bluuebunny/crossref_metadata_2025 huggingface dataset:
Msx1 (Hox-7.1) in the adult mouse uterus: cellular interactions underlying regulation of expression
Authors: Pavlova Anna , Boutin Eugenie , Cunha Gerald , Sassoon David
Published: April 2021

We report here that Msx1 (formerly Hox-7.1) is expressed at high levels in uterine epithelial cells of the non-pregnant adult. These cells undergo pronounced changes in morphology in response to embryo implantation and show a concomitant decrease in Msx1 levels. While Msx1 is restricted to the uterus in adulthood, we observe Msx1 expression throughout the entire perinatal Müllerian duct epithelium in the prospective uterus, cervix and vagina. Through analysis of tissue recombinants, the expression of Msx1 in the epithelium was shown to be dependent upon an interaction with the underlying mesenchyme of uterine origin. The capacity of uterine mesenchyme to support or induce Msx1 expression in Müllerian epithelium is correlated with mesenchymal expression of Wnt-5a. Whereas Msx1 expression in the epithelium results from interaction with uterine mesenchyme, Wnt-5a expression is an intrinsic property of the uterine mesenchyme and does not depend upon the epithelium. The observation that Msx1 is expressed in the adult uterine epithelium and that conversion of the presumptive vaginal epithelium to uterine epithelium can be elicited only during the first week of postnatal development when Msx1 expression is detected suggests that, in addition to regulating various aspects of uterine epithelial morphology and function (e.g. gestation), this homeobox-containing gene plays a role in maintaining the uterus in a morphogenic and developmentally responsive state prerequisite for its unique function.
The daughterless gene functions together with Notch and Delta in the control of ovarian follicle development in Drosophila
Authors: Cummings Craig A. , Cronmiller Claire
Published: April 2021

The daughterless (da) gene in Drosophila encodes a broadly expressed transcriptional regulator whose specific functions in the control of sex determination and neurogenesis have been extensively examined. We describe here a third major developmental role for this regulatory gene: follicle formation during oogenesis. A survey of da RNA and protein distribution during oogenesis reveals a multiphasic expression pattern that includes both germline and soma. Whereas the germline expression reflects da’s role in progeny sex determination, the somatic ovary expression of da correlates with the gene’s role during egg chamber morphogenesis. Severe, but viable, hypomorphic da mutant genotypes exhibit dramatic defects during oogenesis, including aberrantly defined follicles and loss of interfollicular stalks. The follicular defects observed in da mutant ovaries are qualitatively very similar to those described in Notch (N) or Delta (Dl) mutant ovaries. Moreover, in the ovary da− alleles exhibit dominant synergistic interactions with N or Dl mutations. We propose that all three of these genes function in the same regulatory pathway to control follicle formation.
no tail (ntl) is the zebrafish homologue of the mouse T (Brachyury) gene
Authors: Schulte-Merker S. , Eeden F. J. M. van , Halpern M. E. , Kimmel C. B. , Nüsslein-Volhard C.
Published: April 2021

The mouse T (Brachyury) gene is required for normal mesoderm development and the extension of the body axis. Recently, two mutant alleles of a zebrafish gene, no tail (ntl), have been isolated (Halpern, M. E., Ho., R. K., Walker, C. and Kimmel, C. B. (1993) Cell 75, 99-111). ntl mutant embryos resemble mouse T/T mutant embryos in that they lack a differentiated notochord and the caudal region of their bodies. We report here that this phenotype is caused by mutation of the zebrafish homologue of the T gene. While ntl embryos express mutant mRNA, they show no nuclear protein product. Later, expression of mRNA in mutants, but not in wild types, is greatly reduced along the dorsal midline where the notochord normally forms. This suggests that the protein is required for maintaining transcription of its own gene.
Profilin mutations disrupt multiple actin-dependent processes during Drosophila development
Authors: Verheyen Esther M. , Cooley Lynn
Published: April 2021

The chickadee gene of Drosophila encodes profilin, a small actin binding protein. We present the first analysis of the effects of profilin deletion in a multicellular organism. Genomic deletions of the chickadee locus result in a late embryonic lethal phenotype indicating that profilin is essential in flies. In addition, viable alleles of chickadee with defects in oogenesis, spermatogenesis and bristle formation provide insight into profilin function in a variety of cell types. Defects in oogenesis include the previously described failure to assemble nurse cell actin filament bundles in addition to abnormal regulation of mitosis, binucleate cells and stalled cell migration. Malformed bristles are a result of aberrant actin assembly. Monoclonal antibodies against Drosophila profilin were generated to study profilin’s cellular and subcellular localization.
The Drosophila SRF homolog is expressed in a subset of tracheal cells and maps within a genomic region required for tracheal development
Authors: Affolter Markus , Montagne Jacques , Walldorf Uwe , Groppe Jay , Kloter Urs , LaRosa Marilina , Gehring Walter J.
Published: April 2021

The Drosophila homolog of the vertebrate serum response factor (SRF) was isolated by low stringency hybridization. Nucleotide sequence analysis revealed that the Drosophila SRF homolog (DSRF) codes for a protein that displays 93% sequence identity with human SRF in the MADS domain, the region required for DNA binding, dimerization and interaction with accessory factors. The DSRF gene is expressed during several phases of embryonic development. In the egg, both the RNA and the protein are maternal in origin and slowly decrease in amount during gastrulation. After germ band retraction, high levels of zygotic expression are observed in a distinct subset of peripheral tracheal cells distributed throughout the embryo. Many of these cells are at the tip of tracheal branches and are in direct contact with the target tissues.

The DSRF gene was mapped to position 60C on the second chromosome, and overlapping deficiencies which remove the gene were identified. Analysis of tracheal development in embryos carrying these deletions revealed a degeneration of most of the major branches of the tracheal system. Although the initial migration of tracheal cells was not affected in those deficient embryos, many tracheal cells appeared not to maintain their correct position and continued to migrate. Thus, the DSRF gene might play a role in the proper formation and maintenance of the trachea.
Muscle pattern diversification in Drosophila is determined by the autonomous function of homeotic genes in the embryonic mesoderm
Authors: Michelson Alan M.
Published: April 2021

Muscle diversification in the Drosophila embryo is manifest in a stereotyped array of myofibers that exhibit distinct segment-specific patterns. Here it is shown that the homeotic genes of the Bithorax complex control the identities of abdominal somatic muscles and their precursors by functioning directly in cells of the mesoderm. Whereas Ultrabithorax (Ubx) and abdominal-A (abd-A) have equivalent functions in promoting the formation of particular muscle precursors in the anterior abdominal segments, Abdominal-B (Abd-B) suppresses the development of these same myogenic cells in the posterior region of the abdomen. When expressed in the same mesodermal cells, however, either UBX or ABD-A can override the inhibitory influence of ABD-B, suggesting that these factors may compete in the regulation of common downstream genes. Furthermore, targeted ectopic expression of Ubx or abd-A indicates that these homeotic genes influence muscle cell fates by autonomous action in mesodermal cells. Muscle identity also appears to be sensitive to the level of UBX in myogenic precursors. Finally, these experiments reveal that homeotic cues specific to both the mesoderm and the ectoderm cooperate to specify the pattern of muscle attachment sites.
A chicken achaete-scute homolog (CASH-1) is expressed in a temporally and spatially discrete manner in the developing nervous system
Authors: Jasoni C.L. , Walker M. B. , Morris M. D. , Reh T. A.
Published: April 2021

We have identified a basic helix-loop-helix encoding cDNA from embryonic chicken retina which shares sequence similarity with the achaete-scute family of genes of Drosophila. The deduced amino acid sequence of this chicken achaete-scute homolog (CASH-1) is identical, over the region encoding the basic helix-loop-helix domain, to the recently identified mammalian achaete-scute homolog (MASH-1) and to the Xenopus homolog (XASH1), and 70% identical, over the same region, to Drosophila achaete-scute complex members. The expression of CASH-1 is restricted to subsets of neuronal progenitor cells in the developing chicken nervous system, similar in distribution to that reported for MASH-1 and XASH1. In addition, in situ localization in the retina reveals a dynamic character of expression of the gene in a particular region of the CNS, and suggests that the expression of CASH-1 may be important in defining a particular stage in the progenitor cell necessary for the differentiation of particular neuronal phenotypes.
Expression of zebrafish goosecoid and no tail gene products in wild-type and mutant no tail embryos
Authors: Schulte-Merker S. , Hammerschmidt M. , Beuchle D. , Cho K. W. , Robertis E. M. De , Nüsslein-Volhard C.
Published: April 2021

goosecoid is an immediate early gene expressed at the dorsal blastoporal lip of the Xenopus gastrula. Microinjec- tion experiments have suggested a direct role for goosecoid in organizing the dorsoventral axis of the frog embryo. Here we characterize the zebrafish homologue of goosecoid (gsc) and compare its expression to that of Brachyury or no tail (ntl), another immediate early gene required in developing mesoderm. We show that gsc exhibits two indepen- dent phases of expression: an early one in cells anterior to the presumptive notochord, but not in cells of the notochord itself, and a later one in neural crest derivatives in the larval head. Zygotic gsc transcripts are detected soon after the midblastula transition, and at the blastula stage form a gradient with a maximum at the dorsal side. Use of gsc as a dorsal marker allowed us to demonstrate that ntl expression is initially activated at the dorsal side of the blastula. At this early stage, gsc and ntl show overlapping domains of expression and are co-expressed in cells at the dorsal midline of the early gastrula. However, gsc- and ntl- expressing cells become separated in the course of gastrulation, with gsc being expressed in the axial hypoblast (pre- chordal plate) anterior to the ntl-expressing presumptive notochord cells. Studies with mutant embryos suggest that gsc is independent of ntl function in vivo.
Autonomy and non-autonomy in Drosophila mesoderm determination and morphogenesis
Authors: Leptin Maria , Roth Siegfried
Published: April 2021

The mesoderm in Drosophila invaginates by a series of characteristic cell shape changes. Mosaics of wild-type cells in an environment of mutant cells incapable of making mesodermal invaginations show that this morphogenetic behaviour does not require interactions between large numbers of cells but that small patches of cells can invaginate independent of their neighbours’ behaviour. While the initiation of cell shape change is locally autonomous, the shapes the cells assume are partly determined by the individual cell’s environment. Cytoplasmic transplantation experiments show that areas of cells expressing mesodermal genes ectopically at any position in the egg form an invagination. We propose that ventral furrow formation is the consequence of all prospective mesodermal cells independently following their developmental program. Gene expression at the border of the mesoderm is induced by the apposition of mesodermal and non-mesodermal cells.
The Drosophila fusome, a germline-specific organelle, contains membrane skeletal proteins and functions in cyst formation
Authors: Lin Haifan , Yue Lin , Spradling Allan C.
Published: April 2021

