The establishment of the hepatic architecture is a prerequisite for the development of a lobular pattern of gene expression

Development ◽  
1996 ◽  
Vol 122 (1) ◽  
pp. 321-332
Author(s):  
R.G. Notenboom ◽  
P.A. de Boer ◽  
A.F. Moorman ◽  
W.H. Lamers
Keyword(s):  
Serum Proteins ◽  
Expression Patterns ◽  
In Situ Hybridisation ◽  
Three Dimensional ◽  
Immunohistochemical Analysis ◽  
Normal Liver ◽  
Rat Hepatocytes ◽  
Liver Development ◽  
Multistep Process

We have studied the expression patterns of ammonia-metabolising enzymes and serum proteins in intrasplenically transplanted embryonic rat hepatocytes by in situ hybridisation and immunohistochemical analysis. The enzymic phenotype of individually settled hepatocytes was compared with that of hepatocytes being organised into a three-dimensional hepatic structure. Our results demonstrate that development towards the terminally differentiated state with zonal differences in enzyme content requires the incorporation of hepatocytes into lobular structures. Outside such an architectural context, phenotypic maturation becomes arrested and hepatocytes linger in the protodifferentiated state. These features identify the foetal period as a crucial time for normal liver development and show that the establishment of the terminally differentiated hepatocellular phenotype, beginning with the differentiation of hepatocytes from the embryonic foregut, is realised via a multistep process.

Regulation of expression of the retinoic acid-synthesising enzymes retinaldehyde dehydrogenases in the uteri of ovariectomised mice after treatment with oestrogen, gestagen and their combination

2006 ◽  
Vol 18 (3) ◽  
pp. 339 ◽  
Author(s):  
Ralph Rühl ◽  
Britta Fritzsche ◽  
Julien Vermot ◽  
Karen Niederreither ◽  
Ulrike Neumann ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Early Pregnancy ◽  
Expression Patterns ◽  
In Situ Hybridisation ◽  
Oestrous Cycle ◽  
Regulation Of Expression ◽  
Mouse Uterus ◽  
Blastocyst Implantation ◽  
Polymerase Chain

The active metabolite of vitamin A, retinoic acid (RA), plays an important role in the female reproductive system. The synthesis of RA is tightly regulated by the activity of retinaldehyde dehydrogenases (Raldh). Among these, Raldh1 and Raldh2 exhibit specific temporal and spatial expression patterns in the mouse uterus, both during the oestrous cycle and early pregnancy. In the present study, we have assessed whether oestradiol and progesterone directly influence the uterine expression of Raldh1 and Raldh2 in ovariectomised mice. We investigated the effect of gestagen (promegestone 0.3 mg kg−1 bodyweight), oestrogen (oestradiol 3 µg kg−1 bodyweight) and their combination on the uterine expression of Raldh2. Expression was analysed using in situ hybridisation and quantified using real-time detection reverse transcription–polymerase chain reaction. The results show that the expression of Raldh2 is rapidly (within 1–4 h) induced in stromal cells by oestrogen, but not by gestagen, treatment, whereas combined oestrogen + gestagen treatment leads to a more prolonged (48 h) response. In contrast, oestrogen, but not progesterone, treatment downregulates (within 4–24 h) Raldh1 expression in the uterine glandular epithelium. We conclude that the uterine RA concentrations are regulated by oestrogens via an effect on the expression of the Raldh synthesising enzymes. Such a regulation is consistent with the natural fluctuations of Raldh expression during the oestrous cycle, early pregnancy and blastocyst implantation.

Immunological mechanisms to establish embryo tolerance in early bovine pregnancy

2011 ◽  
Vol 23 (5) ◽  
pp. 619 ◽  
Author(s):  
A. E. Groebner ◽  
K. Schulke ◽  
J. C. Schefold ◽  
G. Fusch ◽  
F. Sinowatz ◽  
...  
Keyword(s):  
In Situ Hybridisation ◽  
Immunohistochemical Analysis ◽  
Kynurenine Pathway ◽  
Endometrial Cells ◽  
Maternal Rejection ◽  
Immunological Mechanisms ◽  
Bovine Pregnancy ◽  
Coculture Model

