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Nematocyte differentiation in hydra: commitment to nematocyte type occurs at the beginning of the pathway.

In hydra the four types of nematocytes arise by differentiation from the multipotent stem cells among the interstitial cells. It has been unclear where along the nematocyte pathway commitment to type occurs. Some evidence suggests that this commitment occurs at the beginning of the pathway, while other data suggest that it occurs at the terminal cell cycle midway through the pathway. Upon reduction of cell population sizes of the interstitial cell lineage by treatment with hydroxyurea, interstitial cells entering nematocyte pathways frequently undergo an amplification division. A nearest-neighbor analysis of pairs of nematoblast nests in such depleted animals has shown that the fraction of the nearest-neighbor pairs that are matched pairs, in which both nests are of the same type, is higher than predicted. A very high fraction of the matched pairs were identical pairs in which the number of cells in each nest was the same. Also, in a large majority of the identical pairs the nests were shown to be in the same stage of development. The simplest interpretation of these results is that the two daughters of the amplification division giving rise to the matched pair were committed to nematocyte type before the division occurred. In another experiment we show that the length of the G2 phase of the next-to-terminal cell cycle differs between desmonemes and stenoteles. This indicates that differences in the differentiation pathways of these two types exist before the terminal cell cycle. This result also supports the idea that commitment to type occurs at the beginning of the pathway. A means of reconciling the view that commitment occurs early with the view that it occurs late in teh differentiation pathway is discussed.

Animals↗

Nestin expression in ganglioglioma.

It has been suggested that gangliogliomas represent a neoplastic transformation of a dysplastic focus or heterotopia. Other theories propose that gangliogliomas arise from multipotent stem cells with the ability to differentiate along glial and neuronal cell lines. Our goal was to characterize the expression of nestin, a neuroepithelial precursor/stem cell antigen, in gangliogliomas along with other pathological and clinical features of this entity. The clinical and operative features of 18 recent cases meeting the histological criteria for ganglioglioma were reviewed. The expression of nestin, microtubule-associated protein 2 (MAP2), neurofilament, and glial fibrillary acidic protein (GFAP) was assessed by immunohistochemistry and confocal scanning laser microscopy. Abundant MAP2- and nestin-positive neuronal cells were found by immunohistochemistry in all 18 gangliogliomas. GFAP staining was found in reactive and lesional astrocytes but not in cells of neuronal morphology. Confocal microscopy demonstrated colocalization of nestin and MAP2 in select neuronal cells. The true lineage of gangliogliomas remains controversial. Our findings confirm the presence of cells within these lesions that harbor a persistent stem cell cytoskeletal protein (nestin). Further insight into the cytoskeletal derangement of nestin-positive neuronal cells may shed further light on the pathogenesis of gangliogliomas and its associated epilepsy.

Adolescent↗

A classification of acute leukaemia for the 1990s.

The need for reproducibility in the classification of acute leukaemia has made it necessary to incorporate information derived from new techniques which have become essential for the study of these disorders. In addition to classic morphology and cytochemistry (FAB proposals), it is necessary to add immunology and cytogenetics (MIC proposals), as well as to investigate further the biological and diagnostic significance of molecular events. As a result of these investigations a new group of leukaemias merit recognition as distinct entities. These include three types of ALL with specific chromosome abnormalities, namely, i) t (9;22), ii) t (4;11) and iii) t (1;19) and four subtypes of AML, i) with minimal differentiation or AML-M0, ii) with basophilic precursors or M2Baso, iii) AML (M4/M5) with t (8;16) and iv) AML with trilineage myelodysplasia. Biphenotypic acute leukaemia constitutes also a distinct entity with features of ALL and AML and represents a malignancy probably affecting multipotent stem cells. We propose an objective evaluation system for biphenotypic leukaemias based on a score in which the various lineage markers are graded according to their known specificity.

Humans↗

Myelo-lymphopoiesis in long-term bone marrow culture.

In vitro analysis of early haemopoietic events has been hampered by the absence of culture systems allowing long-term maintainance and proliferation of self-renewing multipotent stem cells. Recently, a liquid culture system has been established which allows in vitro proliferation of haemopoietic stem cells and of haemopoietic precursor and mature end cells for several months. There is good evidence that this long-term proliferation is due to a micro-environment established by bone-marrow-derived stromal cells. This review attempts to summarize recent results in analysing haemopoietic control mechanisms, leukaemogenesis and haematological disorders by applying this culture system using both murine and human bone marrow cells.

Animals↗

Stem cell plasticity in the hematopoietic system.

Bone marrow (BM) contains hematopoietic stem cells, which differentiate into all mature blood cells, and marrow stromal cells that provide the microenvironment for hematopoietic stem/progenitor cells along with the capability to differentiate into mature cells of multiple mesenchymal tissues including fat, bone, and cartilage. Recent studies indicate that adult BM also contains cells that can differentiate into nonhematopoietic cells of ectodermal, mesodermal, and endodermal tissues other than hematopoietic tissues, including liver, pancreas, kidney, lung, skin, gastrointestinal tract, heart, skeletal muscles, and neural tissues. Studies reporting the multipotentiality of BM cells have become a focus of interest because they suggest that clinical applications could be at hand using easily obtainable cells in the treatment of tissue damage or degenerative diseases. Presently, however, definitive evidence explaining the mechanism of this multipotentiality of BM stem cells is lacking. In this review, we summarize recent progress and controversies in investigation of the multipotentiality of adult BM-derived stem cells to differentiate into nonhematopoietic tissues.

Adult↗

Stem cell plasticity, beyond alchemy.

Cell plasticity is a central issue in stem cell biology. Differentiated somatic nuclei have the flexibility to dedifferentiate when transferred into oocytes or when fused to pluripotent embryonic stem cells. Recent publications also claim that somatic stem cells can convert into developmentally unrelated cell types both in vivo and ex vivo without such drastic cell manipulations. Some of these claims are still controversial, making it difficult for us to determine the reality of somatic stem cell plasticity. Indeed, we have heard enough about the "potentials" of cell plasticity; how much do we know about mechanisms? A fundamental issue in current stem cell biology is to understand the mechanisms underlying cell plasticity. In this short review, we overview three research fields related to cell plasticity: nuclear transfer, transdifferentiation, and cell fusion, with an emphasis on studies of molecular mechanisms underlying cell plasticity.

Adult↗

Expression and evolutionary conservation of nanos-related genes in Hydra.

The Drosophila gene nanos encodes two particular zinc finger motifs which are also found in germline-associated factors from nematodes to vertebrates. We cloned two nanos (nos)-related genes, Cnnos1 and Cnnos2 from Hydra magnipapillata. Using whole-mount in situ hybridization, the expression of Cnnos1 and Cnnos2 was examined. Cnnos1 was specifically expressed in multipotent stem cells and germline cells, but not in somatic cells. Cnnos2 was weakly expressed in germline cells and more specifically in the endoderm of the hypostome where it appears to be involved in head morphogenesis. In addition to structural conservation in the zinc finger domain of nanos-related genes, functional conservation of Cnnos1 was also demonstrated by the finding that a Cnnos1 transgene can partially rescue the nosRC phenotype that is defective in the egg production of Drosophila. Thus, the function of nanos-related genes in the germline appears to be well conserved from primitive to highly evolved metazoans.

Amino Acid Sequence↗

How to make a Barrett esophagus: pathophysiology of columnar metaplasia of the esophagus.

Barrett esophagus is defined as a specialized intestinal replacing the squamous epithelium of the esophageal mucosa in response to gastroesophageal reflux. Barrett metaplasia is a healing process that develops to protect the esophagus from further damage. Although mechanisms by which Barrett metaplasia evolves toward dysplasia and adenocarcinoma have been extensively studied, the process by which squamous epithelium is replaced by specialized intestinal metaplasia is poorly understood. Barrett esophagus develops when defense mechanisms in the esophageal mucosa (luminal secretion of mucus, bicarbonate, growth factors, etc.) are overwhelmed by an ongoing cycle of mucosal injury and repair. Hydrogen ion, pepsin, trypsin, and bile acids are considered harmful agents that synergistically invade the esophageal mucosa. Areas of destroyed squamous epithelium are then progressively reepithelized by a columnar epithelium that may originate from multipotent stem cells located within the basal layer of the normal esophageal mucosa or in the ducts of submucosal glands.