Oogenesis in Drosophila takes place within germline cysts that support polarized transport through ring canals inter-connecting their 15 nurse cells and single oocyte. Developing cystocytes are spanned by a large cytoplasmic structure known as the fusome that has been postulated to help form ring canals and determine the pattern of nurse cell-oocyte interconnections. We identified the adducin-like hts product and α-spectrin as molecular components of fusomes, discovered a related structure in germline stem cells and documented regular associations between fusomes and cystocyte centrosomes. hts mutations completely eliminated fusomes, causing abnormal cysts containing a reduced number of cells to form. Our results imply that Drosophila fusomes are required for ovarian cyst formation and suggest that membrane skeletal proteins regulate cystocyte divisions.
Regulation of Pax-3 expression in the dermomyotome and its role in muscle development
Authors: Goulding Martyn , Lumsden Andrew , Paquette Alice J.
Published: April 2021

The segmented mesoderm in vertebrates gives rise to a variety of cell types in the embryo including the axial skeleton and muscle. A number of transcription factors containing a paired domain (Pax proteins) are expressed in the segmented mesoderm during embryogenesis. These include Pax-3 and a closely related gene, Pax-7, both of which are expressed in the segmental plate and in the dermomyotome. In this paper, we show that signals from the notochord pattern the expression of Pax-3, Pax-7 and Pax-9 in somites and the subsequent differentiation of cell types that arise from the somitic mesoderm. We directly assess the role of the Pax-3 gene in the differentiation of cell types derived from the dermomyotome by analyzing the development of muscle in splotch mouse embryos which lack a functional Pax-3 gene. A population of Pax-3-expressing cells derived from the dermomyotome that normally migrate into the limb are absent in homozygous splotch embryos and, as a result, limb muscles are lost. No abnormalities were detected in the trunk musculature of splotch embryos indicating that Pax-3 is necessary for the development of the limb but not trunk muscle.
Overexpression of a cellular retinoic acid binding protein (xCRABP) causes anteroposterior defects in developing Xenopus embryos
Authors: Dekker Erik-Jan , Vaessen Marie-Josée , van den Berg Claudia , Timmermans Ans , Godsave Sue , Holling Tjadine , Nieuwkoop Pieter , van Kessel Ad Geurts , Durston Antony
Published: April 2021

We have isolated the first Xenopus laevis cDNA coding for a cellular retinoic acid binding protein (xCRABP). xCRABP contains a single open reading frame, coding for an approximately 15×103Mr protein. Northern blot analysis shows that this cDNA hybridizes to a mRNA that is expressed both maternally and zygotically and which already reaches maximal expression during gastrulation (much earlier than previously described CRABP genes from other species). In situ hybridisation showed that at the onset of gastrulation, xCRABP mRNA is localised at the dorsal side of the embryo, in the ectoderm and in invaginating mesoderm. xCRABP expression then rapidly resolves into two domains; a neural domain, which becomes localised in the anterior hindbrain, and a posterior domain in neuroectoderm and mesoderm. These two domains were already evident by the mid-gastrula stage. We investigated the function of xCRABP by injecting fertilized eggs with an excess of sense xCRABP mRNA and examined the effects on development. We observed embryos with clear anteroposterior defects, many of which resembled the effects of treating Xenopus gastrulae with all-trans retinoic acid. Notably, the heart was deleted, anterior brain structures and the tail were reduced, and segmentation of the hindbrain was inhibited. The effects of injecting xCRABP transcripts are compatible with the idea that xCRABP overexpression modulates the action of an endogenous retinoid, thereby regulating the expression of retinoid target genes, such as Hox genes. In support of this, we showed that the expression of two Xenopus Hoxb genes, Hoxb-9 and Hoxb-4, is strongly enhanced by xCRABP over-expression. These results suggest that xCRABP expression may help to specify the anteroposterior axis during the early development of Xenopus laevis.
Negative regulation of the P
o gene in Schwann cells: suppression of Po mRNA and protein induction in cultured Schwann cells by FGF2 and TGFβ1, TGFβ2 and TGFβ3
Authors: Morgan Louise , Jessen Kristján R. , Mirsky Rhona
Published: April 2021

During the development of peripheral nerves, Schwann cells are induced to form myelin sheaths round the larger axons. This process involves a complex series of events and the nature of the molecular signals that regulate and control myelin formation in Schwann cells is not well understood.

Our previous experiments on rat Schwann cells in vitro, using serum-free defined medium, showed that a myelinrelated protein phenotype could be induced in early postnatal Schwann cells in culture by elevation of intracellular cyclic AMP levels in the absence of growth factors, conditions under which the cells are not dividing. Cells with this phenotype expressed the major myelin glycoprotein Po and expression of p75 NGF receptor, N-CAM, GFAP and A5E3 proteins was down-regulated. These changes are all characteristics associated with myelination in vivo. In contrast, when cyclic AMP levels were elevated in the presence of serum, suppression of cyclic AMP-induced differentiation resulted and DNA synthesis was induced.

