Gata3 Hypomorphic Mutant Mice Rescued with a Yeast Artificial Chromosome Transgene Suffer a Glomerular Mesangial Cell Defect

Abstract

bGATA3 is a zinc finger transcription factor that plays a crucial role in embryonic kidney development, while its precise functions in the adult kidney remain largely unexplored. Here, we demonstrate that GATA3 is specifically expressed in glomerular mesangial cells and plays a critical role in the maintenance of renal glomerular function. Newly generated Gata3 hypomorphic mutant mice exhibited neonatal lethality associated with severe renal hypoplasia. Normal kidney size was restored by breeding the hypomorphic mutant with a rescuing transgenic mouse line bearing a 662-kb Gata3 yeast artificial chromosome (YAC), and these animals (termed G3YR mice) survived to adulthood. However, most of the G3YR mice showed degenerative changes in glomerular mesangial cells, which deteriorated progressively during postnatal development. Consequently, the G3YR adult mice suffered severe renal failure. We found that the 662-kb Gata3 YAC transgene recapitulated Gata3 expression in the renal tubules but failed to direct sufficient GATA3 activity to mesangial cells. Renal glomeruli of the G3YR mice had significantly reduced amounts of platelet-derived growth factor receptor (PDGFR), which is known to participate in the development and maintenance of glomerular mesangial cells. These results demonstrate a critical role for GATA3 in the maintenance of mesangial cells and its absolute requirement for prevention of glomerular disease.

Induction of erythropoietin gene expression in epithelial cells by chemicals identified in GATA inhibitor screenings

“…Of the six GATA factors, we examined the expression levels of Gata2 , Gata3 , Gata4 and Gata6 mRNAs in the kidneys and lungs compared with those in the hearts and livers. Both Gata2 and Gata3 were abundantly expressed in the kidneys, in agreement with the previous reports (Moriguchi et al., ; Yu et al., ), and also in the lungs (Figure ). Gata6 was also expressed in the lungs.…”

GATA3 Abundance Is a Critical Determinant of T Cell Receptor β Allelic Exclusion

“…In concert with those observations, hypomorphic Gata3 g/g homozygous thymocytes were shown to exhibit a second peak of GFP fluorescence starting at the DN2 stage in thymocytes that were generated after adoptive transfer of fetal liver hematopoietic stem cells (HSC) and progenitors (see Fig. 3 in reference 49) (Gata3 g/g mutants could not be examined directly since the homozygous mutation leads to perinatal lethality [50]). In initially curious contrast, GFP fluorescence was never observed at the ETP, DN2, or DN3 stage in Gata3 g/ϩ heterozygous mutants (32,49).…”

“…Gata3 g (32,50), GATA3 Tg Cd2-GATA3 transgenic line 720 (36), TCR␣␤ transgenic OT-II.2 (53) (purchased from Jackson Laboratory), and TCR␤ transgenic (52) (generously provided by Karen Hathcock at National Institutes of Health, Baltimore, MD) mice were described previously and were maintained on a congenic C57BL/6J background. C57BL/6J inbred mice used for backcrossing and FVB/NJ mice were purchased from Jackson Laboratory.…”

“…Since the Gata3 monoallelic-to-biallelic transcriptional switch (49) is first observed at around the same developmental stage (DN2/DN3) as when the Tcrb loci initiate VDJ genomic DNA rearrangement, we hypothesized that Gata3 monoallelic expression might be functionally associated with allelic exclusion at the Tcrb locus. We previously generated a GATA3-enhanced green fluorescent protein (eGFP) fusion protein knock-in allele (Gata3 g ) by embryonic stem (ES) cell gene targeting (32,50), and by both RNA-fluorescence in situ hybridization (FISH) and RT-PCR single nucleotide variant (SNV) sequencing of single thymocytes, we recently demonstrated that a Gata3 monoallelic-to-biallelic transcriptional switch initiates at around the DN2 stage (49). In concert with those observations, hypomorphic Gata3 g/g homozygous thymocytes were shown to exhibit a second peak of GFP fluorescence starting at the DN2 stage in thymocytes that were generated after adoptive transfer of fetal liver hematopoietic stem cells (HSC) and progenitors (see Fig.…”

A Novel Role for GATA3 in Mesangial Cells in Glomerular Development and Injury

“…14 More recently, HDR has been associated with nephrotic syndrome and isolated cases of GN, 15,16 whereas Gata3 hypomorphic mutant mice rescued with a yeast artificial chromosome Gata3 transgene, whose expression was limited to renal tubules and not glomeruli, have been shown to suffer glomerular mesangial cell (MC) defects. 17 In genome-wide transcriptome studies, GATA3 has been identified as a highly enriched transcript in MCs, 18,19 but the functional significance of GATA3 expression in MCs in health and disease has not been explored. Moreover, GN in HDR could reflect the effect of GATA3 deficiency on immunity because overexpression in T cells protects against lupus nephritis in BXSB/ MpJ-Yaa mice, 20 and on ILC2s, which are reported to be central to renal repair.…”