scholarly journals Whole-mount Immunohistochemical Analysis for Embryonic Limb Skin Vasculature: a Model System to Study Vascular Branching Morphogenesis in Embryo

10.3791/2620 ◽  
2011 ◽  
Author(s):  
Wenling Li ◽  
Yoh-suke Mukouyama
Keyword(s):  
Immunohistochemical Analysis ◽  
Branching Morphogenesis ◽  
Model System ◽  
Whole Mount ◽  
Embryonic Limb ◽  
Vascular Branching

scholarly journals Whole-Mount Confocal Microscopy for Vascular Branching Morphogenesis

2011 ◽  
pp. 69-78 ◽  
Author(s):  
Yoh-suke Mukouyama ◽  
Jennifer James ◽  
Joseph Nam ◽  
Yutaka Uchida
Keyword(s):  
Confocal Microscopy ◽  
Branching Morphogenesis ◽  
Whole Mount ◽  
Vascular Branching

scholarly journals Whole-Mount Adult Ear Skin Imaging Reveals Defective Neuro-Vascular Branching Morphogenesis in Obese and Type 2 Diabetic Mouse Models

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Tomoko Yamazaki ◽  
Wenling Li ◽  
Ling Yang ◽  
Ping Li ◽  
Haiming Cao ◽  
...  
Keyword(s):  
Mouse Models ◽  
Branching Morphogenesis ◽  
Diabetic Mouse ◽  
Skin Imaging ◽  
Whole Mount ◽  
Type 2 Diabetic ◽  
Vascular Branching

FGF10 is an inducer and Pax6 a competence factor for lacrimal gland development

Development ◽  
2000 ◽  
Vol 127 (12) ◽  
pp. 2563-2572 ◽  
Author(s):  
H.P. Makarenkova ◽  
M. Ito ◽  
V. Govindarajan ◽  
S.C. Faber ◽  
L. Sun ◽  
...  
Keyword(s):  
Lacrimal Gland ◽  
Normal Development ◽  
Branching Morphogenesis ◽  
Model System ◽  
Specific Marker ◽  
Fibroblast Growth ◽  
Secretory Function ◽  
Competence Factor ◽  
Branched Structure ◽  
Gland Development

We investigated the mechanism of tissue induction and specification using the lacrimal gland as a model system. This structure begins its morphogenesis as a bud-like outgrowth of the conjunctival epithelium and ultimately forms a branched structure with secretory function. Using a reporter transgene as a specific marker for gland epithelium, we show that the transcription factor Pax6 is required for normal development of the gland and is probably an important competence factor. In investigating the cell-cell signaling required, we show that fibroblast growth factor (FGF) 10 is sufficient to stimulate ectopic lacrimal bud formation in ocular explants. Expression of FGF10 in the mesenchyme adjacent to the presumptive lacrimal bud and absence of lacrimal gland development in FGF10-null mice strongly suggest that it is an endogenous inducer. This was supported by the observation that inhibition of signaling by a receptor for FGF10 (receptor 2 IIIb) suppressed development of the endogenous lacrimal bud. In explants of mesenchyme-free gland epithelium, FGF10 stimulated growth but not branching morphogenesis. This suggested that its role in induction is to stimulate proliferation and, in turn, that FGF10 combines with other factors to provide the instructive signals required for lacrimal gland development.

scholarly journals Matrix GLA protein modulates branching morphogenesis in fetal rat lung

2004 ◽  
Vol 286 (6) ◽  
pp. L1179-L1187 ◽  
Author(s):  
Kirk A. Gilbert ◽  
Stephen R. Rannels
Keyword(s):  
Mrna Expression ◽  
Time Course ◽  
Immunohistochemical Analysis ◽  
Fetal Lung ◽  
Branching Morphogenesis ◽  
Matrix Gla Protein ◽  
Antibody Treatment ◽  
Developmentally Regulated

The regulation of matrix γ-carboxyglutamic acid protein (MGP) expression during the process of lung branching morphogenesis and development was investigated. MGP mRNA expression was determined over an embryonic and postnatal time course and shown to be developmentally regulated. Immunohistochemical analysis revealed increased staining for MGP in peripheral mesenchyme surrounding distal epithelial tubules. Fetal lung explants were used as an in vitro growth model to examine expression and regulation of MGP during branching morphogenesis. MGP mRNA expression over the culture interval mimicked the in vivo time course. Explants cultured in the presence of antibodies against MGP showed gross dilation and reduced terminal lung bud counts, accompanied by changes in MGP, sonic hedgehog, and patched mRNA expression. Similarly, antifibronectin antibody treatment resulted in explant dilation and reduced MGP expression, providing evidence for an interaction with MGP and fibronectin. Conversely, intraluminal microinjection of anti-MGP antibodies had no effect either on explant growth or MGP expression, supporting the hypothesis that MGP exerts its effects through the mesenchyme. Taken together, the results suggest that MGP plays a role in lung growth and development, likely via temporally and spatially specific interactions with other branching morphogenesis-related proteins to influence growth processes.

scholarly journals Retinal myeloid cells regulate tip cell selection and vascular branching morphogenesis via Notch ligand Delta-like 1

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Fabian Haupt ◽  
Kashyap Krishnasamy ◽  
L. Christian Napp ◽  
Michael Augustynik ◽  
Anne Limbourg ◽  
...  
Keyword(s):  
Myeloid Cells ◽  
Branching Morphogenesis ◽  
Cell Selection ◽  
Notch Ligand ◽  
Tip Cell ◽  
Vascular Branching

scholarly journals Immunocytochemical detection of regional protein changes in rat brain sections using computer-assisted image analysis.

1996 ◽  
Vol 44 (9) ◽  
pp. 981-987 ◽  
Author(s):  
X Huang ◽  
S Chen ◽  
E I Tietz
Keyword(s):  
Image Analysis ◽  
Rat Brain ◽  
Immunohistochemical Analysis ◽  
Brain Homogenate ◽  
Model System ◽  
Computer Assisted ◽  
Antigen Concentration ◽  
Immunocytochemical Detection ◽  
Low Degree ◽  
Computer Assisted Image

We used several approaches to assess the reliability and sensitivity of computer-assisted densitometry to detect regional changes in tissue antigen content as a function of immunohistochemical staining density. We designed a model system to mimic variations in antigen concentration in postfixed, slide-mounted rat brain sections by varying the ratios of conjugated (biotinylated) to unconjugated secondary antibody. Antigen concentration was also varied in tissue discs made from mixing rat brain homogenate with increasing amounts of tissue embedding compound. The monoclonal antibody bd-17 to the beta2/3 subunit of the GABAA receptor was used as the primary antibody. Immunostaining density was visualized with diaminobenzidine (DAB). There was a significant, positive linear relationship (r = 0.97-0.99) between immunostaining intensity and antigen concentration. With this approach, changes in antigen content of less than 10%, as reflected in immunostaining intensity, were detectable in brain sections. The low degree of variability in measures of regional variation in immunostaining in sections from naive rats (n = 7) suggested that the method was suitable for quantitative analysis and indicated the reliability of the method. This systematic study of the utility of computer-assisted image analysis for semiquantitative immunohistochemical analysis found the method to be both reliable and sensitive.

Immunolocalisation of collagenase in rabbit periosteal tissue explants and extraction of the enzyme. The effect of the cytokines IL-1 alpha and EGF

1994 ◽  
Vol 107 (4) ◽  
pp. 1047-1053 ◽  
Author(s):  
E. van der Zee ◽  
V. Everts ◽  
K. Hoeben ◽  
W. Beertsen
Keyword(s):  
Extracellular Matrix ◽  
In Vitro Model ◽  
Interleukin 1 ◽  
Immunohistochemical Analysis ◽  
Model System ◽  
High Concentration ◽  
Active Collagenase ◽  
Epidermal Growth ◽  
Vitro Model

The effect of interleukin-1 alpha (IL-1 alpha) and murine epidermal growth factor (EGF) on incorporation of endogenously produced collagenase in the extracellular matrix of soft connective tissue was studied in an in vitro model system using periosteal explants obtained from rabbit calvariae. Immunohistochemical analysis indicated the highest level of collagenase in explants cultured for 72 hours with IL-1 alpha in combination with EGF. Most enzyme appeared to be associated with the extracellular matrix, but labeling was also found in numerous fibroblast-like cells. Explants cultured in the presence of IL-1 alpha alone contained less enzyme and in periostea treated without cytokines, or with EGF alone, only a faint label, if any, was seen. Freshly isolated, non-cultured periostea contained no detectable enzyme. Extraction of collagenase from periostea revealed that: (1) non-cultured periosteum did not contain detectable levels of enzyme. (2) The amount of total activatable enzyme synergistically increased (10-fold) under the influence of IL-1 alpha and EGF, whereas IL-1 alpha alone showed a 4-fold enhancement compared to control or EGF-incubated explants. (3) The latent fraction of the enzyme was synergistically increased (up to 100-fold or more) in periostea cultured in the presence of IL-1 alpha + EGF (21.17 mU/explant versus 0.05 mU/explant in controls). (4) Active collagenase, on the other hand, appeared to be present in a relatively high concentration in explants cultured without cytokines (2.45 mU/explant versus 0.36 mU/explant in IL-1 alpha + EGF-treated explants).(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Ott1 (Rbm15) Is Essential for Placental Vascular Branching Morphogenesis and Embryonic Development of the Heart and Spleen