In this paper, we have used this model system and extended our analysis to explore the relationship between defined growth factors and suppression of myelination. We have used pure recombinant growth factors normally present in peripheral nerves, i.e. FGF1 and FGF2 and TGFβ1, TGFβ2, and TGFβ3 and shown that, like serum, they can strongly suppress the forskolin-mediated induction of the Po gene, both at the level of mRNA and protein synthesis. For both growth factor families, the suppression of Po gene expression is dose-dependent and takes place in serum-starved cells that are mitotically quiescent. In the case of FGF2, however, even more complete suppression is obtained when the cells are simultaneously allowed to enter the cell cycle by inclusion of high concentrations of insulin in the culture medium. The present results raise the possibility that, in addition to the positive axonal signals that are usually envisaged to control the onset of myelination, growth factors present in the nerve may exert negative regulatory signals during development and thus help control the time of onset and the rate of myelination in peripheral nerves.
Anti-apogens and anti-engulfens: monoclonal antibodies reveal specific antigens on apoptotic and engulfment cells during chicken embryonic development
Authors: Rotello Rocco J. , Fernandez Pierre-Alain , Yuan Junying
Published: April 2021

We have isolated a group of monoclonal antibodies that specifically recognize either apoptotic or engulfment cells in the interdigit areas of chicken hind limb foot plates, and throughout the embryo. Ten of these antibodies (anti-apogens) detect epitopes on dying cells that colocalize to areas of programmed cell death, characterized by the presence of apoptotic cells and bodies with typical cellular and nuclear morphology. Our results indicate that cells destined to die, or that are in the process of dying, express specific antigens that are not detectable in or on the surface of living cells. The detection of these apoptotic cell antigens in other areas of programmed cell death throughout the chick embryo indicates that different cell types, which form specific tissues and organs, may utilize similar cell death mechanisms. Six of the monoclonal antibodies (anti -engulfens) define a class of engulfment cells which contain various numbers of apoptotic cells and/or apoptotic bodies in areas of programmed cell death. The immunostaining pattern of the anti-engulfen R15F is similar to that of an antibody against a common leukocyte antigen, suggesting the participation of cells from the immune system in the removal of apoptotic cell debris. These novel monoclonal antibody markers for apoptotic and engulfment cells will provide new tools to assist the further understanding of developmental programmed cell death in vertebrates.
Myogenic specification in somites: induction by axial structures
Authors: Buffinger Nicholas , Stockdale Frank E.
Published: April 2021

Specification of the myogenic phenotype in somites was examined in the early chick embryo using organotypic explant cultures stained with monoclonal antibodies to myosin heavy chain. It was found that myogenic specification (formation of muscle fibers in explants of somites or segmental plates cultured alone) does not occur until Hamburger and Hamilton stage 11 (12-14 somites). At this stage, only the somites in the rostral half of the embryo are myogenically specified. By Hamburger and Hamilton stage 12 (15-17 somites), the three most caudal somites were not specified to be myogenic while most or all of the more rostral somites are specified to myogenesis. Somites from older embryos (stage 13-15, 18-26 somites) showed the same pattern of myogenic specification — all but the three most caudal somites were specified. We investigated the effects of the axial structures, the notochord and neural tube, on myogenic specification. Both the notochord and neural tube were able to induce myogenesis in unspecified somites. In contrast, the neural tube, but not the notochord, was able to induce myogenesis in explants of segmental plate, a structure which is not myogenic when cultured alone. When explants of specified somites were stained with antibodies to slow or fast MyHC, it was found that myofiber diversity (fast and fast slow fibers) was established very early in development (as early as Hamburger and Hamilton stage 11). We also found fiber diversity in explants of unspecified somites (the three most caudal somites from stage 11 to 15) when they were recombined with notochord or neural tube. We conclude that myogenic specification in the embryo results in diverse fiber types and is an inductive process which is mediated by factors produced by the neural tube and notochord.
Evidence for a mitogenic effect of Wnt-1 in the developing mammalian central nervous system
Authors: Dickinson Mary E. , Krumlauf Robb , McMahon Andrew P.
Published: April 2021

The analysis of mutant alleles at the Wnt-1 locus has demonstrated that Wnt-1-mediated cell signalling plays a critical role in development of distinct regions of the embryonic central nervous system (CNS). To determine how these signals participate in the formation of the CNS, we have ectopically expressed this factor in the spinal cord under the control of the Hoxb-4 Region A enhancer. Ectopic Wnt-1 expression causes a dramatic increase in the number of cells undergoing mitosis in the ventricular region and a concomitant ventricular expansion. Although this leads to consistent changes in the relative proportions of dorsal and ventral regions, Wnt-1 does not appear to act as a primary patterning signal. Rather, our experiments indicate that Wnt-1 can act as a mitogen in the developing CNS.
A functional analysis of imprinting in parthenogenetic embryonic stem cells
Authors: Allen Nicholas D. , Barton Sheila C. , Hilton Kathy , Norris Mike L. , Surani M. Azim
Published: April 2021

A detailed analysis of the developmental potential of parthenogenetic embryonic stem cells (PGES) was made in vivo and in vitro, and a comparison was made with the development of cells from parthenogenetic embryos (PG). In vivo, in chimeras with normal host cells (N), PGES cells showed a restricted tissue distribution consistent with that of PG cells, suggesting faithful imprinting in PGES cells with respect to genes involved in lineage allocation and differentiation. Restricted developmental potential was also observed in teratomas formed by ectopic transfer under the kidney capsule. In contrast, the classic phenotype of growth retardation normally observed in PG⟷N chimeras was not seen, suggesting aberrant regulation in PGES cells of genes involved in growth regulation.

