Search PubMed⌕ Search

Biomedical subjects

S S Tan

Publications and source records attributed to S S Tan.

At least 37 records · Page 2Linked to original sources

Multiple cadherin mRNA expression and developmental regulation of a novel cadherin in the developing mouse eye.

Cadherins form a large family of transmembrane glycoproteins whose members include the classical cadherins, the desmosomal cadherins, and the protocadherins. The classical cadherins mediate homophilic cell-cell adhesion and are key regulators of many morphogenetic processes. More than a dozen classical cadherins are expressed in both the developing and the mature central nervous system. Although individual cadherins have been identified in the retina of various species, we wished to determine the range of cadherins expressed at distinct developmental stages in the mouse retina. Using a PCR-based cloning strategy, we detected 10 different classical cadherin mRNAs of both type I and type II subtypes. The most abundant cDNA was that encoding the type II cadherin, Cadherin-11. The other type II cadherins detected were VE- and T2-cadherin and Cadherin-6 and -12. Four type I cadherins, N-, R-, P-, and E-cadherin, were also present. One cadherin cDNA encoded a novel cadherin, called EY-cadherin for cloned from eye. EY-cadherin is most closely related to human Cadherin-14 (93% identical). EY-cadherin mRNA was detected in the adult mouse eye, brain, and testis with a 20-fold increase in expression levels in the embryonic head from E11 to E19 and a 50-fold increase in expression levels in the postnatal eye from PN1 to PN16. Multiple cadherin gene expression is consistent with the hypothesis that different cadherins regulate morphogenetic processes, such as neuronal migration and lamination, and determine the specific interneuronal connections found in the mature retina.

Amino Acid Sequence↗

Cloning and expression of mouse Cadherin-7, a type-II cadherin isolated from the developing eye.

We report the molecular cloning of Cadherin-7 from the embryonic mouse eye. The deduced amino acid sequence shows it to be a type-II cadherin similar to Xenopus F-cadherin and chick Cadherin-7. The mouse Cadherin-7 gene maps to chromosome 1, outside the conserved linkage group of cadherin genes on chromosome 8. Cadherin-7 is expressed throughout the entire period of neural development and mRNA levels are developmentally regulated in both the embryonic and the postnatal central nervous system (CNS). In adult mice, Cadherin-7 expression is restricted to the CNS, with highest levels in the retina. In the developing eye, Cadherin-7 mRNA is found only in the neural retina. It is expressed by all retinal neuroblasts from E11 onward, but becomes progressively restricted to neurons in the inner neuroblast and developing ganglion cell layers (GCL). In the adult retina it is confined to subpopulations of cells in the GCL and to amacrine cells in the inner part of the inner nuclear layer. This expression pattern suggests a role for Cadherin-7 in mouse retinal development, particularly in the formation and maintenance of the GCL.

Aging↗

Clonal expansion and cell dispersion in the developing mouse retina.

The present study has used two different approaches for labelling progenitor cells at the optic vesicle stage in order to examine patterns of clonal expansion and cellular dispersion within the developing retina. X-inactivation transgenic mice and chimeric mice expressing the lacZ reporter transgene were examined during development and in adulthood to study the radial and tangential dispersion of proliferating neuroepithelial cells and postmitotic retinal cells of known identities. Chimeric retinas were used to measure tangential dispersion distances, while transgenic retinas were used to assess the frequency of tangential dispersion for individual populations of retinal neurons. Tangential dispersion is shown to be a universal feature of particular retinal cell types, being contrasted with the strictly radial dispersion of other cells. Tangential dispersion is a relatively short-distance phenomenon, with distinct dispersion distances characteristic for cone, horizontal, amacrine and ganglion cells. Embryonic and postnatal retinas show that tangential dispersion occurs at different times for these distinct cell types, associated with their times of differentiation rather than their neurogenetic periods. These developmental results rule out the possibility that tangential dispersion is due to a passive displacement produced by the proliferation of later-born cells, or to the lateral dispersion of a dividing sibling; rather, they are consistent with the hypothesis that tangential dispersion plays a role in the establishment of the orderly spatial distribution of retinal mosaics.

Aging↗

Granule cell dispersion is restricted across transverse boundaries in mouse chimeras.