Barrett Esophagus↗

Deletion of the acetylcholinesterase locus at 7q22 associated with myelodysplastic syndromes (MDS) and acute myeloid leukaemia (AML).

The genes for acetylcholinesterase (ACHE) and butyrylcholinesterase (BCHE) are located within regions subject to non-random chromosomal abnormalities in the myelodysplastic syndromes (MDS) and acute myeloid leukaemia (AML). Acetylcholinesterase is mapped to 7q22, within the critical deleted region presumed to contain a myeloid specific tumour suppressor gene. Butyrylcholinesterase is mapped to 3q26: abnormalities at this region are associated with sub-types of MDS and AML with thrombocytopenia, or with increased platelet counts. Both ACHE and BCHE have been implicated as playing a role in megakaryopoiesis and thrombopoiesis, and these genes have been observed to be co-amplified in acute myeloid leukaemia. Recent findings suggest a more significant role for the ACHE gene in haemopoiesis by regulating multipotent stem cell proliferation, and apoptosis in cells undergoing erythroid and myeloid differentiation. This led us to investigate gene copy-number alterations at these genes in MDS and AML. Samples were screened by slot-blot hybridization, and if changes were observed, by Southern blotting. A total of 42 samples from 31 de novo AML patients, 10 samples from eight cases of post-MDS AML and 85 samples from 67 MDS patients were analysed with probes for ACHE, BCHE, c-MYC, MDR-1 and globin control. Changes in ACHE and/or BCHE were observed in 9/31 de novo AML patients, and in 7/67 MDS patients: 1/37 cases of refractory anaemia (RA), 1/10 cases of refractory anaemia with excess blasts (RAEB) and 5/20 chronic myelomonocytic leukaemia (CMML) patients. The amplification events observed generated copy numbers no greater than 10, showed normal restriction patterns and had no clear correlation with megakaryopoiesis or thrombopoiesis. Loss of signal at the ACHE locus was observed: haploid signal intensity was seen in seven samples: one RA with thrombocytopenia, three CMML, one AML-M5a (no karyotypic abnormalities of chromosome 7), one AML-M4 (monosomy 7), and one case of AML-M7 (karyotype unknown). Homozygous deletion was observed at relapse of an additional patient with AML-M4. These data reinforce the possibility that ACHE may play a role as a myeloid tumour suppressor gene.

Acetylcholinesterase↗

Chromosome 1q+ in erythroid and granulocyte-monocyte precursors in a patient with essential thrombocythemia.

A 55-year-old man with essential thrombocythemia had multiplication of bands q21 to q32 of chromosome 1 in all studied mitoses from bone marrow, from unstimulated blood, and from erythroid and granulocyte-monocyte colonies grown in vitro. The multiplication was in the form of triplication in 16 out of 20 mitoses from the marrow and in 4 of 6 mitoses from the blood; the rest showed a duplication of this region. All 20 mitoses from erythroid and granulocyte-monocyte colonies showed the abnormality in the form of duplication. These findings indicate most probably a clonal evolution, the triplication having arisen in the clone with the duplication. This may be associated with early leukemic transformation. The detection of the 1q+ aberration in two different types of hematopoietic colonies indicates the involvement of multipotent stem cells in at least this patient with essential thrombocythemia.

Adult↗

Granulocyte colony-stimulating factor stimulates neurogenesis via vascular endothelial growth factor with STAT activation.

