In situ hybridization analysis of TGF beta 3 RNA expression during mouse development: comparative studies with TGF beta 1 and beta 2

Development ◽  
1990 ◽  
Vol 110 (2) ◽  
pp. 609-620 ◽  
Author(s):  
R.W. Pelton ◽  
M.E. Dickinson ◽  
H.L. Moses ◽  
B.L. Hogan
Keyword(s):  
Intervertebral Discs ◽  
The Other ◽  
Rna Expression ◽  
Tgf Beta ◽  
Mouse Development ◽  
Olfactory Bulbs ◽  
Embryonic Tissues ◽  
Organ Systems ◽  
Beta 2

To date, three closely-related TGF beta genes have been found in the mouse; TGF beta 1, TGF beta 2 and TGF beta 3. Previous experiments have indicated that TGF beta 1 and TGF beta 2 may play important roles during mouse embryogenesis. The present study now reports the distribution of transcripts of TGF beta 3 in comparison to the other two genes and reveals overlapping but distinct patterns of RNA expression. TGF beta 3 RNA is expressed in a diverse array of tissues including perichondrium, bone, intervertebral discs, mesenteries, pleura, heart, lung, palate, and amnion, as well as in central nervous system (CNS) structures such as the meninges, choroid plexus and the olfactory bulbs. Furthermore, in several organ systems, TGF beta 3 transcripts are expressed during periods of active morphogenesis suggesting that the protein may be an important factor for the growth and differentiation of many embryonic tissues.

Protein Accumulation in Early Chick Embryos Grown under Different Conditions of Explantation

Development ◽  
1959 ◽  
Vol 7 (1) ◽  
pp. 66-72
Author(s):  
L. Gwen Britt ◽  
Heinz Herrmann
Keyword(s):  
Chick Embryo ◽  
Slow Rate ◽  
The Other ◽  
Protein Accumulation ◽  
Chick Embryos ◽  
Developmental Processes ◽  
Embryonic Tissues ◽  
Somite Formation ◽  
Explanted Chick Embryo

The recent development of techniques originally devised by Waddington (1932) for the maintenance of the explanted chick embryo (Spratt, 1947; New, 1955; Wolff & Simon, 1955) has opened the possibility of determining quantitatively some parameters of the developmental processes occurring in embryonic tissues under these conditions. As a result of such measurements, protein accumulation in explanted embryos was found to be much smaller than in embryos developing in the egg. On the other hand, the progress of somite formation was found to take place at similar rates in embryos developing as explants or in situ (Herrmann & Schultz, 1958). The slow rate of protein accumulation in the explanted embryos made it seem desirable to investigate whether under some other conditions of explantation protein accumulation would approach more closely the rate of protein formation observed in the naturally developing embryo.

Regulated expression and growth inhibitory effects of transforming growth factor-beta isoforms in mouse mammary gland development

Development ◽  
1991 ◽  
Vol 113 (3) ◽  
pp. 867-878 ◽  
Author(s):  
S.D. Robinson ◽  
G.B. Silberstein ◽  
A.B. Roberts ◽  
K.C. Flanders ◽  
C.W. Daniel
Keyword(s):  
Gene Expression ◽  
Mammary Gland ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Slow Release ◽  
Mouse Mammary Gland ◽  
Tgf Beta ◽  
Growth Factor Beta ◽  
Beta 2

Transforming Growth Factor-beta 1 (TGF-beta 1) was previously shown to inhibit reversibly the growth of mouse mammary ducts when administered in vivo by miniature slow-release plastic implants. We now report a comparative analysis of three TGF-beta isoforms with respect to gene expression and localization of protein products within the mouse mammary gland. Our studies revealed overlapping expression patterns of TGF-beta 1, TGF-beta 2 and TGF-beta 3 within the epithelium of the actively-growing mammary end buds during branching morphogenesis, as well as within the epithelium of growth-quiescent ducts. However, TGF-beta 3 was the only isoform detected in myoepithelial progenitor cells (cap cells) of the growing end buds and myoepithelial cells of the mature ducts. During pregnancy, TGF-beta 2 and TGF-beta 3 transcripts increased to high levels, in contrast to TGF-beta 1 transcripts which were moderately abundant; TGF-beta 2 was significantly transcribed only during pregnancy. Molecular hybridization in situ revealed overlapping patterns of expression for the three TGF-beta isoforms during alveolar morphogenesis, but showed that, in contrast to the patterns of TGF-beta 1 and TGF-beta 2 expression, TGF-beta 3 is expressed more heavily in ducts than in alveoli during pregnancy. Developing alveolar tissue and its associated ducts displayed striking TGF-beta 3 immunoreactivity which was greatly reduced during lactation. All three isoforms showed dramatically reduced expression in lactating tissue. The biological effects of active, exogenous TGF-beta 2 and TGF-beta 3 were tested with slow-release plastic implants. These isoforms, like TGF-beta 1, inhibited mammary ductal elongation in situ by causing the disappearance of the proliferating stem cell layer (cap cells) and rapid involution of ductal end buds. None of the isoforms were active in inhibiting alveolar morphogenesis. We conclude that under the limited conditions of these tests, the three mammalian isoforms are functionally equivalent. However, striking differences in patterns of gene expression and in the distribution of immunoreactive peptides suggest that TGF-beta isoforms may have distinct roles in mammary growth regulation, morphogenesis and functional differentiation.

Differential expression of TGF beta 1, beta 2 and beta 3 genes during mouse embryogenesis

Development ◽  
1991 ◽  
Vol 111 (1) ◽  
pp. 117-130 ◽  
Author(s):  
P. Schmid ◽  
D. Cox ◽  
G. Bilbe ◽  
R. Maier ◽  
G.K. McMaster
Keyword(s):  
Fetal Liver ◽  
In Situ Hybridisation ◽  
Regulatory Elements ◽  
Northern Analysis ◽  
Tgf Beta ◽  
Mouse Embryogenesis ◽  
Root Sheath ◽  
Cell Layers ◽  
Beta 2

We have examined by Northern analysis and in situ hybridisation the expression of TGF beta 1, beta 2 and beta 3 during mouse embryogenesis. TGF beta 1 is expressed predominantly in the mesodermal components of the embryo e.g. the hematopoietic cells of both fetal liver and the hemopoietic islands of the yolk sac, the mesenchymal tissues of several internal organs and in ossifying bone tissues. The strongest TGF beta 2 signals were found in early facial mesenchyme and in some endodermal and ectodermal epithelial cell layers e.g., lung and cochlea epithelia. TGF beta 3 was strongest in prevertebral tissue, in some mesothelia and in lung epithelia. All three isoforms were expressed in bone tissues but showed distinct patterns of expression both spatially and temporally. In the root sheath of the whisker follicle, TGF beta 1, beta 2 and beta 3 were expressed simultaneously. We discuss the implication of these results in regard to known regulatory elements of the TGF beta genes and their receptors.

Expression of transforming growth factor beta 2 RNA during murine embryogenesis

Development ◽  
1989 ◽  
Vol 106 (4) ◽  
pp. 759-767 ◽  
Author(s):  
R.W. Pelton ◽  
S. Nomura ◽  
H.L. Moses ◽  
B.L. Hogan
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Post Partum ◽  
Mouse Embryos ◽  
Tgf Beta ◽  
Growth Factor Beta ◽  
Beta 2 ◽  
Temporal And Spatial

We have studied the temporal and spatial expression of transforming growth factor beta 2 (TGF beta 2) RNA in mouse embryos from 10.5 days post coitum (p.c.) to 3 days post partum (p.p.) by in situ hybridization analysis. TGF beta 2 RNA is expressed in a variety of tissues including bone, cartilage, tendon, gut, blood vessels, skin and fetal placenta, and is in general found in the mesenchymal component of these tissues. The expression of TGF beta 2 RNA changes during development in a manner consistent with a role for the gene product in mediating mesenchymal-epithelial interactions.

scholarly journals The Collagen Type XVIII Endostatin Domain Is Co-localized with Perlecan in Basement Membranes in Vivo

2003 ◽  
Vol 51 (3) ◽  
pp. 285-296 ◽  
Author(s):  
Nicolai Miosge ◽  
Timo Simniok ◽  
Patricia Sprysch ◽  
Rainer Herken
Keyword(s):  
Collagen Type ◽  
Biological Properties ◽  
Basement Membranes ◽  
Mouse Development ◽  
Electron Microscopic ◽  
Double Labeling ◽  
Organ Systems ◽  
Almost All

The C-terminal globular endostatin domain of collagen type XVIII is anti-angiogenic in a variety of experimental tumor models, and clinical trials to test it as an anti-tumor agent are already under way. In contrast, many of its cell biological properties are still unknown. We systematically localized the mRNA of collagen type XVIII with the help of in situ hybridization (ISH) and detected it in epithelial and mesenchymal cells of almost all organ systems throughout mouse development. Light and electron microscopic immunohistochemistry (IHC) revealed that the endostatin domain is a widespread component of almost all epithelial basement membranes in all major developing organs, and in all basement membranes of capillaries and blood vessels. Furthermore, quantitative immunogold double labeling demonstrated a co-localization of 50% of the detected endostatin domain together with perlecan in basement membranes in vivo. We conclude that the endostatin domain of collagen type XVIII plays a role, even in early stages of mouse development, other than regulating angiogenesis. In the adult, the endostatin domain could well be involved in connecting collagen type XVIII to the basement membrane scaffolds. At least in part, perlecan appears to be an adaptor molecule for the endostatin domain in basement membranes in vivo.

