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C Ziller

Publications and source records attributed to C Ziller.

At least 19 recordsLinked to original sources
PubMed2001

The expression of Quox 1, a homeodomain-containing protein, in sympathetic ganglion cells is regulated in vitro by growth factors.

Quox 1, a quail homeobox gene, is the first vertebrate Antp-type homeobox gene to be described that is expressed in the forebrain. We have already shown that the Quox 1 protein is specifically expressed in post-mitotic sensory neurons. A subpopulation of sympathetic ganglion cells was also found to be labelled by anti-Quox 1 in vitro, but it is not clear whether this protein is expressed in sympathetic ganglion cells in vivo and, if so, the conditions which regulate its expression in vitro. In the present study, we used immunocytochemistry to find out whether Quox 1 expression in sympathetic ganglion cells in vitro is regulated by environmental signals. We found that several peptide growth factors can regulate Quox 1 expression in cultured sympathetic ganglion cells, and that they do so at physiological concentration and in a variety of ways. Basic fibroblast growth factor (FGF-2) induces Quox 1 protein expression, whereas insulin and human insulin-like growth factor-I (IGF-I) down-regulate Quox 1 expression.

Animals↗
PubMed2000

The developmental potentials of the caudalmost part of the neural crest are restricted to melanocytes and glia.

The avian spinal cord is characterized by an absence of motor nerves and sensory nerves and ganglia at its caudalmost part. Since peripheral sensory neurons derive from neural crest cells, three basic mechanisms could account for this feature: (i) the caudalmost neural tube does not generate any neural crest cells; (ii) neural crest cells originating from the caudal part of the neural tube cannot give rise to dorsal root ganglia or (iii) the caudal environment is not permissive for the formation of dorsal root ganglia. To solve this problem, we have first studied the pattern of expression of ventral (HNF3beta) and dorsal (slug) marker genes in the caudal region of the neural tube; in a second approach, we have recorded the emergence of neural crest cells using the HNK1 monoclonal antibody; and finally, we have analyzed the developmental potentials of neural crest cells arising from the caudalmost part of the neural tube in avian embryo in in vitro culture and by means of heterotopic transplantations in vivo. We show here that neural crest cells arising from the neural tube located at the level of somites 47-53 can differentiate both in vitro and in vivo into melanocytes and Schwann cells but not into neurons. Furthermore, the neural tube located caudally to the last pair of somites (i.e. the 53rd pair) does not give rise to neural crest cells in any of the situations tested. The specific anatomical aspect of the avian spinal cord can thus be accounted for by limited developmental potentials of neural crest cells arising from the most caudal part of the neural tube.

Animals↗
PubMed2000

Ovarian carcinoma cells are effectively transfected by polyethylenimine (PEI) derivatives.

As a prerequisite to nonviral gene therapy approaches of ovarian carcinoma, we evaluated the possibility of transfecting established tumor cell lines (SKOV3, IGROV1) as well as primary mesothelial and tumor cells by various polyethylenimine (PEI) derivatives. Several PEI-based vectors were able to effectively transfect these cells, as shown by high luciferase expression levels (10(8) to 10(9) relative light units per milligram of cell protein) that corresponded with 25-50% of green fluorescent protein-positive cells after 24 hours. However, unpredictable differences were observed among the vectors and cell types that a posteriori justified the screening procedure. We also showed that cells that were not transfected after the first experiment remained transfectable in a subsequent transfection experiment to a level similar to that of the initial population. This experiment does not support the emergence of a transfection-resistant cell population and opens the door to multiple therapeutic gene deliveries. Although efficacy and cell targeting still remain to be improved, PEI derivatives appear to be promising molecules for the development of nonviral gene therapy of ovarian carcinoma.

Adenocarcinoma↗
PubMed1999

Regulation of adenovirus-mediated transgene expression by the viral E4 gene products: requirement for E4 ORF3.

