المؤلفون: Steven M. Cohn, Kevin A. Roth, E H Birkenmeier, T. C. Simon, Jeffrey I. Gordon

المصدر: The Journal of Cell Biology

مصطلحات موضوعية: Colon, Duodenum, Recombinant Fusion Proteins, Crypt, Molecular Sequence Data, Fluorescent Antibody Technique, Mice, Transgenic, Nerve Tissue Proteins, Biology, Regulatory Sequences, Nucleic Acid, Fatty Acid-Binding Proteins, Epithelium, Mice, Intestinal mucosa, Enhancer binding, Gene expression, medicine, Animals, Intestinal Mucosa, Regulation of gene expression, Base Sequence, Fatty Acids, Nuclear Proteins, Cell Biology, Articles, Blotting, Northern, Molecular biology, Intestinal epithelium, Neoplasm Proteins, DNA-Binding Proteins, medicine.anatomical_structure, Gene Expression Regulation, Regulatory sequence, Growth Hormone, Paneth cell, CCAAT-Enhancer-Binding Proteins, Carrier Proteins, Fatty Acid-Binding Protein 7, Transcription Factors

الوصف: The mouse intestinal epithelium is able to establish and maintain complex lineage-specific, spatial, and temporal patterns of gene expression despite its rapid and continuous renewal. A multipotent stem cell located near the base of each intestinal crypt gives rise to progeny which undergo amplification and allocation to either enterocytic, Paneth cell, goblet cell, or enteroendocrine cell lineages. Differentiation of these four lineages occurs during their geographically ordered migration along the crypt-villus axis. Gut stem cells appear to have a "positional address" which is manifested by differences in the differentiation programs of their lineal descendants along the duodenal-colonic (cephalocaudal) axis. We have used the intestinal fatty acid binding protein gene (Fabpi) as a model to identify cis-acting elements which regulate cell- and region-specific patterns of gene expression in the gut. Nucleotides -1178 to +28 of rat Fabpi direct a pattern of expression of a reporter (human growth hormone [hGH]) which mimics that of mouse Fabpi (a) steady-state levels of hGH mRNA are highest in the distal jejunum of adult transgenic mice and fall progressively toward both the duodenum and the mid-colon; and (b) hGH is confined to the enterocytic lineage and first appears as postmitotic, differentiating cells exit the crypt and migrate to the base of small intestinal villi or their colonic homologs, the surface epithelial cuffs. Nucleotides -103 to +28, which are highly conserved in rat, mouse and human Fabpi, are able to correctly initiate transgene expression in late fetal life, restrict hGH to the enterocytic lineage, and establish an appropriate cephalocaudal gradient of reporter expression. This cephalocaudal gradient is also influenced by cis-acting elements located between nucleotides -1178 and -278, and -277 and -185 that enhance and suppress (respectively) expression in the ileum and colon and by element(s) located upstream of nucleotide -277 that are needed to sustain high levels of hGH production after weaning. Nucleotides -277 to -185 contain part of a domain conserved between the three orthologous Fabpi genes (nucleotides -240 to -159), a 24-bp element (nucleotides -212 to -188) that binds nuclear factors present in colonic but not small intestinal epithelial cells, and a portion of a CCAAT/enhancer binding protein footprint (C/EBP alpha, nucleotides -188 to -167). Removal of nucleotides -277 to -185 (yielding I-FABP-184 to +28/hGH+3) results in inappropriate expression of hGH in proliferating and nonproliferating epithelial cells located in the mid and upper portions of duodenal, jejunal, ileal, and colonic crypts without affecting the "shape" of the cephalocaudal gradient of transgene expression.(ABSTRACT TRUNCATED AT 400 WORDS)

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a97a2b3ae338cebe770a9a643712cf96Test
http://europepmc.org/articles/PMC2289640Test

المؤلفون: Kevin A. Roth, J I Gordon, A R Moser, W F Dove

المصدر: The Journal of Cell Biology
Scopus-Elsevier

مصطلحات موضوعية: Adenoma, Serotonin, Cellular differentiation, Enteroendocrine cell, Mice, Inbred Strains, Nerve Tissue Proteins, Biology, medicine.disease_cause, Fatty Acid-Binding Proteins, Epithelium, Mice, Intestinal Neoplasms, medicine, Animals, Intestinal Mucosa, Crosses, Genetic, Epithelial cell differentiation, Fatty Acids, Mucins, Cell Differentiation, Cell Biology, Articles, medicine.disease, Molecular biology, Mice, Mutant Strains, Gut Epithelium, Neoplasm Proteins, medicine.anatomical_structure, Phenotype, Multipotent Stem Cell, Immunology, Mutation, Neoplastic Stem Cells, Muramidase, Carcinogenesis, Carrier Proteins, Fatty Acid-Binding Protein 7

