bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential reaction of crossing and non-crossing rat retinal axons on cell membrane preparations from the chiasm midline: an in vitro study
Authors: Wizenmann A. , Thanos S. , Boxberg Y. v. , Bonhoeffer F.
Published: April 2021

In the rat, a small subpopulation of retinal ganglion cell axons forms a persistent projection to the ipsilateral half of the brain. These fibres originate almost exclusively from the ventrotemporal margin of the retina. In contrast to all other retinal axons they seem to be deflected from the midline of the optic chiasm and thereby led into the ipsilateral optic tract.

In order to analyse the interactions between growing fibres and chiasm midline, we have developed the following in vitro model. Axons of the embryonic rat retina are grown on a carpet of tectal cell membranes used as a general growth-permissive substratum. At a certain distance from the explant (200-450 μ m), the advancing fibres are confronted with two stripes of cell membranes prepared from the chiasm midline. Such chiasm membranes are shown to act as a barrier for the presumptive non-crossing axons, while they do not influence growth of fibres originating from any other regions of the retina, including the dorsotemporal part. The repulsion of non-crossing fibres by chiasm membranes is observed in vitro only when retinal explants from embryonic day (E) 17/18 and chiasm preparations from E14/15 are used. Fibres and tissue from different regions of the brain as well as from different developmental ages, and even from different species, can be combined in this assay system. In a first attempt to characterize the molecular basis of the repulsive effect of chiasm membranes on ventrotemporal fibres, similar assays were performed with membranes derived from other regions of the central nervous system midline, some of which are known to have repulsive properties against certain axon populations. Since these cell membranes did not act as a barrier for the ventrotemporal retinal axons, we suggest that the guidance cues at the chiasm are very specific.

Our results are consistent with the hypothesis that certain cells at the chiasm midline (very likely radial glial cells) express ‘repulsive or inhibitory’ molecules, which act in a specific way on ipsilaterally projecting axons.
Sex determination in Drosophila: sis-b, a major numerator element of the X:A ratio in the soma, does not contribute to the X:A ratio in the germ line
Authors: Steinmann-Zwicky Monica
Published: April 2021

In soma and germ cells of Drosophila, the X:A ratio builds a primary signal for sex determination, and in both tissues Sex-lethal (Sxl) function is required for cells to enter the female pathway.

In somatic cells of XX animals, the products of X-chromosomal elements of the X:A ratio activate Sxl. Here I show that sisterless-b (sis-b), which is the X-chro-mosomal element of the somatic X:A ratio that has best been analysed, is not required for oogenesis. I also present evidence that Sxl function might not be suffi-cient to direct germ cells into the female pathway. These results show that the elements forming the X:A ratio in the germ line are different from the elements forming the X:A ratio in the soma and they suggest that, in the germ line, Sxl might not be regulated by the X:A ratio.
A role for subplate neurons in the patterning of connections from thalamus to neocortex
Authors: Ghosh Anirvan , Shatz Carla J.
Published: April 2021

During cerebral cortical development, ingrowing axons from different thalamic nuclei select and invade their cortical targets. The selection of an appropriate target is first evident even before thalamic axons grow into the cortical plate: initially axons accumulate and wait below their cortical target area in a zone called the subplate. This zone also contains the first postmitotic neurons of the cerebral cortex, the subplate neurons. Here we have investigated whether subplate neurons are involved in the process of target selection by thalamic axons by ablating them from specific cortical regions at the onset of the waiting period and examining the subsequent thalamocortical axon projection patterns.

Subplate neurons were ablated at the onset of the waiting period by intracortical injections of kainic acid. The effect of the ablation on the thalamocortical projection from visual thalamus was examined by DiI-labeling of the LGN days to weeks following the lesion. At two to four weeks post-lesion, times when LGN axons would have normally invaded the cortical plate, the axons remained below the cortical plate and grew past their appropriate cortical target in an anomalous pathway. Moreover, examination of LGN axons at one week post-lesion, a time when they would normally be waiting and branching within the visual subplate, indicated that the axons had already grown past their correct destination. These observations suggest that visual subplate neurons are involved in the process by which LGN axons select and subsequently grow into visual cortex. In contrast, subplate neurons do not appear to play a major role in the initial morphological development of the LGN itself. Subplate ablations did not alter dendritic growth or shapes of LGN projection neurons during the period under study, nor did it prevent the segregation of retinal ganglion cell axons into eye-specific layers. However, the overall size of the LGN was reduced, suggesting that there may be increased cell death of LGN neurons in the absence of subplate neurons.

