Expression Database

GUDMAP:21886

Gene:	Akr1b7, aldo-keto reductase family 1, member B7
Stage:	Theiler Stage 23
Tissue:	urinary system
Species:	Mus musculus
Assay Type:	TG mouse marker strain

Images

	Wholemount TagRFP-T detection in 15.5dpc Akr1b7TagRFP-T/+ UGS. Strong TagRFP-T fluorescence was visible in the adrenal glands from dissected UGSs. Expression in the medullary vasculature was more apparent after removal of the adrenal glands. TagRFP-T expression was limited to the Akr1b7 domain.
	TagRFP-T signal is detected strongly in the adrenal glands and in the medullary vasculature of Akr1b7TagRFP-T/+ embryos. 15.5 dpc UGS from Akr1b7TagRFP-T/+ embryos probed with anti-TagRFP-T, anti-Flk 1(endothelial cell), cytokeratin antibodies, DAPI nuclear staining.
	TagRFP-T positve cells are closely associated but distinct from PECAM positive medullary arterioles in the kidneys of 15.5 dpc Akr1b7TagRFP-T/+ embryos. Kidneys from Akr1b7TagRFP-T/+ embryos probed with anti-TagRFP-T, and anti-PECAM (endothelial cell) antibodies show close apposition of the TagRFP-T positive cells with PECAM positive endothelial cells.
	TagRFP-T signal is associated with the medullary vasculature in the kidneys of 15.5 dpc Akr1b7TagRFP-T/+ embryos. Kidneys from Akr1b7TagRFP-T/+ and wildtype embryos probed with anti-TagRFP-T, anti-Flk 1 (endothelial cells) and anti-Cytokeratin antibodies show a close association of the TagRFP-T expressing cells with Flk 1 positive endothelial cells of the medullary arterioles.
	TagRFP-T signal is detected in close association with endothelial cells of the medullary vasculature in the kidney of 15.5 dpc Akr1b7TagRFP-T/+ embryos. Kidneys from Akr1b7TagRFP-T/+ and wildtype embryos probed with anti-TagRFP-T, anti-PECAM ( endothelial cell) and anti-Smooth Muscle Actin antibodies show a coexpression of the TagRFP-T and Smooth Muscle Actin in non endothelial cells closely associated with the medullary arterioles.

Results Notes

Submitter findings: Analysis has confirmed expression of TagRFP-T cells in the adrenal glands and in association with the intrarenal arteries of the kidney. Potential expression in non-kidney components of the urogenital system were not characterized but have been reported (Taragnet C et al, 1988, J Reprod Fertil 83:835-842). Renal vasculature associated expression is observed in Smooth Muscle Actin positive cells that are closely associated with Flk 1 and PECAM positive intrarenal arteries. The expression of the Akr1b7-TagRFP-T is very similar to the expression of the Renin gene described in (Jones CA et als, 2000 Physiol Genomics 4:75-81) and may indicate a co-localization in the expression domains of the two genes. This possibility is being followed up.

Mouse marker strain entries are unique GUDMAP entries that can contain data for multiple probes at multiple stages of development (more info).

Submitters

Principal Investigator(s):	Andrew P McMahon, Keck School of Medicine, Los Angeles, CA, USA, amcmahon@med.usc.edu
Contributors:	Andrew P McMahon, M Todd Valerius, Jinjin Guo, Jill McMahon
Submitted By:	Jill McMahon, Keck School of Medicine, Los Angeles, CA, USA
Batch ID:	555
Submission ID:	21886

Expression Mapping

Expression Strengths Key:

	Present (unspecified strength)
	Present (strong)
	Present (moderate)
	Present (weak)
	Uncertain
	Not Detected

Expression Patterns Key:

	Homogeneous
	Graded
	Regional
	Spotted
	Ubiquitous

	Restricted
	Single cell

Nerve Density:

Relative to Total:

	High
	Medium
	Low

Relative to P0/Adult:

	Increase, large
	Increase, small
	Decrease, large
	Decrease, small

	Contains note

View annotated components as a list

Show annotation under groups

Specimen

Stage:

TS23

Other Staging System:

15.5 dpc

Tissue:

urinary system

Strain:

mixed background

Genotype:

Non-wild type Allele:
Gene		Akr1b7
MGI ID		MGI:4830367
MGI Symbol or lab name		Tg(Akr1b7-RFP)9Amc
Type		Transgenic, (Reporter)
Allele First Chromatid		Tg(Akr1b7-RFP)9Amc
Allele Second Chromatid		wild type
Notes:		Mouse strain is available from the Jackson Lab, see here: http://jaxmice.jax.org/strain/013137.html

Sex:

both

Specimen Preparation:

mouse marker strain

Fixation Method:

unspecified

Experiment Notes:

Immunohistochemistry was performed to examine whether the TagRFP-T allele was expressed in the expected Akr1b7 domain. UGS samples from 15.5 dpc Akr1b7 TagRFP-T +/- embryos and wildtype litter mates were examined. Whole UGSs were fixed in 4% paraformaldehyde at 4ºC for 2 hours, washed 3 times in PBS, equilibrated in 30% sucrose overnight and then embedded in OCT and flash frozen on dry-ice. The UGSs were sectioned at 16um and stained with either; rabbit anti-tRFP/rat-anti-PECAM/mouse-anti-Cytokeratin, rabbit-anti-tRFP/rat-anti-Flk1/mouse anti-Cytokeratin, or rabbit-anti-tRFP/mouse-anti-Actin, α-Smooth Muscle/ chicken-anti-Laminin. Anti-tRFP (Rabbit, Evrogen AB234,1:500); anti-Actin, α-Smooth Muscle (Mouse IgG2a, Sigma, A5228, 1: 1000); anti-Flk1 (Rat, BD Pharmingen, 555307,1:1000); anti-PECAM (Rat, BD Pharmingen, 553370, 1:1000);anti- Laminin ( Chicken, Abcam, ab14055, 1:500); anti-Cytokeratin (Mouse IgG1, Sigma, C 2562, 1:500) were incubated overnight at 4ºC and detected with secondary antibodies Alexafluor 488, 568, 633, and 647 (Molecular probes).

Transgene Visualisation

Reporter:	RFP
Direct method for visualising reporter:	Fluorescence

Acknowledgements

Comprehensive PDFs containing allele verification and characterisation can be downloaded from the GUDMAP mouse strains page (http://www.gudmap.org/Resources/MouseStrains/Mouse_Strains_Summary.html). GUDMAP:21886 entry compiled from these PDFs by GUDMAP Editorial Office.