We also analysed the expression of known imprinted genes after ES cell differentiation. Igf2, H19 and Igf2r were all appropriately expressed in the PGES derived cells following induction of differentiation in vitro with all-trans retinoic acid or DMSO, when compared with control (D3) and androgenetic ES cells (AGES). Interestingly, H19 was found to be expressed at high levels following differentiation of the AGES cells. Due to the unexpected normal growth regulation of PGES⟷N chimeras we also examined Igf2 expression in PGES derived cells differentiated in vivo and found that this gene was still repressed.

Our studies show that PGES cells provide a valuable in vitro model system to study the effects of imprinting on cell differentiation and they also provide invaluable material for extensive molecular studies on imprinted genes. In addition, the aberrant growth phenotype observed in chimeras has implications for mechanisms that regulate the somatic establishment and maintenance of some imprints. This is of particular interest as aberrant imprinting has recently been invoked in the etiology of some human diseases.
Expression of the LIM class homeobox gene Xlim-1 in pronephros and CNS cell lineages of Xenopus embryos is affected by retinoic acid and exogastrulation
Authors: Taira Masanori , Otani Hiroki , Jamrich Milan , Dawid Igor B.
Published: April 2021

The LIM class homeobox gene Xlim-1 is expressed in Xenopus embryos in the lineages leading to (i) the notochord, (ii) the pronephros, and (iii) certain cells of the central nervous system (CNS). In its first expression phase, Xlim-1 mRNA arises in the Spemann organizer region, accumulates in prechordal mesoderm and notochord during gastrulation, and decays in these tissues during neurula stages except that it persists in the posterior tip of the notochord. In the second phase, expression in lateral mesoderm begins at late gastrula, and converges to the pronephros at tailbud stages. Expression in a central location of the neural plate also initiates at late gastrula, expands anteriorly and posteriorly, and becomes established in the lateral regions of the spinal cord and hindbrain at tailbud stages. Thus Xlim-1 expression precedes morphogenesis, suggesting that it may be involved in cell specification in these lineages. Enhancement of Xlim-1 expression by retinoic acid (RA) was first detectable in the dorsal mesoderm at initial gastrula. During gastrulation and early neurulation, RA strongly enhanced Xlim-1 expression in all three lineages and also expanded its expressing domains; this overexpression correlated well with RA phenotypes such as enlarged pronephros and hindbrain-like structure. Exogastrulation reduced Xlim-1 expression in the lateral mesoderm and ectoderm but not in the notochord, suggesting that the second phase of Xlim-1 expression requires mesoderm/ectoderm interactions. RA treatment of exogastrulae did not revert this reduction.
Competition and position-dependent targeting in the development of the Drosophila R7 visual projections
Authors: Ashley Jennifer A. , Katz Flora N.
Published: April 2021

The R7 photoreceptor neuron projections form a retino - topic map in the medulla of the Drosophila optic lobe. The more inner photoreceptors mutation, an allele of gap1, results in the differentiation of excess R7s in the eye, whose axons invade the brain and establish functional connections. We have used this hyperinnervation phenotype to explore the roles of photoreceptor-target regulation, competitive interactions, and chemoaffinity in map formation. We show that the extra axons are supported in a wild-type brain, with all R7s from a single ommatidium sharing a single termination site, and thus there is no evidence that the target regulates the size of the presynaptic population. In mosaic eyes, in which ommatidia containing extra R7s are surrounded by ommatidia lacking all R7 cells, R7 axons still target to appropriate retinotopic locations in a largely empty R7 terminal field. Axons at the edges of the projection, however, send collaterals into vacant areas of the field, suggesting they are normally restrained to share single termination sites by competitive interactions. In contrast, no sprouts are seen when the vacant sites are juxtaposed with singly innervated sites. In the third instar, R7 and R8 axons transiently display halos of filopodia that overlap adjacent terminals and provide a means to assess occupancy at adjacent sites. Finally, in sine oculislarvae in which only a small number of ommatidia develop, the R7/R8 axons target to predicted dorsoventral portions of the medulla despite the absence of their neighbors, suggesting that position in the eye field determines their connectivity in the brain. We suggest that the mechanisms used to set up this insect map are formally similar to strategies used by vertebrates. The availability of a genetic model for these events should facilitate studies aimed at understanding the molecular bases of retinotopic map development.
Expression of a mouse Zfy-1/lacZ transgene in the somatic cells of the embryonic gonad and germ cells of the adult testis
Authors: Zambrowicz Brian P. , Zimmermann James W. , Harendza Christopher J. , Simpson Elizabeth M. , Page David C. , Brinster Ralph L. , Palmiter Richard D.
Published: April 2021