الوصف: Min is a fully penetrant dominant mutation that leads to the development of multiple intestinal adenomas throughout the duodenal-to-colonic axis. Min/+ C57BL6/J mice have an average life-span of 120 d. Multi-label immunocytochemical studies of these lesions demonstrate patches of differentiated enterocytes, and scattered enteroendocrine, goblet and Paneth cells. Expression of endogenous marker genes within these differentiated cells can be directly correlated with the position occupied by the adenoma along the duodenal-to-colonic axis and mirrors the regional differentiation of the normal gut epithelium. The presence of multiple lineages in adenomas together with their retention of spatial information suggests that tumorigenesis in Min/+ mice may be initiated in a multipotent stem cell normally located at the base of intestinal crypts. To study the time-dependent properties of these tumors, genetic conditions were sought in which Min/+ animals could survive for up to 300 d. Min is fully penetrant in hybrids with either AKR/J or MA/MyJ. However, the hybrids demonstrate a reduction in the number of intestinal adenomas. Preliminary backcross analysis is consistent with a single major modifier locus unlinked to Min in both the AKR/J and MA/MyJ strains. The increased lifespan of the hybrid animals is also associated with the development of invasive tumors. New tumors do not arise continuously over the lifespan of these animals; instead all adenomas appear to be established by 100 d of age or sooner. These studies indicate that the Min/+ mouse is a powerful model system for analyzing the mechanisms that establish and maintain a balance between proliferation and differentiation in the continuously renewing gut epithelium and for an assessment of the multi-step hypothesis of intestinal neoplasia.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8109f8cc98a9ae05d700ca78742c452fTest
http://europepmc.org/articles/PMC2289373Test

المؤلفون: Kevin A. Roth, Sungsu Kim, S Harris, S Pease, DJ Ahnen, Sherrie M. Hauft, JR Hansbrough, GH Schmidt, Steven M. Cohn, S Rees

المصدر: The Journal of Cell Biology

مصطلحات موضوعية: Cell division, Cellular differentiation, Antigens, Polyomavirus Transforming, Gene Expression, Mice, Transgenic, Nerve Tissue Proteins, Biology, Regulatory Sequences, Nucleic Acid, Fatty Acid-Binding Proteins, digestive system, Mice, Proliferating Cell Nuclear Antigen, Intestine, Small, medicine, Animals, Neoplastic transformation, RNA, Messenger, Intestinal Mucosa, Cell growth, digestive, oral, and skin physiology, Cell Cycle, Nuclear Proteins, Cell Differentiation, Cell Biology, Articles, Cell cycle, Intestinal epithelium, Molecular biology, Neoplasm Proteins, medicine.anatomical_structure, Cell Transformation, Neoplastic, Multipotent Stem Cell, Paneth cell, Carrier Proteins, Fatty Acid-Binding Protein 7

الوصف: The mouse intestinal epithelium represents a unique mammalian system for examining the relationship between cell division, commitment, and differentiation. Proliferation and differentiation are rapid, perpetual, and spatially well-organized processes that occur along the crypt-to-villus axis and involve clearly defined cell lineages derived from a common multipotent stem cell located near the base of each crypt. Nucleotides -1178 to +28 of the rat intestinal fatty acid binding protein gene were used to establish three pedigrees of transgenic mice that expressed SV-40 large T antigen (TAg) in epithelial cells situated in the uppermost portion of small intestinal crypts and in already committed, differentiating enterocytes as they exited these crypts and migrated up the villus. T antigen production was associated with increases in crypt cell proliferation but had no apparent effect on commitment to differentiate along enterocytic, enteroendocrine, or Paneth cell lineages. Single- and multilabel-immunocytochemical studies plus RNA blot hybridization analyses suggested that the differentiation programs of these lineages were similar in transgenic mice and their normal littermates. This included enterocytes which, based on the pattern of [3H]thymidine and 5-bromo-2'-deoxyuridine labeling and proliferating nuclear antigen expression, had reentered the cell cycle during their migration up the villus. The state of cellular differentiation and/or TAg production appeared to affect the nature of the cell cycle; analysis of the ratio of S-phase to M-phase cells (collected by metaphase arrest with vincristine) and of the intensities of labeling of nuclei by [3H]thymidine indicated that the duration of S phase was longer in differentiating, villus-associated enterocytes than in the less well-differentiated crypt epithelial cell population and that there may be a block at the G2/M boundary. Sustained increases in crypt and villus epithelial cell proliferation over a 9-mo period were not associated with the development of gut neoplasms--suggesting that tumorigenesis in the intestine may require that the initiated cell have many of the properties of the gut stem cell including functional anchorage.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::74a1aa0140d1c9549fc9389eca7ec65cTest
https://pubmed.ncbi.nlm.nih.gov/1349609Test

