Search PubMed⌕ Search

Biomedical subjects

C Ziller

Publications and source records attributed to C Ziller.

At least 37 records · Page 2Linked to original sources
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↗
PubMed1993

Plasticity in neural crest cell differentiation.

The neural crest is a pluripotent population of cells that are endowed with migratory capacities. It has long been known that the differentiation pathway taken by cells derived from the neural crest is largely controlled by the microenvironment to which they home after their migration phase, indicating a high degree of plasticity in their developmental fate. Recent progress has been made concerning the factors which influence survival, growth and differentiation of selected sets of precursors in each embryonic site colonised by derivatives of the neural crest.

Adrenal Medulla↗
PubMed1992

Developmental potentialities of cells derived from the truncal neural crest in clonal cultures.

The developmental potentialities of single truncal neural crest derived cells were analysed in clonal cultures. The clone-forming ability and differentiation potential of crest cells migrating through the somitic mesoderm of 3-day-old embryos (E3) and of non-neuronal cells of dorsal root ganglia taken at E6-14 were compared. Since most of the cells present in the sclerotomal and rostral parts of the somite at E3 become later on incorporated into the spinal ganglia, one can consider that these two cell populations represent the same derivatives of the trunk neural crest at different developmental stages. After 10 days in vitro, the size of clones and their phenotypic composition varied noticeably, revealing a certain heterogeneity in the founder cell populations in terms of developmental potencies. Clones obtained from migrating neural crest cells at E3 were often large (greater than 1000 cells) and many of them contained neuronal and non-neuronal cells. Dorsal root ganglion cells produced mostly small clones (less than 100 cells) in which only non-neuronal (i.e. glial) phenotypes were expressed. Therefore, both the capacity for proliferation and the differentiation ability of cloned neural crest derived cells decrease considerably with increasing embryonic age. This is even more striking if these results are compared with those obtained previously in our laboratory with single cells cultures of E2 cephalic neural crest. In the latter case, both clone sizes and cellular diversity within the colonies were much higher than with E3 truncal crest and dorsal root ganglia (DRG) non-neuronal cells. The second result of the present work concerns the differentiation of the dormant autonomic neuronal precursors of the DRG. It has been established previously that the non-neuronal cells of the DRG include adrenergic precursors than can differentiate in mass culture of dissociated DRG cells. We show that these cells never differentiate in clonal cultures but depend upon the cell density of the culture. This suggests that cell to cell interaction between crest derived cells are critical in eliciting the differentiation of the adrenergic phenotype.

Animals↗
PubMed1992

The neural tube/notochord complex is necessary for vertebral but not limb and body wall striated muscle differentiation.

The aim of this work was to investigate the role played by the axial organs, neural tube and notochord, on the differentiation of muscle cells from the somites in the avian embryo. Two of us have previously shown that neuralectomy and notochordectomy is followed by necrosis of the somites and consecutive absence of vertebrae and of most muscle cells derived from the myotomes while the limbs develop normally with muscles. Here we have focused our attention on muscle cell differentiation by using the 13F4 mAb that recognizes a cytoplasmic antigen specific of all types of muscle cells. We show that differentiation of muscle cells of myotomes can occur in the absence of notochord and neural tube provided that the somites from which they are derived have been in contact with the axial organs for a defined period of time, about 10 hours for the first somites formed at the cervical level, a duration that progressively reduces caudalward (i.e. for thoracic and lumbar somites). Either one or the other of the two axial organs, the neural tube or the notochord can prevent somitic cell death and fulfill the requirements for myotomal muscle cell differentiation. Separation of the neural tube/notochord complex from the somites by a surgical slit on one side of the embryo gave the same results as extirpation of these organs and provided a perfect control on the non-operated side. A striking finding was that limb and body wall muscles, although derived from the somites, differentiated in the absence of the axial organs. However, limb muscles that develop after excision of the neural tube started to degenerate from E10 onward due to lack of innervation. In vitro explantation of somites from different axial levels confirmed and defined precisely the chronology of muscle cell commitment in the myotomes as revealed by the in vivo experiments.