The granular layer of the developing and adult cerebellum is marked by the presence of several transverse boundaries, revealed in gene expression patterns or as a consequence of genetic mutations. It is unclear whether these boundaries represent fundamental differences between granule cell populations or if they are a secondary response to regional differences in the underlying Purkinje cells. One possibility is that boundaries mark different spatial domains of granule cells in a lineage-dependent fashion. To test this hypothesis, we have analysed a series of murine embryonic stem cell chimeras marked by the constitutive expression of beta-galactosidase in donor granule cells. The chimeras show a consistent spatial restriction boundary, located in the granular layer of lobule VI in the vermis and extending laterally into crus I of the hemispheres. A second boundary was found separating lobules IX and X in the vermis. No correlation was found between the genotypes of molecular layer interneurons and the underlying granule cells, suggesting that they arise independently. No transverse boundaries were observed for the molecular layer interneurons, consistent with the hypothesis that they are not generated from precursors in the external granular layer. These results indicate that the granular layer of the cerebellum comprises cellular domains with different histories separated by consistent spatial restriction boundaries.

Animals↗

Lineage and clonal development of gastric glands.

Individual gastric glands of the stomach are composed of cells of different phenotypes. These are derived from multipotent progenitor stem cells located at the isthmus region of the gland. Previous cell lineage analyses suggest that gastric glands, as in the colon and small intestine, are invariably monoclonal by adult stages. However, little is known about the ontogenetic progression of glandular clonality in the stomach. To examine this issue, we employed an in situ cell lineage marker in female mice heterozygous for an X-linked transgene. We found that stomach glands commence development as polyclonal units, but by adulthood (6 weeks), the majority progressed to monoclonal units. Our analysis suggests that at least three progenitor cells are required to initiate the development of individual gastric glands if they are analyzed just after birth. Hence, unlike the colon and small intestine, stomachs showed a significant fraction (10-25%) of polyclonal glands at adult stages. We suggest that these glands persist from polyclonal glands present in the embryonic stomach and hypothesize that they represent a subpopulation of glands with larger numbers of self-renewing stem cells.

Animals↗

Retarded postimplantation development of X0 mouse embryos: impact of the parental origin of the monosomic X chromosome.

About 12-17% of the embryos obtained by mating mice carrying the In(X)1H or Paf mutations are of the 39,X (X0) genotype. Depending on the mutant mice used for mating, the monosomic X chromosome can be inherited from the paternal (XP) or the maternal (XM) parent. The XP0 embryos display developmental retardation at gastrulation and early organogenesis. XP0 embryos also display poor development of the ectoplacental cone, which is significantly smaller in size and contains fewer trophoblasts than XX siblings. In contrast, XM0 embryos develop normally and are indistinguishable from XX littermates. In both types of X0 embryos, an X-linked lacZ transgene is expressed in nearly all cells in both the embryonic and the extraembryonic tissues, suggesting that X inactivation does not occur when only one X is present. Of particular significance is the maintenance of an active XP chromosome in the extraembryonic tissues where normally the paternal X chromosome is preferentially inactivated in XX embryos. The differential impact of the inheritance of X chromosomes from different parents on the development of the X0 embryos raises the possibility that the XP is less capable than the XM in providing the appropriate dosage of X-linked activity that is necessary to support normal development of the embryo and the ectoplacental cone. Alternatively, the development of the XP0 embryo may be compromised by the lack of activity of one or several X-linked genes which are expressed only from the maternal X chromosome. Without the activity of these genes, embryonic development may be curtailed even though all other loci on the XP chromosome are actively transcribed.

Animals↗

Clonal boundary analysis in the developing retina using X-inactivation transgenic mosaic mice.

Transgenic mice harboring the lacZ reporter gene on one X chromosome have been used to mark 50% of all retinal progenitors. The distribution of clones arising from this population of marked progenitors reveals a conspicuous columnar segregation of clonally related cells, indicating that most retinal neuroblasts migrate exclusively radially. Against this columnar background of the transgenic retina, single cone, horizontal, amacrine and ganglion cells are observed to transgress clonal borders, mixing freely with cells derived from different precursors. This tangential dispersion is due to the lateral movement of postmitotic neuroblasts around the time of their differentiation, rather than to the dispersion of a proliferative sibling at the time of cell birth. Tangential dispersion is suggested to play a significant role in creating the functional architecture of the mature retina, being the means by which the orderly spacing, or regularity, of retinal mosaics is established during development.

Animals↗

Separate progenitors for radial and tangential cell dispersion during development of the cerebral neocortex.

Cell lineage analyses suggest that cortical neuroblasts are capable of undertaking both radial and tangential modes of cell movement. However, it is unclear whether distinct progenitors are committed to generating neuroblasts that disperse exclusively in either radial or tangential directions. Using highly unbalanced mouse stem cell chimeras, we have identified certain progenitors that are committed to one mode of cell dispersion only. Radially dispersed neurons expressed glutamate, the neurochemical signature of excitatory pyramidal cells. In contrast, tangential progenitors gave rise to widely scattered neurons that are predominantly GABAergic. These results suggest lineage-based mechanisms for early specification of certain progenitors to distinct dispersion pathways and neuronal phenotypes.