A well-balanced immunological interaction between mother and the semi-allogenic embryo is of particular importance. The objective of the present study was to analyse mechanisms of immune tolerance in bovine pregnancy during peri-implantation. Simmental heifers inseminated with either cryopreserved spermatozoa or seminal plasma were killed 12, 15 or 18 days after oestrus. Uteri were flushed for the recovery of conceptuses and the ipsilateral intercaruncular endometrium was sampled for gene expression analysis. Indoleamine 2,3-dioxygenase (IDO) mRNA, coding for the initial enzyme of the kynurenine pathway, was 18-fold (P < 0.001) more abundant in the endometrium of Day 18 pregnant v. non-pregnant animals. Tandem mass spectrometry revealed a decrease of endometrial l-tryptophan (P = 0.0008), but an increase of l-kynurenine concentration (P = 0.005) from Day 12 to Day 18, suggesting increasing IDO activity (P < 0.03). An in vitro coculture model of endometrial cells showed an induction of IDO expression following interferon-τ exposure primarily in stroma cells, which was confirmed by in situ hybridisation localising IDO mRNA mainly in deep stroma cells. Immunohistochemical analysis revealed fewer CD45-positive leucocytes in the zona basalis of pregnant animals. Elevated IDO activity may reduce the presence of leucocytes in the pregnant endometrium, providing a possible mechanism for protecting the semi-allogenic conceptus from maternal rejection.

scholarly journals Cytochrome P450 Expression and Chemical Metabolic Activity before Full Liver Development in Zebrafish

2020 ◽  
Vol 13 (12) ◽  
pp. 456
Author(s):  
Tasuku Nawaji ◽  
Natsumi Yamashita ◽  
Haruka Umeda ◽  
Shuangyi Zhang ◽  
Naohiro Mizoguchi ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
In Situ Hybridization ◽  
Metabolic Activity ◽  
Expression Patterns ◽  
Xenobiotic Metabolism ◽  
Liver Development ◽  
P450 Expression ◽  
Whole Mount ◽  
Metabolic Activities

Zebrafish are used widely in biomedical, toxicological, and developmental research, but information on their xenobiotic metabolism is limited. Here, we characterized the expression of 14 xenobiotic cytochrome P450 (CYP) subtypes in whole embryos and larvae of zebrafish (4 to 144 h post-fertilization (hpf)) and the metabolic activities of several representative human CYP substrates. The 14 CYPs showed various changes in expression patterns during development. Many CYP transcripts abruptly increased at about 96 hpf, when the hepatic outgrowth progresses; however, the expression of some cyp1s (1b1, 1c1, 1c2, 1d1) and cyp2r1 peaked at 48 or 72 hpf, before full liver development. Whole-mount in situ hybridization revealed cyp2y3, 2r1, and 3a65 transcripts in larvae at 55 hpf after exposure to rifampicin, phenobarbital, or 2,3,7,8-tetrachlorodibenzo-p-dioxin from 30 hpf onward. Marked conversions of diclofenac to 4′-hydroxydiclofenac and 5-hydroxydiclofenac, and of caffeine to 1,7-dimethylxanthine, were detected as early as 24 or 50 hpf. The rate of metabolism to 4’-hydroxydiclofenac was more marked at 48 and 72 hpf than at 120 hpf, after the liver had become almost fully developed. These findings reveal the expression of various CYPs involved in chemical metabolism in developing zebrafish, even before full liver development.

Elucidating the role of long intergenic non-coding RNA 339 in human endometrium and endometriosis

2021 ◽  
Author(s):  
Sarah J Holdsworth-Carson ◽  
Molly Churchill ◽  
Jacqueline F Donoghue ◽  
Sally Mortlock ◽  
Jenny N Fung ◽  
...  
Keyword(s):  
Rna Sequencing ◽  
Cell Lines ◽  
Expression Patterns ◽  
In Situ Hybridisation ◽  
Immune Defense ◽  
Human Endometrium ◽  
Altered Expression ◽  
Non Coding Rna

Abstract Endometriosis is a complex disease, influenced by genetic factors. Genetic markers associated with endometriosis exist at chromosome 1p36.12 and lead to altered expression of the long intergenic non-coding RNA 339 (LINC00339), however the role of LINC00339 in endometriosis pathophysiology remains unknown. The aim of this work was to characterise the expression patterns of LINC00339 mRNA in endometrium and endometriotic lesions in situ and to determine the functional role of LINC00339 in human endometrium. We employed RNA-sequencing, quantitative RT-PCR and in situ hybridisation to investigate the abundance of LINC00339 transcripts in endometrium and endometrial cell lines and to describe the pattern and localisation of LINC00339 expression in endometrium and endometriotic lesions. LINC00339 mRNA expression was manipulated (overexpressed and silenced) in endometrial stomal cell lines and RNA-sequencing data from overexpression models were analysed using online bioinformatics platforms (STRING and Ingenuity Pathway Analysis) to determine functional processes. We demonstrated the expression of LINC00339 in endometriotic lesions for the first time; we found LINC00339 expression was restricted to the lesion foci and absent in surrounding non-lesion tissue. Furthermore, manipulation of LINC00339 expression in endometrial stromal cell lines significantly impacted the expression of genes involved in immune defense pathways. These studies identify a novel mechanism for LINC00339 activity in endometrium and endometriosis, paving the way for future work, which is essential for understanding the pathogenesis of endometriosis.

