Biomedical subjects

S B Carroll

Publications and source records attributed to S B Carroll.

At least 19 recordsLinked to original sources

PubMed1997

The origin and evolution of animal appendages.

Animals have evolved diverse appendages adapted for locomotion, feeding and other functions. The genetics underlying appendage formation are best understood in insects and vertebrates. The expression of the Distal-less (Dll) homeoprotein during arthropod limb outgrowth and of Dll orthologs (Dlx) in fish fin and tetrapod limb buds led us to examine whether expression of this regulatory gene may be a general feature of appendage formation in protostomes and deuterostomes. We find that Dll is expressed along the proximodistal axis of developing polychaete annelid parapodia, onychophoran lobopodia, ascidian ampullae, and even echinoderm tube feet. Dll/Dlx expression in such diverse appendages in these six coelomate phyla could be convergent, but this would have required the independent co-option of Dll/Dlx several times in evolution. It appears more likely that ectodermal Dll/Dlx expression along proximodistal axes originated once in a common ancestor and has been used subsequently to pattern body wall outgrowths in a variety of organisms. We suggest that this pre-Cambrian ancestor of most protostomes and the deuterostomes possessed elements of the genetic machinery for and may have even borne appendages.

Amino Acid Sequence↗

PubMed1996

daughterless is required for Drosophila photoreceptor cell determination, eye morphogenesis, and cell cycle progression.

Initiation of Drosophila peripheral nervous system (PNS) development requires the achaete-scute complex (AS-C) and the atonal (ato) genes. The AS-C and ato encode basic helix-loop-helix (bHLH) transcription factors that dimerize in vitro with another bHLH protein, daughterless (da). da has many functions during Drosophila embryonic development, as it is required for proper sex determination, oogenesis, and neurogenesis. Here, we examine the expression and function of da within the developing Drosophila eye. The use of a monoclonal antibody to the Da protein revealed that Da levels are modulated across the developing eye disc. Within the morphogenetic furrow (MF) and photoreceptor cell R8, there is a cell-by-cell correspondence between high levels of Da protein expression and Ato protein expression. Mosaic analysis of adult tissue demonstrates that da function is cell autonomous and required within R2, R3, R4, R5, and R8. Examination of gene expression in da- imaginal disc clones reveals that da regulates Ato expression in the MF, affects the progression of the MF, and is necessary for the reestablishment of the G2 and M phases of the synchronized cell cycle posterior to the MF.

Animals↗

PubMed1996

Integration of positional signals and regulation of wing formation and identity by Drosophila vestigial gene.

Appendage formation is organized by signals from discrete sources that presumably act upon downstream genes to control growth and patterning. The Drosophila vestigial gene is selectively required for wing-cell proliferation, and is sufficient to induce outgrowths of wing tissue from eyes, legs and antennae. Different signals activate separate enhancers to control vestigial expression: first, in the dorsal/ventral organizer through the Notch pathway, and subsequently, in the developing wing blade by decapentaplegic and a signal from the dorsal/ventral organizer. Signal integration must be a general feature of genes like vestigial, that regulate growth or patterning along more than one axis.

Animals↗

PubMed1996

Polyembryonic development: insect pattern formation in a cellularized environment.

THe polyembryonic wasp Copidosoma floridanum produces up to 2000 individuals from a single egg. During the production of individual embryos the original anteroposterior axis of the egg is lost and axial patterning must subsequently be reestablished within each embryo. The mechanism by which this occurs is unknown. In most insects, egg polarity is established during oogenesis and early development takes place in a syncytium. In Drosophila melanogaster, the syncytium is considered essential for establishing the morphogenetic gradients that initiate segmental patterning. However, we found that development of C. floridanum occurs almost exclusively in a cellularized environment. To determine whether the D. melanogaster patterning cascade is conserved in the absence of a syncytium, we analyzed the expression of Even-skipped, Engrailed and Ultrabithorax/Abdominal-A during polyembryonic development. Here we show that in spite of the absence of a syncytium, the elements of the D. melanogaster segmentation hierarchy are conserved. The segment-polarity gene Engrailed and the homeotic genes Ultrabithorax/Abdominal-A are expressed in a conserved pattern relative to D. melanogaster. However, we detect an alteration in the expression of the Even-skipped antigen. Even-skipped is initially expressed in segmentally reiterated stripes and not in the pair-rule pattern as it is in D. melanogaster. We also observe that the expression of these regulatory proteins does not occur during the early proliferative phases of polyembryony. Our results indicate that a syncytium is not required for segmental patterning in this insect.