Animals↗
PubMed1989

A monoclonal antibody directed against quail tyrosine hydroxylase: description and use in immunocytochemical studies on differentiating neural crest cells.

Catecholamine (CA) synthesis is one of the phenotypic traits expressed by some neural crest-derived cells in vivo and in vitro. In the present study, we have evidenced, in quail embryos, the expression of the first enzyme of CA metabolism, tyrosine hydroxylase (TOH), using a monoclonal antibody raised against the quail enzyme. This antibody also recognizes TOH from chick and pleurodele, but not from several mammalian species (rat, human). We have also investigated the extent to which TOH-positive cells, differentiated in neural crest cultures, express structural neuronal markers and display vasoactive intestinal polypeptide (VIP) and substance P (SP) immunoreactivity. Double-immunolabeling experiments show that, in vitro, half of the population of TOH-positive cells exhibits tetanus toxin binding sites but none of them are recognized by a neurofilament antibody. On the other hand, some TOH-positive cells contain VIP or SP. These observations suggest that under our culture conditions autonomic neural crest precursors differentiate only into immature sympathoblasts, but are able to synthesize peptides in addition to CA.

Animals↗
PubMed1988

A monoclonal antibody recognizing a common antigen on neurons and fibroblasts in chicken and quail.

A monoclonal antibody, FiN1, obtained by immunization of a mouse with homogenates of embryonic quail nodose ganglia, was found to react with a surface antigenic determinant, both in quail and chick, present on practically all neurons of the spinal cord and of the peripheral nervous system and on a subpopulation of fibroblasts. An ontogenetic study performed on tissue sections, cell suspensions and cultures showed that FiN1 defines a differentiation marker which appears relatively late in development, during the second half of embryonic life, and persists after hatching. The onset and evolution of its expression during development varies in a tissue-specific manner.

Animals↗
PubMed1988

A surface protein expressed by avian myelinating and nonmyelinating Schwann cells but not by satellite or enteric glial cells.

Searching for specific markers of neural crest-derived cell lineages, we immunized mice with glycoproteins purified from adult quail peripheral myelin. We obtained a monoclonal antibody that reacts with myelin and peripheral glial cells. This antibody, to Schwann cell myelin protein (SMP), is specific for the membranes of all Schwann cells, irrespective of whether they are associated with myelinated nerves. SMP persists on Schwann cells in long-term cultures in vitro, but is absent from satellite cells of peripheral ganglia, both in vivo and in vitro. The antigen (a protein doublet of Mr 75,000-80,000) is present in, but not restricted to, the myelin lamellae, since it is distributed along the whole myelinating Schwann cell membrane. In the CNS, SMP appears as a single band of Mr 80,000. SMP is first detectable by immunofluorescence at E6 in the quail, which is at least 6 days earlier than the first appearance of already described markers related to myelination.

Animals↗
PubMed1987

Cell lineages in peripheral nervous system ontogeny: medium-induced modulation of neuronal phenotypic expression in neural crest cell cultures.