2008 ◽  
Vol 29 (2) ◽  
pp. 333-341 ◽  
Author(s):  
Glen D. Raffel ◽  
Gerald C. Chu ◽  
Jonathan L. Jesneck ◽  
Dana E. Cullen ◽  
Roderick T. Bronson ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Cardiac Development ◽  
Germ Line ◽  
Global Gene Expression ◽  
Branching Morphogenesis ◽  
Conditional Allele ◽  
Recognition Motifs ◽  
Regulatory Effects ◽  
Incomplete Closure ◽  
Vascular Branching

ABSTRACT The infant leukemia-associated gene Ott1 (Rbm15) has broad regulatory effects within murine hematopoiesis. However, germ line Ott1 deletion results in fetal demise prior to embryonic day 10.5, indicating additional developmental requirements for Ott1. The spen gene family, to which Ott1 belongs, has a transcriptional activation/repression domain and RNA recognition motifs and has a significant role in the development of the head and thorax in Drosophila melanogaster. Early Ott1-deficient embryos show growth retardation and incomplete closure of the notochord. Further analysis demonstrated placental defects in the spongiotrophoblast and syncytiotrophoblast layers, resulting in an arrest of vascular branching morphogenesis. The rescue of the placental defect using a conditional allele with a trophoblast-sparing cre transgene allowed embryos to form a normal placenta and survive gestation. This outcome showed that the process of vascular branching morphogenesis in Ott1-deficient animals was regulated by the trophoblast compartment rather than the fetal vasculature. Mice surviving to term manifested hyposplenia and abnormal cardiac development. Analysis of global gene expression of Ott1-deficient embryonic hearts showed an enrichment of hypoxia-related genes and a significant alteration of several candidate genes critical for cardiac development. Thus, Ott1-dependent pathways, in addition to being implicated in leukemogenesis, may also be important for the pathogenesis of placental insufficiency and cardiac malformations.

scholarly journals Hoxa11andHoxd11regulate branching morphogenesis of the ureteric bud in the developing kidney

Development ◽  
2001 ◽  
Vol 128 (11) ◽  
pp. 2153-2161 ◽  
Author(s):  
Larry T. Patterson ◽  
Martina Pembaur ◽  
S. Steven Potter
Keyword(s):  
Gene Expression ◽  
Kidney Development ◽  
Growth Failure ◽  
Immunohistochemical Analysis ◽  
Branching Morphogenesis ◽  
Metanephric Mesenchyme ◽  
Ureteric Bud ◽  
Metanephric Kidney ◽  
Normal Gene ◽  
Ureteric Buds

Hoxa11 and Hoxd11 are functionally redundant during kidney development. Mice with homozygous null mutation of either gene have normal kidneys, but double mutants have rudimentary, or in extreme cases, absent kidneys. We have examined the mechanism for renal growth failure in this mouse model and find defects in ureteric bud branching morphogenesis. The ureteric buds are either unbranched or have an atypical pattern characterized by lack of terminal branches in the midventral renal cortex. The mutant embryos show that Hoxa11 and Hoxd11 control development of a dorsoventral renal axis. By immunohistochemical analysis, Hoxa11 expression is restricted to the early metanephric mesenchyme, which induces ureteric bud formation and branching. It is not found in the ureteric bud. This suggests that the branching defect had been caused by failure of mesenchyme to epithelium signaling. In situ hybridizations with Wnt7b, a marker of the metanephric kidney, show that the branching defect was not simply the result of homeotic transformation of metanephros to mesonephros. Absent Bf2 and Gdnf expression in the midventral mesenchyme, findings that could by themselves account for branching defects, shows that Hoxa11 and Hoxd11 are necessary for normal gene expression in the ventral mesenchyme. Attenuation of normal gene expression along with the absence of a detectable proliferative or apoptotic change in the mutants show that one function of Hoxa11 and Hoxd11 in the developing renal mesenchyme is to regulate differentiation necessary for mesenchymal-epithelial reciprocal inductive interactions.

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