Download The Effect of the BAF Chromatin Regulator in Heart Valve Maturation

THE EFFECT OF THE
BAF CHROMATIN
REGULATOR IN
HEART VALVE
MATURATION
OF MUS MUSCULUS
Kelsey Ward
Kryn Stankunas, P.I.
Brynn Simek, Mentor
CONGENITAL HEART DISEASE (CHD)
• Most common type of birth defect
•1 in 125 births
EPIGENETICS AND CHROMATIN REMODELING
VALVULOGENESIS
Chromatin Remodeling by the
BAF Complex
Brg1
VALVULOGENESIS
BRG1 DELETION IN ALL ENDOCARDIAL CELLS
IS EMBRYONIC LETHAL
Stankunas, K.
Methodology
♂
SCL-Cre-ER(T);
Brg1F/F
♀
Brg1 F/F
Genotype
embryos
Imaging
whole
embryos
Characterize valve
defects and consider
new time points and
crosses
• Check Plugs
• Administer Tamoxifen
(2x40mg/kg)
• Harvest embryos
•
•
•
•
•
Preserve in PFA
Transfer to PBS
Dehydrate to 100% EtOH
Replace with xylenes
Replace with paraffin and fix in
blocks
• Section at 7 microns
• H&E stain
• Image sections
VALVULOGENESIS
TAMOXIFEN ADMINISTRATION AT E12.5
A
B
C
D
IS BRG1 ACTUALLY DELETED?
Pulmonic Valve
Mitral Valve
HOW QUICKLY DOES DELETION OCCUR?
VENTRICULAR SEPTAL DEFECTS
79.8
3.5
HOW EFFICIENT IS DELETION?
CONCLUSIONS AND FUTURE DIRECTIONS
Brg1 deletion has no phenotypic effect on later
valve maturation.
 Characterization of Scl-cre line and optimization
of Tamoxifen dosage.

Earlier Brg1 deletion leads to cushion defects and
ventricular defects.
 Allowance of a longer growth period before
harvesting.
 Describing the molecular and cellular basis for
the phenotypes.

ACKNOWLEDGEMENTS

Brynn Simek, Mentor

Kryn Stankunas, P.I.

Stankunas Lab
•Alex
•Andrew
•Fern
•Matt
•Scott

SPUR