Abstract
Glial cells missing homolog 2 (GCM2) is a transcription factor that is expressed predominately in the pharyngeal pouches and, at later stages, in the developing and mature parathyroid glands. In humans, loss of GCM2 function, either through recessive apomorphic mutations or dominant inhibitor mutations in the human GCM2 gene, leads to isolated hypoparathyroidism. In mice, homozygous disruption of Gcm2 by conventional gene targeting results in parathyroid aplasia and hypoparathyroidism. In this study, we report the generation and functional characterization of mice encoding a conditional null allele of Gcm2. We demonstrate the functional integrity of the conditional Gcm2 allele and report successful in vivo deletion of exon 2 using Cre recombinase. The mice with conditional deletion of Gcm2 displayed phenotypes similar to those previously described for a conventional Gcm2 knockout, including perinatal lethality, hypocalemia, low or undetectable serum levels of parathyroid hormone, and absent parathyroid glands. The production of a conditional mutant allele for Gcm2 represents a valuable resource for the study of the temporal- and spatial-specific roles for Gcm2, and for understanding the postnatal activities of GCM2 protein.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akiyama Y, Hosoya T, Poole AM, Hotta Y (1996) The gcm-motif: a novel DNA-binding motif conserved in Drosophila and mammals. Proc Natl Acad Sci USA 93(25):14912–14916
Altshuller Y, Copeland NG, Gilbert DJ, Jenkins NA, Frohman MA (1996) Gcm1, a mammalian homolog of Drosophila glial cells missing. FEBS Lett 393(2–3):201–204
Baumber L, Tufarelli C, Patel S, King P, Johnson CA, Maher ER, Trembath RC (2005) Identification of a novel mutation disrupting the DNA binding activity of GCM2 in autosomal recessive familial isolated hypoparathyroidism. J Med Genet 42(5):443–448
Canaff L, Zhou X, Mosesova I, Cole DE, Hendy GN (2009) Glial cells missing-2 (GCM2) transactivates the calcium-sensing receptor gene: effect of a dominant-negative GCM2 mutant associated with autosomal dominant hypoparathyroidism. Hum Mutat 30(1):85–92. doi:10.1002/humu.20827
Chotard C, Leung W, Salecker I (2005) Glial cells missing and gcm2 cell autonomously regulate both glial and neuronal development in the visual system of Drosophila. Neuron 48(2):237–251
Cohen SX, Moulin M, Schilling O, Meyer-Klaucke W, Schreiber J, Wegner M, Muller CW (2002) The GCM domain is a Zn-coordinating DNA-binding domain. FEBS Lett 528(1–3):95–100
Cohen SX, Moulin M, Hashemolhosseini S, Kilian K, Wegner M, Muller CW (2003) Structure of the GCM domain–DNA complex: a DNA-binding domain with a novel fold and mode of target site recognition. EMBO J 22(8):1835–1845
Collins MT, Lindsay JR, Jain A, Kelly MH, Cutler CM, Weinstein LS, Liu J, Fedarko NS, Winer KK (2005) Fibroblast growth factor-23 is regulated by 1alpha,25-dihydroxyvitamin D. J Bone Miner Res 20(11):1944–1950
Ding C, Buckingham B, Levine MA (2001) Familial isolated hypoparathyroidism caused by a mutation in the gene for the transcription factor GCMB. J Clin Invest 108(8):1215–1220
Doyle D, Kirwin SM, Sol-Church K, Levine MA (2012) A novel mutation in the GCM2 gene causing severe congenital isolated hypoparathyroidism. J Pediatr Endocrinol Metab 25(7–8):741–746
Fraser RA, Kronenberg HM, Pang PK, Harvey S (1990) Parathyroid hormone messenger ribonucleic acid in the rat hypothalamus. Endocrinology 127(5):2517–2522