In a previous study we showed that multiple deletions of the adenoviral regulatory E1/E3/E4 or E1/E3/E2A genes did not influence the in vivo persistence of the viral genome or affect the antiviral host immune response (Lusky et al., J. Virol. 72:2022-2032, 1998). In this study, the influence of the adenoviral E4 region on the strength and persistence of transgene expression was evaluated by using as a model system the human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA transcribed from the cytomegalovirus (CMV) promoter. We show that the viral E4 region is indispensable for persistent expression from the CMV promoter in vitro and in vivo, with, however, a tissue-specific modulation of E4 function(s). In the liver, E4 open reading frame 3 (ORF3) was necessary and sufficient to establish and maintain CFTR expression. In addition, the E4 ORF3-dependent activation of transgene expression was enhanced in the presence of either E4 ORF4 or E4 ORF6 and ORF6/7. In the lung, establishment of transgene expression was independent of the E4 gene products but maintenance of stable transgene expression required E4 ORF3 together with either E4 ORF4 or E4 ORF6 and ORF6/7. Nuclear run-on experiments showed that initiation of transcription from the CMV promoter was severely reduced in the absence of E4 functions but could be partially restored in the presence of either ORF3 and ORF4 or ORFs 1 through 4. These results imply a direct involvement of some of the E4-encoded proteins in the transcriptional regulation of heterologous transgenes. We also report that C57BL/6 mice are immunologically weakly responsive to the human CFTR protein. This observation implies that such mice may constitute attractive hosts for the in vivo evaluation of vectors for cystic fibrosis gene therapy.

Adenoviridae↗
PubMed1998

Endothelin 3 selectively promotes survival and proliferation of neural crest-derived glial and melanocytic precursors in vitro.

Genetic data in the mouse have shown that endothelin 3 (ET3) and its receptor B (ETRB) are essential for the development of two neural crest (NC) derivatives, the melanocytes and the enteric nervous system. We report here the effects of ET3 in vitro on the differentiation of quail trunk NC cells (NCC) in mass and clonal cultures. Treatment with ET3 is highly mitogenic to the undifferentiated NCC population, which leads to expansion of the population of cells in the melanocytic, and to a lesser extent, the glial lineages. The effect of ET3 on these two NC derivatives was confirmed by the quantitative analysis of clones derived from individual NCC subjected to ET3: we found a large increase in the survival and proliferation of unipotent and bipotent precursors for glial cells and melanocytes, with no significant effect on multipotent cells generating neurons. ET3 first stimulates expression of both ETRB and ETRB2 by cultured NCC. Then, under prolonged exposure to ET3, ETRB expression decreases and switches toward an ETRB2-positive melanogenic cell population. We therefore propose that the present in vitro experiments (long-lasting exposure to a high concentration of ET3) mimic the environment encountered by NCC in vivo when they migrate to the skin under the ectoderm that expresses ET3.

3T3 Cells↗
PubMed1998

Quox 1 homeobox protein is expressed in postmitotic sensory neurons of dorsal root ganglia

The expression of vertebrate homeoproteins has been extensively studied in a variety of normal and cancerous tissues, but little is known on the role of vertebrate homeoproteins in the proliferation and differentiation of cells from these tissues. In the present study, we investigate the relationship between Quox 1 protein (a quail homeodomain containing protein) expression and the proliferation and differentiation of quail dorsal root ganglia (DRG) and neural crest cells. In vivo [3H]TdR labeling experiments demonstrate that the postmitotic sensory neuroblasts appear before the formation of the ganglion, and that more than half of sensory neuroblasts from DRG have already terminated their proliferation in embryos of 2 days of incubation (E2). All DRG neurons have completely ceased to proliferate from E6.5 onwards. By means of immunocytochemistry, we observe that Quox 1 protein is accumulated exclusively in all bipolar neurons in culture of DRG from E9-E11, and in all postmitotic sensory-like neuroblasts during in vitro cell differentiation of the neural crest. The Quox 1 immunoreactive neurons express simultaneously neurofilaments or substance P, and they are never labeled by anti-bromodeoxyuridine. These observations together with the morphology of Quox 1 positive cells, demonstrate that Quox1 protein is expressed in the postmitotic sensory neurons of DRG. Our previous experiments have shown that between E4 and E6, the accumulation of Quox 1 protein increases in DRG in vivo, but decreases in the central nervous system in which cell proliferation decreases (Xue et al., (1993) Mech. Dev. 43, 149-158). Taken together, our results show that the accumulation of Quox 1 protein in DRG is tightly linked to the increase in the number of postmitotic neurons, whereas in the central nervous system the level of expression of Quox 1 seems concomitant with the extent of cell proliferation.

Journal Article↗
PubMed1998

Quox 1 homeobox protein is expressed in postmitotic sensory neurons of dorsal root ganglia.

