In Class I mutant larvae (belladonna, detour, you-too, iguana, umleitung, blowout) axons grow directly to the ipsilateral tectal lobe after leaving the eye. Class II mutant larvae (chameleon, bashful) have ipsilaterally projecting axons and, in addition, pathfinding mistakes are seen within the eye. In Class III mutant larvae (esrom, tilsit, tofu) fewer axons than normal cross the midline, but some axons do reach the contralateral tectal lobe. Class IV mutant larvae (boxer, dackel, pinscher) have defects in axon sorting after the midline and retinal axons occasionally make further pathfinding errors upon reaching the contralateral tectal lobe. Finally, Class V mutant larvae (bashful, grumpy, sleepy, cyclops, astray) have anterior-posterior axon trajectory defects at or after the midline.

The analysis of these mutants supports several conclusions about the mechanisms of retinal axon pathfinding from eye to tectum. A series of sequential cues seems to guide retinal axons to the contralateral tectal lobe. Pre-existing axon tracts seem not to be necessary to guide axons across the midline. The midline itself seems to play a central role in guiding retinal axons. Axons in nearby regions of the brain seem to use different cues to cross the ventral midline. Mutant effects are not all- or-none, as misrouted axons may reach their target, and if they do, they project normally on the tectum. The retinotectal pathfinding mutants reveal important choice points encountered by neuronal growth cones as they navigate between eye and tectum.
dino and mercedes, two genes regulating dorsal development in the zebrafish embryo
Authors: Hammerschmidt Matthias , Pelegri Francisco , Mullins Mary C. , Kane Donald A. , Eeden Fredericus J. M. van , Granato Michael , Brand Michael , Furutani-Seiki Makoto , Haffter Pascal , Heisenberg Carl-Philipp , Jiang Yun-Jin , Kelsh Robert N. , Odenthal Jörg , Warga Rachel M. , Nüsslein-Volhard Christiane
Published: April 2021

We describe two genes, dino and mercedes, which are required for the organization of the zebrafish body plan. In dino mutant embryos, the tail is enlarged at the expense of the head and the anterior region of the trunk. The altered expression patterns of various marker genes reveal that, with the exception of the dorsal most marginal zone, all regions of the early dino mutant embryo acquire more ventral fates. These alterations are already apparent before the onset of gastrulation. mercedes mutant embryos show a similar but weaker phenotype, suggesting a role in the same patterning processes. The phenotypes suggests that dino and mercedes are required for the establishment of dorsal fates in both the marginal and the animal zone of the early gastrula embryo. Their function in the patterning of the ventrolateral mesoderm and the induction of the neu-roectoderm is similar to the function of the Spemann organizer in the amphibian embryo.
Hedgehog organises the pattern and polarity of epidermal cells in the Drosophila abdomen
Authors: Struhl Gary , Barbash Daniel A. , Lawrence Peter A.
Published: April 2021

The abdomen of adult Drosophila, like that of other insects, is formed by a continuous epithelium spanning several segments. Each segment is subdivided into an anterior (A) and posterior (P) compartment, distinguished by activity of the selector gene engrailed (en) in P but not A compartment cells. Here we provide evidence that Hedgehog (Hh), a protein secreted by P compartment cells, spreads into each A compartment across the anterior and the posterior boundaries to form opposing concentration gradients that organize cell pattern and polarity. We find that anteriorly and posteriorly situated cells within the A compartment respond in distinct ways to Hh: they express different combinations of genes and form different cell types. They also form polarised structures that, in the anterior part, point down the Hh gradient and, in the posterior part, point up the gradient — therefore all structures point posteriorly. Finally, we show that ectopic Hh can induce cells in the middle of each A compartment to activate en. Where this happens, A compartment cells are transformed into an ectopic P compartment and reorganise pattern and polarity both within and around the transformed tissue. Many of these results are unexpected and lead us to reassess the role of gradients and compartments in patterning insect segments.
NeuroM, a neural helix-loop-helix transcription factor, defines a new transition stage in neurogenesis
Authors: Roztocil Tomas , Matter-Sadzinski Lidia , Alliod Christine , Ballivet Marc , Matter Jean-Marc
Published: April 2021