To examine whether subplate neurons beneath other neocortical areas play a similar role in the formation of thalamocortical connections, subplate neurons were deleted beneath auditory cortex at the onset of the waiting period for auditory thalamic axons. Subsequent DiI labeling revealed that in these animals the majority of MGN axons had grown past auditory cortex instead of innervating it. Taken together these observations underscore a general requirement for subplate neurons throughout neocortex in the process of cortical target selection and ingrowth by thalamic axons. Moreover, they imply that the cortical plate alone has insufficient information to direct the formation of specific sets of thalamocortical connections: interactions involving subplate neurons must also be present.
A unique pattern of expression of the four muscle regulatory factor proteins distinguishes somitic from embryonic, fetal and newborn mouse myogenic cells
Authors: Smith Timothy H. , Block, Nancy E. , Rhodes, Simon J. , Miller Jeffrey Boone
Published: April 2021

A unique pattern of expression of the four muscle regulatory factor (MRF) proteins was found to distinguish early somitic from embryonic, fetal and newborn limb myogenic cells in vitro. Expression of the myosin heavy chain (MHC), MyoD, myogenin, Myf-5, and MRF4 proteins was examined by immunocytochemistry in cultures of four distinct types of mouse myogenic cells: somitic (E8.5), embryonic (E11.5), fetal (E16.5) and newborn limb. In embryonic, fetal and newborn cultures, the MRF proteins were expressed in generally similar patterns: MyoD was the first MRF expressed; MyoD and myogenin were expressed by more cells than Myf-5 or MRF4; and each of the four MRFs was found both in cells that expressed MHC and in cells that did not express MHC. In cultures of somitic cells, in contrast, Myf-5 was expressed first and by more cells than MyoD or myogenin; MRF4 was not detected; and the MRFs were never found to be coexpressed with MHC in the same cell. Thus, some somitic cells had the unexpected ability to maintain MHC expression in the absence of detectable MRF protein expression. The different myogenic programs of embryonic, fetal and newborn myogenic cells are not, therefore, a simple result of qualitatively different MRF expression patterns, whereas myogenesis by somitic cells does include a unique pattern of MRF expression.
Tight junction protein cingulin is expressed by maternal and embryonic genomes during early mouse development
Authors: Javed Qamar , Fleming Tom P. , Hay Mark , Citi Sandra
Published: April 2021

The expression of the tight junction peripheral membrane protein, cingulin (140 103Mr), was investigated in mouse eggs and staged preimplantation embryos by immunoblotting and immunoprecipitation. Polyclonal antibody to chicken brush border cingulin detected a single 140 103Mr protein in immunoblots of unfertilised eggs and all preimplantation stages. Relative protein levels were high in eggs and early cleavage stages, declined during later cleavage and increased again in expanding blastocysts. Quantitative immunoprecipitation of metabolically labelled eggs and staged embryos also revealed a biphasic pattern for cingulin synthesis with relative net levels being high in unfertilised eggs, minimal during early cleavage, rising 2.3-fold specifically at the onset of compaction (8-cell stage, when tight junction formation begins), and increasing further at a linear rate during morula and blastocyst stages. Cingulin synthesis in eggs is not influenced by fertilisation (or aging, if unfertilised), but this level declines sharply after first cleavage. These results indicate that cingulin is expressed by both maternal and embryonic genomes. The turnover of maternal cingulin (unfertilised eggs) and embryonic cingulin at a stage before tight junction formation begins (4-cell stage) is higher (t ∼4 hours) than cingulin synthesised after tight junction formation (blastocysts; t ∼10 hours). This increase in cingulin stability is reversed in the absence of extracellular calcium. Cingulin synthesis is also tissue-specific in blastocysts, being up-regulated in trophectoderm and down-regulated in the inner cell mass. Taken together, the results suggest that (i) cingulin may have a role during oogenesis and (ii) cell-cell contact patterns regulate cingulin biosynthesis during early morphogenesis, contributing to lineage-specific epithelial maturation.
Developmental role of endogenous retinoids in the determination of morphallactic field in budding tunicates
Authors: Kawamura Kazuo , Hara Kenji , Fujiwara Shigeki
Published: April 2021

We have extracted retinoids from the budding tunicate Polyandrocarpa misakiensis and, using HPLC, identified some major peaks as cis-retinal, all-trans-retinal and all-trans-retinoic acid, of which cis-retinal was most abundant (∼2 M). In developing buds, the amount of cis-retinal was about one-fifth that of the adult animals. In those buds, aldehyde dehydrogenase, which could metabolize retinal in vitro, was expressed in epithelial cells and then in mesenchymal cells at the proximal extremity, that is, the future developmental field of the bud. Exogenous retinoic acid comparable to the endogenous level could induce an additional field at the distal end of the bud, resulting in a double monster. The induction always accompanied an ectopic expression of aldehyde dehydrogenase. The results of this work suggest that retinoic acid or related molecule(s) act as an endogenous trigger of morphallactic development of Polyandrocarpa buds.
Cell lineage analysis of the expression of an engrailed homolog in leech embryos
Authors: Lans Deborah , Wedeen Cathy J. , Weisblat David A.
Published: April 2021

ht-en is an engrailed-class gene that is expressed during early development and neurogenesis in embryos of the leech Helobdella triserialis. During the early development of this annelid (stages 7-9), ht-en is expressed in each of the ectodermal and mesodermal teloblast lineages that contributes progeny to the definitive segments. ht-en is expressed transiently by individually identified cells within the segmentally iterated primary blast cell clones. Its expression is correlated with the age of the primary blast cell clone. After consegmental primary blast cell clones from the different teloblast lineages have come into segmental register, cells that express ht-en during stages 7-9 are clearly confined to a transverse region corresponding to the posterior portion of the segmental anlage, but not all cells within this region express ht-en. Only a minority of the identified cells that express ht-en during terminal differentiation of the segmental ganglia and body wall (stages 10-11) are descendants of cells that express ht-en in early development (stages 7-9).
Characterization of the unusually rapid cell cycles during rat gastrulation
Authors: Auley Alasdair Mac , Werb Zena , Mirkes Philip E.
Published: April 2021

