Expression Database

GUDMAP:22105

Gene:	Crym, crystallin, mu
Stage:	Theiler Stage 23
Tissue:
Species:	Mus musculus
Assay Type:	TG mouse marker strain

Images

	Expression of EGFP in the kidneys of Crym-EGFP BAC transgenic mice. Top) Bright field microscopy image detailing the expression of Crym using whole-mount in situ hybridization in the kidneys of E12.5 embryos (Image courtesy of GUDMAP:7810, Little Group). Middle) Fluorescent microscopy image showing Crym-EGFP expression in the kidney from E18.5 embryos. Note the expression of GFP expression in the cap mesencyme. Lower) Image represents a close-up detailing GFP expression in the developing cap mesenchyme.
	Confocal image details the expression of Crym-EGFP, which can be seen in the cap mesenchyme. The tubules of the kidney were labeled with E-cadherin. Crym (green) and E-cadherin (blue).
	Confocal movie showing expression of Crym-EGFP in the developing kidney of E18.5 embryos. Strong Crym-EGFP expression can be detected in the developing cap mesenchyme. The tubules of the kidney were labeled with E-cadherin Crym (green) and E-cadherin (blue).

Results Notes

Analysis suggests that the Crym-EGFP transgenic mice may be a useful tool to study the development of the cap mesenchyme and renal vesicle.

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):	Steve Potter, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA, steve.potter@cchmc.org
Contributors:	Eric W Brunskill, Steven Potter
Submitted By:	Eric Brunskill, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA, brune0@cchmc.org
Batch ID:	566
Submission ID:	22105

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:

Strain:

unspecified

Genotype:

Non-wild type Allele:
Gene		Crym
MGI ID		MGI:4846969
MGI Symbol or lab name		Tg(Crym-EGFP)GF82Gsat
Type		Transgenic, (Reporter)
Allele First Chromatid		Tg(Crym-EGFP)GF82Gsat
Allele Second Chromatid		wild type
Notes:		Mouse strain generated during NIH-funded project, GENSAT (http://www.gensat.org/index.html). The strain can be obtained from MMRRC (https://www.mmrrc.org/catalog/sds.php?mmrrc_id=12003).

Sex:

unknown

Specimen Preparation:

mouse marker strain

Fixation Method:

unspecified

Experiment Notes:

Tissue processing for confocal microscopy Kidneys were dissected in phosphate buffered saline (PBS). The kidneys or the organ explants were rocked for 1–2 h in 2% paraformaldehyde in PBS, washed twice with PBS, and then rocked for 1–2 h in 100% methanol. The tissues were washed twice with cold PBS containing 0.05% Tween-20 (PBT). Kidneys were bisected. Primary antibodies, diluted to 1:250 to 1:400, were added to the tissues in 400 μL of PBT containing 2% goat serum and incubated overnight with rocking. Tissues were washed with 5 exchanges of PBT over 8 h with rocking. The secondary antibodies, diluted to 1:400 in PBT containing 2% goat serum, were added and incubated overnight. The tissues were again washed with 5 exchanges of PBT over 8 h. The tissue was washed for 5–10 min and mounted in a depression slide in PBT before they were examined by confocal microscopy. The entire procedure was performed at 4 °C with pre- cooled reagents. The following primary antibodies were utilized: anti-WT1 (c-19, Santa Cruz), anti- Uvomorulin (E-cadherin, Sigma). The secondary antibodies were Alexa 555-conjugated anti-rabbit and Alexa 633-conjugated anti-rat secondary antibodies (Molecular Probes). Confocal imaging The tissues were imaged with a Zeiss LSM510 equipped with an Argon (488 nm) and two HeNe lasers (543 nm and 633 nm). We used a multi-track configuration, refractive index correction, and automatic gain control. Approximately 2 μm thick optical sections were obtained every 5 μm to a depth of at least 80 μm. The sections began at the surface of the kidney and were on a plane tangential to it.

Transgene Visualisation

Reporter:	GFP
Direct method for visualising reporter:	fluorescence

Acknowledgements

Comprehensive PDF containing allele characterisation can be downloaded from the GUDMAP mouse strains page (http://www.gudmap.org/Resources/MouseStrains/Mouse_Strains_Summary.html). GUDMAP:22105 entry compiled from this PDF by GUDMAP Editorial Office.