The expression of vertebrate homeoproteins has been extensively studied in a variety of normal and cancerous tissues, but little is known on the role of vertebrate homeoproteins in the proliferation and differentiation of cells from these tissues. In the present study, we investigate the relationship between Quox 1 protein (a quail homeodomain containing protein) expression and the proliferation and differentiation of quail dorsal root ganglia (DRG) and neural crest cells. In vivo [3H]TdR labeling experiments demonstrate that the postmitotic sensory neuroblasts appear before the formation of the ganglion, and that more than half of sensory neuroblasts from DRG have already terminated their proliferation in embryos of 2 days of incubation (E2). All DRG neurons have completely ceased to proliferate from E6.5 onwards. By means of immunocytochemistry, we observe that Quox 1 protein is accumulated exclusively in all bipolar neurons in culture of DRG from E9-E11, and in all postmitotic sensory-like neuroblasts during in vitro cell differentiation of the neural crest. The Quox 1 immunoreactive neurons express simultaneously neurofilaments or substance P, and they are never labeled by anti-bromodeoxyuridine. These observations together with the morphology of Quox 1 positive cells, demonstrate that Quox 1 protein is expressed in the postmitotic sensory neurons of DRG. Our previous experiments have shown that between E4 and E6, the accumulation of Quox 1 protein increases in DRG in vivo, but decreases in the central nervous system in which cell proliferation decreases (Xue et al., (1993) Mech. Dev. 43, 149-158). Taken together, our results show that the accumulation of Quox 1 protein in DRG is tightly linked to the increase in the number of postmitotic neurons, whereas in the central nervous system the level of expression of Quox 1 seems concomitant with the extent of cell proliferation.

Animals↗
PubMed1998

The avian embryo as a model in developmental studies: chimeras and in vitro clonal analysis.

The avian embryo is a model in which techniques of experimental embryology and cellular and molecular biology can converge to address fundamental questions of development biology. The first part of the chapter describes two examples of transplantation and cell labeling experiments performed in ovo. Thanks to the distinctive histologic and immunocytochemical characteristics of quail and chick cells, the migration and development of definite cells are followed in suitably constructed chimeric quail-chick embryos. Isotopic transplantations of neural tube portions between quail and chick, combined with in situ hybridization with a nucleic probe specific for a quail oligodendrocyte marker, allowed study of the origin and migration of oligodendroblasts in the spinal cord. Heterotopic transplantations of rhombomeres were performed to establish the degree of plasticity of these segments of the hindbrain regarding Hox gene expression, which was revealed by labeling with chick-specific nucleic probes. The second part describes in vitro cell cloning experiments devised to investigate cell lineage segregation and diversification during development of the NC. An original cloning procedure and optimal culture conditions permitted analysis of the developmental potentials of individual NC cells taken at definite migration stages. The results revealed a striking heterogeneity of the crest cell population, which appeared to be composed of precursors at different states of determination. Clonal cultures also provide a means to identify subsets of cells that are the target of environmental factors and to understand how extrinsic signals influence the development of responsive cells.

Animals↗
PubMed1997

HERG- and IRK-like inward rectifier currents are sequentially expressed during neuronal development of neural crest cells and their derivatives.

Quail neural crest cells were cultured in a differentiative medium to study the inward K+ channel profile in neuronal precursors at various stages of maturation. Between 12 and 24 h of culture, neural crest-derived neurons displayed, in addition to the previously described outward depolarization-activated K+ currents, an inward current enhanced in high K+ medium. A biophysical and pharmacological analysis led us to conclude that this inward K+ current is identical to that previously demonstrated in mouse and human neuroblastoma cell lines (I[IR]). This current (quail I[IR] or ql[IR]), which is active at membrane potentials positive to -35 mV, was blocked by Cs+ and by class III antiarrhythmic drugs, thus resembling the K+ current encoded by the human ether-a-gò-gò-related gene (HERG). At later stages of incubation (>48 h), neural crest-derived neurons underwent morphological and biochemical differentiation and expressed fast Na+ currents. At this stage the cells lost qI[IR], displaying instead a classical inward rectifier K+ (IRK) current (quail I[IRK] = qI[IRK]). This substitution was reflected in the resting potential (VREST), which became hyperpolarized by >20 mV compared with the 24 h cells. Neurons were also harvested from peripheral ganglia and other derivatives originating from the migration of neural crest cells, viz. ciliary ganglia, dorsal root ganglia, adrenal medulla and sympathetic chain ganglia. After brief culture following harvesting from young embryos, ganglionic neurons always expressed qI(IR). On the other hand, when ganglia were explanted from older embryos (7-12 days), briefly cultured neurons displayed the IRK-like current. Again, in all the above derivatives the qI(IR) substitution by qI(IRK) was accompanied by a 20 mV hyperpolarization of VREST. Together, these data indicate that the VREST of normal neuronal precursors is sequentially regulated by HERG- and IRK-like currents, suggesting that HERG-like channels mark an immature and transient stage of neuronal differentiation, probably the same stage frozen in neuroblastomas by neoplastic transformation.