The onset of gastrulation in rodents is associated with the start of differentiation within the embryo proper and a dramatic increase in the rate of growth and proliferation. We have determined the duration of the cell cycle for mesodermal and ectodermal cells of rat embryos during gastrulation (days 8.5 to 9.5 of gestation) using a stathmokinetic analysis. These embryonic cells are the most rapidly dividing mammalian cells yet described. Most cells of the ectoderm and mesoderm had a cell cycle time of 7 to 7.5 hours, but the cells of the primitive streak divided every 3 to 3.5 hours. Total cell cycle time was reduced by shortening S and G2, as well as G1, in contrast to cells later in development, when cell cycle duration is modulated largely by varying the length of G1. In the ectoderm and mesoderm, G1 was 1.5 to 2 hours, S was 3.5 to 4 hours, and G2 was 30 to 40 minutes. G1, S and G2 were shortened even further in the cells of the primitive streak: G1 was less than 30 minutes, S was 2 to 2.75 hours, and G2 was less than 20 minutes. Thus, progress of cells through all phases of the cell cycle is extensively modified during rodent embryogenesis. Specifically, the increased growth rate during gastrulation is associated with radical changes in cell cycle structure and duration. Further, the commitment of cells to become mesoderm and endoderm by entering the primitive streak is associated with expression of a very short cell cycle during transit of the primitive streak, such that developmental decisions determining germ layer fate are reflected in differences in cell cycle regulation.
Gene expression during imaginal disc regeneration detected using enhancer-sensitive P-elements
Authors: Brook William J. , Ostafichuk Lisa M. , Piorecky Jana , Wilkinson Mark D. , Hodgetts Daralyn J. , Russell Michael A.
Published: April 2021

When imaginal disc fragments from Drosophila are cultured in adult female hosts, they either duplicate the part of the pattern specified by the fate map, or regenerate to replace the missing part. The new tissue is added by proliferation of a small number of cells from the cut edge, brought together when the wound heals to form a regeneration blastema. Specification of the new pattern has been explained by assuming interactions among cells of different positional value in the regeneration blastema. In order to identify genes which might mediate these events, we screened over eight hundred independently isolated autosomal insertions of an enhancer-sensitive P-element, for altered lac-z expression in regenerating discs following cell death induced by a temperature-sensitive cell-lethal mutation. Two further screens divided the positive lines into four groups based on appropriate timing of the lac-z response in the cell-lethal mutant background and the expected response to an alternate source of cell death. Expression in wing disc fragments cultured in vivo was most frequent in the target class defined by the screens. In this direct test, lac-z expression was found in 23 lines and in most cases was spatially and temporally correlated with the formation of the regeneration blastema. Our results suggest a very substantial transcriptional response during the early stages of imaginal disc regeneration. lac-z expression in control imaginal discs, embryos and adult ovaries of the positive lines was also assayed. The selected insertions included: a small class expressed only in discs undergoing regeneration and apparently not at any other stage, possibly representing genes active exclusively in regeneration; a larger class expressed in the embryo or during oogenesis, but not normally in imaginal discs, as expected for functions recruited from earlier stages of the developmental program; and finally a class with spatially patterned expression in normal discs. This class included several insertions with expression associated with compartment boundaries, including one at the decapentaplegic (dpp), and one at the crumbs (crb) locus, a growth factor homologue, and an EGF-repeat gene respectively. Some of the expression patterns observed in cultured disc fragments provide evidence for cell communication in the regeneration blastema.
The cell adhesion molecule M-cadherin is specifically expressed in developing and regenerating, but not denervated skeletal muscle
Authors: Moore Robert , Walsh Frank S.
Published: April 2021

The spatiotemporal distribution of M-cadherin mRNA has been determined by in situ hybridization in the mouse embryo and in adult skeletal muscle following experimental regeneration and denervation. M-cadherin mRNA is highly tissue specific and is found only in developing skeletal muscle. In contrast, N-cadherin mRNA has a broader tissue distribution in the embryo, being found on both neural elements and skeletal and cardiac muscle. M-cadherin is expressed in the myotomes shortly after they form, along with the myo-genic regulatory factor myogenin. M-cadherin is expressed in muscles derived from the myotomes and is detected in forelimb bud precursor cells at embryonic day 11.5. In the latter case M-cadherin expression appears co-ordinately with that of myogenin and cardiac-actin. Shortly before birth, M-cadherin expres-sion is down regulated. M-cadherin can, however, be re-expressed following experimental regeneration of skele-tal muscle. Here M-cadherin is transiently expressed on regenerating myoblasts but not myotubes. Following muscle denervation no evidence was found for re-expression of M-cadherin under conditions where there was strong expression of the nicotinic acetylcholine receptor on myofibres.