المؤلفون: Jeffrey I. Gordon, E H Birkenmeier, Kevin A. Roth, Deborah C. Rubin

المصدر: The Journal of Cell Biology

مصطلحات موضوعية: Genetically modified mouse, medicine.medical_specialty, Cellular differentiation, Crypt, Gene Expression, Enteroendocrine cell, Mice, Inbred Strains, Mice, Transgenic, Nerve Tissue Proteins, Fatty Acid-Binding Proteins, digestive system, Mice, Nude mouse, Fetal Tissue Transplantation, Internal medicine, Intestine, Small, medicine, Animals, Humans, Epithelial cell differentiation, Cholecystokinin, biology, Tumor Suppressor Proteins, digestive, oral, and skin physiology, Fatty Acids, Cell Differentiation, Epithelial Cells, Cell Biology, Articles, biology.organism_classification, Gut Epithelium, Cell biology, Neoplasm Proteins, Rats, Transplantation, Isogeneic, Endocrinology, Jejunum, Liver, Growth Hormone, Carrier Proteins, Fatty Acid-Binding Protein 7

الوصف: Transgenes consisting of segments of the rat liver fatty acid-binding protein (L-FABP) gene's 5' non-transcribed domain linked to the human growth hormone (hGH) gene (minus its regulatory elements) have provided useful tools for analyzing the mechanisms that regulate cellular and spatial differentiation of the continuously renewing gut epithelium. We have removed the jejunum from normal and transgenic fetal mice before or coincident with, cytodifferentiation of its epithelium. These segments were implanted into the subcutaneous tissues of young adult CBY/B6 nude mouse hosts to determine whether the bipolar, migration-dependent differentiation pathways of gut epithelial cells can be established and maintained in the absence of its normal luminal environment. Immunocytochemical analysis of isografts harvested 4-6 wk after implantation revealed that activation of the intact endogenous mouse L-FABP gene (fabpl) in differentiating enterocytes is perfectly recapitulated as these cells are translocated along the crypt-to-villus axis. Similarly, Paneth and goblet cells appear to appropriately differentiate as they migrate to the crypt base and villus tip, respectively. The enteroendocrine cell subpopulations present in intact 4-6-wk-old jejunum are represented in these isografts. Their precise spatial distribution along the crypt-to-villus axis mimics that seen in the intact gut. A number of complex interrelationships between enteroendocrine subpopulations are also recapitulated. In both "intact" and isografted jejunum, nucleotides -596 to +21 of the rat L-FABP gene were sufficient to direct efficient expression of the hGH reporter to enterocytes although precocious expression of the transgene occurred in cells located in the upper crypt, before their translocation to the villus base. Inappropriate expression of hGH occurred in a high percentage (greater than 80%) of secretin, gastrin, cholecystokinin, and gastric inhibitory peptide producing enteroendocrine cells present in the intact jejunum of 4-6-wk-old L-FABP-596 to +21/hGH transgenics. Addition of nucleotides -597 to -4,000 reduced the percentage of cells co-expressing this reporter four- to eightfold in several of the subpopulations. Jejunal isografts from each transgenic pedigree studied contained a lower percentage of hGH positive enteroendocrine cells than in the comparably aged intact jejunum.(ABSTRACT TRUNCATED AT 400 WORDS)

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4537cd2a8f268c345c50c9f98c8f0c22Test
https://pubmed.ncbi.nlm.nih.gov/2040647Test