PUBLICATION
Visualizing the Interrenal Steroidogenic Tissue and Its Vascular Microenvironment in Zebrafish
- Authors
- Chou, C.W., Lin, J., Hou, H.Y., Liu, Y.W.
- ID
- ZDB-PUB-170108-9
- Date
- 2016
- Source
- Journal of visualized experiments : JoVE (118): (Journal)
- Registered Authors
- Liu, Yi-wen
- Keywords
- Developmental Biology, 3-β-hydroxysteroid dehydrogenase, zebrafish, steroidogenic, adrenocortical, interrenal, kidney, blood vessels, microenvironment, vibratome sectioning, immunohistochemistry
- MeSH Terms
-
- 3-Hydroxysteroid Dehydrogenases/metabolism*
- Animals
- Animals, Genetically Modified
- Interrenal Gland/cytology*
- Staining and Labeling/methods*
- Zebrafish*
- Zebrafish Proteins/metabolism*
- PubMed
- 28060344 Full text @ J. Vis. Exp.
Citation
Chou, C.W., Lin, J., Hou, H.Y., Liu, Y.W. (2016) Visualizing the Interrenal Steroidogenic Tissue and Its Vascular Microenvironment in Zebrafish. Journal of visualized experiments : JoVE. (118).
Abstract
This protocol introduces how to detect differentiated interrenal steroidogenic cells through a simple whole-mount enzymatic activity assay. Identifying differentiated steroidogenic tissues through chromogenic histochemical staining of 3-β-Hydroxysteroid dehydrogenase /Δ5-4 isomerase (3β-Hsd) activity-positive cells is critical for monitoring the morphology and differentiation of adrenocortical and interrenal tissues in mammals and teleosts, respectively. In the zebrafish model, the optical transparency and tissue permeability of the developing embryos and larvae allow for whole-mount staining of 3β-Hsd activity. This staining protocol, as performed on transgenic fluorescent reporter lines marking the developing pronephric and endothelial cells, enables the detection of the steroidogenic interrenal tissue in addition to the kidney and neighboring vasculature. In combination with vibratome sectioning, immunohistochemistry, and confocal microscopy, we can visualize and assay the vascular microenvironment of interrenal steroidogenic tissues. The 3β-Hsd activity assay is essential for studying the cell biology of the zebrafish interrenal gland because to date, no suitable antibody is available for labeling zebrafish steroidogenic cells. Furthermore, this assay is rapid and simple, thus providing a powerful tool for mutant screens targeting adrenal (interrenal) genetic disorders as well as for determining disruption effects of chemicals on steroidogenesis in pharmaceutical or toxicological studies.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping