Expression Database

GUDMAP:22103

Gene:	Hes5, hairy and enhancer of split 5 (Drosophila)
Stage:	Theiler Stage 23
Tissue:
Species:	Mus musculus
Assay Type:	TG mouse marker strain

Images

	A) Bright field microscopy image detailing the expression of Hes5 using whole-mount in situ hybridization in the kidneys of E15.5 embryos (Image courtesy of GUDMAP:5928, McMahon Group). B) Fluorescent microscopy image showing Hes5-EGFP expression in the kidney from E15.5 embryos. Note the punctate GFP expression recapitulates the endogenous Hes5 expression.
	A) Bright-field microscopy image showing a cryo-section (8?M section) through an E15.5 Hes5-EGFP transgenic kidney. B) Fluorescent microscopy image showing Hes5-EGFP expression in the developing kidney. Note the expression of GFP in the developing S-shape body.
	Expression of Hes5-EGFP, which can be seen in the medial region of the developing S-shape body. This is consistent with the known expression pattern of Hes5 in the kidney. Arrows indicate the expression of Hes5 in the S-shaped body. The tubules of the kidney were labeled with E-cadherin, and the mesenchyme and developing glomeruli labeled by WT-1 expression. Hes5 (green), Ecadherin (blue), WT-1 (red).
	Confocal movie showing strong Hes5-EGFP expression detected in the medial region of the developing S-shape body. In addition, the expression of Hes5 can also be sub-region of the renal vesicle. The expression from Hes5- GFP transgenic mice is consistent with the known expression pattern of Hes5 in the kidney. The tubules of the kidney were labeled with E-cadherin, and the mesenchyme and developing glomeruli labeled by WT-1 expression. Hes5 (green), E-cadherin (blue), WT-1 (red).

Results Notes

Analysis suggests that Hes5-EGFP transgenic mice may be useful to further studies regarding the developing renal vesicle and S-shaped bodies.

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:	22103

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		Hes5
MGI ID		MGI:3840943
MGI Symbol or lab name		Tg(Hes5-EGFP)CV50Gsat
Type		Transgenic, (Reporter)
Allele First Chromatid		Tg(Hes5-EGFP)CV50Gsat
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=316).

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:22103 entry compiled from this PDF by GUDMAP Editorial Office.