Embryonic gene expression patterns of TGF beta 1, beta 2 and beta 3 suggest different developmental functions in vivo

Development ◽  
1991 ◽  
Vol 111 (1) ◽  
pp. 131-143 ◽  
Author(s):  
F.A. Millan ◽  
F. Denhez ◽  
P. Kondaiah ◽  
R.J. Akhurst
Keyword(s):  
Growth Factors ◽  
Developmental Process ◽  
Expression Patterns ◽  
In Situ Hybridisation ◽  
Tgf Beta ◽  
Genes Encoding ◽  
Beta 2 ◽  
Valve Formation

We have compared the expression of the genes encoding transforming growth factors beta 1, beta 2 and beta 3 during mouse embryogenesis from 9.5 to 16.5 days p.c. using in situ hybridisation to cellular RNAs. Each gene has a different expression pattern, which gives some indication of possible biological function in vivo. All three genes appear to be involved in chondroossification, though each is expressed in a different cell type. Transcripts of each gene are also present in embryonic epithelia. Epithelial expression of TGF beta 1, beta 2 and beta 3 RNA is associated with regions of active morphogenesis involving epithelial-mesenchymal interactions. In addition, widespread epithelial expression of TGF beta 2 RNA can be correlated with epithelial differentiation per se. The localisation of TGF beta 2 RNA in neuronal tissue might also be correlated with differentiation. Finally both TGF beta 1 and beta 2 transcripts are seen in regions actively undergoing cardiac septation and valve formation, suggesting some interaction of these growth factors in this developmental process.

RNA and protein localisations of TGF beta 2 in the early mouse embryo suggest an involvement in cardiac development

Development ◽  
1993 ◽  
Vol 117 (2) ◽  
pp. 625-639 ◽  
Author(s):  
M.C. Dickson ◽  
H.G. Slager ◽  
E. Duffie ◽  
C.L. Mummery ◽  
R.J. Akhurst
Keyword(s):  
Cardiac Development ◽  
In Situ Hybridisation ◽  
Outflow Tract ◽  
Tgf Beta ◽  
Atrioventricular Canal ◽  
Whole Mount ◽  
Beta 2 ◽  
Low Levels ◽  
Early Mouse

We have performed a detailed analysis of the localisations of RNAs for TGF beta 2 and beta 3, and of TGF beta 2 protein in mouse embryos from 6.5 to 9.5 days post coitum, using in situ hybridisation and immunohistochemistry on serial sections, and whole-mount in situ hybridisation to complete embryos. TGF beta 3 RNA was not seen in any of the tissue sections, but very low levels of the RNA were seen by whole-mount in situ hybridisation around the outflow tract of the heart at 8.5 days post coitum. TGF beta 2 RNA is expressed at high levels in all cells with the potential to differentiate into cardiomyocytes. Additionally, the foregut endoderm, juxtaposed to the heart, and the neuroepithelium at the rostral extremity of the foregut, express very high levels of TGF beta 2 RNA, between 8.5 and 9.5 days post coitum. As cardiomyogenesis proceeds, TGF beta 2 RNA levels diminishes within the myocytes, with a concomitant increase in staining for TGF beta 2 protein. TGF beta 2 protein staining of cardiomyocytes persists throughout development and in the adult, in the absence of detectable levels of the corresponding RNA. Superimposed upon this myocardial pattern of expression, there is an upregulation of TGF beta 2 RNA in the myocardium of the outflow tract and atrioventricular canal between 8.5 and 9.5 days post coitum, which returns to low levels by 11.5 days post coitum. The results are discussed in terms of a potential role of TGF beta 2 in controlling cardiomyogenesis and in inductive interactions leading to cardiac cushion tissue formation.

