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P M Iannaccone

Publications and source records attributed to P M Iannaccone.

At least 19 recordsLinked to original sources
PubMed2001

Nodal regulates trophoblast differentiation and placental development.

Nodal has been thought to be an embryo-specific factor that regulates development, but nodal is also expressed in the mouse placenta beginning at midgestation, specifically in the spongiotrophoblasts. In an insertional null nodal mutant, not only is embryonic development disrupted, but mouse placental development is also grossly altered with the loss of the diploid spongiotrophoblasts and labyrinth and an expansion of the polyploid giant cell layer. A hypomorphic mutation in nodal results in an expansion of the giant cell and spongiotrophoblast layers, and a decrease in labyrinthine development. Expression of nodal in trophoblast cell cultures is sufficient to inhibit trophoblast giant cell differentiation, demonstrating that nodal can act directly on trophoblasts. The mechanism of nodal action includes the inhibition of junB gene transcription. These results suggest that nodal may be involved in redirecting trophoblast fate towards the midgestational expansion of the labyrinth region while maintaining the thin layer of trophoblast giant cells and the underlying layer of spongiotrophoblasts that form the boundary between the maternal and extraembryonic compartments.

Animals↗
PubMed2001

GLI1 localization in the germinal epithelial cells alternates between cytoplasm and nucleus: upregulation in transgenic mice blocks spermatogenesis in pachytene.

The zinc finger transcription factor GLI1 is the mediator of signaling by members of the Hedgehog (Hh) family. Male mice in which Desert hedgehog (Dhh), an Hh homologue expressed in Sertoli cells of the testis, was knocked out are sterile, suggesting that the Dhh/GLI1 pathway plays a role in spermatogenesis. Using an antiserum raised against human GLI1, we found that during the first round of spermatogenesis, GLI1 expression is initially cytoplasmic, then shifts to the nuclei of Sertoli and germ cells, and finally shifts back to the cytoplasm. In the adult mouse testis, GLI1 expression localized to the nuclei of germ cells, beginning with pachytene cells and persisting through round spermatids. Localization of GLI1 in elongating spermatids shifted from the nucleus to the cytoplasm and became associated with microtubules. We also examined a line of transgenic mice that overexpressed human GLI1. Male mice in this line were sterile. Spermatogenesis was blocked at the pachytene stage, and a subset of the morphologically indistinguishable pachytene cells underwent apoptosis. Patched-2, which is a Dhh receptor, and Fused, another component of the signal transduction pathway, are expressed in Leydig cells and in primary and secondary spermatocytes. Expression of GLI1 in the same cell types as Patched-2 and Fused and the disruption of spermatogenesis by GLI1 overexpression suggest that GLI1 is the mediator of the Dhh signal in the testis, and that it may be a regulator of spermatogenesis.

Animals↗
PubMed2000

Nodal and bone morphogenetic protein 5 interact in murine mesoderm formation and implantation.

Mice mutant for the TGF-beta family member, nodal, lack mesoderm and die between E8.5 and E9.5. The short ear-lethal (se(l) ) mutation, a deletion that eliminates Bmp-5, causes a strikingly similar gastrulation defect. Here we analyze se(l);nodal compound mutants and find a dosage effect. Embryos homozygous for one mutation show distinct gastrulation stage defects that depend on whether they are heterozygous or homozygous for the other mutation. Embryos mutant for nodal or se(l);nodal compound mutants fail to execute an antigenic shift indicative of mesoderm differentiation and ectoderm cells are shunted into an apoptotic pathway. Furthermore, we find a novel phenotype in se(l);nodal double mutant litters, in which two to four genetically different embryos are contained within the same deciduum. Both the gastrulation and implantation phenotypes can also arise in short ear-viable (se(v) ) and se(v); nodal mutant mice. These data indicate that loss of Bmp-5 may underlie the se(l) gastrulation phenotype and suggest that nodal and Bmp-5 interact during murine mesoderm formation. Our data also reveal an unsuspected role for Bmp-5 in implantation and the decidual response in the mouse.

Animals↗
PubMed2000

Modeling of mosaic patterns in chimeric liver and adrenal cortex: algorithmic organogenesis?

If organogenesis were a completely deterministic process, then the amount of information required to store the spatial position and fate of every cell in vertebrate organisms would be larger than the total information that could be contained in their genomes. This suggests that the instructions of developmental mechanisms involved in organogenesis, coded in DNA, must be at least in part procedural or algorithmically based. Chimeric mosaic patterns in rat livers have been shown to be isotropic and to have fractal profiles (D approximately 1.3) whereas adrenal gland mosaics show a less irregular radial pattern, with lower fractal dimension (D approximately 1.2) than in the liver. These findings suggested a possible model of parenchyma generation. We propose that during organogenesis in both liver and adrenal cortex, the same basic mechanism is directed to organ mass enlargement, whereas the differences observed in mosaic patterns between the organs could be due to the control of a single parameter, namely, a form of contact inhibition. Computer simulations in two dimensions returned comparable results in both the fractal dimension value of mosaic patches and appearance of the mosaic 'tissues', as observed histologically in chimeras. This suggests that position information and locomotion of cells would not be required to produce the mosaic pattern observed in chimeras.