The Zfy-1 and Zfy-2 genes, which arose by gene duplication, map to the mouse Y chromosome and encode nearly identical zinc-finger proteins. Zfy-1 is expressed in the genital ridge and adult testis and likely encodes a transcription activator. Although potential roles in sex determination and spermatogenesis have been hotly debated, the biological functions ofZfy-1remain unknown. To study the gene’s regulation, transgenes with 21-28 kb of Zfy-1 5′ flanking DNA placed upstream of lacZ were constructed in plasmids or created by homologous recombination of coinjected DNA molecules. The resulting transgenic mice expressed β-galactosidase in the genital ridge of both males and females starting between embryonic day 10 and 11 (E10-E11), peaking at E12-E13 and then declining to low levels by E15, a pattern that matches Zfy-1 mRNA as detected by RT-PCR. This lacZ expression in genital ridge was confined to somatic cells as demonstrated by its absence from the alkaline phosphatase-positive germ cells. It had been reported previously that Zfy-1 mRNA was absent from the embryonic gonad of homozygous Weembryos, which virtually lack germ cells. By contrast, we observed normal expression of the Zfy-1/lacZ transgene when introduced into the We background, suggesting that germ cells are not necessary for expression. In the adult, the Zfy-1/lacZtransgene is expressed abundantly in developing germ cells. Extragonadal (kidney, meninges, arteries, choroid plexus) expression of the transgene was also observed in embryos. A smaller transgene with only 4.3 kb of Zfy-1 5′ flanking DNA was expressed only in germ cells of adult mice. These results suggest that an enhancer for germ cell expression in the adult lies near the Zfy-1 promoter and that an enhancer for expression in the somatic cells of the embryonic gonad is located further 5′.
Early phenotypic choices by neuronal precursors, revealed by clonal analysis of the chick embryo hindbrain
Authors: Lumsden Andrew , Clarke Jonathan D. W. , Keynes Roger , Fraser Scott
Published: April 2021

The mechanisms that generate diverse neuronal phenotypes within the central nervous system are thought to involve local cues or cell-cell interactions acting late in neurogenesis, perhaps as late as the last precursor cell division. We describe here a clonal analysis of neuronal development in the chick hindbrain, using an intracellular tracer to mark single precursor cells, that suggests the operation of an alternative strategy. The majority of clones, ranging from 1 to 46 cells, contained neurons of only one of several possible phenotypes. These single-phenotype clones were not positionally restricted within a rhombomere but were interspersed with other clones containing distinct phenotypes. The assignment of neuronal phenotype in this brain region may, therefore, be made in early precursors and remembered through several rounds of mitotic expansion and dispersal.
Sensory ganglia require neurotrophin-3 early in development
Authors: Gaese Franziska , Kolbeck Roland , Barde Yves-Alain
Published: April 2021

The role played by neurotrophin-3 during the development of quail sensory ganglia was investigated using a mono-clonal antibody that specifically blocks the biological activity of this neurotrophin. Neutralisation of neu-rotrophin-3 was initiated during completion of gangliogen-esis. Neuronal cell counts indicate that about 30% of the neurons normally present in either the placode-derived ganglion nodosum or in a leg-innervating, neural crest-derived dorsal root ganglion are eliminated by the antibody treatment. In both ganglia, this reduction is seen early in development, and the results obtained with the ganglion nodosum indicate that neurotrophin-3 plays an essential role already during gangliogenesis. Neuronal numbers are also compared with those obtained after treatment with a monoclonal antibody to nerve growth factor, used either alone or in combination with the neurotrophin-3 antibody.
Expression and methylation of imprinted genes during in vitro differentiation of mouse parthenogenetic and androgenetic embryonic stem cell lines
Authors: Szabó Piroska , Mann Jeff R.
Published: April 2021

Messenger RNA and methylation levels of four imprinted genes, H19, Igf2r, Igf-2 and Snrpn were examined by northern and Southern blotting in mouse parthenogenetic, androgenetic and normal or wild-type embryonic stem cell lines during their differentiation in vitro as embryoid bodies. In most instances, mRNA levels in parthenogenetic and androgenetic embryoid bodies differed from wild type as expected from previously determined patterns of monoallelic expression in midgestation embryos and at later stages of development. These findings implicate aberrant mRNA levels of these genes in the abnormal development of parthenogenetic and androgenetic embryos and chimeras. Whereas complete silence of one of the parental alleles has previously been observed in vivo, we detected some mRNA in the corresponding embryonic stem cell line. This ‘leakage’ phenomenon could be explained by partial erasure, bypass or override of imprints, or could represent the actual activity status at very early stages of development. The mRNA levels of H19, Igf2r and Igf-2 and the degree of methylation at specific associated sequences were correlated according to previous studies in embryos, and thereby are consistent with suggestions that the methylation might play a role in controlling transcription of these genes. Paternal-specific methylation of the H19 promoter region is absent in sperm, yet we observed its presence in undifferentiated androgenetic embryonic stem cells, or before the potential expression phase of this gene in embryoid bodies. As such methylation is likely to invoke a repressive effect, this finding raises the possibility that it is part of the imprinting mechanism of H19, taking the form of a secondary imprint or postfertilization epigenetic modification necessary for repression of the paternal allele.
wingless acts through the shaggy/zeste-white 3 kinase to direct dorsal-ventral axis formation in the Drosophila leg
Authors: Diaz-Benjumea Fernando J. , Cohen Stephen M.
Published: April 2021