Animals↗

PubMed1995

Cell recognition, signal induction, and symmetrical gene activation at the dorsal-ventral boundary of the developing Drosophila wing.

Appendage formation in insects and vertebrates depends upon signals from both the anterior-posterior and dorsal-ventral (DV) axes. In Drosophila, wing formation is organized symmetrically around the DV boundary of the growing wing imaginal disc and requires interactions between dorsal and ventral cells. Compartmentalization of the wing disc, dorsal cell behavior, and the expression of two dorsally expressed putative signaling molecules, fringe (fng) and Serrate (Ser), are regulated by the apterous selector gene. Here, we demonstrate that fng and Ser have distinct roles in a novel cell recognition and signal induction process. fng serves as a boundary-determining molecule such that Ser is induced wherever cells expressing fng and cells not expressing fng are juxtaposed. Ser in turn triggers the expression of genes involved in wing growth and patterning on both sides of the DV boundary.

Animals↗

PubMed1995

Homeotic genes and the evolution of arthropods and chordates.

Clusters of homeotic genes sculpt the morphology of animal body plans and body parts. Different body patterns may evolve through changes in homeotic gene number, regulation or function. Recent evidence suggests that homeotic gene clusters were duplicated early in vertebrate evolution, but the generation of arthropod and tetrapod diversity has largely involved regulatory changes in the expression of conserved arrays of homeotic genes and the evolution of interactions between homeotic proteins and the genes they regulate.

Animals↗

PubMed1995

Specification of neuroblast identity in the Drosophila embryonic central nervous system by gooseberry-distal.

The Drosophila central nervous system develops from a segmentally reiterated array of 30 unique neural precursors, called neuroblasts. Each neuroblast goes through a stereotyped cell lineage to produce an invariant clone of neural progeny. It is critical to identify the genes that specify neuroblast identity as these genes control the time of formation, gene expression profile, and cell lineage characteristics of each neuroblast. Here we show that the Pax-type gooseberry-distal gene specifies row 5 neuroblast identity. Initially, four rows of neuroblasts form per segment (1, 3, 5, 7) and gooseberry-distal is expressed in row 5 neuroblasts. By using 10 molecular markers, and by following the number and orientation of neuroblast divisions, we show that lack of gooseberry-distal transforms row 5 neuroblasts into row 3 neuroblasts, whereas ubiquitous gooseberry-distal generates the reciprocal transformation. Thus, gooseberry-distal is necessary and sufficient to specify row 5 neuroblast identity autonomously. The 10 genes coordinately regulated by gooseberry-distal are prime candidates for controlling specific aspects of neuroblast identity.

Animals↗

PubMed1995

Homeotic genes and the regulation and evolution of insect wing number.

The evolution of wings catalysed the radiation of insects which make up some 75 per cent of known animals. Fossil evidence suggests that wings evolved from a segment of the leg and that early pterygotes bore wings on all thoracic and abdominal segments. The pterygote body plan subsequently diverged producing orders bearing three, two or just one pair of thoracic wings. We have investigated the role of homeotic genes in pterygote evolution by examining their function in Drosophila wing development and their expression in a primitive apterygote. Wing formation is not promoted by any homeotic gene, but is repressed in different segments by different homeotic genes. We suggest here that wings first arose without any homeotic gene involvement in an ancestor with a homeotic 'groundplan' similar to modern winged insects and that wing formation subsequently fell under the negative control of individual homeotic genes at different stages of pterygote evolution.

Animals↗

PubMed1995

Hairy and emc negatively regulate morphogenetic furrow progression in the Drosophila eye.