Genes encoding transcription factors of the helix-loop-helix family are essential for the development of the nervous system in Drosophila and vertebrates. Screens of an embryonic chick neural cDNA library have yielded NeuroM, a novel neural-specific helix-loop-helix transcription factor related to the Drosophila proneural gene atonal. The NeuroM protein most closely resembles the vertebrate NeuroD and Nex1/MATH2 factors, and is capable of trans-activating an E-box promoter in vivo. In situ hybridization studies have been conducted, in conjunction with pulse-labeling of S-phase nuclei, to compare NeuroM to NeuroD expression in the developing nervous system. In spinal cord and optic tectum, NeuroM expression precedes that of NeuroD. It is transient and restricted to cells lining the ventricular zone that have ceased proliferating but have not yet begun to migrate into the outer layers. In retina, NeuroM is also transiently expressed in cells as they withdraw from the mitotic cycle, but persists in horizontal and bipolar neurons until full differentiation, assuming an expression pattern exactly complementary to NeuroD. In the peripheral nervous system, NeuroM expression closely follows cell proliferation, suggesting that it intervenes at a similar developmental juncture in all parts of the nervous system. We propose that availability of the NeuroM helix-loop-helix factor defines a new stage in neurogenesis, at the transition between undifferentiated, premigratory and differentiating, migratory neural precursors.
Fgfr2 and osteopontin domains in the developing skull vault are mutually exclusive and can be altered by locally applied FGF2
Authors: Iseki S. , Wilkie A. O. M. , Heath J. K. , Ishimaru T. , Eto K. , Morriss-Kay G. M.
Published: April 2021

Mutations in the human fibroblast growth factor receptor type 2 (FGFR2) gene cause craniosynostosis, particularly affecting the coronal suture. We show here that, in the fetal mouse skull vault, Fgfr2 transcripts are most abundant at the periphery of the membrane bones; they are mutually exclusive with those of osteopontin (an early marker of osteogenic differentiation) but coincide with sites of rapid cell proliferation. Fibroblast growth factor type 2 (FGF2) protein, which has a high affinity for the FGFR2 splice variant associated with craniosynostosis, is locally abundant; immunohistochemical detection showed it to be present at low levels in Fgfr2 expression domains and at high levels in differentiated areas. Implantation of FGF2-soaked beads onto the fetal coronal suture by ex utero surgery resulted in ectopic osteopontin expression, encircled by Fgfr2 expression, after 48 hours. We suggest that increased FGF/FGFR signalling in the developing skull, whether due to FGFR2 mutation or to ectopic FGF2, shifts the cell proliferation/differentiation balance towards differentiation by enhancing the normal paracrine down-regulation of Fgfr2.
The mouse Ulnaless mutation deregulates posterior HoxD gene expression and alters appendicular patterning
Authors: Peichel Catherine L. , Prabhakaran Bindu , Vogt Thomas F.
Published: April 2021