Animals↗

Pattern formation in the cerebellum of murine embryonic stem cell chimeras.

The cerebellar cortex is subdivided into an elaborate, stereotyped array of transverse zones and parasagittal stripes. It has been speculated that (i) all Purkinje cells derive from 10 to 20 precursors allocated early in embryogenesis and (ii) that pattern formation is based on cell lineage restriction in the founder pool. These hypotheses have been tested by clonal analysis of embryonic stem cell chimeras. Neither speculation is supported: the analysis suggests that Purkinje cells derive from a founder population of > 102 precursors, and that neither cerebellar transverse developmental boundaries nor parasagittal stripes have a clonal origin. We conclude that early lineage restriction plays no role in cerebellar pattern formation.

Animals↗

Translocation breakpoint possibly predisposes to nonrandom X-chromosome inactivation in mouse embryos bearing Searle's T(X;16)16H translocation.

To clarify the sequence of events that ultimately achieves the nonrandom inactivation of the paternally inherited X chromosome in postpartum female mice heterozygous for T(X;16)16H, we set out to examine the expression of Xist alleles and the X-linked HMG-lacZ transgene in embryos recovered at the egg cylinder stage. Lack of expression of the Xist(b) allele on the 16X translocation chromosome in the embryonic region of 7.5 d postcoitum (dpc) X16/X(n)Xist(a);16(X)Xist(b)/16 embryos strongly suggested the occurrence of nonrandom inactivation in favor of the normal X chromosome. The simplest explanation would be biased choice, followed by postinactivation selection against genetically unbalanced cells. However, the frequency and distribution of beta-galactosidase-positive cells in X16/X(n)lacZ;16X/16 embryos at 6.5 and 7.5 dpc, together with earlier cytogenetic data, raised an intriguing possibility that the majority of 16X chromosomes were prevented from completing the inactivation process, when they had been chosen to be silenced. Phenotypes of female mice carrying a spontaneous recombination between Xn and 16X in the segment defined by the T16H breakpoint and the X-linked Ta locus suggested that the nonrandomness was brought about by disruption of an X-chromosomal sequence or structure at the translocation breakpoint.

Animals↗

Anaesthesia for tracheoplasty: case report and review of anaesthesia techniques.

Congenital tracheal stenosis is an uncommon cause of stridor and can be associated with other anomalies. It may present anytime during childhood depending on the severity of the lesion. The anaesthetist may be involved in all stages of management of this problem; from diagnostic investigations to surgical correction. Various methods of tracheoplasty have been described, some requiring cardiopulmonary bypass support and others may be carried out with conventional ventilatory techniques.

Anesthesiology↗

A survey of patients' knowledge of anaesthesia and perioperative care.

AIM: The present study was undertaken to assess patients' knowledge of anaesthesia and perioperative care as well as their perception of anaesthetics and their role. METHODOLOGY: A questionnaire survey of 23 items was developed and completed pre-operatively by 132 patients. Included in this study were 45 men and 87 women; 80% with at least secondary education with a mean age of 39.9 and 35.6 years, respectively. RESULTS: Results indicated that only 56.8% of patients understood that anaesthetists are doctors. There were significant misconceptions though in general, knowledge was good. Although all patients realised the importance of fasting, however only 74.2% of patients realised it meant limitation of both solids and liquids from a specified time. Pain was the major concern among the patients surveyed (39.4%), followed by fear of not waking up after surgery (18.9%). Twenty-one per cent of patients believed that postoperative pain was a necessary part of the healing process and 28% thought that pain was something that they had to put up with in the postoperative period. Only 23.5% correctly identified the anaesthetist as being responsible for analgesia in the recovery period. A majority (75.8%) wished to have more information about anaesthesia. CONCLUSION: When forthcoming surgery and anaesthesia are discussed, it is important that patients realise that they are being cared for by anaesthetists who are doctors and efforts must be taken to educate the profession and public on the anaesthetists' role in perioperative care. This should enhance the professional image of anaesthesia and more importantly, improve patient confidence and quality of care.

Adult↗

The expression patterns of guidance receptors, DCC and Neogenin, are spatially and temporally distinct throughout mouse embryogenesis.

