PubMed Health⌕ Search

Biomedical subjects

Lu Teng

Publications and source records attributed to Lu Teng.

10 recordsLinked to original sources

Expression and shifting subcellular localization of the transcription factor, Foxd3, in embryonic and adult pancreas.

Multipotent progenitor cells self renew throughout an animal's lifetime and can differentiate to give rise to different cell types. Before we can fully understand the developmental potential of progenitor cells and control their differentiation both in vivo and in vitro as stem cells, identification and characterization of the genes that control stem cell fate must first be obtained. Foxd3, a member of the forkhead family of transcriptional regulators, is required for the maintenance of embryonic stem cells and trophoblast stem cells of the early mouse embryo. We describe here the expression of this protein in the developing pancreas. Foxd3 is expressed in most beta cells and infrequently in alpha and PP cells but is not expressed in somatostatin cells. The subcellular localization of Foxd3 varies with fat content in the diet; with a high fat diet the protein is found primarily in the cytoplasm while a low fat diet results in nuclear localization. Foxd3 is differentially localized in a rat model of diabetes: it is nuclear in ZDF rats but cytoplasmic in their lean counterparts. Foxd3 is nuclear in Lep(Ob/Ob) mice.

Animals↗

Neural crest stem cells.

Stem cells are defined by their ability to both self-renew and give rise to multiple lineages in vivo and/or in vitro. As discussed in other chapters in this volume, the embryonic neural crest is a multipotent tissue that gives rise to a plethora of differentiated cell types in the adult organism and is unique to vertebrate embryos. From the point of view of stem cell biology, the neural crest is an ideal source for multipotent adult stem cells. Significant advances have been made in the past few years isolating neural crest stem cell lines that can be maintained in vitro and can give rise to many neural crest derivatives either in vitro or when placed back into the context of an embryo. The initial work identifying these stem cells was carried out with premigratory neural crest from the embryonic neural tube. Later, neural crest stem cells were isolated from postmigratory neural crest, presumably more restricted in developmental potential. More recently it has been demonstrated that neural crest stem cell progenitors persist in the adult in at least two differentiated tissues, the enteric nervous system of the gut and the whisker follicles of the facial skin. In all cases, the properties of the stem cells derived reflect their tissue of origin and the potential of the progenitors becomes more restricted with age. In this chapter we will review this work and speculate on future possibilities with respect to combining our knowledge of neural crest gene function in the embryo and the manipulation of adult neural crest stem cells in vitro and eventually in vivo.

Animals↗

Newly recognized physiologic and pathophysiologic actions of the angiotensin-converting enzyme.

Despite several decades of research into the renin-angiotensin system, new aspects of this endocrine system are elucidated every few years, expanding its role not only in hypertension but also in diabetes, oncology, and cardiology. In this review, we describe newly recognized physiologic actions of the angiotensin-converting enzyme (ACE). These include the role of local versus systemic ACE in maintaining blood pressure, the physiology of bradykinin accumulation during ACE inactivation, and the role of alternate "non-angiotensin" substrates and potential non-enzymatic properties of ACE.

Angiotensin-Converting Enzyme Inhibitors↗

The use of knockout mouse technology to achieve tissue selective expression of angiotensin converting enzyme.

The resin angiotensin system (RAS) plays an essential role in blood pressure regulation and electrolyte homeostasis. The effecter peptide of the RAS, angiotensin II, is produced by angiotensin converting enzyme (ACE) in multiple tissues. Genetic deletion of ACE in mice resulted a phenotype of low blood pressure, anemia and kidney defects. However, it is not clear whether the lack of the systemic or the local production of angiotensin II caused these defects. To understand the role of local angiotensin II production, we developed a method to achieve tissue specific ACE expression through homologous recombination. In this review, we discuss mouse models in which endothelial ACE was eliminated and replaced by hepatic ACE. These studies suggest that both circulating angiotensin II and local angiotensin II production play a role in angiotensin II generation; the elimination of local angiotensin II generation up-regulates systemic production and maintains physiologic homeostasis.

Angiotensin II↗

Circulating versus local angiotensin II in blood pressure control: lessons from tissue-specific expression of angiotensin-converting enzyme (ACE).

The renin angiotensin system (RAS) is a central player in blood pressure control. Its effector peptide, angiotensin II, regulates blood pressure through coordinated actions in multiple tissues. The RAS is generally considered to be an endocrine system, and angiotensin II to be a circulating hormone. In recent years, however, a role for locally produced angiotensin II has been proposed. The major site for angiotensin II production is endothelium, where angiotensin-converting enzyme (ACE) is abundantly expressed. To elucidate the relative importance of circulating angiotensin II versus locally produced angiotensin II, one approach is to create a mouse model in which ACE is expressed in a tissue-specific manner. In this review, we discuss strategies to create such a model. In a mouse model we generated using a novel promoter-swapping technique, the endothelial ACE is eliminated and replaced by ectopic production of ACE in the liver. This model specifically addresses the question of whether local production of angiotensin II is essential for RAS function.

Angiotensin II↗

[Establishing mouse embryonic stem cell line carrying a fluorescent undifferentiated marker].

To label mouse ES cells,a cell line derived from the inner cell mass of 3.5-day blastocysts,with enhanced green fluorescent protein (EGFP), the vector of pRex-1-EGFP was transferred into mouse ES cells by electroporation. The expressions of Rex-1 in undifferentiated and differentiated ES cells were detected by the microscopic observation of EGFP and by RT-PCR. The results showed that the EGFP gene was transferred into the mouse ES cell line, and the transfected cells in undifferentiated state showed high levels of EGFP expression. When the cells began to differentiate, the EGFP expressions were gradually reduced. A mouse ES cell line expressing EGFP under the control of Rex-1 gene promoter was generated. The cell line provides a powerful approach for the research of the process of mammalian development and for the screening of small molecules that can regulate this process.