Adrenal Cortex↗
PubMed1999

Developmental pathways: Sonic hedgehog-Patched-GLI.

Developmental pathways are networks of genes that act coordinately to establish the body plan. Disruptions of genes in one pathway can have effects in related pathways and may result in serious dysmorphogenesis or cancer. Environmental exposures can be associated with poor pregnancy outcomes, including dysmorphic offspring or children with a variety of diseases. An important goal of environmental science should be reduction of these poor outcomes. This will require an understanding of the genes affected by specific exposures and the consequence of alterations in these genes or their products, which in turn will require an understanding of the pathways critical in development. The ligand Sonic hedgehog, the receptors Patched and Smoothened, and the GLI family of transcription factors represent one such pathway. This pathway illustrates several operating principles important in the consideration of developmental consequences of environmental exposures to toxins.

Abnormalities, Drug-Induced↗
PubMed1997

Cloning and developmental expression of the murine homolog of the acute leukemia proto-oncogene AF4.

AF4 is the 4q21 gene involved in the acute lymphoblastic leukemia associated t(4;11)(q21;q23) where it forms a fusion gene with MLL. In order to gain insight into AF4's role in leukemogenesis we have studied its functional domains and expression pattern during murine development. We have cloned the murine homolog, Af4. We have demonstrated that 5' half of Af4 encodes a region with transcriptional transactivation activity which is disrupted by the t(4;11) in human leukemias. We have also localized the murine AF4 protein to the nucleus supporting a role for AF4 in transcription. The developmental expression pattern of Af4 was determined in situ hybridization and suggests Af4 plays an important role in the development of the hematopoietic, cardiovascular, skeletal and central nervous systems. A repeating pattern of Af4 expression in development is down-regulation with differentiation of a tissue. Among the cell types where this pattern of down-regulation is noted are B-lymphocytes. These findings raise the possibility that the disruption of normal AF4 function by the translocation may contribute to leukemogenesis.

Amino Acid Sequence↗
PubMed1997

Transgenic technology: an overview of approaches useful in surgical research.

Advances in transgenic science have created powerful tools for the investigation of both genetic and protein regulatory systems. Recently, transgenic animals have been utilized in several vascular and transplantation research laboratories. The ability to specifically mutate genes important in oncologic and cardiovascular research is leading to a greater understanding of the role of gene and protein regulatory systems in cancer and cardiovascular disease. The expanding use of transgenic animals will undoubtedly increase our insight into complex problems in surgical research. This review briefly describes the various techniques utilized to create transgenic animals including: transgene design, gene-transfer utilizing transfection, microinjection and retroviral infection, as well as the use of embryonic stem cells, and methods for screening transgenic offspring.

Animals↗
PubMed1997

Expression of human GLI in mice results in failure to thrive, early death, and patchy Hirschsprung-like gastrointestinal dilatation.

BACKGROUND: GLI is an oncodevelopmental gene in the vertebrate hedgehog/patched signaling pathway that is spatiotemporally regulated during development and is amplified in a subset of human cancers. GLI is the prototype for the Gli-Kruppel family of transcription factors, which includes the Drosophila segment polarity gene ci, the C. elegans sex-determining gene tra-1, and human and mouse GLI3, all of which contain a conserved domain of five C2-H2 zinc fingers. GLI3 mutations have been implicated in the mouse mutant extra toes, as well as in human Greig cephalopolydactaly syndrome and the autosomal dominant form of Pallister-Hall syndrome. As such, GLI and the vertebrate hedgehog/patched signaling pathway appear to play important roles in both normal development and neoplasia. MATERIALS AND METHODS: Since it is not known whether aberrant GLI expression is similarly linked to developmental disorders, we developed gain-of-function transgenic mice which express human GLI ectopically. RESULTS: Affected transgenic mice exhibit a phenotype of failure to thrive, early death, and Hirschsprung-like patches of gastrointestinal dilatation. The colons of affected mice have greatly attenuated smooth muscle layers and abnormal overlying epithelium. The density of myenteric plexuses is reduced in the colonic walls. The severity of the phenotype is related to the level of transgene expression. CONCLUSIONS: The transgenic mouse model supports a role for GLI in gastrointestinal development. As part of the vertebrate hedgehog/patched signaling pathway, GLI is essential to mesoderm and CNS ectoderm development and transgenic GLI expression affects neuronal, muscular, and epithelial cell differentiation in the gut. Expression of human GLI in mice results in impairment of enteric neuronal development and a Hirschsprung-like phenotype.

