Expression Database

GUDMAP:21869

Gene:	Cyp11a1, cytochrome P450, family 11, subfamily a, polypeptide 1
Stage:	Theiler Stage 23
Tissue:	urinary system
Species:	Mus musculus
Assay Type:	TG mouse marker strain

Images

	Cre- B-gal activity in Cyp11a1GC/+;R26RlacZ/+ UGSs. ?-gal activity was detected in all of the Cyp11a1GC/+;R26RlacZ/+ UGSs. Not in R26RlacZ/+ littermates UGGs. Upper two rows: Cyp11a1GC/+;R26RlacZ/+ and R26RlacZ/+ littermates UGSs were stained with Xgal, (X-gal Blue), at low magnification. Middle row: Cyp11a1GC/+;R26RlacZ/+ and R26RlacZ/+ littermates UGSs were stained with X-gal, (Xgal Blue), at high magnification. Lower two rows: Cyp11a1GC/+;R26RlacZ/+ and R26RlacZ/+ littermates UGS of the sections were stained with X-gal, (X-gal Blue).
	GFP protein detected by antibody staining . GFP and Bgal protein was detected in Cyp11a1GC/+;R26RlacZ/+ kidneys, no GFP and Bgal was detected in R26RlacZ/+ litter mates control kidneys. Upper four rows: Cyp11a1GC/+;R26RlacZ/+ kidneys and R26RlacZ/+ littermates control kidneys are stained with anti- b-gal and anti-GFP, (b-gal red, GFP Green). Lower two rows: Cyp11a1GC/+;R26RlacZ/+ kidneys and R26RlacZ/+ littermates control kidneys are stained with anti- Laminin and anti-GFP, (Laminin red, GFP Green).
	GFP protein detected by antibody staining (Confocal). GFP and Bgal protein was detected in Cyp11a1GC/+;R26RlacZ/+ kidneys, no GFP and Bgal was detected in R26RlacZ/+ littermates control kidneys. Upper four rows: Cyp11a1GC/+;R26RlacZ/+ kidneys and R26RlacZ/+ littermates control kidneys are stained with anti- b-gal and anti-GFP, (b-gal red, GFP Green). Lower two rows: Cyp11a1GC/+;R26RlacZ/+ kidneys and R26RlacZ/+ littermates control kidneys are stained with anti- Laminin and anti-GFP, (Laminin red, GFP Green).
	GFP protein detected by antibody staining. GFP was detected in the Cyp11a1GC/+;R26RlacZ/+ testis; cytochrome P450 was detected in both of the Cyp11a1GC/+; R26RlacZ/+ and R26RlacZ/+ littermate control testis. GFP and cytochrome P450 were colocalized in the Leydig cells of the Cyp11a1GC/+;R26RlacZ/+ testis. The upper two rows were collected by confocal microscopy. The lower to rows were collected by epifluorescence.

Results Notes

Submitter finding: This allele is expressing GFP-Cre in the expected cell populations and Cre-dependent R26R-LacZ expression is observed in those tissues. We conclude this allele does exhibit the expected activity in leydig cells of testis and adrenal gland.

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, Pumin Zhang
Submitted By:	M. Todd Valerius, Harvard University, Cambridge, MA, USA, tvalerius@mcb.harvard.edu
Batch ID:	555
Submission ID:	21869

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		Cyp11a1
MGI ID		MGI:4442187
MGI Symbol or lab name		Cyp11a1^{tm1(GFP/cre)Pzg}
Type		Targeted (knock-in)
Allele First Chromatid		Cyp11a1^{tm1(GFP/cre)Pzg}
Allele Second Chromatid		wild type
Notes:		Mouse strain is available from the Jackson Lab, see here: http://jaxmice.jax.org/strain/010988.html

Sex:

both

Specimen Preparation:

mouse marker strain

Fixation Method:

unspecified

Experiment Notes:

Rosa26RlacZ/+ (R26R) female mice were crossed with Cyp11a1GC/+ males to obtain Cyp11a1GC/+;R26RlacZ/+ embryos. The embryos were dissected on E15.5 to collect the urogenital system (UGS). Immunohistochemistry was performed to examine if the GC allele was expressed in the expected Cyp11a1 domain. Two of each male and female of Cyp11a1GC/+;R26RlacZ/+ experimentals and R26RlacZ/+ littermate controls from two litters were assayed, respectively. GFP protein was assayed by staining with chicken-anti-GFP. To test for Cre function, β-gal and GFP expression were examined by double staining with rabbitanti- b-gal and chicken-anti-GFP. To test for the GFP-Cre cassette expressed in the expected Cyp11a1 domain, GFP and Cyp11a1 (i.e. P450) protein were examined by double staining with rabbit-anti- rat cytochrome P450 side chain cleavage antibody and chicken-anti-GFP. Whole UGSs were fixed in 4% paraformaldehyde at 4ºC 2 hours, washed 3 times in PBS, equilibrated in 30% sucrose overnight, embedded in OCT and flash frozen on dry ice. The UGSs were sectioned at 20um and stained with rabbit-anti-b-gal/chicken-anti- GFP/mouse-anti-Cytokeratin, rabbit-anti-Laminin/chicken-anti-GFP, and rabbit-anticytochrome p450/chicken-anti-GFP, respectively. GFP (Chicken, Aves Labs, Inc, GFP- 1020, 1:500); beta-gal (Rabbit, MP Biomedicals, LLC, 55976, 1: 20000); cytochrome P450 side chain cleavage antibody ( Chemicon International, AB 1294, 1:200); Laminin (Rabbit, Sigma, L9393, 1:300) Cytokeratin (Mouse IgG1, Sigma, C 2562, 1:500) were incubated overnight at 4ºC and detected with secondary antibodies Alexafluor 488, 568, 647 (Molecular probes) as indicated in the figures.

Transgene Visualisation

Reporter:	GFP
Direct method for visualising reporter:	Fluorescence

Acknowledgements

Comprehensive PDFs containing allele verification (provided by Zhang lab) and allele characterisation (provided by McMahon lab) can be downloaded from the GUDMAP mouse strains page (http://www.gudmap.org/Resources/MouseStrains/Mouse_Strains_Summary.html). GUDMAP:21869 entry compiled from these PDFs by GUDMAP Editorial Office.