The semi-dominant mouse mutation Ulnaless alters patterning of the appendicular but not the axial skeleton. Ulnaless forelimbs and hindlimbs have severe reductions of the proximal limb and less severe reductions of the distal limb. Genetic and physical mapping has failed to separate the Ulnaless locus from the HoxD gene cluster (Peichel, C. L., Abbott, C. M. and Vogt, T. F. (1996) Genetics 144, 1757-1767). The Ulnaless limb phenotypes are not recapitulated by targeted mutations in any single HoxD gene, suggesting that Ulnaless may be a gain-of-function mutation in a coding sequence or a regulatory mutation. Deregulation of 5′ HoxD gene expression is observed in Ulnaless limb buds. There is ectopic expression of Hoxd-13 and Hoxd-12 in the proximal limb and reduction of Hoxd-13, Hoxd-12 and Hoxd-11 expression in the distal limb. Skeletal reductions in the proximal limb may be a consequence of posterior prevalence, whereby proximal misexpression of Hoxd-13 and Hoxd-12 results in the transcriptional and/or functional inactivation of Hox group 11 genes. The Ulnaless digit phenotypes are attributed to a reduction in the distal expression of Hoxd-13, Hoxd-12, Hoxd-11 and Hoxa-13. In addition, Hoxd-13 expression is reduced in the genital bud, consistent with the observed alterations of the Ulnaless penian bone. No alterations of HoxD expression or skeletal phenotypes were observed in the Ulnaless primary axis. We propose that the Ulnaless mutation alters a cis-acting element that regulates HoxD expression specifically in the appendicular axes of the embryo.
Ulnaless (Ul), a regulatory mutation inducing both loss-of-function and gain- of-function of posterior Hoxd genes
Authors: Hérault Yann , Fraudeau Nadine , Zákány József , Duboule Denis
Published: April 2021

Ulnaless (Ul), an X-ray-induced dominant mutation in mice, severely disrupts development of forearms and forelegs. The mutation maps on chromosome 2, tightly linked to the HoxD complex, a cluster of regulatory genes required for proper morphogenesis. In particular, 5′-located (posterior) Hoxd genes are involved in limb development and combined mutations within these genes result in severe alterations in appendicular skeleton. We have used several engineered alleles of the HoxD complex to genetically assess the potential linkage between these two loci. We present evidence indicating that Ulnaless is allelic to Hoxd genes. Important modifications in the expression patterns of the posterior Hoxd-12 and Hoxd-13 genes at the Ul locus suggest that Ul is a regulatory mutation that interferes with a control mechanism shared by multiple genes to coordinate Hoxd function during limb morphogenesis.
Genetic control of brain morphogenesis through Otx gene dosage requirement
Authors: Acampora Dario , Avantaggiato, Virginia , Tuorto Francesca , Simeone Antonio
Published: April 2021

Understanding the genetic mechanisms that control patterning of the vertebrate brain represents a major challenge for developmental neurobiology. Previous data suggest that Otx1 and Otx2, two murine homologs of the Drosophila orthodenticle (otd) gene, might both contribute to brain morphogenesis. To gain insight into this possibility, the level of OTX proteins was modified by altering in vivo the Otx gene dosage. Here we report that Otx genes may cooperate in brain morphogenesis and that a minimal level of OTX proteins, corresponding either to one copy each of Otx1 and Otx2, or to only two copies of Otx2, is required for proper regionalization and subsequent patterning of the developing brain. Thus, as revealed by anatomical and molecular analyses, only Otx1−/−; Otx2+/− embryos lacked mesencephalon, pretectal area, dorsal thalamus and showed an heavy reduction of the Ammon’s horn, while the metencephalon was dramatically enlarged occupying the mesencencephalic area. In 8.5 days post coitum (d.p.c.) Otx1−/−; Otx2+/− embryos, the expression patterns of mesencephalic-metencephalic (mes-met) markers such as En-1 and Wnt-1 confirmed the early presence of the area fated to give rise to mesencephalon and metencephalon while Fgf-8 transcripts were improperly localized in a broader domain. Thus, in Otx1−/−; Otx2+/− embryos, Fgf-8 misexpression is likely to be the consequence of a reduced level of specification between mes-met primitive neuroepithelia that triggers the following repatterning involving the transformation of mesencephalon into metencephalon, the establishment of an isthmic-like structure in the caudal diencephalon and, by 12.5 d.p.c., the telencephalic expression of Wnt-1 and En-2. Taken together these findings support the existence of a molecular mechanism depending on a precise threshold of OTX proteins that is required to specify early regional diversity between adjacent mes-met territories and, in turn, to allow the correct positioning of the isthmic organizer.
Induction of Drosophila eye development by Decapentaplegic
Authors: Pignoni Francesca , Zipursky S. Lawrence
Published: April 2021