The highly specific tissue distribution and unique developmental profile distinguishes M-cadherin from other cadherins and suggests a role in cell surface events during early myogenesis.
Multiple extracellular signals are required for long-term oligodendrocyte survival
Authors: Barres B. A. , Schmid R. , Sendnter M. , Raff M. C.
Published: April 2021

We showed previously that oligodendrocytes and their precursors require continuous signalling by protein trophic factors to avoid programmed cell death in culture. Here we show that three classes of such trophic factors promote oligodendrocyte survival in vitro: (1) insulin and insulin-like growth factors (IGFs), (2) neurotrophins, particularly neurotrophin-3 (NT-3), and (3) ciliary-neurotrophic factor (CNTF), leukemia inhibitory factor (LIF) and interleukin 6 (IL-6). A single factor, or combinations of factors within the same class, promote only short-term survival of oligodendrocytes and their precursors, while combinations of factors from different classes promote survival additively. Long-term survival of oligodendrocytes in vitro requires at least one factor from each class, suggesting that multiple signals may be required for long-term oligodendrocyte survival in vivo. We also show that CNTF promotes oligodendrocyte survival in vivo, that platelet-derived growth factor (PDGF) can promote the survival of oligodendrocyte precursors in vitro by acting on a novel, very high affinity PDGF receptor, and that, in addition to its effect on survival, NT-3 is a potent mitogen for oligodendrocyte precursor cells.
The spatial organization of epidermal structures: hairy establishes the geometrical pattern of Drosophila leg bristles by delimiting the domains of achaete expression
Authors: Orenic Teresa V. , Held Lewis I. , Paddock Stephen W. , Carroll Sean B.
Published: April 2021

The spatial organization of Drosophila melanogaster epidermal structures in embryos and adults constitutes a classic model system for understanding how the two dimensional arrangement of particular cell types is generated. For example, the legs of the Drosophila melanogaster adult are covered with bristles, which in most segments are arranged in longitudinal rows. Here we elucidate the key roles of two regulatory genes, hairy and achaete, in setting up this periodic bristle pattern. We show that achaete is expressed during pupal leg development in a dynamic pattern which changes, by approximately 6 hours after puparium formation, into narrow longitudinal stripes of 3–4 cells in width, each of which represents a field of cells (proneural field) from which bristle precursor cells are selected. This pattern of gene expression foreshadows the adult bristle pattern and is established in part through the function of the hairy gene, which also functions in patterning other adult sense organs. In pupal legs, hairy is expressed in four longitudinal stripes, located between every other pair of achaete stripes. We show that in the absence of hairy function achaete expression expands into the interstripe regions that normally express hairy, fusing the two achaete stripes and resulting in extra-wide stripes of achaete expression. This misexpression of achaete, in turn, alters the fields of potential bristle precursor cells which leads to the misalignment of bristle rows in the adult. This function of hairy in patterning achaete expression is distinct from that in the wing in which hairy suppresses late expression of achaete but has no effect on the initial patterning of achaete expression. Thus, the leg bristle pattern is apparently regulated at two levels: a global regulation of the hairy and achaete expression patterns which partitions the leg epidermis into striped zones (this study) and a local regulation (inferred from other studies on the selection of neural precursor cells) that involves refinement steps which may control the alignment and spacing of bristle cells within these zones.
Multiple cell interactions are required for fate specification during male spicule development in Caenorhabditis elegans
Authors: Chamberlin Helen M. , Sternberg Paul W.
Published: April 2021

The B blast cell divides postembryonically in C. elegans males to produce 47 progeny that include all of the cells of the copulatory spicules. During the early development of the B lineage, the anterior daughter of B, B.a, generates eight cells. These cells migrate to form four pairs of cells that flank the developing cloaca (ventral, dorsal, and two identical lateral pairs). For each pair, the more anterior cell produces a distinct lineage (‘anterior fate’) from the posterior cell (‘posterior fate’). For the ventral and dorsal pairs, either cell can migrate to the anterior position and produce the anterior lineage, and the other cell migrates posterior and produces the posterior lineage (Sulston and Horvitz, 1977, Dev. Biol. 56, 110–156). The migration is variable, although the resultant fate pattern is invariant. In the two lateral pairs, both the migration and fate pattern are invariant. Using a laser microbeam to selectively ablate neighboring cells we have found that the cells of the lateral pair also respond to positional cues. For all four pairs other male-specific blast cells provide extracellular cues. In general, F and U promote anterior fates, Y promotes some posterior fates, and the B.a progeny promote posterior fates. Several of these cues are redundant. By ablating combinations of cells we have deduced how these signals may act in concert to specify the fates of the B.a progeny. We propose that fate specification in these pairs depends on three general classes of extracellular cues: positional cues, modulators of positional cues, and lateral signals. The B lineage thus provides an opportunity to study with single cell resolution the integration of multiple intercellular signals.
Genetic analysis of laminin A reveals diverse functions during morphogenesis in Drosophila
Authors: Henchcliffe Claire , García-Alonso Luis , Tang Joyce , Goodman Corey S.
Published: April 2021

