Gata3-deficient mice develop parathyroid abnormalities due to dysregulation of the parathyroid-specific transcription factor Gcm2
Irina V. Grigorieva, … , Frank Grosveld, Rajesh V. Thakker
Irina V. Grigorieva, … , Frank Grosveld, Rajesh V. Thakker
Published May 17, 2010
Citation Information: J Clin Invest. 2010;120(6):2144-2155. https://doi.org/10.1172/JCI42021.
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Research Article Endocrinology

Gata3-deficient mice develop parathyroid abnormalities due to dysregulation of the parathyroid-specific transcription factor Gcm2

Abstract

Heterozygous mutations of GATA3, which encodes a dual zinc-finger transcription factor, cause hypoparathyroidism with sensorineural deafness and renal dysplasia. Here, we have investigated the role of GATA3 in parathyroid function by challenging Gata3+/– mice with a diet low in calcium and vitamin D so as to expose any defects in parathyroid function. This led to a higher mortality among Gata3+/– mice compared with Gata3+/+ mice. Compared with their wild-type littermates, Gata3+/– mice had lower plasma concentrations of calcium and parathyroid hormone (PTH) and smaller parathyroid glands with a reduced Ki-67 proliferation rate. At E11.5, Gata3+/– embryos had smaller parathyroid-thymus primordia with fewer cells expressing the parathyroid-specific gene glial cells missing 2 (Gcm2), the homolog of human GCMB. In contrast, E11.5 Gata3–/– embryos had no Gcm2 expression and by E12.5 had gross defects in the third and fourth pharyngeal pouches, including absent parathyroid-thymus primordia. Electrophoretic mobility shift, luciferase reporter, and chromatin immunoprecipitation assays showed that GATA3 binds specifically to a functional double-GATA motif within the GCMB promoter. Thus, GATA3 is critical for the differentiation and survival of parathyroid progenitor cells and, with GCM2/B, forms part of a transcriptional cascade in parathyroid development and function.

Authors

Irina V. Grigorieva, Samantha Mirczuk, Katherine U. Gaynor, M. Andrew Nesbit, Elena F. Grigorieva, Qiaozhi Wei, Asif Ali, Rebecca J. Fairclough, Joanna M. Stacey, Michael J. Stechman, Radu Mihai, Dorota Kurek, William D. Fraser, Tertius Hough, Brian G. Condie, Nancy Manley, Frank Grosveld, Rajesh V. Thakker

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Figure 2

Parathyroid histology and proliferation studies in Gata3+/– and Gata3–/– mice.

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Parathyroid histology and proliferation studies in Gata3+/– and Gata3–/–...
(A) Parathyroid size in Gata3+/+ and Gata3+/– mice after 12 days, after weaning, on control or low calcium/vitamin D diets. H&E-stained parathyroid sections (top). Scale bars: 50 μm. Parathyroid size assessed by cross-sectional area analysis of longitudinally cut serial sections (6 microns) from 5–6 animals per group (n) and corrected for body weight (μm2/g) in Gata3+/+ (black bars) and Gata3+/– (gray bars) mice. (B) Parathyroid proliferation rates assessed by Ki-67 immunostaining. Nuclear Ki-67 immunostaining in parathyroid cells (top). Scale bars: 50 μm. Insets show details at higher magnification (×100), and arrows point to nuclear staining. Ki-67 proliferation rate (Ki-67–positive cells/total number of cells) in Gata3+/+ (black bars) and Gata3+/– (gray bars) mice. *P < 0.05. (C) Studies of parathyroid-thymus primordia in E12.5 embryos. H&E staining (top panel) and corresponding serial section immunostained with CaSR antibody (bottom panel). Scale bars: 50 μm. P, parathyroid; tm, thymus; vnt, vagus nerve trunk. (D) Sagittal sections of pharyngeal pouches and pharynx (phx) from E11.5 embryos hybridized with Gcm2 riboprobe to identify the parathyroid domain of the common parathyroid-thymus primordium (arrows) arising from the third pharyngeal pouch. Scale bars: 100 μm (top panels); 50 μm (bottom panels). h, heart. (E) Volume of third pharyngeal pouch in E11.5 embryos (n = 6 of each genotype) and the proportion of Gcm2-expressing cells. *P < 0.02; **P < 0.001; ***P < 0.002. P values calculated using Student’s t test. Error bars represent SEM.