The Drosophila decapentaplegic (dpp) gene, encoding a secreted protein of the transforming growth factor-β (TGFβ) superfamily, controls proliferation and patterning in diverse tissues, including the eye imaginal disc. Pattern formation in this tissue is initiated at the posterior edge and moves anteriorly as a wave; the front of this wave is called the morphogenetic furrow (MF). Dpp is required for proliferation and initiation of pattern formation at the posterior edge of the eye disc. It has also been suggested that Dpp is the principal mediator of Hedgehog function in driving progression of the MF across the disc. In this paper, ectopic Dpp expression is shown to be sufficient to induce a duplicated eye disc with normal shape, MF progression, neuronal cluster formation and direction of axon outgrowth. Induction of ectopic eye development occurs preferentially along the anterior margin of the eye disc. Ectopic Dpp clones situated away from the margins induce neither proliferation nor patterning. The Dpp signalling pathway is shown to be under tight transcriptional and post-transcriptional control within different spatial domains in the developing eye disc. In addition, Dpp positively controls its own expression and suppresses wingless transcription. In contrast to the wing disc, Dpp does not appear to be the principal mediator of Hedgehog function in the eye.
Murine gastrulation requires HNF-4 regulated gene expression in the visceral endoderm: tetraploid rescue of Hnf-4−/− embryos
Authors: Duncan Stephen A. , Nagy Andras , Chan Wendy
Published: April 2021

Immediately prior to gastrulation the murine embryo consists of an outer layer of visceral endoderm (VE) and an inner layer of ectoderm. Differentiation and migration of the ectoderm then occurs to produce the three germ layers (ectoderm, embryonic endoderm and mesoderm) from which the fetus is derived. An indication that the VE might have a critical role in this process emerged from studies of Hnf-4− /− mouse embryos which fail to undergo normal gastrulation. Since expression of the transcription factor HNF-4 is restricted to the VE during this phase development, we proposed that HNF-4-regulated gene expression in the VE creates an environment capable of supporting gastrulation. To address this directly we have exploited the versatility of embryonic stem (ES) cells which are amenable to genetic manipulation and can be induced to form VE in vitro. Moreover, embryos derived solely from ES cells can be generated by aggregation with tetraploid morulae. Using Hnf-4− /− ES cells we demonstrate that HNF-4 is a key regulator of tissue-speci?c gene expression in the VE, required for normal expression of secreted factors including alphafetoprotein, apolipoproteins, transthyretin, retinol binding protein, and transferrin. Furthermore, speci?c complementation of Hnf-4− /− embryos with tetraploid-derived Hnf-4+/+ VE rescues their of early developmental arrest, showing conclusively that a functional VE is mandatory for gastrulation.
α-Spectrin is required for ovarian follicle monolayer integrity in Drosophila melanogaster
Authors: Lee John K. , Brandin Eric , Branton Daniel , Goldstein Lawrence S. B.
Published: April 2021

To understand the role of the spectrin-based membrane skeleton in generating epithelial polarity, we characterized the distribution of membrane skeletal components in Drosophila ovarian follicle cells and in somatic clones of mutant cells that lack αspectrin. Immunolocalization data reveal that wild-type follicle cells contain two populations of spectrin heterodimers: a network of αβ heterodimers concentrated on the lateral plasma membrane and an αβH population targeted to the apical surface. Induction of somatic clones lacking αspectrin leads to follicle cell hyperplasia. Surprisingly, elimination of αspectrin from follicle cells does not appear to prevent the assembly of conventional βspectrin and ankyrin at the lateral domain of the follicle cell plasma membrane. However, the α-subunit is essential for the correct localization of βH-spectrin to the apical surface. As a consequence of disrupting the apical membrane skeleton a distinct sub population of follicle cells undergoes unregulated proliferation which leads to the loss of monolayer organization and disruption of the anterior-posterior axis of the oocyte. These results suggest that the spectrin-based membrane skeleton is required in a developmental pathway that controls follicle cell monolayer integrity and proliferation.