The initial steps of pattern formation in the developing Drosophila eye involve the coordination of cell cycles, changes in cell shape, and the specification of the R8 photoreceptor cell. These events begin several cell rows ahead of the morphogenetic furrow and are positively regulated by secreted signaling proteins and the proneural HLH transcription factor atonal (ato). Two HLH regulatory proteins that function to suppress neuronal development in other tissues, extra macrochaetae (emc) and hairy (h), are expressed ahead of the morphogenetic furrow. While neither h nor emc is required for photoreceptor cell determination, in emc-h-clones the morphogenetic furrow and differentiated eye field advance up to eight ommatidial rows ahead of adjacent wild-type tissue. This indicates that morphogenetic furrow progression and neuronal differentiation are negatively regulated by a combination of anteriorly expressed HLH regulatory proteins.

Animals↗

PubMed1995

Spontaneous premature chromosome condensation, micronucleus formation, and non-apoptotic cell death in heated HeLa S3 cells. Ultrastructural observations.

Hyperthermia is an efficient means of inducing cell death in vivo and in vitro. Among human neoplastic cells, HeLa S3 cells are susceptible to heat injury when exposed to long duration moderate hyperthermia (41.5 C), conditions that are reproducible and sustainable in the clinical setting. Hence, HeLa S3 cells are a useful substrate for evaluation of hyperthermic injury in human neoplasia. Previous studies have demonstrated a consistent response of HeLa S3 cells to moderate hyperthermia: spontaneous premature condensation of chromosomes during heat exposure in S phase followed by apparent nuclear fragmentation and, inevitably, cell death. To further characterize the morphological features of this process, HeLa S3 cells grown in suspension at 37 C were heated for 4, 8, 12, or 16 hours at 41.5 C and harvested in glutaraldehyde for electron microscopic evaluation. Compared with untreated controls, heated samples exhibited a characteristic pattern of chromosome condensation that mimicked mitotic prophase but was followed by haphazard asymmetric segregation of chromatid clusters in abnormal metaphase/anaphase and premature reformation of nuclear membrane, resulting not in nuclear fragmentation, but in multiple micronuclei. This pattern of nuclear morphology was not observed in controls. The fraction of cells with micronuclear morphology increased with time in heated samples (from 3.6% at 4 hours to 16.6% at 16 hours), consistent with previous light microscopic analyses of nuclear fragmentation. Cells with multiple micronuclei subsequently exhibited features similar to necrotic cell death. Apoptosis was never observed. Moderate hyperthermia appears to induce a novel morphological pattern of cell injury and death in HeLa S3 cell lines that may be useful as a means of screening cell lines for nonmorphological analyses of hyperthermic injury.

Cell Death↗

PubMed1994

The role of the Distal-less gene in the development and evolution of insect limbs.

BACKGROUND: Arthropod diversity is apparent in the variations in limb number, type, and position along the body axis. Among the insects, for example, butterflies and moths (Lepidoptera) develop larval abdominal and caudal appendages ('prolegs'), whereas flies (Diptera) do not. Comparative studies of the expression and regulation during development of limb-patterning genes, such as Distal-less (Dll), may provide insights into arthropod evolution. RESULTS: We report the cloning of a Dll homolog from the butterfly Precis coenia, and present data showing that it is expressed in all developing limbs (except the mandible), including the prolegs; the relationship between Dll and wingless expression observed in Drosophila is conserved in Precis among all limbs. However, Dll is deployed in distinct spatial and temporal patterns within each limb type. CONCLUSIONS: These data suggest that Dll function, suppressed in the abdomen early in insect evolution, has been derepressed in Lepidoptera, and also suggest that there is a common mechanism underlying the formation of all insect appendages. The limb-type-specific patterns of Dll expression (and its exclusion from the mandible) indicate that regulation of Dll expression may be critical to limb morphology, and are inconsistent with Dll functioning in a simple distal-to-proximal concentration gradient.

Amino Acid Sequence↗

PubMed1994

Expression of an endogenous asialoglycoprotein receptor in a human intestinal epithelial cell line, Caco-2.