The secreted glycoproteins encoded by Wnt genes are thought to function as intercellular signaling molecules which convey positional information. Localized expression of Wingless protein is required to specify the fate of ventral cells in the developing Drosophila leg. We report here that Wingless acts through inactivation of the shaggy/zeste white 3 protein kinase to specify ventral cell fate in the leg. Ectopic expression of Wingless outside its normal ventral domain has been shown reorganize the dorsal-ventral axis of the leg in a non-autonomous manner. Using genetic mosaics, we show that cells that lack shaggy/zeste white 3 activity can influence the fate of neighboring cells to reo-r ganize dorsal-ventral pattern in the leg, in the same manner as Wingless-expressing cells. Therefore, clones of cells that lack shaggy/zeste white 3activity exhibit all of the organizer activity previously attributed to Wingless-expressing cells, but do so without expressingwingless. We also show that the organizing activity of ventral cells depends upon the location of the clone along the dorsalventral axis. These findings suggest that Wingless protein does not function as a morphogen in the dorsal-ventral axis of the leg.
Pleiotropic effects of the mouse lethal yellow (A
y) mutation explained by deletion of a maternally expressed gene and the simultaneous production of agouti fusion RNAs
Authors: Duhl David M. J. , Stevens Mary E. , Vrieling Harry , Saxon Paul J. , Miller Miles W. , Epstein Charles J. , Barsh Gregory S.
Published: April 2021

Heterozygosity for the mouse lethal yellow(Ay) mutation leads to obesity, increased tumor susceptibility and increased activity of the agouti coat color gene; homozygosity for Ay results in embryonic death around the time of implantation. Although these pleiotropic effects have not been separated by recombination, previous studies have suggested that the dominant and recessive effects result from distinct genetic lesions. Here we use a combination of genomic and cDNA cloning experiments to demonstrate that the Ay mutation is caused by a 120 kb deletion which lies centromere-proximal to the agouti coat color gene. The deletion removes coding but not 5′ untranslated sequences for a ubiquitously expressed gene predicted to encode a protein similar in sequence to an RNA-binding protein, which we named Merc, for maternally expressed hnRNP C-related gene, but have renamed Raly, since the gene is nearly identical to one reported recently by Michaud et al. (Gene Dev. 7, 1203-1213, 1993). The Ay deletion results in the splicing of Merc/Raly 5′ untranslated sequences to agouti protein-coding sequences, which suggests that ectopic expression of the normal agouti protein by the Ay fusion RNA is responsible for the pleiotropic effects associated with heterozygosity for Ay.

We find that Merc/Raly RNA is present in the unfertilized egg and is also transcribed in preimplantation embryos. Using a PCR-based assay to determine the genotype of individual embryos from an Ay/a × Ay/a intercross, we show that, in the absence of zygoticMerc/Raly expression, Ay/Ay embryos develop to the blastocyst stage, but do not hatch from the zona pellucida or form trophoblastic outgrowths. Injection of a Merc/Raly antisense oligonucleotide into non-mutant embryos blocks development prior to the blastocyst stage, and can be rescued by coinjection of a Merc/Raly transgene. These results suggest that maternal expression of Merc/Raly plays an important role in preimplantation development and that its deletion of is sufficient to explainAy-associated embryonic lethality.
Bacterial symbionts induce host organ morphogenesis during early postembryonic development of the squid Euprymna scolopes
Authors: Montgomery Mary K. , McFall-Ngai Margaret
Published: April 2021

The mutualistic association between the squid Euprymna scolopes and the bacterium Vibrio fischeri is an emerging experimental system for the study of the influence of bacteria on animal development. Taking advantage of the ability to raise both this host and its microbial partner independently under laboratory conditions, we describe the effects of bacterial interactions on morphogenesis of the juvenile host symbiotic organ. Our results show that bacteria are essential for normal postembryonic development of the symbiotic organ, which involves changes in both the surface epithelium and the epithelial tissue within the organ where the bacterial culture will take up residence. Cell death induced by exposure to symbiotic V. fischeri results in the regression of a complex ciliated surface epithelium, a tissue that apparently functions to facilitate inoculation of the juvenile organ with the appropriate specific bacterial species. Regression of this tissue begins within hours of exposure to symbiosis-competent bacteria and progresses over the next 5 days, at which time full regression is complete, resulting in a symbiotic organ whose epithelial surface resembles that of the fully mature organ. Moreover, symbiosis-competent bacteria induce modification of the epithelial cells of the crypts that will house these symbionts; these cells undergo significant changes in shape and size in response to interactions with symbiotic V. fischeri. In contrast, we find that when these tissues are not exposed to the proper bacterial symbionts they remain in a state of arrested morphogenesis, a condition that can be rescued by interactions with symbionts. The results of these studies are the first experimental data demonstrating that a specific bacterial symbiont can play an inductive role in animal development.
Rhombomere rotation reveals that multiple mechanisms contribute to the segmental pattern of hindbrain neural crest migration
Authors: Sechrist John , Scherson Talma , Bronner-Fraser Marianne
Published: April 2021

