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Biomedical subjects

S S Tan

Publications and source records attributed to S S Tan.

At least 73 records · Page 4Linked to original sources
PubMed1993

Radial mosaicism and tangential cell dispersion both contribute to mouse neocortical development.

The mammalian neocortex is generated by waves of migrating cells originating from the ventricular zone. Radial migration along radial glia has been proposed as the dominant mechanism for this process. The radial unit hypothesis is poorly supported by retroviral lineage studies, however, and although some clones show limited radial organization, the emphasis appears to be on widespread tangential dispersion. Here we investigate the pattern of cortical cell dispersion using transgenic mice in which roughly half of the brain cells are coloured by a transgene. We find that the neocortex is randomly divided into diffused bands, the majority of cells within each band have the same colour, and their radial orientation suggests radial dispersion. Superimposed upon this was a significant contribution by tangentially dispersed cells that did not respect clonal borders. These observations indicate that cortical specification is not dependent upon a single mechanism of cell allocation, but that both radial mosaicism and tangential cell migration are involved.

Animals↗
PubMed1993

Adhesion molecules in neural crest development.

Peripheral nerve cells, various endocrine and pigment cells and cranial connective tissue cells of vertebrates stem mainly from the embryonic neural crest. This originates with the central nervous system, but the crest cells detach from this tissue, via a decrease of cell-cell adhesion involving, particularly, a reduction of the adherens junction cell adhesive molecule A-CAM. This epithelio-mesenchymal transformation allows crest cells to migrate along pathways that are defined partly by the distribution of substrate adhesion molecules, the archetype being fibronectin, an extracellular matrix molecule recognized by integrin receptors on crest cells. Many other molecules, however, may act in the same way. In contrast, some molecules may define migration pathways by reducing adhesion; chondroitin sulfate proteoglycan is a candidate for this role. Pathway selection is most likely achieved by balanced combinations of molecules that promote and reduce adhesion. Cessation of migration, in the case of the nervous ganglia, correlated with re-expression of cell-cell adhesion molecules like A-CAM and others, consistent with an adhesive basis, although functional tests have not yet been performed. The development of the neural crest system provides a useful model that emphasizes the role of adhesion in morphogenesis.

Amino Acid Sequence↗
PubMed1993

X-chromosome inactivation occurs at different times in different tissues of the post-implantation mouse embryo.

During mammalian development, one of the two X chromosomes in female embryos is randomly inactivated in the somatic cell in order to achieve gene dosage compensation. But is X inactivation established simultaneously or is it accomplished over time in a lineage-dependent fashion? We have examined this question in mouse embryos carrying an X-linked lacZ transgene. This transgene is subject to inactivation and the loss of beta-galactosidase activity provides a direct indication of X inactivation in individual cells. We find that X inactivation proceeds with different schedules in different somatic tissues, and the notochord, the heart, cranial mesoderm and the hindgut are among the last tissues to undergo X inactivation.

Animals↗
PubMed1993

Murine complement receptor gene expression: Cr2 gene transcripts are depressed during a high dose microbial challenge.

The murine Cr2 gene encodes mRNA that produce two protein products predicted to be approximately 145,000 M(r) (Cr2-145) and 190,000 M(r) (Cr2-190). All cells examined which express the Cr2 gene produce transcripts encoding both the Cr2-145 and Cr2-190 proteins: both transcripts are constitutively expressed by mature B cells. To determine if Cr2 expression could be altered by activating splenic B cells, splenic cultures were incubated with lipopolysaccharide (LPS) and cell surface Ig chains were cross-linked with anti-mu. In the presence of LPS and anti-mu both Cr2 and Oct-2 transcripts were diminished while the control beta-actin transcript levels remained unchanged. However, when LPS alone was added, only the Cr2 transcript levels were diminished. To test if these findings could be reproduced in vivo, animals were provided with a peritoneal injection of either Escherichia coli or Listeria monocytogenes and transcript levels analysed. The quantities of both Cr2 transcripts, as well as those encoding Oct-2, were substantially reduced in splenocytes and peripheral lymphatic tissues obtained from these infected mice while those encoding the mouse Crry protein, the B-cell marker CD19 and beta-actin remained unchanged. These data suggest that when confronted with a major bacterial infection, murine B cells respond by shutting down synthesis of transcripts encoding the Cr2 and Oct-2 gene products.

Animals↗
PubMed1992

Identification of sites for distinct DNA binding proteins including Oct-1 and Oct-2 in the Cr2 gene.

The murine Cr2 gene produces two distinct products in a variety of murine cell types. Both of these transcripts appear to initiate from the same position within the gene but vary from one another via an alternative splicing event within the coding exons. An analysis of those gene sequences that might control the cell specific expression of the Cr2 gene has identified a region of Cr2 5' of the transcription start site that is conserved in both the murine Cr2 and human CR2 genes. When this region was examined using the gel shift assay with nuclear extracts from cells expressing Cr2 (B cells) and those that do not (T cells and fibroblasts), at least four distinct proteins were identified that bound to at least three distinct sites. The DNA sequence recognized by two of these proteins is the octamer sequence recognized by a family of transcriptional regulators including the B cell specific Oct-2 protein. During an acute bacterial infection, the levels of Oct-2 and Cr2 mRNA are both depressed. This suggests that the Oct-2 protein directly controls the transcriptional activity of the Cr2 gene and that during such an infection, the levels of Ag receptors on B cells (Ig and complement receptors) are diminished.

Animals↗
PubMed1992

Development of a sensitive reverse transcriptase PCR assay, RT-RPCR, utilizing rapid cycle times.

We describe a procedure to analyze rare gene transcripts quantitatively using a reverse transcriptase-polymerase chain (RT-PCR) reaction. RNA purified from cells and tissues is reverse-transcribed using random hexamer primers and is amplified using very short cycle times. The products are trace-labeled with [32P[dCTP, which allows for the quantitation of the products by gel electrophoresis and excision of bands. The quantity of the PCR product is directly proportional to the quantity of cDNA added and is reproducible from a single cDNA preparation or from samples derived from separate preparations. Because cDNA synthesis is primed with random oligonucleotides, the same cDNA sample preparation can be examined for many different gene products.

Animals↗
PubMed1992

The somitogenetic potential of cells in the primitive streak and the tail bud of the organogenesis-stage mouse embryo.

The developmental potency of cells isolated from the primitive streak and the tail bud of 8.5- to 13.5-day-old mouse embryos was examined by analyzing the pattern of tissue colonization after transplanting these cells to the primitive streak of 8.5-day embryos. Cells derived from these progenitor tissues contributed predominantly to tissues of the paraxial and lateral mesoderm. Cells isolated from older embryos could alter their segmental fate and participated in the formation of anterior somites after transplantation to the primitive streak of 8.5-day host embryo. There was, however, a developmental lag in the recruitment of the transplanted cells to the paraxial mesoderm and this lag increased with the extent of mismatch of developmental ages between donor and host embryos. It is postulated that certain forms of cell-cell or cell-matrix interaction are involved in the specification of segmental units and that there may be age-related variations in the interactive capability of the somitic progenitor cells during development. Tail bud mesenchyme isolated from 13.5-day embryos, in which somite formation will shortly cease, was still capable of somite formation after transplantation to 8.5-day embryos. The cessation of somite formation is therefore likely to result from a change in the tissue environment in the tail bud rather than a loss of cellular somitogenetic potency.

Animals↗
PubMed1991

Cytotactin expression in somites after dorsal neural tube and neural crest ablation in chicken embryos.

The spatiotemporal expression of the extracellular matrix protein cytotactin/tenascin during somitogenesis suggests that it plays a role in the morphogenetic events that give rise to the pattern of neural crest (NC) development. In the present study, the spatial distribution and molecular forms of cytotactin in somites were examined using in situ hybridization, Western blotting, and immunohistochemistry during normal development and after injury. In situ hybridization showed that prior to NC cell invasion cytotactin mRNA was restricted to the caudal half of the newly formed epithelial somites. As each epithelial somite matured, giving rise to a sclerotome and dermamyotome, the mRNA was first restricted to the dermamyotome and later restricted to the rostral protion of the sclerotome, consistent with the previously reported protein distribution. Immunocytochemical analysis of the distribution of cytotactin and NC cells in embryos with ablations that removed NC cells, or with simple wounds that left NC cells in place, demonstrated that the presence of NC cells is neither necessary nor sufficient for the correct positioning of cytotactin. Immunoblotting analysis showed that cytotactin synthesized by sclerotomes in the absence of NC cells was of similar molecular mass to that produced in their presence. These findings are in accord with the notion that the abnormalities of cytotactin distribution are related to the wounding process. We conclude that, contrary to the suggestion of Stern et al. [Stern, C. D., Norris, W. E., Bronner-Fraser, M., Carlson, G. J., Faissner, A., Keynes, R. J. & Schachner, M. (1989) Development 107, 309-319], there is no causal link between the presence of NC cells and the distribution and molecular mass of sclerotomal cytotactin.

Animals↗
PubMed1991

Liver-specific and position-effect expression of a retinol-binding protein-lacZ fusion gene (RBP-lacZ) in transgenic mice.

Transgenic mice carrying a liver-specific promoter fused to a nuclear-targeted lacZ reporter gene were generated. Three separate lines of mice showed liver expression in the adult and no expression elsewhere in the animal. These results show that a 1.7 kb 5'-flanking region in the retinol-binding protein gene contains necessary and sufficient transcriptional signals for expression in adult livers. A fourth line (R197) did not express the transgene in the liver; instead, constitutive lacZ expression was seen during postimplantation stages of development from Day 9.5 onwards and appeared to be associated with segmented structures including somites, branchial arches, and hindbrain rhombomeres until late fetal stages. The beta-galactosidase positive cells in R197 were later seen to give rise to facial and flank musculature, and to other region-specific subpopulations of the jaws, neocortex, and brain stem. Northern blot analysis for the host retinol-binding protein RNA transcript did not show overlapping tissue expression with the reporter gene and suggests that transgenic phenotype seen in segmented embryonic structures of R197 and other extra-hepatic sites is from novel cis-acting transcriptional specificity. RNase protection assays of the R197 mRNA indicate that the lacZ sequences are appropriately transcribed downstream of the RBP canonical TATA box, even though the RBP promoter is itself silent. This result would suggest host flanking sequences with enhancer activity may have either activated the lacZ reporter gene or cooperated with the RBP promoter to create novel region-specific transcription. Breeding experiments have so far failed to produce offsprings that are homozygous for the transgene, and mating of transgenic F1 siblings routinely produce reduced litter sizes. Embryos that are homozygous for the transgene appear to be unable to survive beyond the egg cylinder stages of development. Thus, disruption of the host genome by insertional mutation appears to be manifest at an earlier stage than when position-effect expression of the transgene is first apparent. These experiments demonstrate that the component parts of a transgene may be subject to differential activation or suppression by host genomic flanking sequences and that even a strong, tissue-specific promoter may be overridden by host genes.

Animals↗
PubMed1991

Removal of a laryngeal foreign body using high frequency jet ventilation.

Aspiration of a foreign body into the respiratory tract is a common and serious accident in childhood. Laryngotracheal foreign bodies, although less common than bronchial foreign bodies, are potentially more dangerous. Removal is commonly achieved using a rigid ventilating bronchoscope. We report a 16-month-old boy who had an open safety pin impacted in his larynx. This was removed through a tracheostomy, using high frequency jet ventilation to maintain gaseous exchange. We believe that this is the first case in which this method of removal has been reported.

Bronchoscopy↗
PubMed1990

Expression of chicken liver cell adhesion molecule fusion genes in transgenic mice.

The tissue-specific expression of the chicken liver cell adhesion molecule (L-CAM) was studied by generating transgenic mice. The rat insulin II promoter was fused to a chicken L-CAM cDNA or to chicken genomic L-CAM sequences. Mice carrying the cDNA showed no expression of L-CAM. Mice carrying L-CAM genomic sequences showed expression in the beta cells of the pancreas, suggesting that sequences in introns or in flanking regions are required for expression. Murine L-CAM was undetectable in the beta cells of the pancreas of those transgenic mice expressing chicken L-CAM and thus appeared to be down-regulated, but expression of the mouse protein was not altered at other sites. Chicken L-CAM was also found in extrapancreatic tissues such as skin, kidney, liver, lung, intestine, blood vessels, and the choroid plexus and leptomeninges of the central nervous system. These findings raised the possibility that the chicken L-CAM gene contains cis regulatory elements that interfere with the specificity of a tissue-specific promoter such as the rat insulin promoter. To test this hypothesis, transgenic mice were produced with a construct containing the murine neurofilament promoter fused to genomic chicken L-CAM sequences. Chicken L-CAM was expressed in the brain and spinal cord, where L-CAM is not normally found, but it was also found in some nonneural tissues (kidney, liver, intestine, lung) in which L-CAM is normally expressed. The combined results suggest that tissue-specific cis-acting elements in the chicken L-CAM gene, when combined with heterologous promoters/enhancers, can generate novel patterns of gene expression.

Animals↗
PubMed1989

Cytotactin and its proteoglycan ligand mark structural and functional boundaries in somatosensory cortex of the early postnatal mouse.

The expression of the extracellular matrix molecules cytotactin, which is synthesized by glia, and cytotactin-binding (CTB) proteoglycan, which is synthesized by neurons, was examined in the developing brain of the mouse, specifically in the cortical barrel field, using highly specific polyclonal antibodies to the purified molecules. Both molecules appeared early in the development of the cortex but were excluded from the centers of the developing barrels at the time of entry and arborization of thalamocortical axons. Of the two major forms of cytotactin (220 and 200 kDa), the larger form predominated during development of the mouse brain and also predominated in mixed neuron-glia cultures but not in pure glial cultures. Both cytotactin and CTB proteoglycan were recognized by various lectins that have been shown in other studies to demarcate the barrel field: both molecules were recognized by lentil lectin and concanavalin A and CTB proteoglycan was also recognized by peanut and wheat germ agglutinins. The HNK-1 carbohydrate antigen, present on cytotactin, CTB proteoglycan, and other adhesion molecules, was also found in the barrel walls and diminished in the barrel hollows. Cytotactin and CTB proteoglycan were preferentially expressed in barrel walls through P12. After this time, their expression became uniform even though the histological pattern of barrel walls and hollows was maintained. The fusion of a row of barrels which results from peripheral damage to a row of whiskers was accompanied by the loss of patterned expression of both molecules following electrocauterization of a row of whisker follicles at P1.5. We conclude that activity from the periphery is important not only to development of anatomical pattern but also of the molecular pattern and that the expression of both glial and neuronal proteins can respond to such activity. The results are consistent with previous studies showing that incoming thalamocortical axons play a primary role in barrel field formation. They also suggest that both the migration of cortical neurons on glia and the refinement of the mapping between the peripheral whisker field and its cortical representation may depend upon the distribution of substrate adhesion molecules.

Age Factors↗
PubMed1987

Asymmetric expression in somites of cytotactin and its proteoglycan ligand is correlated with neural crest cell distribution.

The development of the vertebrate neural crest presents a particularly challenging problem in pattern formation. Several studies have revealed that a population of neural crest cells penetrates the sclerotomal mesenchyme of the somite only in its rostral half. In a search for molecular correlates of this pattern, we have observed that cytotactin and a chondroitin sulfate proteoglycan, two interactive extracellular matrix molecules, show a specialized distribution within the sclerotome. Cytotactin was localized in the rostral half of the sclerotome at about the time of neural crest cell invasion. The proteoglycan was initially diffuse throughout the sclerotome but became restricted to the caudal half after the appearance of cytotactin and invasion of neural crest cells in the rostral half. These distributions were crest cell-independent; they occurred on the same schedule even when all crest cells were removed by surgical extirpation of the neural tube. Furthermore, in tissue culture, somite cells synthesized high levels of both molecules. In vitro, crest cells rounded up in the presence of these molecules and cell migration assays revealed that neither cytotactin nor proteoglycan alone was as good a substratum for crest cell migration as fibronectin. In combination with fibronectin, however, cytotactin or proteoglycan only restricted cell movement but did not prevent it. Taken together, these observations support the hypothesis that cytotactin and the chondroitin sulfate proteoglycan may contribute to pattern formation during embryogenesis by means of their site-restricted distribution, their ability to alter migration on other substrates such as fibronectin, and their ability to induce cell-surface modulation.

Animals↗
PubMed1987

Multilocular renal cyst. A hamartoma with potential for neoplastic transformation?

Multilocular renal cyst is a pathologically distinctive lesion of uncertain pathogenesis that usually presents as a large abdominal mass in male infants or middle-aged women. We performed detailed light and electron microscopic studies of three nephrectomy specimens that contained multilocular cysts and attempted to correlate their morphology with various theories of pathogenesis. In one of our cases, a microscopic focus of clear cells that resembled renal cell adenocarcinoma was observed in the wall of one locule. Although such a finding has been reported previously, the lack of well-documented cases with metastases suggests that the prognosis for patients with this lesion remains uniformly excellent.

Adult↗
PubMed1986

Analysis of cranial neural crest cell migration and early fates in postimplantation rat chimaeras.

Rat embryos were grown in vitro during the period of cranial neural crest cell migration. In order to study the pathways and positional fates of cells from different regions of the neural crest, labelled premigratory crest cells from donor embryos were microinjected orthotopically into host embryos of the same developmental stage except for area 1 (forebrain) grafts which were, for technical reasons, injected into area 2. After various periods of time in whole embryo culture, the embryos were examined by immunohistochemical staining in order to determine the new positions of the labelled cells, and a map of their migration pathways was constructed. The observed patterns of migration were consistent with predictions from morphological studies in mammals and with extrapolations from transplantation studies in birds. However, crest cell migratory behaviour in rat and chick embryos was not identical; possible reasons for this are discussed.

Animals↗
PubMed1985

The development and distribution of the cranial neural crest in the rat embryo.

The head region of rat embryos was investigated by scanning electron microscopy after removal of the surface ectoderm with adhesive tape. Observations were made in embryos from 6-somite to 11-somite stages of development, in order to determine: (1) the sequence of emigration of neural crest cells from the different regions of the future brain; (2) the appearance of crest cells before, during, and after their conversion from an epithelial to a mesenchymal form; (3) the migration pathways. Emigration occurs first from the midbrain, and next from the rostral hindbrain; crest cells from these two regions migrate into the first visceral arch. Subsequently cells emigrate from the caudal hindbrain, but not in a rostrocaudal sequence. At the time of crest cell emigration, the neural fold morphology varies from a slightly convex, widely open plate (midbrain) to a closed tube (caudal hindbrain). Thus the timing of emigration is related neither to age (as reflected in rostrocaudal levels) nor to morphology of the neural epithelium.

Animals↗