Animals↗
PubMed1995

Overexpression of human glutathione S-transferase pi protects NIH 3T3 cells against (+/-)anti BPDE cytotoxicity but not tumor formation.

In order to explore the protective function of human glutathione S-transferase pi (GST-pi) in vitro and in vivo, transfected NIH 3T3 clones were examined in cytotoxicity assays using the carcinogen (+/-)anti-benzo(a)pyrene 7,8-diol-9,10-epoxide (BPDE) or inoculated into nude mice and treated with the carcinogen benzo(a)pyrene (BP) to induce tumor formation. The human GST-pi cDNA under the control of the murine alpha 2(I)collagen promoter was transfected into NIH 3T3 cells and G418 resistant clones were analyzed by Southern, northern, western, and two-dimensional analysis. Clone A2 stably expressed human GST-pi and has 2.5-fold greater activity toward the substrate 1-chloro-2,4-dinitrobenzene and a 1.7-fold increase in LD50 for BPDE in vitro when compared to control-transfected clone G3. This increase in protection, however, did not prevent the formation of BP-induced tumors in vivo.

3T3 Cells↗
PubMed1994

An upstream region of the enoyl-coenzyme A hydratase/3-hydroxyacyl-coenzyme A dehydrogenase gene directs luciferase expression in liver in response to peroxisome proliferators in transgenic mice.

Peroxisome proliferators, which are structurally diverse nonmutagenic agents, induce hepatocarcinogenesis in rats and mice. Exposure to these xenobiotics leads to a rapid and coordinated transcriptional activation of the genes for the peroxisomal beta-oxidation enzyme system pathway in the liver. We have previously identified a peroxisome proliferator-responsive element in the 5'-flanking region of the rat peroxisomal hydratase/dehydrogenase (PBE) gene, the second enzyme in the beta-oxidation pathway. The peroxisome proliferator-responsive element in the PBE gene was shown to direct the induction of a luciferase reporter gene in vitro. We have now used this 3.2-kilobase 5'-flanking region of the PBE gene fused to the coding region of luciferase to generate transgenic mice. Three independent lines of transgenic mice expressed luciferase in response to ciprofibrate, a peroxisome proliferator. The induction of luciferase is specific to the liver; this agrees with the tissue-specific induction of PBE. Two other hypolipidemic drugs, nafenopin and Wy-14,643, were also capable of inducing luciferase activity in the liver. This study suggests that the PBE upstream element can be used to direct and modulate the expression of cloned genes by changing the levels of peroxisome proliferators. Also, the PBE-luciferase transgenic mouse should be an excellent model system for screening xenobiotics for potential peroxisome proliferator property.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed1994

Pluripotent embryonic stem cells from the rat are capable of producing chimeras.

Embryonic stem cells have been enormously important in the production of targeted mutations in mice used in the study of gene function and biological aspects of disease states. The use of these cells for mouse studies is now wide-spread but the production of animals from similar cell lines derived from other species has not been previously reported. We demonstrate here the derivation of diploid rat embryonic stem cells (RESC-01). RESC-01 cells are SSEA-1 and alkaline phosphatase positive, grow best on primary rat embryonic fibroblasts, and can differentiate extensively in vivo. RESC-01 cells form cystic embryoid bodies capable of rhythmic contractions. Rat blastocysts injected with RESC-01 cells form chimeras. The results indicate that the successful in vitro propagation and chimera production with embryonic stem cells is not limited to the mouse. The long-term culture of rat ES cells will provide an important resource for the study of normal physiology and disease models where rat is the species of choice.

Animals↗
PubMed1994

Fractal geometry in rat chimeras demonstrates that a repetitive cell division program may generate liver parenchyma.