Transplanted reporter cells help in defining onset of hepatocyte proliferation during the life of F344 rats

2000 ◽  
Vol 279 (3) ◽  
pp. G631-G640 ◽  
Author(s):  
Rana P. Sokhi ◽  
Pankaj Rajvanshi ◽  
Sanjeev Gupta
Keyword(s):  
Proliferative Activity ◽  
Expression Patterns ◽  
Rat Hepatocytes ◽  
Liver Parenchyma ◽  
Hepatocyte Proliferation ◽  
Liver Development ◽  
Fischer 344 ◽  
Old Rats ◽  
Cell And Gene Therapy ◽  
F344 Rats

Transplanted hepatocytes integrate in the liver parenchyma and exhibit gene expression patterns that are similar to adjacent host hepatocytes. To determine the fate of genetically marked hepatocytes in the context of hepatocellular proliferation throughout the rodent life span, we transplanted Fischer 344 (F344) rat hepatocytes into syngeneic dipeptidyl peptidase IV-deficient rats. The proliferative activity in transplanted hepatocytes was studied in animals ranging in age from a few days to 2 yr. Transplanted hepatocytes proliferated during liver development between 1 and 6 wk of age, each dividing an estimated two to five times. DNA synthesis in occasional cells was demonstrated by localizing bromodeoxyuridine incorporation. There was no evidence for transplanted cell proliferation between 6 wk and 1 yr of age. Subsequently, transplanted cells proliferated again, with increased sizes of transplanted cell clusters at 18 and 24 mo of age. The proliferative activity of transplanted cells was greater in rats entering senescence compared with during postnatal liver development. In old rats, some liver lobules were composed entirely of transplanted cells. We conclude that hepatocyte proliferation in the livers of very young and old F344 rats is regulated in a temporally determined, biphasic manner. The findings will be relevant to mechanisms concerning liver development, senescence, and oncogenesis, as well as to cell and gene therapy.

scholarly journals ClusterMap: multi-scale clustering analysis of spatial gene expression

2021 ◽  
Author(s):  
Yichun He ◽  
Xin Tang ◽  
Jiahao Huang ◽  
Haowen Zhou ◽  
Kevin Chen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dimensional Space ◽  
Expression Patterns ◽  
Three Dimensional ◽  
Point Pattern Analysis ◽  
Point Pattern ◽  
Computational Framework ◽  
Spatially Resolved ◽  
Tissue Organization

AbstractQuantifying RNAs in their spatial context is crucial to understanding gene expression and regulation in complex tissues. In situ transcriptomic methods generate spatially resolved RNA profiles in intact tissues. However, there is a lack of a unified computational framework for integrative analysis of in situ transcriptomic data. Here, we present an unsupervised and annotation-free framework, termed ClusterMap, which incorporates physical proximity and gene identity of RNAs, formulates the task as a point pattern analysis problem, and thus defines biologically meaningful structures and groups. Specifically, ClusterMap precisely clusters RNAs into subcellular structures, cell bodies, and tissue regions in both two- and three-dimensional space, and consistently performs on diverse tissue types, including mouse brain, placenta, gut, and human cardiac organoids. We demonstrate ClusterMap to be broadly applicable to various in situ transcriptomic measurements to uncover gene expression patterns, cell-cell interactions, and tissue organization principles from high-dimensional transcriptomic images.

ZebraFISH: Fluorescent In Situ Hybridization Protocol and Three-Dimensional Imaging of Gene Expression Patterns

Zebrafish ◽  
2006 ◽  
Vol 3 (4) ◽  
pp. 465-476 ◽  
Author(s):  
Monique C.M. Welten ◽  
Simon B. de Haan ◽  
Niels van den Boogert ◽  
Jasprien N. Noordermeer ◽  
Gerda E.M. Lamers ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Expression Patterns ◽  
Three Dimensional ◽  
Gene Expression Patterns ◽  
Three Dimensional Imaging ◽  
Hybridization Protocol
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