Loss of Fam20c causes defects in the acinar and duct structure of salivary glands in mice

Miao,Nan; Zhan,Yuanbo; Xu,Yingying; Yuan,Haoze; Qin,Chunlin; Lin,Feng; Xie,Xiaohua; Mu,Sen; Yuan,Mengtong; Mu,Haibin; Guo,Shouli; Li,Ying; Zhang,Bin

doi:10.3892/ijmm.2019.4126

Loss of Fam20c causes defects in the acinar and duct structure of salivary glands in mice

Authors:
- Nan Miao
- Yuanbo Zhan
- Yingying Xu
- Haoze Yuan
- Chunlin Qin
- Feng Lin
- Xiaohua Xie
- Sen Mu
- Mengtong Yuan
- Haibin Mu
- Shouli Guo
- Ying Li
- Bin Zhang
View Affiliations

Affiliations: Institute of Hard Tissue Development and Regeneration, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Periodontology and Oral Mucosa, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China, Department of Biomedical Sciences, Texas A&M University College of Dentistry, Dallas, TX 75246, USA, Department of General Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: March 6, 2019 https://doi.org/10.3892/ijmm.2019.4126
Pages: 2103-2117
Copyright: © Miao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Family with sequence similarity 20‑member C (FAM20C), a recently characterized Golgi kinase, performs numerous biological functions by phosphorylating more than 100 secreted proteins. However, the role of FAM20C in the salivary glands remains undefined. The present study demonstrated that FAM20C is mainly located in the cytoplasm of duct epithelial cells in the salivary glands. Fam20cf/f; Mmtv‑Cre mice were created in which Fam20c was inactivated in the salivary gland cells and observed that the number of ducts and the ductal cross‑sectional area increased significantly, while the number of acinar cells was reduced. The granular convoluted tubules (GCTs) exhibited an accumulation of aberrant secretory granules, along with a reduced expression and altered distribution patterns of β nerve growth factor, α‑amylase and bone morphogenetic protein (BMP) 4. This abnormality suggested that the GCT cells were immature and exhibited defects in developmental and secretory functions. In accordance with the morphological alterations and the reduced number of acinar cells, FAM20C deficiency in the salivary glands significantly decreased the salivary flow rate. The Na+, Cl– and K+ concentrations in the saliva were all significantly increased due to dysfunction of the ducts. Furthermore, Fam20c deficiency significantly increased BMP2 and BMP7 expression, decreased BMP4 expression, and attenuated the canonical and noncanonical BMP signaling pathways in the salivary glands. Collectively, the results of the present study demonstrate that FAM20C is a key regulator of acinar and duct structure and duct maturation and provide a novel avenue for investigating novel therapeutic targets for oral diseases including xerostomia.