We have previously shown that rat asialoglycoprotein receptor expressed in the intestine and liver differ in mRNA size, cell surface distribution, and ratio of compositional protein subunits. In this study, we examined a well characterized intestinal epithelial cell line, Caco-2, as a potential model for studying endogenous receptor in a polarized cell line. Both subunits H1 and H2 of human asialoglycoprotein receptor were detected in Caco-2 cells by Western blots using subunit-specific antisera raised against the hepatic receptor. Antigenic receptor level in fully differentiated Caco-2 cells was approx. 1/3 to 1/2 the level of hepatic HepG2 cells H1 was the dominant subunit in both cell lines. The apparent size of H1 and H2 in Caco-2 cells was not the same as that in HepG2 cells, due to differences in N-linked glycosylation. Consistent with this finding, Northern blot analysis showed that receptor mRNA in the two cell types was of identical size. In pulse-chase experiments H1 was first detected as a 'high-mannose' precursor (40 kDa) in Caco-2 cells that was converted to mature H1 (43 kDa) with a half-life of approx. 60 min. Antigenic levels of H1 and H2 in undifferentiated Caco-2 cells were low, but increased rapidly during cell differentiation, reaching a peak level at 7 days after confluence. Immunocytochemical staining and domain-selective cell surface biotinylation assays showed that the ASGP-R was predominantly localized in the basolateral domain. The receptor in Caco-2 cells was capable of mediating specific uptake and degradation of [125I]asialoorosomucoid. The ligand uptake capacity of the basolateral surface of was approx. 10-fold higher than the apical. These characteristics (H1 subunit and basolateral predominance) of the receptor in Caco-2 cells, resembles the hepatic receptor. We conclude that Caco-2 cells endogenously express in ectopic hepatic-type functional asialoglycoprotein receptor.

Asialoglycoprotein Receptor↗

PubMed1994

Pattern formation and eyespot determination in butterfly wings.

Butterfly wings display pattern elements of many types and colors. To identify the molecular processes underlying the generation of these patterns, several butterfly cognates of Drosophila appendage patterning genes have been cloned and their expression patterns have been analyzed. Butterfly wing patterns are organized by two spatial coordinate systems. One system specifies positional information with respect to the entire wing field and is conserved between fruit flies and butterflies. A second system, superimposed on the general system and involving several of the same genes, operates within each wing subdivision to elaborate discrete pattern elements. Eyespots, which form from discrete developmental organizers, are marked by Distal-less gene expression. These circular pattern elements appear to be generated by a process similar to, and perhaps evolved from, proximodistal pattern formation in insect appendages.

Amino Acid Sequence↗

PubMed1994

Organization of wing formation and induction of a wing-patterning gene at the dorsal/ventral compartment boundary.

The appendages of arthropods and vertebrates possess a third, proximodistal patterning axis that is established after the primary anteroposterior and dorsoventral body axes by mechanisms that are largely unknown. The vestigial gene is required for formation of the entire Drosophila wing, and the dorsal/ventral boundary is shown to organize wing formation and vestigial gene expression. Interactions between dorsal and ventral cells in the growing imaginal disc induce vestigial gene expression through a discrete, extraordinarily conserved imaginal disc-specific enhancer. The link between dorsal/ventral compartmentalization and wing formation distinguishes the development of this sheet-like appendage from that of legs and antennae.

Animals↗

PubMed1994

The achaete-scute complex: generation of cellular pattern and fate within the Drosophila nervous system.

In developing embryos, cells receive and interpret positional information as they become organized into discrete patterns and structures. One excellent model for understanding the genetic regulatory mechanisms that pattern cellular fields is the regulation and function of the achaete-scute complex (AS-C) in the developing nervous system of the fruit fly, Drosophila melanogaster. Three structurally homologous proneural genes--achaete (ac), scute (sc), and lethal of scute (l'sc)--are required for neural stem cell formation. In Drosophila, the AS-C genes are initially expressed in patterns of cell clusters at reproducible anteroposterior (AP) and dorsoventral (DV) coordinates that foreshadow where neural precursors arise. In the embryonic central nervous system (CNS), the gene products of AP and DV axis-patterning genes act combinatorially via a large array of cis-regulatory regions scattered throughout the AS-C to generate a segmentally repeated pattern of proneural clusters. Within each cluster (an equivalence group), one cell then retains proneural gene expression and is singled out as the neural stem cell (neuroblast). The neuroblast inhibits the surrounding cells from adopting neural fates (lateral inhibition) through a signaling pathway that is mediated via the action of the proneural and neurogenic genes. The proneural genes therefore represent a nodal point in the patterning of the nervous system. They receive global positional information, transduce it to discrete sets of cells, and trigger local cell interactions that mediate cell fate decisions.