Expression of TGF-beta isoforms during first trimester human embryogenesis

Development ◽  
1990 ◽  
Vol 110 (2) ◽  
pp. 445-460 ◽  
Author(s):  
D. Gatherer ◽  
P. Ten Dijke ◽  
D.T. Baird ◽  
R.J. Akhurst
Keyword(s):  
First Trimester ◽  
Human Embryos ◽  
Regulatory Sequences ◽  
Tgf Beta ◽  
Human Embryogenesis ◽  
Genes Encoding ◽  
Gene Regulatory ◽  
Beta 2 ◽  
Beta Gene

We have studied the expression of the genes encoding transforming growth factors (TGFs) beta 1, beta 2 and beta 3 in human embryos ranging from 32 to 57 days post-coitum, using in situ hybridization. The spatial and temporal pattern of expression of each gene is distinct, though each occasionally overlaps. TGF-beta 1 is expressed in haematopoietic, endothelial and osteogenic tissues. TGF-beta 2 and TGF-beta 3 are expressed in a wide variety of mesenchymal tissues including areas of chondrogenic activity. TGF-beta 2 is also found in several epithelial and in the ventral nervous system. The differential transcript distributions are broadly similar to those seen in mouse embryos suggesting that there is conservation of TGF-beta gene regulatory sequences and developmental function across this species boundary.

Interactions between retinoids and TGF beta s in mouse morphogenesis

Development ◽  
1992 ◽  
Vol 115 (1) ◽  
pp. 67-74 ◽  
Author(s):  
R. Mahmood ◽  
K.C. Flanders ◽  
G.M. Morriss-Kay
Keyword(s):  
Retinoic Acid ◽  
Mesenchymal Cells ◽  
Neural Plate ◽  
Tgf Beta ◽  
Down Regulation ◽  
Initial Exposure ◽  
Embryonic Tissues ◽  
Beta 2 ◽  
Cranial Neural Crest Cells ◽  
Early Mouse

Using immunocytochemical methods we describe the distribution of different TGF beta isoforms and the effects of excess retinoic acid on their expression during early mouse embryogenesis (8 1/2 - 10 1/2 days of development). In normal embryos at 9 days, intracellular TGF beta 1 is expressed most intensely in neuroepithelium and cardiac myocardium whereas extracellular TGF beta 1 is expressed in mesenchymal cells and in the endocardium of the heart. At later stages, intracellular TGF beta 1 becomes very restricted to the myocardium and to a limited number of head mesenchymal cells; extracellular TGF beta 1 continues to be expressed widely in cells of mesenchymal origin, particularly in head and trunk mesenchyme, and also in endocardium. TGF beta 2 is widely expressed at all stages investigated while TGF beta 3 is not expressed strongly in any tissue at the stages examined. Exposure of early neural plate stage embryos to retinoic acid caused reduced expression of TGF beta 1 and TGF beta 2 proteins but had no effect on TGF beta 3. Intracellular TGF beta 1 expression was reduced in all tissues except in the myocardium, while extracellular TGF beta 1 was specifically reduced in neuroepithelium and cranial neural crest cells at early stages. TGF beta 2 was reduced in all embryonic tissues. The down-regulation of intracellular TGF beta 1 was observed up to 48 hours after initial exposure to retinoic acid while some down-regulation of TGF beta 2 was still seen up to 60 hours after initial exposure.(ABSTRACT TRUNCATED AT 250 WORDS)

Macrophages, but not dendritic cells, accumulate colloidal carbon following administration in situ

1993 ◽  
Vol 105 (4) ◽  
pp. 965-973
Author(s):  
M.D. Witmer-Pack ◽  
M.T. Crowley ◽  
K. Inaba ◽  
R.M. Steinman
Keyword(s):  
Dendritic Cells ◽  
Cell System ◽  
The Other ◽  
Scavenging Activity ◽  
Tissue Sections ◽  
Colloidal Carbon ◽  
Beta 2 ◽  
Endocytic Activity ◽  
Antigen Presenting

The dendritic cell system operates in situ to capture and present antigens in a form that is immunogenic to T cells. It is likely that dendritic cells require endocytic activity in order to process antigens. On the other hand, macrophages are considered to be the principal cells that internalize substrates in situ. We therefore investigated the phenotype of cells that scavenge the indigestible endocytic tracer, colloidal carbon, by phenotyping the endocytic cells with monoclonal antibodies that help distinguish macrophages from dendritic cells. Of some importance was the monoclonal N418, an antibody to the p150/90 leukocyte beta 2 integrin. FACS analyses on isolates from blood, spleen and peritoneal cavity showed that N418 reacts primarily with dendritic cells. N418 also stained dendritic profiles strongly in tissue sections of liver and spleen, but most of the cells that actively endocytosed carbon in both organs showed little or no N418 staining. Likewise, carbon could not be identified in cells that react with M342, which stains intracellular granules of dendritic cells. In contrast, the carbon-labeled cells in both liver and spleen were labeled with antibodies (SER-4, F4/80, FA11) that bind primarily to isolated macrophages. Therefore the clearance of colloidal carbon in situ reflects the scavenging activity of macrophages and not the endocytic activity that underlies the antigen presenting function of dendritic cells.

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