A method for examining differential mRNA expression along the crypt–villus axis of the human small intestine

1998 ◽  
Vol 95 (2) ◽  
pp. 171-177
Author(s):  
Eddy CHUA ◽  
Qiong WANG ◽  
Paul O'TOOLE ◽  
Martin LOMBARD
Keyword(s):  
Small Intestine ◽  
Messenger Rna ◽  
Thymidine Incorporation ◽  
Cell Types ◽  
Functional Differentiation ◽  
Small Samples ◽  
Human Small Intestine ◽  
Small Intestinal ◽  
Cell Subpopulations ◽  
Different Cell Types

1.The aim of this study was to devise a method of segregating crypt and villus cell subpopulations from endoscopic human small intestinal biopsies which might be used to examine changes associated with functional differentiation at the molecular level. 2.Routine endoscopic biopsies from the human small intestine were subjected to a modified protocol of mechanical disruption and chelation to yield subpopulations of different cell types. The purity and character of the cell populations isolated was assessed by measuring enzyme activity and thymidine incorporation and by histology. A guanidinium isothiocyanate method was adapted for small samples to extract RNA from the isolated subpopulations, and probes for RNA with a known predilection for crypt and villus cells were used to further investigate the application and usefulness of the technique. 3.Sequential histological examination during the segregation protocol demonstrated that different cell types were removed serially from the biopsy samples. Cell-type enrichment of the segregated subpopulations was demonstrated by differential alkaline phosphatase activity and by differences in thymidine incorporation in the samples isolated. Sufficient quantities of RNA could be extracted from the segregated subpopulations for Northern blot analysis and the differential expression of mRNA for sucrase-isomaltase and transferrin receptor was demonstrated in the villus and crypt subpopulations respectively. 4.Messenger RNA can be successfully extracted from different cell types segregated from routine human endoscopic small intestinal biopsies. This technique should prove useful for investigating the mechanisms regulating the functional differentiation of epithelial cells in the small intestine and the regulatory mechanisms governing absorption of macromolecules.

scholarly journals A breakdown in communication? Understanding the effects of aging on the human small intestine epithelium

2015 ◽  
Vol 129 (7) ◽  
pp. 529-531 ◽  
Author(s):  
Neil A. Mabbott
Keyword(s):  
Small Intestine ◽  
Barrier Function ◽  
Intestinal Barrier ◽  
Intestinal Barrier Function ◽  
Human Small Intestine ◽  
Small Intestinal ◽  
Effects Of Aging ◽  
Insight Into

A new study by Man and colleagues provides further insight into the effects of aging on small intestinal barrier function in humans. Here, their findings are briefly summarised and the wider implications discussed.

scholarly journals Mitochondrial Retrograde Signaling Contributes to Metabolic Differentiation in Yeast Colonies

2021 ◽  
Vol 22 (11) ◽  
pp. 5597
Author(s):  
Vítězslav Plocek ◽  
Kristýna Fadrhonc ◽  
Jana Maršíková ◽  
Libuše Váchová ◽  
Alexandra Pokorná ◽  
...  
Keyword(s):  
Cell Types ◽  
Colony Development ◽  
Mitochondrial Activity ◽  
Mitochondrial Translation ◽  
Metabolic Intermediates ◽  
Nutritional Conditions ◽  
Mitochondrial Ribosome ◽  
Development And Differentiation ◽  
Cell Subpopulations ◽  
Different Cell Types

During development of yeast colonies, various cell subpopulations form, which differ in their properties and specifically localize within the structure. Three branches of mitochondrial retrograde (RTG) signaling play a role in colony development and differentiation, each of them activating the production of specific markers in different cell types. Here, aiming to identify proteins and processes controlled by the RTG pathway, we analyzed proteomes of individual cell subpopulations from colonies of strains, mutated in genes of the RTG pathway. Resulting data, along with microscopic analyses revealed that the RTG pathway predominantly regulates processes in U cells, long-lived cells with unique properties, which are localized in upper colony regions. Rtg proteins therein activate processes leading to amino acid biosynthesis, including transport of metabolic intermediates between compartments, but also repress expression of mitochondrial ribosome components, thus possibly contributing to reduced mitochondrial translation in U cells. The results reveal the RTG pathway’s role in activating metabolic processes, important in U cell adaptation to altered nutritional conditions. They also point to the important role of Rtg regulators in repressing mitochondrial activity in U cells.

scholarly journals Age-related histochemical investigations of small intestinal goblet cells in bronze turkeys (Meleagris gallopavo gallopavo)

2021 ◽  
Vol 24 (2) ◽  
pp. 176-183
Author(s):  
D. Yovchev ◽  
G. Penchev
Keyword(s):  
Small Intestine ◽  
Age Groups ◽  
Negative Relationship ◽  
Meleagris Gallopavo ◽  
Cell Types ◽  
Goblet Cells ◽  
Age Related ◽  
Pas Staining ◽  
Small Intestinal ◽  
Acid Mucins

The aim of the study was to investigate the goblet cell types and their density in the small intestine of bronze turkey (Meleagris meleagris gallopovo), by means of Alcian blue-PAS staining. Sixty birds from 10 age groups were used. In the duodenum and jejunum, goblet cells produced acid, neutral and mixed mucins, while in the jejunum - acid mucins. A negative relationship was observed between cell density and either duodenum or jejunum lengths; such a correlation was not established in the ileum.