In order to dissect the functions of laminin A in vivo, we have undertaken a molecular and genetic characterization of the laminin A subunit (lamA) gene in Drosophila. Sequence analysis predicts a multidomain structure similar to mammalian homologs. We generated a series of complete and partial loss-of-function mutant alleles of the lamA gene; complete loss-of-function mutations lead to late embryonic lethality. Certain combinations of partial loss-of-function lamA alleles give rise to escaper adults, which have rough eyes associated with changes in cell fate and pattern, misshapen legs and defects in wing structure. These phenotypes suggest that laminin A has diverse functions during morphogenesis in Drosophila.
Ectopic expression and function of the Antp and Scr homeotic genes: the N terminus of the homeodomain is critical to functional specificity
Authors: Zeng Wenlin , Andrew Deborah J. , Mathies Laura D. , Horner Michael A. , Scott Matthew P.
Published: April 2021

The transcription factors encoded by homeotic genes determine cell fates during development. Each homeotic protein causes cells to follow a distinct pathway, presumably by differentially regulating downstream ‘target’ genes. The homeodomain, the DNA-binding part of homeotic proteins, is necessary for conferring the specificity of each homeotic protein’s action. The two Drosophila homeotic proteins encoded by Antennapedia and Sex combs reduced determine cell fates in the epidermis and internal tissues of the posterior head and thorax. Genes encoding chimeric Antp/Scr proteins were introduced into flies and their effects on morphology and target gene regulation observed. We find that the N terminus of the homeodomain is critical for determining the specific effects of these homeotic proteins in vivo, but other parts of the proteins have some influence as well. The N-terminal part of the homeodomain has been observed, in crystal structures and in NMR studies in solution, to contact the minor groove of the DNA. The different effects of Antennapedia and Sex combs reduced proteins in vivo may depend on differences in DNA binding, protein-protein interactions, or both.
Function and spatial distribution in developing chick retina of the laminin receptor α6β1 and its isoforms
Authors: Curtis Ivan de , Reichardt Louis F.
Published: April 2021

We have recently shown that the laminin-binding integrin receptor, α 6 β 1, is prominently expressed in the developing chick retina, and its expression and activity are regulated during development on both retinal ganglion cells and other neural retinal cells. In the present study, we show that antibodies specific for the extracellular portion of the chick α6 subunit dramatically inhibit interactions in vitro between embryonic day 6 neural retinal cells and laminin, showing that α 6 β 1 functions as an important laminin receptor on developing retinal neurons. In previous work, we showed that α6 mRNA levels on retinal ganglion cells decrease dramatically after E6 during the period that RGC axons innervate the optic tectum. In the present study, we show decreases in α6 mRNA are not prevented by ablation of the optic tectum, indicating that tectal contact is not the major cause of this decrease. Within the embryonic retina, the α 6 subunit is codistributed, in part, with laminin, suggesting that it functions as a laminin receptor during retina development in vivo. Furthermore, two isoforms of the α 6 protein with distinct cytoplasmic domains generated by differential splicing have quite different distribution patterns in the retina, suggesting that these two isoforms may have different functions during retinal development.
Zebrafish primary neurons initiate expression of the LIM homeodomain protein Isl-1 at the end of gastrulation
Authors: Korzh V. , Edlund T. , Thor S.
Published: April 2021

Isl-1 has previously been established as the earliest marker of developing chicken spinal motor neurons where it is regulated by inductive signals from the floorplate and notochord. We now report that, in zebrafish, the expression of Isl-1 is initiated in Rohon-Beard cells, primary motor neurons, interneurons and cranial ganglia, hours before the neural tube itself is formed. The expression is initiated simultaneously in the RohonBeard cells and the primary motor neurons, at the axial level of the presumptive first somite. The Isl-1-expressing motor neurons appear on either side of the ventral midline whereas the interneurons and Rohon-Beard cells initiate expression while located at the edge of the germinal shield. Isl-1 expression is initiated in these cells before the formation of a differentiated notochord.

Isl-1 is expressed in the various functional classes of primary neurons at 24 hours postfertilization. This selective expression of a homeodomain protein in the primary neurons implies that these neurons share a common program of early development and that they have evolved and been selected for as a coordinated system. One of the functions of the primary neurons is to send long axons which pioneer the major axon tracts in the zebrafish embryo. An evolutionary conserved functional role for Isl-1 in the expression of the pioneering phenotype of the primary neurons is suggested.
Selective dispersal of avian rhombomere cells in orthotopic and heterotopic grafts
Authors: Guthrie Sarah , Prince Victoria , Lumsden Andrew
Published: April 2021