Animals↗
PubMed1996

Endothelin 3 promotes neural crest cell proliferation and mediates a vast increase in melanocyte number in culture.

Mutations in the endothelin 3 (EDN3) gene severely affect the development of neural crest-derived melanocytes. In this paper, we report the action of EDN3 on neural crest cells in vitro. The presence of EDN3 leads to a large increase in the number of cells, the majority of which eventually differentiate into melanocytes that aggregate to form a reproducible pigmentation pattern. Quantitative analysis of the effect of different culture conditions revealed that EDN3 initially promotes neural crest cell proliferation. This phase of expansion, which can be prolonged for a few weeks if the cells are replaced regularly, is followed by both a decrease in cell proliferation and the onset of melanocytic differentiation. Therefore, EDN3 is a potent mitogen for early neural crest cell precursors that can give rise to melanocytes.

Animals↗
PubMed1994

Developmental potentials of enteric neural crest-derived cells in clonal and mass cultures.

The aim of this work was to test the differentiation capacities of neural crest-derived cells that had already migrated to the gut and were in the process of forming the intrinsic gut plexuses, when they were withdrawn from the gut wall environment. The experiments were performed on the quail bowel of embryos at 4 to 12 days of incubation (E4 to E12), which was dissociated into a single cell suspension. The crest cells invading the gut carry the NC1/HNK1 epitope. This allowed us to select them as singlets under the control of a uv-illuminated inverted microscope. Crest-derived cells from the gizzard were then clonally cultured on a feeder layer of growth-inhibited 3T3 fibroblasts, as described before for cephalic and truncal neural crest cells. The clonal efficiency of these cells reached about 28% from E4 to E7 and decreased sharply at E8. The size of the clones generated by cells taken between E4 to E8 varied considerably. Large clones (> 10(3)) were numerous from E4 and E5 gizzard-derived cells, whereas E7 and E8 cells produced only very small colonies. The phenotypic diversity of the clones decreased similarly during the time period under scrutiny. Two phenotypes never encountered in the enteric plexuses were found in the cultures: adrenergic cells and glial cells expressing SMP, a marker exclusively expressed by Schwann cells in vivo and normally absent in enteric glia. The capacity to yield adrenergic cells was present up to E6 in clonal cultures of gizzard neural crest-derived cells. Mass cultures of cells dissociated from the gizzard and bowel, including gut mesenchymal and epithelial cells in addition to cells of neural crest origin, revealed that both adrenergic and SMP+ cells could arise under these conditions even in a defined culture medium devoid of serum and chick embryo extract. The number of TH+ cells developing in bowel cultures reached a peak at E7, while at this stage no TH+ cells could be obtained from gizzard cultures. A craniocaudal gradient of disappearance of adrenergic precursors in the gut could thus be demonstrated. Finally we point out that these and previous experiments from other laboratories show that precursor cells of the sympathoadrenal lineage are present in all types of avian sensory and autonomic ganglia during development even though, in most of them, these cells never express the adrenergic phenotype.

Animals↗
PubMed1994

The tyrosine hydroxylase gene is expressed in endoderm and pancreas of early quail embryos.

The initial expression of the gene encoding tyrosine hydroxylase (TH) was studied in the trunk of quail embryos by in situ hybridization. We detected the presence of quail TH mRNA on embryonic day 3.5 (E3.5) in the sympathetic ganglia and aortic plexus, both neural crest derived structures. In contrast, the TH gene was expressed much earlier in the endodermal layer of E2 embryos, i.e. from the 8-somite stage onwards. TH mRNA was found also in the pancreatic bud, an endoderm-derived structure. The TH protein and catecholamines were subsequently looked for in these structures. TH immunoreactivity was found in cells of E2 explanted endoderm, but no catecholamine histofluorescence was observed before or after a few days in culture. TH-positive cells were also detected in cultures of pancreatic rudiments, explanted from E3 to E6 quail embryos. We suggest that the TH-positive cells of the endoderm are the progenitors of the catecholaminergic cells of the pancreas and of the enterochromaffin cells of the gut. The hypothesis that the TH-positive cells of the endoderm are involved in the expression of the catecholaminergic phenotype by neural crest cells is discussed.

Animals↗
PubMed1994

Genetic and epigenetic control in neural crest development.