Animals↗

The labeling of C57BL/6j derived embryonic stem cells with enhanced green fluorescent protein.

OBJECTIVE: To labele MESPU35, a embryonic stem (ES) cell line derived from C57BL/6j mouse, with enhanced green fluorescent protein (EGFP) for further application. METHODS: The EGFP gene was controlled by the hybrid CA promoter/enhancer (CMV enhancer/chicken beta-actin promoter/beta-actin intron) to construct the vector of the transgene, pCA-EGFP. The vector was transfected into MESPU35 by electroporation. RESULTS: We generated EGFP expressing ES cells demonstrating normal properties. The green fluorescence of EGFP expressing cells was maintained in propagation of the ES cells for more than 30 passages as well as in differentiated cells. Cultured in suspension, the "green" ES cells aggregated, and formed embryoid bodies maintaining the green fluorescence at varying developmental stages. The "green" embryoid bodies could expand and differentiate into various types of cells, exhibiting ubiquitous green fluorescence. CONCLUSIONS: The hybrid CA promoter/enhancer used to control the EGFP expressing ES cells, resulted in more intense and ubiquitous activity. The EGFP transfected cells yield bright green fluorescence, which can be visualized in real time and in situ. In addition, the ES cells, MESPU35, are derived from C57BL/6j mice, which are the most widely used in oncology, physiology and genetics. Compared to 129 substrains, C57BL/6j mice avoid a number of potential problems apparent in the other strains.

Animals↗

Labeling embryonic stem cells with enhanced green fluorescent protein on the hypoxanthineguanine phosphoribosyl transferase locus.

OBJECTIVE: To label embryonic stem (ES) cells with enhanced green fluorescent protein (EGFP) on the hypoxanthineguanine phosphoribosyl transferase (HPRT) gene locus for the first time to provide a convenient and efficient way for cell tracking and manipulation in the studies of transplantation and stem cell therapy. METHODS: Homologous fragments were obtained by polymerase chain reaction (PCR), from which the gene targeting vector pHPRT-EGFP was constructed. The linearized vector was introduced into ES cells by electroporation. The G418(r)6TG(r) cell clones were obtained after selection with G418 and 6TG media. The integration patterns of these resistant cell clones were identified with Southern blotting. RESULTS: EGFP expressing ES cells on the locus of HPRT were successfully generated. They have normal properties, such as karyotype, viability and differentiation ability. The green fluorescence of EGFP expressing cells was maintained in propagation of the ES cells for more than 30 passages and in differentiated cells. Cultured in suspension, the "green" ES cells aggregated and formed embryoid bodies, retaining the green fluorescence at varying developmental stages. The "green" embryoid bodies could expand and differentiate into various types of cells, exhibiting ubiquitous green fluorescence. CONCLUSIONS: This generation of "green" targeted ES cells is described in an efficient protocol for obtaining the homologous fragments by PCR. Introducing the marker gene in the genome of ES cells, we should be able to manipulate them in vitro and use them as vehicles in cell-replacement therapy as well as for other biomedical and research purposes.

Animals↗

[Researches on the methodology of establishment of BALB/c murine embryonic stem cell lines].

Several methods and processes of establishment and culture of BALB/c mouse ES cell lines were discussed detailedly. A new method to establish and culture ES cell lines derived from BALB/c mouse was set up successfully using mouse embryonic fibroblast feeder layer and rat-heart-cell-conditioned medium (RH-CM). These culture conditions not only maintain the undifferentiated state and normal diploid karyotypes of BALB/c mouse ES cells effectively, but also maintain a series of their characteristics of murine stem cells. two different kinds of digestive methods and Two kinds of digestive juice with different concentrations were designed to dissociate proliferous inner cell mass (ICM) and ES cell colonies derived from dissociative ICM. two different kinds of digestive methods are "single time dissociation method" and "several times dissociation method", two kinds of digestive juice are 0.25% Trypsin-0.04% EDTA and 0.05% Trypsin-0.008% EDTA. At the same time, appropriate dissociated occasion of ICM and the effect of RH-CM on establishment and culture of BALB/c mouse ES cell lines were discussed. The results suggested that it is a reasonable method to establish BALB/c mouse ES cell lines using low concentration digestive juice and "several times dissociation method" to dissociate ICM after 4 days' proliferation. Judged by the form of ES cells and its colonies, proliferous capability, karyotypes examine, alkaline phosphatase activity assay and differentiation capability in vitro and in vivo, the 9 ES cell lines that we established satisfied the all traits of murine ES cell line.

Animals↗

Effect of emodin on proliferation and differentiation of 3T3-L1 preadipocyte and FAS activity.

OBJECTIVE: To study the effects of emodin on proliferation and differentiation of 3T3-L1 preadipocyte and the possible mechanism. METHODS: Cell proliferation was determined by MTT spectrophotometry, cell differentiation was determined by Oil Red O staining,and fatty acid synthase (FAS) activity was determined by spectrophotometry. RESULTS: Emodin promoted proliferation of 3T3-L1 preadipocyte at low concentration and inhibited the proliferation at high concentration in a dose-related manner. In contrast, it inhibited cell differentiation into adipocyte at low concentration in a dose-related manner. In vitro emodin inhibited the activity of FAS in a dose-related manner. CONCLUSIONS: The effects of emodin on 3T3-L1 cell's proliferation and differentiation are dose dependent. Emodin inhibits the activity of FAS. Our results suggest that emodin should have a potential to serve as a fat-reducing drug.

3T3 Cells↗