During hindbrain development, cells become segregated into segmental groups, rhombomeres, by mechanisms that are presently unknown. One contributory factor early in development may be an alternating periodicity in cell surface properties down the neuraxis. This possibility was previously suggested by experiments in which tissue from different segmental levels was apposed in the absence of a boundary. New boundaries were regenerated only when rhombomeres from adjacent positions or positions three rhombomeres distant from one another were apposed. Combinations of two odd-numbered or two even-numbered rhombomeres usually failed to generate a boundary. In order to pursue this phenomenon to the cellular level, we have used two approaches, both involving donor-to-host transplantation. First, quail rhombomeres were grafted at various hindbrain levels of a chick host. Apposition of rhombomere 4 (r4) with r3 was concomitant with negligible cell mixing across the interface. By contrast, combinations of r3 with r5 or with r3 tissue led to cell mixing that was more extensive in combinations of identical rhombomeres (r3 with r3) than between two alternate ones (r3 with r5). Secondly, we grafted small pieces of fluorescently prelabelled chick rhombomere tissue at various hindbrain levels of chick hosts. In most cases, cells dispersed widely when transplanted orthopically or two segments distant from that of their origin. Cells transplanted into an adjacent segment, however, showed a tendency to remain undispersed. Among the different graft combinations, furthermore, there was a variation in the extent of dispersal that showed an additional level of complexity not revealed in boundary regeneration experiments. The possibility is raised that the early partitioning of rhombomeres involves a hierarchy in the adhesive preferences of cell-cell interactions along the neuraxis.
Catenins in Xenopus embryogenesis and their relation to the cadherin-mediated cell-cell adhesion system
Authors: Schneider Stephan , Herrenknecht Kurt , Butz Stefan , Kemler Rolf , Hausen Peter
Published: April 2021

In the course of an analysis of cell-cell adhesion in the Xenopus embryo, antibodies directed against α and β catenin were applied to investigate their relation to the cadherins occurring early in this system. The results demonstrate that α and β-catenin are provided maternally and increase in amount throughout embryogenesis. Immunoprecipitations indicate that both of the catenins are complexed to U-cadherin in the early phase of embryogenesis and to β-cadherin, when it appears during gastrulation. An excess of α-catenin occurs in free form in the early embryo, whereas all of the catenin seems to be complexed to cadherin. Synthesis of the two components throughout early embryogenesis and their binding to newly synthesized cadherins were demonstrated by metabolic labelling.

The spatial distribution of α-catenin was analysed by immunohistology. During cleavage β-catenin is deposited evenly along the plasma membranes within the embryo, while the cell peripheries at the surface of the embryo remain devoid of α-catenin. At later stages, the pattern of α-catenin distribution becomes more complex. Quantitative differences in the intensity of staining along the plasma membranes in the different regions of the embryo can be distinguished. Particularly the appearance of β-cadherin in the gastrula ectoderm is accompanied by conspicous depositions of α-catenin along the respective plasma membranes in this layer. All cells in the later embryo, apart from the neural crest cells, carry α-catenin on their plasma membranes indicating the universal character of cadherin-mediated cell-cell adhesion in the Xenopus embryo.
Later embryogenesis: regulatory circuitry in morphogenetic fields
Authors: Davidson Eric H.
Published: April 2021

The subject of this review is the nature of regulatory processes underlying the spatial subdivision of morphogenetic regions in later embryogenesis. I have applied a non-classical definition of morphogenetic field, the progenitor field, which is a region of an embryo composed of cells whose progeny will constitute a given morphological structure. An important feature of such fields is that they have sharp spatial boundaries, across which lie cells whose progeny will express different fates. Two examples of the embryonic specification and development of such fields are considered. These are the formation of the archenteron in the sea urchin embryo and the formation of dorsal axial mesoderm in the Xenopus embryo. From these and a number of additional examples, from vertebrate, Drosophila, Caenorhabditis elegans and sea urchin embryos, it is concluded that the initial formation of the boundaries of morphogenetic progenitor fields depends on both positive and negative transcription control functions. Specification of morphogenetic progenitor fields, organization of the boundaries and their subsequent regionalization or subdivision are mediated by intercellular signaling. Genes encoding regionally expressed transcription factors that are activated in response to intercell signaling, and that in turn mediate signaling changes downstream, appear as fundamental regulatory circuit elements. Such [signal →transcription factor gene → signal] circuit elements appear to be utilized, often repetitively, in many different morphogenetic processes.
The gene tinman is required for specification of the heart and visceral muscles in Drosophila
Authors: Bodmer Rolf
Published: April 2021

The homeobox-containing gene tinman (msh-2, Bodmer et al., 1990Development 110, 661-669) is expressed in the mesoderm primordium, and this expression requires the function of the mesoderm determinant twist. Later in development, as the first mesodermal subdivisions are occurring, expression becomes limited to the visceral mesoderm and the heart. Here, I show that the function of tinman is required for visceral muscle and heart development. Embryos that are mutant for the tinman gene lack the appearance of visceral mesoderm and of heart primordia, and the fusion of the anterior and posterior endoderm is impaired. Even though tinman mutant embryos do not have a heart or visceral muscles, many of the somatic body wall muscles appear to develop although abnormally. When the tinman cDNA is ubiquitously expressed in tinman mutant embryos, via a heatshock promoter, formation of heart cells and visceral mesoderm is partially restored. tinman seems to be one of the earliest genes required for heart development and the first gene reported for which a crucial function in the early mesodermal subdivisions has been implicated.
Rescue of the T-associated maternal effect in mice carrying null mutations in Igf-2 and Igf2r, two reciprocally imprinted genes
Authors: Filson Adele J. , Louvi Angeliki , Efstratiadis Argiris , Robertson Elizabeth J.
Published: April 2021