Neural crest, taken from cephalic and trunk levels of quail embryos, was grown in vitro in conventional tissue culture medium (Dulbecco's modified Eagle's medium containing 15% fetal calf serum and either 2 or 15% chick embryo extract (CEE] or in a chemically defined serum- and CEE-free medium. Depending on the conditions employed, different types of neuronal or neuronlike cells developed in the cultures. Thus, in medium containing 15% CEE, adrenergic cells (identified by tyrosine hydroxylase immunoreactivity and catecholamine histofluorescence) emerged after 5-6 days. These cells lacked tetanus toxin binding sites and did not react with an antibody directed against 70-kDa neurofilament protein. In the fully defined medium, a neuronal cell type exhibiting neurofilament and substance P (SP) immunoreactivity differentiated from noncycling precursors within 1 or 2 days of culture. If serum was added to the medium, the neurites disintegrated and the neuronal cells ultimately died. By sequentially culturing neural crest, first in the wholly synthetic medium for 1-3 days and then in the conventional medium supplemented with serum and 15% CEE, the disappearance of the SP-positive neurons was followed, several days later, by the emergence of adrenergic cells. The majority of these cells and/or their precursors were found to undergo cell division in culture. We conclude that the cells expressing the adrenergic phenotype (characteristic of the sympathetic nervous system) and those displaying SP immunoreactivity, comparable to a category of neurons in dorsal root and cranial sensory ganglia, derive from distinct sets of precursors. Our results reinforce the contention, deduced from in ovo transplantation experiments (see N. M. Le Douarin, (1984) In Cellular and Molecular Biology of Neuronal Development (I. Black, Ed.), pp. 3-28. Plenum, New York), that at least two lineages, from which sensory and autonomic cell types are derived respectively, are segregated early during neural crest ontogeny and have extremely different survival and trophic requirements.

Acetylcholine↗
PubMed1987

A monoclonal antibody specific for avian early myogenic cells and differentiated muscle.

A monoclonal antibody raised in mouse in response to homogenates of Remark ganglia and dorsal mesentery of chicken embryos was found to exhibit a unique reactivity towards myogenic cells, heart, striated muscles, and smooth muscles in chicken and quail. Indirect immunofluorescence assays were performed at different stages of chicken and quail embryonic development and, after hatching, on tissue sections and cultured cells. They revealed that the cytoplasmic marker recognized by 13F4 is expressed in early embryonic heart, in somitic myotome (from stage 14 onward), in the skeletal muscles in limbs and trunk, in all muscles in the head and the branchial arches, in the smooth muscles of the digestive tract and blood vessels. In myofibrils of striated muscles, the antigen is localized in the Z lines. The antigenicity of the molecule recognized by 13F4 is not associated with a glycolipid or a glycoprotein. It is of peptidic nature and its molecular weight is 54 kDa. We stress the value of this cell-type-specific marker in studies on ontogenesis and differentiation of all muscular structures, namely, of myocardium and striated muscles, which express 13F4 antigenicity from an early developmental stage.

Animals↗
PubMed1987

In vivo and in vitro expression of vasoactive intestinal polypeptide-like immunoreactivity by neural crest derivatives.

Qualitative and quantitative in vivo studies were performed on the development of the neuropeptide vasoactive intestinal polypeptide (VIP) in the peripheral nervous system of quail embryos. VIP-like immunoreactivity (VIPLI) was found by radioimmunoassay (RIA) from the sixth day of embryonic life onward in the sympathetic chain, the esophagus and duodenum, and from day 15 of incubation onward in the adrenal glands and the nodose ganglia. By using immunocytochemistry, we identified cells expressing VIPLI in sensory spinal ganglia of 13- to 15-day-old embryos. In neural crest cultures, cells expressing the VIP phenotype differentiated constantly under various culture conditions, in contrast to other phenotypes which had specific medium requirements, i.e. adrenergic cells or substance P-containing neurons.

Animals↗
PubMed1986

Heterogeneity in migrating neural crest cells revealed by a monoclonal antibody.

A monoclonal antibody, GlN1, obtained by immunization with extracts of the 14 d embryonic quail nodose ganglion, is described. GlN1 recognizes an antigenic determinant present in virtually all the satellite cells of the peripheral ganglia, all Schwann cells of the peripheral nerves, and in subpopulations of sensory and autonomic neurons of embryonic and adult quails and chickens. The molecular weight of the antigen(s) revealed by GlN1 in embryonic day 12 quail dorsal root ganglion (DRG) cultures is around 80 kDa. In the neural crest, GlN1 determinant is found as soon as the crest cells leave the neural primordium. Only a proportion (25%) of the migrating neural crest cells carry the antigen. This demonstrates that the neural crest is composed of a heterogeneous population of cells from its early migratory stages. Being selectively distributed on neural crest cells and its derivatives, the GlN1 determinant may be considered as a "differentiation antigen" that will be useful in further studies on cell-line segregation during the ontogeny of the PNS.