The adult brain harbors multipotent stem cells, which reside in specialized niches that support self-renewal. Granulocyte colony-stimulating factor (G-CSF) induces bone marrow stem cells proliferation and mobilization from their niche, and activates endothelial cell proliferation, which might help to establish a vascular niche for neural stem cells (NSCs). Here, we show that G-CSF induced receptor-mediated proliferation and differentiation of neural precursors in human NSCs cultures and in adult rat brain in vivo. In human NSCs cultures, G-CSF activated STAT3 and 5, and increased VEGF and its receptor, VEGFR2 (Flk-1) expression, and VEGFR2 tyrosine kinase inhibitor blocked the neurogenesis stimulated by G-CSF. G-CSF also activated endothelial cell proliferation in adult rat brain in vivo. Our results indicate that G-CSF stimulates neurogenesis through reciprocal interaction with VEGF and STAT activation.

Brain↗

Early inactivation of p53 tumor suppressor gene cooperating with NF1 loss induces malignant astrocytoma.

Malignant astrocytoma, the most prevalent primary brain tumor, is resistant to all known therapies and frequently harbors mutations that inactivate p53 and activate Ras signaling. We have generated mouse strains that lack p53 and harbor a conditional allele of the NF1 tumor suppressor that negatively regulates Ras signaling. The mice develop malignant astrocytomas with complete penetrance. The majority of tumors display characteristics of glioblastoma multiforme with concomitant alteration of signaling pathways previously described in the human counterparts of this neoplasm. We find that the sequence of tumor suppressor inactivation influences tumorigenicity and that earliest evidence of tumor formation localizes to regions of the brain that contain a multipotent stem cell population capable of in vivo differentiation into neurons and glia.

Animals↗

Live and let die in the intestinal epithelium.

The intestinal epithelium is a relatively simple developmental system and a prime example of tissue renewal from a source of multipotent stem cells. Throughout adulthood, intestinal epithelial proliferation, cell-fate specification and differentiation are coupled to migration in discrete units known as crypts of Lieberkühn. Physically guided by Eph receptors and their ligands, the ephrins, stem cell progeny transit through the proliferation/differentiation switch, and Notch diversifies their subsequent fates. Wnt signalling appears to control most of these events.

Animals↗

Stem cell origin of cancer and differentiation therapy.

Our forefathers in pathology, on observing cancer tissue under the microscope in the mid-19th century, noticed the similarity between embryonic tissue and cancer, and suggested that tumors arise from embryo-like cells [Recherches dur le Traitement du Cancer, etc. Paris. (1829); Editoral Archiv fuer pathologische Anatomie und Physiologie und fuer klinische Medizin 8 (1855) 23]. The concept that adult tissues contain embryonic remnants that generally lie dormant, but that could be activated to become cancer was later formalized by Cohnheim [Path. Anat. Physiol. Klin. Med. 40 (1867) 1-79; Virchows Arch. 65 (1875) 64] and Durante [Arch. Memori ed Osservazioni di Chirugia Practica 11 (1874) 217-226], as the "embryonal rest" theory of cancer. An updated version of the embryonal rest theory of cancer is that cancers arise from tissue stem cells in adults. Analysis of the cellular origin of carcinomas of different organs indicates that there is, in each instance, a determined stem cell required for normal tissue renewal that is the most likely cell of origin of carcinomas [Lab. Investig. 70 (1994) 6-22]. In the present review, the nature of normal stem cells (embryonal, germinal and somatic) is presented and their relationships to cancer are further expanded. Cell signaling pathways shared by embryonic cells and cancer cells suggest a possible link between embryonic cells and cancer cells. Wilm's tumors (nephroblastomas) and neuroblastomas are presented as possible tumors of embryonic rests in children. Teratocarcinoma is used as the classic example of the totipotent cancer stem cell which can be influenced by its environment to differentiate into a mature adult cell. The observation that "promotion" of an epidermal cancer may be accomplished months or even years after the initial exposure to carcinogen ("initiation"), implies that the original carcinogenic event occurs in a long-lived epithelial stem cell population. The cellular events during hepatocarcinogenesis illustrate that cancers may arise from cells at various stages of differentiation in the hepatocyte lineage. Examples of genetic mutations in epithelial and hematopoietic cancers show how specific alterations in gene expression may be manifested as maturation arrest of a cell lineage at a specific stage of differentiation. Understanding the signals that control normal development may eventually lead us to insights in treating cancer by inducing its differentiation (differentiation therapy). Retinoid acid (RA) induced differentiation therapy has acquired a therapeutic niche in treatment of acute promyelocytic leukemia and the ability of RA to prevent cancer is currently under examination.