In mice, only the paternal allele of the Igf2 gene, encoding insulin-like growth factor II (IGF-II) is expressed due to parental imprinting. Interestingly, the Igf2r gene, which encodes one of the two known receptors (IGF2R) to which IGF-II binds with high affinity is also subject to imprinting, but in a reciprocal fashion. This observation raises the possibility that imprinting of these loci serves to regulate the ratios of the gene products, since IGF2R provides a mechanism for IGF-II turnover. To test this hypothesis, we crossed mice mutant for Igf-2 with animals carrying the Thp chromosomal deletion, which encompasses the Igf2r locus. Inheritance of the Fp chromosome through the maternal germline results in a dominant lethal maternal effect (Tme). However, as we show here, Thp/+ embryos that inherit the Thp maternally are variably rescued to birth if they also lack IGF-II. Based on these data, the Tme phenotype can be viewed as a dominant effect resulting from an overabundance of IGF-II.
A new class of mutations in Arabidopsis thaliana, acaulis1, affecting the development of both inflorescences and leaves
Authors: Tsukaya Hirokazu , Naito Satoshi , Rédei George P. , Komeda Yoshibumi
Published: April 2021

We isolated and analyzed mutants of Arabidopsis thaliana, acaulis, with flower stalks that are almost absent or are much reduced in length. The mutations are divided between two loci, acaulisl (acll) and acaulis2 (acl2). The acll-1 mutation has been assigned to linkage group 4 in the vicinity of locus ap2. The acl1-1 mutant showed premature arrest of the inflorescence meristem after the onset of reproductive development, followed by consequent reduction in the number of flower-bearing phytomers and therefore flowers. The apical meristem of the inflorescences was morphologically normal but its radius was about half that of the wild type. The acl1 mutants are also defective in the development of foliage leaves. Both defects could be rescued by growth at a specific temperature (28°C). The length of the cells in acl1-3 mutant was less than that in the wild type but the numbers of cells in leaves and internodes of acl1 mutants were calculated to be the same as those of the wild type. Thus, the defects in inflorescences and leaves were attributed to defects in the process of elongation (maturation) of these cells. Temperature-shift experiments showed that the Acl1+ product was necessary at all developmental stages. A critical stage was shown to exist for recovery from the cessation of development of inflorescence meristems that was caused by the acl1-1 mutation. Grafting experiments showed that the acl1-1 mutation does not affect diffusible substances. An analysis of double mutants carrying both acl1-1 and one of developmental mutations, ap1, clv1, Ify, or tfl1, showed that ACL1 is a new class of gene.
The expression of the regulatory myosin light chain 2 gene during mouse embryogenesis
Authors: Faerman Alexander , Shani Moshe
Published: April 2021

The fast skeletal muscle myosin light chain 2 (MLC2) gene is expressed specifically in skeletal muscles of new born and adult mice, and has no detectable sequence homology with any of the other MLC genes including the slow cardiac MLC2 gene. The expression of the fast skeletal muscle MLC2 gene during early mouse embryogenesis was studied by in situ hybridization. Serial sections of embryos from 8.5 to 12.5 days post coitum (d.p.c.) were hybridized to MLC2 cRNA and to probes for the myogenic regulatory genes MyoD1 and myogenin. The results revealed different temporal and spatial patterns of hybridization for different muscle groups. MLC2 transcripts were first detected 9.5 d.p.c. in the myotomal regions of rostral somites, already expressing myogenin. Surprisingly, at the same stage, a weak MLC2 signal was also detected in the cardiomyocytes. The cardiac expression was transient and could not be detected at later stages while the myotomal signal persisted and spread to the more caudal somites, very similar to the expression of myogenin.

Beginning from 10.5 d.p.c., several extramyotomal premuscle cells masses have been demarcated by MyoD1 expression. MLC2 transcripts were detected in only one of these cell masses. Although, transcripts of myogenin were detected in all these cell masses, the number of expressing cells was significantly lower than that observed for MyoD1. By 11.5 d.p.c., all three hybridization signals colocalized in most extramyotomal muscle forming regions, with the exception of the diaphragm and the hindlimb buds, where only few cells expressed MLC2 and more cells expressed MyoD1 than myogenin. At 12.5 d.p.c., all three studied genes displayed a similar spatial pattern of expression in most muscle-forming regions. However, in some muscles, the MyoD1 signal spread over more cells compared to myogenin or MLC2. Our results are consistent with the suggestion that multiple myogenic programs exist for myoblasts differentiating in the myotome and extramyotomal regions.
The Drosophila Broad-Complex plays a key role in controlling ecdysone-regulated gene expression at the onset of metamorphosis
Authors: Karim Felix D. , Guild Gregory M. , Thummel Carl S.
Published: April 2021