Animals↗
PubMed1985

Quail neural crest cells transformed by Rous sarcoma virus can be established into differentiating permanent cell cultures.

Quail neural crest cells derived from the truncal neural primordium, infected in vitro by Rous sarcoma virus (RSV) in January 1978, were induced to multiply and have been established into permanent cultures. These cultures contain cells that differentiate into melanocytes, neuron-like cells and flat cells. About 50% of these different cell types are tetanus-toxin positive. Electrophysiological studies have shown that some cells can generate action potentials similar to those reported in quail neural crest primary cultures. Taken together these data show that the RSV-transformed quail neural crest permanent cultures are composed of stem cells which can differentiate into cell types specific for neural crest.

Animals↗
PubMed1983

Early segregation of a neuronal precursor cell line in the neural crest as revealed by culture in a chemically defined medium.

This article addresses the problem of the segregation of cell lines during the development of peripheral nervous system components from the neural crest. We show here that committed precursors of peripheral neurons are present in the crest before the migration of its cells has started. If cultured in a serum-deprived medium, a subpopulation of the crest cells readily differentiates into neurons without dividing. Neuronal markers such as neurofilament proteins and receptor sites for tetanus toxin are not expressed in the committed neuronal precursors, but appear after a few hours in culture. They are coexpressed in neurons with the mesenchymal intermediate filament protein, vimentin, which is common to all neural crest cells regardless of their prospective fate. A strong inhibitory effect of serum factor(s) on neurite outgrowth is demonstrated. We show also that conditions stimulating proliferation of crest cells are incompatible with promotion of neuronal differentiation and vice-versa.

Acetylcholinesterase↗
PubMed1982

Migration and differentiation of neural crest cells and their derivatives: in vivo and in vitro studies on the early development of the avian peripheral nervous system.

After a period of extensive migration through the vertebrate embryo, neural crest cells differentiate into a great variety of cell types, including all the elements of the peripheral nervous system. We have studied crest cell migration in quail-chick chimeras in which quail cells can be identified by means of a stable natural nuclear marker. The results of interspecific grafts of neural primordium, performed systematically at different levels of the neuraxis, have established the sites of origin of the principal peripheral ganglia. In addition, they suggest that the cholinergic and adrenergic phenotypes are not predetermined in the neural crest before migration, but are the result of multiple cellular interactions. Furthermore, the phenotype expression of young differentiating autonomic ganglia is labile and can be modified if the latter are subjected to an appropriate cellular environment by grafting into a younger host embryo. The results of experiments in which fragments of neural crest, sensory and autonomic ganglia were transplanted are presented in terms of a model of crest cell-line segregation. As a step towards the analysis of the cellular interactions occurring during autonomic neuron differentiation, we have also studied neuronal development in tissue cultures of neural crest, taken from the cranial and trunk levels of quail embryos and grown in the presence or absence of other embryonic tissues. The results confirm that both levels of the crest are potentially able to give rise to cells that can make acetylcholine and catecholamines. However, whereas acetylcholine-synthesizing ability is apparently a very early feature of autonomic neuron precursors, the ability to produce catecholamines is acquired later as a result of interactions with other cell types, in particular with mesenchymal derivatives. Although production of both neurotransmitters can be considerably stimulated by associating crest with any of several young embryonic rudiments, only when trunk crest is cultured with the sclerotomal moiety of the somite is biochemical differentiation accompanied by extensive morphological and cytochemical neuronal maturation.

Acetylcholine↗
PubMed1981

[Neuronal differentiation of neural crest cells cultured in a defined medium].

Mesencephalic neural crest cells from Quail embryos, cultivated in a defined serum free medium, exhibit neurite extension and typical neuronal traits. This differentiation is never achieved in the presence of serum. Addition of serum into the culture medium after neuronal differentiation has occurred, induces complete regression of the neurites.

Animals↗