Antineoplastic Agents↗

Sox-2 is expressed by neural progenitors and astroglia in the adult rat brain.

Sox-2 is a transcription factor that is expressed by self-renewing and multipotent stem cells of the embryonic neuroepithelium. Very little is however known about Sox-2 expression in the adult brain and therefore we used immunohistochemistry to examine its distribution and co-localization with specific cell markers. We found that Sox-2 was expressed by actively dividing neural progenitor cells in the neurogenic regions in the adult rat brain, the subventricular zone of the forebrain and the subgranular zone of the dentate gyrus in the hippocampus. Cells expressing immature neuronal markers were essentially Sox-2 immunonegative. Sox-2 was also found to be expressed by glial fibrillary acidic protein immunopositive astroglia, widely distributed in the brain parenchyma. Given the fact that several studies have established the neurogenic capacity of a specialized type of astroglia in the adult brain, the findings of Sox-2 expression in parenchymal astroglia are of potential interest. We conclude that Sox-2 might, in combination with appropriate cell-specific markers, constitute a useful marker to study the in vivo dynamics of the neural progenitor cell compartment also in the adult brain.

Animals↗

Central autonomic control of the bone marrow: multisynaptic tract tracing by recombinant pseudorabies virus.

Bone marrow is the primary place of hematopoiesis, where the development, survival and release of multipotent stem cells, progenitors, precursors and mature cells are under continuous humoral and neural control. Dense network of nerve fibers, containing various neurotransmitters is found in the bone marrow, however, the central neuronal circuit that regulates the activities of the bone marrow through these fibers remained unexplored. Transsynaptically connected neurons were mapped by virus-based transneuronal tracing technique using two isogenic, genetically engineered pseudorabies viruses, Bartha-DupGreen and Ba-DupLac expressing green fluorescent protein and beta-galactosidase, respectively. Bartha-DupGreen was injected into the femoral bone marrow of male rats and the progression of infection was followed 4-7 days post-inoculation. Virus-labeled cells were revealed in ganglia of the paravertebral chain and in the intermediolateral cell column of the lower thoracic spinal cord. Neurons were retrogradely labeled in the C1, A5, A7 catecholaminergic cell groups and several other nuclei of the ventrolateral and ventromedial medulla, the periaqueductal gray matter, the paraventricular and other hypothalamic nuclei, and in the insular and piriform cortex. Nerve transections and double-virus tracing from the bone marrow and the surrounding muscles were used to confirm the specific spreading of the virus. These results provide anatomical evidence for the CNS control of the bone marrow and identify putative brain areas, which are involved in autonomic regulation of the hematopoiesis, the release of progenitor cells, the blood supply and the immune cell function in the bone marrow.

Animals↗

Adult stem cell therapy beyond haemopoietic stem cell transplantation? An update.

The lifesaving potential of haemopoietic stem cell transplantation for the treatment of haematological malignancies and other life threatening disorders of the haemopoietic stem cell is universally accepted. In contrast, the use of adult marrow derived stem cells for tissue repair strategies in degenerative disease or after tissue damage are only in the early stages of evolution. A range of opinion exists within the general public and the scientific community about whether research with human embryonic stem cells is ethically acceptable. Further, the current paucity of human embryonic stem cell data has lead investigators to consider adult marrow as a potential source of stem cells to treat a wide range of degenerative disease and damaged tissues. Target disorders include osteoarthritis, diabetes mellitus, Parkinson's disease, ischaemic heart disease and retinal degeneration. Obvious advantages of this approach, if successful, would be fewer ethical hurdles compared with embryonic stem cells. Treatment with the patients own marrow stem cells would eliminate the possibility of allogeneic rejection.

Adult↗