During Drosophila third instar larval development, one or more pulses of the steroid hormone ecdysone activate three temporally distinct sets of genes in the salivary glands, represented by puffs in the polytene chromosomes. The intermolt genes are induced first, in midthird instar larvae; these genes encode a protein glue used by the animal to adhere itself to a solid substrate for metamorphosis. The intermolt genes are repressed at puparium formation as a high titer ecdysone pulse directly induces a small set of early regulatory genes. The early genes both repress their own expression and activate more than 100 late secondary-response genes. The Broad-Complex (BR-C) is an early ecdysone-inducible gene that encodes a family of DNA binding proteins defined by at least three lethal complementation groups: br, rbp, and l(1)2Bc. We have found that the BR-C is critical for the appropriate regulation of all three classes of ecdysone-inducible genes. Both rbp and l(1)2Bc are required for glue gene induction in mid-third instar larvae. In addition, the l(1)2Bc function is required for glue gene repression in prepupae; in l(1)2Bc mutants the glue genes are re-induced by the late prepupal ecdysone pulse, recapitulating a mid-third instar regulatory response at an inappropriate stage in development. The l(1)2Bc function is also required for the complete ecdysone induction of some early mRNAs (E74A, E75A, and BR-C) and efficient repression of most early mRNAs in prepupae. Like the intermolt secondary-response genes, the late secondary-response genes are absolutely dependent on rbp for their induction. An effect of l(1)2Bc mutations on late gene activity can also be detected, but is most likely a secondary consequence of the submaximal ecdysone-induction of a subset of early regulatory products. Our results indicate that the BR-C plays a key role in dictating the stage-specificity of the ecdysone response. In addition, the ecdysone-receptor protein complex alone is not sufficient for appropriate induction of the early primary-response genes, but requires the prior expression of BR-C proteins. These studies define the BR-C as a key regulator of gene activity at the onset of metamorphosis in Drosophila.
Cell lineage of larval and imaginal thoracic anlagen cells of Drosophila melanogaster, as revealed by single-cell transplantations
Authors: Meise Martin , Janning Wilfried
Published: April 2021

We have analyzed the cell lineage of larval and imaginal cells in the thoracic ectoderm of the early embryo of Drosophila melanogaster, by homotopic transplantation of single cells in the region of 50 –60% egg length. Single cells were isolated prior to transplantation in an in vitro solution. The donors were ‘enhancer-trap’ lines in which the nuclei of all larval and imaginal cells exhibit a uniformly intense expression of the lacZ gene of E. coli. The transplantations were carried out from the blastoderm to the early gastrula stage, as a rule immediately after the onset of gastrulation (stage 6). It was found that at this time the cells of the thoracic ectoderm are not yet committed to form larval or imaginal structures, as indicated by the presence of clones overlapping all structures formed by the thoracic ectoderm, i.e. the nervous system, the larval epidermis, the tracheae and the imaginal discs. The average size of pure epidermal clones was five cells. In clones overlapping either other larval tissues or imaginal discs, the average number of epidermal cells was between three and four. The mean relative clone size was 1/5 of the size of the total structure for leg imaginal discs and 1/7 for the wing imaginal disc. We therefore infer that the precursors for the leg discs and wing disc on one side together number 22 cells in the blastoderm or early gastrula stage. These cells eventually give rise not only to precursors of the imaginal discs but usually also to larval epidermal and nervous-system cells, because most of the imaginal disc clones (80%) overlap larval tissue.

The transplantations were not precisely homotopic; the fact that up to 10 cells were removed from the donor essentially rules out exact homotopy between donor and host sites, because a segment anlage is only about three cells wide. Nevertheless, the clones developed completely normal tissue together with the recipient cells. Although the clones have the capacity to extend over different ectodermal tissues and can include both imaginal discs in a given segment, no clones were found that clearly crossed larval or imaginal segment boundaries.

We propose a model in which the segregation of the cells that are to differentiate into the imaginal tissues does not occur until the second postblastodermal mitosis
Ectopic expression of seven-up causes cell fate changes during ommatidial assembly
Authors: Hiromi Yasushi , Mlodzik Marek , West Steven R. , Rubin Gerald M. , Goodman Corey S.
Published: April 2021

During Drosophila ommatidial development, a single cell is selected within the ommatidial cluster to become the R7 photoreceptor neuron. The seven-up gene has been shown to play a role in this process by preventing four other photoreceptor precursors, R3/R4/R1/R6, from adopting the R7 cell fate. The seven-up gene encodes a steroid receptor-like molecule that is expressed only in those four cells that require seven-up function in the developing Drosophila ommatidium. We have examined the functional significance of the spatially restricted expression of seven-up by misexpressing seven-up isoforms. As expected from the function that seven-up performs in R3/R4/R1/R6, ubiquitous expression of seven-up causes transformation of the R7 cell to an R1-R6 cell fate. In addition, depending on the timing and spatial pattern of expression, various other phenotypes are produced including the loss of the R7 cell and the formation of extra R7 cells. Ubiquitous expression of seven-up close to the morphogenetic furrow interferes with R8 differentiation resulting in failure to express the boss protein, the ligand for the sevenless receptor tyrosine kinase, and the R7 cell is lost consequently. Extra R7 cells are formed by recruiting non-neuronal cone cells as photoreceptor neurons in a sevenless and bride of sevenless independent way. Thus, the spatiotemporal pattern of seven-up expression plays an essential role in controlling the number and cellular origin of the R7 neuron in the ommatidium. Our results also suggest that seven-up controls decisions not only between photoreceptor subtypes, but also between neuronal and non-neuronal fates.