Germain-Lee EL, Schwindinger W, Crane JL, Zewdu R, Zweifel LS, Wand G, Huso DL, Saji M, Ringel MD, Levine MA (2005) A mouse model of Albright hereditary osteodystrophy generated by targeted disruption of exon 1 of the Gnas gene. Endocrinology 146(11):4697–4709
Gordon J, Bennett AR, Blackburn CC, Manley NR (2001) Gcm2 and Foxn1 mark early parathyroid- and thymus-specific domains in the developing third pharyngeal pouch. Mech Dev 103(1–2):141–143
Grigorieva IV, Mirczuk S, Gaynor KU, Nesbit MA, Grigorieva EF, Wei Q, Ali A, Fairclough RJ, Stacey JM, Stechman MJ, Mihai R, Kurek D, Fraser WD, Hough T, Condie BG, Manley N, Grosveld F, Thakker RV (2010) Gata3-deficient mice develop parathyroid abnormalities due to dysregulation of the parathyroid-specific transcription factor Gcm2. J Clin Invest 120(6):2144–2155
Gunther T, Chen ZF, Kim J, Priemel M, Rueger JM, Amling M, Moseley JM, Martin TJ, Anderson DJ, Karsenty G (2000) Genetic ablation of parathyroid glands reveals another source of parathyroid hormone. Nature 406(6792):199–203
Gupta A, Winer K, Econs MJ, Marx SJ, Collins MT (2004) FGF-23 is elevated by chronic hyperphosphatemia. J Clin Endocrinol Metab 89(9):4489–4492
Hashemolhosseini S, Wegner M (2004) Impacts of a new transcription factor family: mammalian GCM proteins in health and disease. J Cell Biol 166(6):765–768
Hitoshi S, Ishino Y, Kumar A, Jasmine S, Tanaka KF, Kondo T, Kato S, Hosoya T, Hotta Y, Ikenaka K (2011) Mammalian Gcm genes induce Hes5 expression by active DNA demethylation and induce neural stem cells. Nat Neurosci 14(8):957–964
Hosoya T, Takizawa K, Nitta K, Hotta Y (1995) Glial cells missing: a binary switch between neuronal and glial determination in Drosophila. Cell 82(6):1025–1036
Jones BW, Fetter RD, Tear G, Goodman CS (1995) Glial cells missing: a genetic switch that controls glial versus neuronal fate. Cell 82(6):1013–1023
Kamitani-Kawamoto A, Hamada M, Moriguchi T, Miyai M, Saji F, Hatamura I, Nishikawa K, Takayanagi H, Hitoshi S, Ikenaka K, Hosoya T, Hotta Y, Takahashi S, Kataoka K (2011) MafB interacts with Gcm2 and regulates parathyroid hormone expression and parathyroid development. J Bone Miner Res 26(10):2463–2472
Kammerer M, Pirola B, Giglio S, Giangrande A (1999) GCMB, a second human homolog of the fly glide/gcm gene. Cytogenet Cell Genet 84(1–2):43–47
Kim J, Jones BW, Zock C, Chen Z, Wang H, Goodman CS, Anderson DJ (1998) Isolation and characterization of mammalian homologs of the drosophila gene glial cells missing. Proc Natl Acad Sci USA 95:12364–12369
Liu Z, Yu S, Manley NR (2007) Gcm2 is required for the differentiation and survival of parathyroid precursor cells in the parathyroid/thymus primordia. Dev Biol 305(1):333–346
Liu Z, Farley A, Chen L, Kirby BJ, Kovacs CS, Blackburn CC, Manley NR (2010) Thymus-associated parathyroid hormone has two cellular origins with distinct endocrine and immunological functions. PLoS Genet 6(12):e1001251
Mannstadt M, Bertrand G, Muresan M, Weryha G, Leheup B, Pulusani SR, Grandchamp B, Juppner H, Silve C (2008) Dominant-negative GCMB mutations cause an autosomal dominant form of hypoparathyroidism. J Clin Endocrinol Metab 93(9):3568–3576
Maret A, Bourdeau I, Ding C, Kadkol SS, Westra WH, Levine MA (2004) Expression of GCMB by intrathymic parathyroid hormone-secreting adenomas indicates their parathyroid cell origin. J Clin Endocrinol Metab 89(1):8–12