Hindbrain neural crest cells adjacent to rhombomeres 2 (r2), r4 and r6 migrate in a segmental pattern, toward the first, second and third branchial arches, respectively. Although all rhombomeres generate neural crest cells, those arising from r3 and r5 deviate rostrally and caudally (J. Sechrist, G. Serbedzija, T. Scherson, S. Fraser and M. Bronner-Fraser (1993) Development 118, 691-703). We have altered the rostrocaudal positions of the cranial neural tube, adjacent ectoderm/mesoderm or presumptive otic vesicle to examine tissue influences on this segmental migratory pattern. After neural tube rotation, labeled neural crest cells follow pathways generally appropriate for their new position after grafting. For example, when r3 and r4 were transposed, labeled r3 cells migrated laterally to the second branchial arch whereas labeled r4 cells primarily deviated caudally toward the second arch, with some cells moving rostrally toward the first. In contrast to r4 neural crest cells, transposed r3 cells leave the neural tube surface in a polarized manner, near the r3/4 border. Surprisingly, some labeled neural crest cells moved directionally toward small ectopic otic vesicles that often formed in the ectoderm adjacent to grafted r4. Similarly, they moved toward grafted or displaced otic vesicles. In contrast, surgical manipulation of the mesoderm adjacent to r3 and r4 had no apparent effects. Our results offer evidence that neural crest cells migrate directionally toward the otic vesicle, either by selective attraction or pathway-derived cues.
Separate pathways for synapse-specific and electrical activity-dependent gene expression in skeletal muscle
Authors: Tang Jicheng , Jo Sangmee Ahn , Burden Steven J.
Published: April 2021

Signaling between nerve and muscle is mediated by multiple mechanisms, including two transcriptional pathways. Signals provided by the nerve terminal activate transcription of acetylcholine receptor (AChR) genes in myofiber nuclei near the synaptic site, and signals associated with myofiber electrical activity inactivate AChR gene expression throughout the myofiber. These opposing effects of innervation are conferred by 1.8 kb of 5′ flanking DNA from the AChR δ subunit gene. These results raise the possibility that synapse-specific and electrical activity-dependent gene expression are mediated by the same DNA sequence and that activation and repression are determined by differential regulation of the same DNA binding protein. We produced transgenic mice carrying AChR δ subunit-hGH gene fusions, and we show here that a binding site (E-box) for myogenic basic helix-loop-helix proteins is required for electrical activity-dependent but not for synapse-specific gene expression of the δ subunit gene. These results indicate that a change in the expression or activity of an E-box binding protein(s) mediates electrical activity-dependent gene regulation and that synapse-specific and electrical activity-dependent gene expression require different DNA sequences. Moreover, we show here that the cis-acting elements for both aspects of innervation-dependent gene regulation are contained in 181 bp of 5′ flanking DNA from the AChR δ subunit gene.
The role of apterous in the control of dorsoventral compartmentalization and PS integrin gene expression in the developing wing of Drosophila
Authors: Blair Seth S. , Brower Danny L. , Thomas John B. , Zavortink Michael
Published: April 2021

During the development of Drosophila appendages from imaginal discs lineage restrictions appear that prevent dividing cells from crossing between regionally distinct compartments. These compartments correspond not only to regions of cell lineage restrictions but also to regions of specific gene expression. When compartments were first discovered, it was proposed that their formation relied on compartment-specific ‘selector’ gene activity; engrailed is thought to play such a role for the early-arising anteriorposterior restriction. Recent results suggest that the dorsally expressed transcription factor encoded by apterous may control dorsoventral identity in the wing. In this study we use mosaic analysis to show that apterous maintains the late-arising dorsoventral lineage restriction in a manner that strongly supports the selector gene hypothesis: loss of apterous function from dorsal cells after the formation of the boundary causes them to cross into the ventral compartment. Moreover, we show that apterous plays a role controlling patterns of gene expression in the developing wing disc. The PS1 and PS2 integrins are normally expressed in primarily dorsal-specific and ventral-specific patterns, respectively. We show that ectopic expression of apterous induces ectopic ventral expression of PS1 integrin and αPS1 mRNA, while loss of apterous can induce the ectopic dorsal expression of PS2 integrin. Thus, apterous plays a selector-like role both in terms of the control of lineage restrictions and the regulation of downstream gene expression.
Embryonic origin of hemocytes and their relationship to cell death in Drosophila
Authors: Tepass Ulrich , Fessler Liselotte I. , Aziz Amina , Hartenstein Volker
Published: April 2021

We have studied the embryonic development of Drosophila hemocytes and their conversion into macrophages. Hemocytes derive exclusively from the mesoderm of the head and disperse along several invariant migratory paths throughout the embryo. The origin of hemocytes from the head mesoderm is further supported by the finding that in Bicaudal D, a mutation that lacks all head structures, and in twist snail double mutants, where no mesoderm develops, hemocytes do not form. All embryonic hemocytes behave like a homogenous population with respect to their potential for phagocytosis. Thus, in the wild type, about 80-90% of hemocytes become macrophages during late development.

In mutations with an increased amount of cell death (knirps; stardust; fork head), this figure approaches 100%. In contrast, in these mutations, the absolute number of hemocytes does not differ from that in wild type, indicating that cell death does not ‘induce’ the formation of hemocytes. Finally, we show that, in the Drosophila embryo, apoptosis can occur independently of macrophages, since mutations lacking macrophages (Bicaudal D; twist snail double mutants; torso4021) show Abundant cell death.