In the development of mammalian organs, a rapid and robust expansion of the parenchymal compartment must occur following allocation of organ primordia. This expansion must be regulated so that sufficient tissue mass is generated for further organization into functional tissues. The discovery that mosaic patches in the liver of rat chimeras are fractal (a geometric form with characteristic complexity) suggests a possible information storage scheme for programs of parenchyma generation. Since fractal objects are produced by the repetitive application of specific rules, it is possible that such a mechanism is responsible for the generation of organ parenchyma. The model cell division program for the generation of organ parenchyma considered here is to choose a cell at random to divide and place the daughter cell in a randomly chosen adjacent position displacing other cells which might occupy the chosen position. The completion of the division creates a new population of cells representing the input conditions for the next division. When this is repeated over and over in a tissue comprising two genetically distinguishable populations of cells, analysis of the geometry of the mosaic pattern obtained should fulfill specific predictions. If cell division occurred in this manner, the complexity of patch boundaries (patches are contiguous aggregates of cells of the same marker lineage in tissue from a chimera) should be independent of the proportion of the two parental cell lineages which make up the chimera's tissue. However, the complexity of the entire patch pattern should be dependent on this proportion. The complexity of the spatial distribution of the patches within a chimera's tissue should also be dependent on the proportion of the two parental lineages. We have measured the complexity of patch boundaries (surface fractal dimension), the complexity of entire fields of patches (mass fractal dimension), and the complexity of the spatial distribution of patches (fractal fragmentation) in rat liver from chimeras. We have established that the surface fractal dimension does not change as the proportion of the two parental lineages in the chimera's tissue changes, that there is a simple relationship between the complexity of entire patches and this proportion, and that the patches are fractally fragmented. These results are consistent with the hypothesis that repetitive application of this simple cell division program accounts for the generation of liver parenchyma.

Animals↗
PubMed1993

Cell-specific expression of a recombinant rat glutathione S-transferase Ya gene in transgenic mice.

Transgenic mice have been generated which carry a cDNA encoding the rat Ya isozyme of glutathione S-transferase (GST) under the transcriptional control of the SV40 early region promoter-enhancer. Expression of the GST transgene was highly tissue-specific, with the highest expression detected in the convoluted tubular epithelium of the mouse kidney cortex. GST Ya mRNA abundance in these cells was greater than that found for GST Ya mRNA in normal rat liver. GST Ya protein was observed in the convoluted tubule cells of the founder mouse as well as an F1 offspring. The transmission of the foreign gene was followed for two generations, and an erratic pattern of inheritance was observed. These animals provide a model for the in vivo study of GST modulation of carcinogenesis and drug toxicity.

Animals↗
PubMed1993

Distribution of expression of cell adhesion molecules in the mid to late gestational mouse fetus.

The control of murine morphogenesis appears to be regulated in part by the expression of the primary cell adhesion molecules, such as the neural cell adhesion molecules (N-CAM). Here we show that the epithelial cell adhesion molecules appear in intestinal epithelium, liver and cartilage, but were absent from intestinal submucosa and neural tissues. N-CAMs on the other hand were present in intestinal submucosa and neural tissues, but absent from intestinal epithelium, liver, and cartilage. Both epithelial cell adhesion molecules and N-CAM were present in intestinal primordium at gestation times (days 12 and 13) when intestinal epithelium and submucosa are not morphologically distinguishable. On day 14 of gestation, when the intestinal epithelium and submucosa are morphologically distinguishable, epithelial cell adhesion molecules are present in intestinal epithelium but not submucosa while N-CAM has the reciprocal pattern of expression. Immunoblots with antibodies to N-CAM revealed two bands of 110-220 and 60 kD which followed specific patterns of expression. As defined by densitometry, the intensity of the larger protein increased from day 12 to 18 in neural tissue groups, but diminished in late gestational intestine and intact fetus and was replaced by a more discrete region of 110-150 kD, suggesting that embryonic to adult conversion of isoform ('E to A conversion') had occurred at this nonneural site.

Animals↗
PubMed1993

Chimeric drift in blood cell populations of chimeric rats constructed between congenic strains.

Chimeric drift is the shift in the proportion over time of the two cell lineages which comprise a chimera (genetic mosaic). Chimeric drift in blood cell populations is determined by both the probability of proliferation from stem cell pools of one or the other of the cell lineages which constitute the chimera and the effects of life span in circulating blood cells. Previous evidence suggests that while chimeric drift occurs in chimeras between genetically disparate strains, it does not occur when the strains used are closely related. No information is available from chimeras between congenic strains. In the present study, chimeric rats were produced between strains with distinguishable class I major histocompatibility complex haplotypes, PVG-RT1a and PVG (which express the haplotype RT1c). PVG-RT1a-specific monoclonal antibodies were used to establish the mosaic patterns in the cell populations of peripheral blood by fluorescein-activated cell sorting. Mosaic cell lineage of red blood cells, white blood cells, lymphocytes, monocytes and neutrophil populations were analyzed weekly over a period of 6 weeks. The ratio of cells of the PVG-RT1a lineage to cells of the PVG lineage shifted either in favor of PVG-RT1a or PVG in cellular components of peripheral blood. The percentage of PVG-RT1a cells in peripheral blood of chimeras changes by as much as 54, 28, 21, 19 and 23% in red blood cell, white blood cell, lymphocyte, monocyte and neutrophil populations, respectively. The shifts in the percentage of PVG-RT1a cells appears to occur in a cyclic fashion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