Maret A, Ding C, Kornfield SL, Levine MA (2008) Analysis of the GCM2 gene in isolated hypoparathyroidism: a molecular and biochemical study. J Clin Endocrinol Metab 93(4):1426–1432
Mirczuk SM, Bowl MR, Nesbit MA, Cranston T, Fratter C, Allgrove J, Brain C, Thakker RV (2010) A missense glial cells missing homolog B (GCMB) mutation, Asn502His, causes autosomal dominant hypoparathyroidism. J Clin Endocrinol Metab 95(7):3512–3516
Nutley MT, Parimi SA, Harvey S (1995) Sequence analysis of hypothalamic parathyroid hormone messenger ribonucleic acid. Endocrinology 136(12):5600–5607
O’Gorman S, Wahl GM (1997) Mouse engineering. Science 277(5329):1025
O’Gorman S, Dagenais NA, Qian M, Marchuk Y (1997) Protamine-Cre recombinase transgenes efficiently recombine target sequences in the male germ line of mice, but not in embryonic stem cells. Proc Natl Acad Sci USA 94(26):14602–14607
Okabe M, Graham A (2004) The origin of the parathyroid gland. Proc Natl Acad Sci USA 101(51):17716–17719
Quinn SJ, Thomsen AR, Pang JL, Kantham L, Brauner-Osborne H, Pollak M, Goltzman D, Brown EM (2013) Interactions between calcium and phosphorus in the regulation of the production of fibroblast growth factor 23 in vivo. Am J Physiol Endocrinol Metab 304(3):E310–E320
Schreiber J, Enderich J, Wegner M (1998) Structural requirements for DNA binding of GCM proteins. Nucleic Acids Res 26(10):2337–2343
Silver J, Naveh-Many T (2012) FGF23 and the parathyroid. Adv Exp Med Biol 728:92–99
Simmonds CS, Karsenty G, Karaplis AC, Kovacs CS (2010) Parathyroid hormone regulates fetal-placental mineral homeostasis. J Bone Miner Res 25(3):594–605
Sticht H, Hashemolhosseini S (2006) A common structural mechanism underlying GCMB mutations that cause hypoparathyroidism. Med Hypotheses 67(3):482–487
Thomee C, Schubert SW, Parma J, Le PQ, Hashemolhosseini S, Wegner M, Abramowicz MJ (2005) GCMB mutation in familial isolated hypoparathyroidism with residual secretion of parathyroid hormone. J Clin Endocrinol Metab 90(5):2487–2492
Wegner M, Riethmacher D (2001) Chronicles of a switch hunt: gcm genes in development. Trends Genet 17(5):286–290
Yamashita H, Yamazaki Y, Hasegawa H, Yamashita T, Fukumoto S, Shigematsu T, Kazama JJ, Fukagawa M, Noguchi S (2007) Fibroblast growth factor-23 (FGF23) in patients with transient hypoparathyroidism: its important role in serum phosphate regulation. Endocr J 54(3):465–470
Yuan Q, Jiang Y, Zhao X, Sato T, Densmore M, Schuler C, Erben RG, McKee MD, Lanske B (2013) Increased osteopontin contributes to inhibition of bone mineralization in FGF23-deficient mice. J Bone Miner Res. 29(3):693–704
Zajac JD, Danks JA (2008) The development of the parathyroid gland: from fish to human. Curr Opin Nephrol Hypertens 17(4):353–356
Acknowledgments
This work was supported, in part, by NIDDK R01DK079970 (M.A.L.), the Cleveland Clinic Lerner Research Institute, the CHOP Research Institute, the Albert Einstein College of Medicine and a generous gift from Linda and Earle Altman (S.K.L.). We acknowledge the generous help and guidance of Dr. Alexander Gorodinsky, Taconic Laboratory.
Conflict of interest
All authors state that they have no conflicts of interest.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yuan, Z., Opas, E.E., Vrikshajanani, C. et al. Generation of mice encoding a conditional null allele of Gcm2 . Transgenic Res 23, 631–641 (2014). https://doi.org/10.1007/s11248-014-9799-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11248-014-9799-7