Gut epithelial IL-27 confers intestinal immunity through the induction of intraepithelial lymphocytes

2021 ◽  
Vol 218 (11) ◽  
Author(s):  
Chia-Hao Lin ◽  
Mei-Chi Chen ◽  
Ling-Li Lin ◽  
David A. Christian ◽  
Booki Min ◽  
...  
Keyword(s):  
Parasitic Infection ◽  
Cell Types ◽  
Functional Differentiation ◽  
Intraepithelial Lymphocytes ◽  
Intestinal Epithelial ◽  
Intestinal Immunity ◽  
Diverse Range ◽  
Pathogen Burden ◽  
Different Cell Types ◽  
Barrier Immunity

IL-27 controls a diverse range of immune responses in many disease settings. Here, we identify intestinal epithelial cells (IECs) as one of the major IL-27 cellular sources in the gut-associated tissue. Unlike IL-27 secreted by innate immune cells, gut epithelial IL-27 is dispensable for T-bet+ regulatory T cell (T reg cell) differentiation or IL-10 induction. Rather, IEC-derived IL-27 specifically promotes a distinct CD8αα+CD4+ intraepithelial lymphocyte (IEL) population that acquires their functional differentiation at the intestinal epithelium. Loss of IL-27 in IECs leads to a selective defect in CD8αα+CD4+ IELs over time. Consequently, mice with IEC-specific IL-27 ablation exhibited elevated pathogen burden during parasitic infection, and this could be rescued by transfer of exogenous CD8αα+CD4+ IELs. Collectively, our data reveal that in addition to its known regulatory properties in preventing immune hyperactivity, gut epithelial IL-27 confers barrier immunity by inducing a specific IEL subset and further suggest that IL-27 produced by different cell types plays distinct roles in maintaining intestinal homeostasis.

THE TRANSLATION OF MESSENGER RNA INJECTED IN LIVING OOCYTES OF XENOPUS LAEVIS

1973 ◽  
Vol 74 (Suppl) ◽  
pp. S225-S243 ◽  
Author(s):  
J. B. Gurdon
Keyword(s):  
Xenopus Laevis ◽  
Messenger Rna ◽  
Cell Types ◽  
Living Cells ◽  
Cell Component ◽  
Globin Mrna ◽  
Spare Capacity ◽  
Specific Component ◽  
Lower Efficiency ◽  
Different Cell Types

ABSTRACT Messenger RNA (mRNA) can be injected in known quantities into oocytes of the frog Xenopus laevis. Haemoglobin mRNA from rabbit, mouse, or duck blood cells is accurately translated, and oocytes can translate messages from different cell-types of all vertebrate groups. Oocytes and eggs have a spare capacity to translate injected messages, even when supplied in excess, and these do not compete with the cells' own messages already in use. The cell component which restricts the translation of injected mRNA above a saturating amount appears not to be KCl-released ribosome factors or transfer RNA. A message-specific component may be responsible for the lower efficiency of translation of α than of β globin mRNA in oocytes. Injected haemoglobin mRNA appears to be translated less frequently in oocytes than in reticulocytes at the same temperature, but continues to be translated in oocytes at nearly the same rate for a period of up to nearly 2 weeks. The injection of mRNA into oocytes is especially valuable for studying the control of protein synthesis in living cells.

Erythropoietin expression in primary rat Sertoli and peritubular myoid cells

Blood ◽  
2001 ◽  
Vol 98 (9) ◽  
pp. 2872-2874 ◽  
Author(s):  
Massimo Magnanti ◽  
Orietta Gandini ◽  
Laura Giuliani ◽  
Paola Gazzaniga ◽  
Hugo H. Marti ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Messenger Rna ◽  
Leydig Cells ◽  
Mrna Level ◽  
Cell Types ◽  
Myoid Cells ◽  
Rt Pcr ◽  
Basal Expression ◽  
Testicular Cells ◽  
Different Cell Types