Animals↗

PubMed1994

Positioning adjacent pair-rule stripes in the posterior Drosophila embryo.

We present a genetic and molecular analysis of two hairy (h) pair-rule stripes in order to determine how gradients of gap proteins position adjacent stripes of gene expression in the posterior of Drosophila embryos. We have delimited regulatory sequences critical for the expression of h stripes 5 and 6 to 302 bp and 526 bp fragments, respectively, and assayed the expression of stripe-specific reporter constructs in several gap mutant backgrounds. We demonstrate that posterior stripe boundaries are established by gap protein repressors unique to each stripe: h stripe 5 is repressed by the giant (gt) protein on its posterior border and h stripe 6 is repressed by the hunchback (hb) protein on its posterior border. Interestingly, Krüppel (Kr) limits the anterior expression limits of both stripes and is the only gap gene to do so, indicating that stripes 5 and 6 may be coordinately positioned by the Kr repressor. In contrast to these very similar cases of spatial repression, stripes 5 and 6 appear to be activated by different mechanisms. Stripe 6 is critically dependent upon knirps (kni) for activation, while stripe 5 likely requires a combination of activating proteins (gap and non-gap). To begin a mechanistic understanding of stripe formation, we locate binding sites for the Kr protein in both stripe enhancers. The stripe 6 enhancer contains higher affinity Kr-binding sites than the stripe 5 enhancer, which may allow for the two stripes to be repressed at different Kr protein concentration thresholds. We also demonstrate that the kni activator binds to the stripe 6 enhancer and present evidence for a competitive mechanism of Kr repression of stripe 6.

Animals↗

PubMed1994

The ventral nervous system defective gene controls proneural gene expression at two distinct steps during neuroblast formation in Drosophila.

Within the Drosophila embryo, the formation of many neuroblasts depends on the functions of the proneural genes of the achaete-scute complex (AS-C): achaete (ac), scute (sc) and lethal of scute (l'sc), and the gene ventral nervous system defective (vnd). Here, we show that vnd controls neuroblast formation, in part, through its regulation of the proneural genes of the AS-C. vnd is absolutely required to activate ac, sc and l'sc gene expression in proneural clusters in specific domains along the medial column of the earliest arising neuroblasts. Using ac-lacZ reporter constructs, we determined that vnd controls proneural gene expression at two distinct steps during neuroblast formation through separable regulatory regions. First, vnd is required to activate proneural cluster formation within the medial column of every other neuroblast row through regulatory elements located 3' to ac; second, through a 5' regulatory region, vnd functions to increase or maintain proneural gene expression in the cell within the proneural cluster that normally becomes the neuroblast. By following neuroblast segregation in vnd mutant embryos, we show that the neuroectoderm forms normally and that the defects in neuroblast formation are specific to particular proneural clusters.

Animals↗

PubMed1993

Pattern formation in a secondary field: a hierarchy of regulatory genes subdivides the developing Drosophila wing disc into discrete subregions.

The legs and wings of insects and vertebrates develop from secondary embryonic fields that arise after the primary body axes have been established. In order to understand how the insect imaginal wing field is patterned, we have examined in detail the temporal and spatial expression patterns of, and epistatic relationships between, four key regulatory genes that are specifically required for wing formation in Drosophila. The wingless protein, in a role surprisingly distinct from its embryonic segment polarity function, appears to be the earliest-acting member of the hierarchy and crucial for distinguishing the notum/wing subfields, and for the compartmentalization of the dorsal and ventral wing surfaces. The wingless product is required to restrict the expression of the apterous gene to dorsal cells and to promote the expression of the vestigial and scalloped genes that demarcate the wing primordia and act in concert to promote morphogenesis.

Animals↗