The neural crest is a fascinating structure of the vertebrate embryo; its ontogeny includes a transient period during which its component cells undergo an epithelio-mesenchymal transition and become migratory. This phase was shown recently to be controlled by the 'Slug' gene which belongs to the 'Snail' family of Drosophila transcription factors. After homing to specific sites in the embryo, the crest-derived cells produce a large variety of phenotypes. Recent advances have shown that during migration most crest cells exhibit various degrees of pluripotentiality, some being already committed to a single and definite fate. Moreover, several lines of evidence point to the existence of totipotent stem cells in the neural crest, the progeny of which become progressively diversified through a combination of intrinsic and extrinsic influences. The latter have been documented by the disruption of several neurotrophin genes, which results in severe deficiencies of selected subsets of neural crest derivatives. The neural crest has also been shown to play an important role in the development of the vertebrate head and hypobranchial region. The genetic control of this process depends on the activity of developmental genes, among which the vertebrate Hox genes are essential, particularly at the rhombencephalic level.

Animals↗
PubMed1994

Effect of the Steel gene product on melanogenesis in avian neural crest cell cultures.

Mutations at the Steel (Sl) and dominant white spotting (W) loci affect three embryonic lineages: primordial germ cells, hemopoietic stem cells and neural-crest-derived melanocytes. The gene products of these loci are a peptide growth factor, called here stem cell factor (SCF), and its tyrosine kinase receptor, the proto-oncogene c-kit. We have studied how chicken recombinant SCF affects the development of melanocytes from quail neural crest cells in secondary culture under defined conditions. We observed that the total number of neural crest cells, of melanocytes and of their precursors was higher in the presence than in the absence of SCF. Labelling with bromodeoxyuridine showed that SCF had a modest and transient mitogenic effect on the neural crest population. SCF also enhanced the differentiation rate of melanocyte precursors, recognized by the "melanocyte early marker" monoclonal antibody (MelEM MAb), and of melanocytes, since the proportion of both subpopulations significantly increased in the presence of SCF. Finally, SCF increased the survival of the neural crest population since in its presence the total number of cells remained stable while it gradually declined in control cultures. Our results support the notion that SCF sustains the survival of the neural crest population and stimulates the rate of the melanogenic differentiation process.

Animals↗
PubMed1993

Patterning of neural crest derivatives in the avian embryo: in vivo and in vitro studies.

In this article we review recent findings from our laboratory on the developmental fate of the neural crest as studied in the avian embryo using the quail-chick marker system in vivo. Quail-chick chimeric experiments carried out at the late neurula stage have revealed the contribution of the neural crest to the skull vault, i.e., the frontal and parietal bones, previously thought to be mesodermally derived. At early developmental stages, the fate of the cephalic mesoderm (free of neural crest cells) could be analyzed. The striking result was that the mesoderm does not contribute to the head and face dermis except in the occipital and otic areas where the skull is derived from the cephalic and somitic mesoderm. Thus, the neural crest forms the dermis, the membrane and cartilage bones of the skull vault, the skull basis, and the face. The limit of the mesoderm-derived skeleton in the skull basis is located in the sella turcica between the basipre- and the basipostsphenoid and coincides with the tip of the notochord. We thus define a "chordal" and an "achordal" skull, the latter being all derived from the neural crest. These results are discussed in the framework of the "New Head" concept of Gans and Northcutt (1983). The second part of this review deals with the role of the environment in the morphogenesis and diversification of neural crest derivatives. The role of the rostrocaudal heterogeneity of the somites in establishing the metameric pattern of the truncal neural crest derivatives is analyzed. The respective contributions of the "in embryo" and in vitro approaches to our understanding of the neural crest cell differentiating potentialities are reviewed. It is pointed out that the search for survival and proliferation factors acting locally on neural crest derivatives when they are wandering and/or settling in various embryonic locations constitutes the new challenge for further understanding their complex patterning and the highly diversified variety of their phenotypes.

Animals↗
PubMed1993

Novel markers of melanocyte differentiation in the avian embryo.

In order to produce markers of melanoblasts and melanocytes, we immunized a mouse with trunk neural crest cells differentiated into melanocytes in culture. We obtained three monoclonal antibodies which react both with pigmented and unpigmented cells in epidermis and in neural crest cultures. These markers are characteristic of distinct differentiation steps. One of them, melanoblast/cyte early marker (MelEM) monoclonal antibody (Mab) detects melanoblasts as soon as they have reached the subectodermal mesenchyme, while the two others, Mel1 and Mel2 Mabs, detect antigens present in melanosomes and promelanosomes later in the differentiation process. Moreover, MelEM is a specific marker of melanoblasts/cytes deriving exclusively from neural crest and not of those from the neural retina. The antigen recognized by MelEM Mab is a protein of M(r) 26,000. The epitopes recognized by Mel1 and Mel2 Mabs are carbohydrate moieties carried by glycoproteins of M(r) 123,000 and 85,000, respectively.

Animals↗