Abstract Kidney and liver are the major organs of erythropoietin (Epo) synthesis. However, Epo messenger RNA (mRNA) has been detected in several organs, such as brain, lung, and testis. Furthermore, functional Epo receptors have been demonstrated on different cell types, including rat Leydig cells. The aim of the study was to identify testicular cells expressing Epo mRNA and to quantitate its levels by competitive reverse transcriptase–polymerase chain reaction (RT-PCR). Besides whole testis, Epo transcripts were found in Sertoli and peritubular myoid cells, while no signal was detected in Leydig cells. Exposure of Sertoli cells to CoCl2 led to an increase of Epo mRNA level. Semiquantitative competitive RT-PCR presented an increase in the level of Epo mRNA in Sertoli cells stimulated by follicle-stimulating hormone, while exposure of peritubular myoid cells cultures to testosterone reduced Epo mRNA expression. Due to the blood-testis barrier, basal expression of Epo suggests a not yet defined function of this hormone in testis.

scholarly journals Identification of cell states using super-enhancer RNA

BMC Genomics ◽  
2021 ◽  
Vol 22 (S3) ◽  
Author(s):  
Yueh-Hua Tu ◽  
Hsueh-Fen Juan ◽  
Hsuan-Cheng Huang
Keyword(s):  
Messenger Rna ◽  
Time Course ◽  
Developmental Process ◽  
Cell Types ◽  
Regulatory Elements ◽  
Cell Type ◽  
Sequencing Data ◽  
Enhancer Rna ◽  
Super Enhancer ◽  
Different Cell Types

Abstract Background A new class of regulatory elements called super-enhancers, comprised of multiple neighboring enhancers, have recently been reported to be the key transcriptional drivers of cellular, developmental, and disease states. Results Here, we defined super-enhancer RNAs as highly expressed enhancer RNAs that are transcribed from a cluster of localized genomic regions. Using the cap analysis of gene expression sequencing data from FANTOM5, we systematically explored the enhancer and messenger RNA landscapes in hundreds of different cell types in response to various environments. Applying non-negative matrix factorization (NMF) to super-enhancer RNA profiles, we found that different cell types were well classified. In addition, through the NMF of individual time-course profiles from a single cell-type, super-enhancer RNAs were clustered into several states with progressive patterns. We further investigated the enriched biological functions of the proximal genes involved in each pattern, and found that they were associated with the corresponding developmental process. Conclusions The proposed super-enhancer RNAs can act as a good alternative, without the complicated measurement of histone modifications, for identifying important regulatory elements of cell type specification and identifying dynamic cell states.

scholarly journals Identification of Cell States Using Super-Enhancer RNA

2017 ◽  
Author(s):  
Yueh-Hua Tu ◽  
Hsueh-Fen Juan ◽  
Hsuan-Cheng Huang
Keyword(s):  
Messenger Rna ◽  
Time Course ◽  
Developmental Process ◽  
Cell Types ◽  
Regulatory Elements ◽  
Cell Type ◽  
Sequencing Data ◽  
Enhancer Rna ◽  
Super Enhancer ◽  
Different Cell Types

AbstractA new class of regulatory elements called super-enhancers, comprised of multiple neighboring enhancers, have recently been reported to be the key transcriptional drivers of cellular, developmental, and disease states. Here, we propose to define super-enhancer RNA as highly expressed enhancer RNAs that are transcribed from a cluster of localized genomic regions. Using the cap analysis of gene expression sequencing data from FANTOM5, we systematically explored the enhancer and messenger RNA landscapes in hundreds of different cell types in response to various environments. Applying non-negative matrix factorization (NMF) to super-enhancer RNA profiles, we found that different cell types were well classified. In addition, through the NMF of individual time-course profiles from a single cell-type, super-enhancer RNAs were clustered into several states with progressive patterns. We further investigated the enriched biological functions of the proximal genes involved in each pattern, and found that they were associated with the corresponding developmental process. The proposed super-enhancer RNAs can act as a good alternative, without the complicated measurement of histone modifications, for identifying important regulatory elements of cell type specification and identifying dynamic cell states.

Absence of synchrony between human small intestinal migrating motor complex and rectal motor complex

1990 ◽  
Vol 258 (1) ◽  
pp. G171-G172 ◽  
Author(s):  
D. Kumar ◽  
P. D. Thompson ◽  
D. L. Wingate
Keyword(s):  
Small Intestine ◽  
Motor Activity ◽  
Healthy Adults ◽  
Migrating Motor Complex ◽  
Human Small Intestine ◽  
Motor Complex ◽  
Small Intestinal

Both the human small intestine and rectum exhibit motor activity in which relatively brief bursts of powerful regular contractions recur with a similar periodicity. We used prolonged ambulant manometry to test the hypothesis that these activities are synchronous. Pressure activity from the duodenojejunum and the rectum was recorded continuously for 24 h in eight freely ambulant healthy adults. A total of 61 migrating motor complexes and 61 rectal motor complexes occurred in the group; the median periodicities of the two rhythms differed significantly (P = 0.025). There was no evidence of synchrony between the two biorhythms. We conclude that they are independent oscillations.

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