To gain a better understanding of the role of DCC and Neogenin in neural and nonneural tissues during vertebrate development we have carried out in situ hybridization studies to determine their expression patterns throughout the mid to late stages of mouse embryogenesis. This analysis revealed striking contrasts in both the spatial and temporal expression patterns of these closely related molecules. While DCC mRNA expression was predominantly restricted to the developing central nervous system (CNS), Neogenin mRNA was detected in a broad spectrum of embryonic tissues. Outside the CNS, Neogenin expression was observed mainly in mesodermal derivatives such as organ primordia and cartilage condensations of many developing embryonic structures. Within the CNS, initiation of DCC expression correlated with the onset of neurogenesis and was maintained at high levels in all regions of the developing CNS actively undergoing neurogenesis. By E18.5, DCC expression was detected only in structures such as the olfactory bulb, the hippocampus, and the cerebellum, that are known to sustain active neurogenesis well into postnatal life. In contrast, Neogenin expression was weak in the early developing CNS but broadened and intensified as neurogenesis proceeded. In summary, these observations indicate that Neogenin is the predominant member of this subfamily in mesodermal tissues, while DCC and Neogenin may play complementary roles in the generation of the fully functional CNS.

Animals↗

Mosaics of islet-1-expressing amacrine cells assembled by short-range cellular interactions.

The nervous system has a modular architecture with neurons of the same type commonly organized in nonrandom arrays or mosaics. Modularity is essential to parallel processing of sensory information and has provided a key element for brain evolution, but we still know very little of the way neuronal mosaics form during development. Here we have identified the immature elements of two retinal mosaics, the choline acetyltransferase (ChAT) amacrine cells, by their early expression of the homeodomain protein Islet-1, and we show that spatial ordering is an intrinsic property of the two Islet-1 mosaics, dynamically maintained while new elements are inserted into the mosaics. Migrating Islet-1 cells do not show this spatial ordering, indicating that they must move tangentially as they enter the mosaic, under the action of local mechanisms. Clonal territory analysis in X-inactivation transgenic mice confirms the lateral displacement of ChAT amacrine cells away from their clonal columns of origin, and mathematical models show how short-range cellular interactions can guide the assemblage of these mosaics via a simple biological rule.

Animals↗

X-chromosome inactivation during the development of the male urogenital ridge of the mouse.

In the mouse, the activity of Sry (sex-determining gene on the Y chromosome) initiates the transformation of the indifferent gonad into a testis. In humans, a partial Xp21 duplication leads to the development of ovaries instead of testes in XY individuals. This observation indicates that sex determination might also be influenced by a gene-dosage compensation mechanism, in addition to a dominant action of the Sry gene. In female mammals, the regulation of X-linked gene dosage at early embryogenesis is achieved through the inactivation of one of the two X chromosomes. Here we have investigated the possibility that inactivation of the X chromosome may play a role in male sex determination. We have shown, using an X-linked lacZ transgenic mouse line, that loss of beta-galactosidase activity occurs in certain somatic cells of the developing male urogenital ridge. When changes associated with apoptosis of mesonephric tubules in the developing urogenital ridges are taken into account, expression of the Xist (X inactive specific transcript) gene correlates with X inactivation revealed by loss of beta-galactosidase activity in very early mesonephric tubule epithelial cells, gonadal interstitial mesenchymal cells and coelomic epithelial cells.

Animals↗

A case of fluid embolism from transcervical endometrial resection.

Hysteroscopic transcervical endometrial resection (TCER) using a urological resectoscope offers an alternative to hysterectomy for some women with menorrhagia. The advantages of this technique are a shorter operating time and a rapid recovery and discharge. However, this procedure is not without risk of complications and one of them is related to dilutional hyponatraemia from absorption of the irrigation solution. This report describes a patient who experienced such a problem.

Adult↗

Leukaemia inhibitory factor or related factors promote the differentiation of neuronal and astrocytic precursors within the developing murine spinal cord.

Previously we have shown that leukaemia inhibitory factor (LIF) potentiates the development of murine spinal cord neurons in vitro, suggesting that it, or related factors, may play an important regulatory role in neuronal development. We have further investigated this role and show here that the generation of neurons in cultures of embryonic day 10 spinal cord cells is inhibited by antibodies to the beta subunit of the LIF receptor. Since there are more undifferentiated precursors in antibody-treated cultures than in control and LIF-treated cultures, it is concluded that the primary action of LIF, or related molecules, is to promote neuronal differentiation, not precursor survival. In addition, the failure of LIF to support neuronal survival in the period immediately following differentiation suggests that the increased numbers of neurons generated with LIF are not attributable to its neurotrophic action. By selecting neuronal precursors on the basis of their inability to express class 1 major histocompatibility complex molecules, it was shown that LIF acted directly upon these cells and not via an intermediary cell. LIF also appears to be involved in regulating the differentiation of astrocytes, since it increases the number of glial fibrillary protein (GFAP)-positive cells present in the cultures and since the spontaneous production of GFAP-positive cells is blocked by antibodies to the LIF beta receptor. These findings suggest that LIF or related factors promote the differentiation of neural precursors in the spinal cord, but that they are not involved in preferentially promoting precursors down a specific differentiation pathway.

Animals↗