View Figures

View References

1	Fox PC: Autoimmune diseases and Sjogren's syndrome: An autoimmune exocrinopathy. Ann NY Acad Sci. 1098:15–21. 2007. View Article : Google Scholar : PubMed/NCBI
2	Shiboski CH, Hodgson TA, Ship JA and Schiødt M: Management of salivary hypofunction during and after radiotherapy. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 103(Suppl): S66.e61–e19. 2007. View Article : Google Scholar
3	Nederfors T: Xerostomia and hyposalivation. Adv Dent Res. 14:48–56. 2000. View Article : Google Scholar
4	Napeñas JJ, Brennan MT and Fox PC: Diagnosis and treatment of xerostomia (dry mouth). Odontology. 97:76–83. 2009. View Article : Google Scholar : PubMed/NCBI
5	Tucker AS: Salivary gland development. Semin Cell Dev Biol. 18:237–244. 2007. View Article : Google Scholar : PubMed/NCBI
6	Patel VN, Rebustini IT and Hoffman MP: Salivary gland branching morphogenesis. Differentiation. 74:349–364. 2006. View Article : Google Scholar : PubMed/NCBI
7	Harunaga J, Hsu JC and Yamada KM: Dynamics of salivary gland morphogenesis. J Dent Res. 90:1070–1077. 2011. View Article : Google Scholar : PubMed/NCBI
8	Heine U, Munoz EF, Flanders KC, Ellingsworth LR, Lam HY, Thompson NL, Roberts AB and Sporn MB: Role of transforming growth factor-beta in the development of the mouse embryo. J Cell Biol. 105:2861–2876. 1987. View Article : Google Scholar : PubMed/NCBI
9	Hoffman MP, Kidder BL, Steinberg ZL, Lakhani S, Ho S, Kleinman HK and Larsen M: Gene expression profiles of mouse submandibular gland development: FGFR1 regulates branching morphogenesis in vitro through BMP- and FGF-dependent mechanisms. Development. 129:5767–5778. 2002. View Article : Google Scholar : PubMed/NCBI
10	Jaskoll T, Zhou YM, Chai Y, Makarenkova HP, Collinson JM, West JD, Hajihosseini MK, Lee J and Melnick M: Embryonic submandibular gland morphogenesis: Stage-specific protein localization of FGFs, BMPs, Pax6 and Pax9 in normal mice and abnormal SMG phenotypes in FgfR2-IIIc(+/Delta), BMP7(−/−) and Pax6(−/−) mice. Cells Tissues Organs. 170:83–98. 2002. View Article : Google Scholar
11	Tagliabracci VS, Engel JL, Wen J, Wiley SE, Worby CA, Kinch LN, Xiao J, Grishin NV and Dixon JE: Secreted kinase phosphorylates extracellular proteins that regulate biomineralization. Science. 336:1150–1153. 2012. View Article : Google Scholar : PubMed/NCBI
12	Hao J, Narayanan K, Muni T, Ramachandran A and George A: Dentin matrix protein 4, a novel secretory calcium-binding protein that modulates odontoblast differentiation. J Biol Chem. 282:15357–15365. 2007. View Article : Google Scholar : PubMed/NCBI
13	Ishikawa HO, Xu A, Ogura E, Manning G and Irvine KD: The Raine syndrome protein FAM20C is a Golgi kinase that phosphorylates bio-mineralization proteins. PLoS One. 7:e429882012. View Article : Google Scholar : PubMed/NCBI
14	George A and Veis A: Phosphorylated proteins and control over apatite nucleation, crystal growth, and inhibition. Chem Rev. 108:4670–4693. 2008. View Article : Google Scholar : PubMed/NCBI
15	Tagliabracci VS, Xiao J and Dixon JE: Phosphorylation of substrates destined for secretion by the Fam20 kinases. Biochem Soc Trans. 41:1061–1065. 2013. View Article : Google Scholar : PubMed/NCBI
16	Tagliabracci VS, Wiley SE, Guo X, Kinch LN, Durrant E, Wen J, Xiao J, Cui J, Nguyen KB, Engel JL, et al: A Single Kinase Generates the Majority of the Secreted Phosphoproteome. Cell. 161:1619–1632. 2015. View Article : Google Scholar : PubMed/NCBI
17	Simpson MA, Hsu R, Keir LS, Hao J, Sivapalan G, Ernst LM, Zackai EH, Al-Gazali LI, Hulskamp G, Kingston HM, et al: Mutations in FAM20C are associated with lethal osteosclerotic bone dysplasia (Raine syndrome), highlighting a crucial molecule in bone development. Am J Hum Genet. 81:906–912. 2007. View Article : Google Scholar : PubMed/NCBI
18	Acevedo AC, Poulter JA, Alves PG, de Lima CL, Castro LC, Yamaguti PM, Paula LM, Parry DA, Logan CV, Smith CE, et al: Variability of systemic and orodental phenotype in two families with non-lethal Raine syndrome with FAM20C mutations. BMC Med Genet. 16:82015. View Article : Google Scholar
19	Seidahmed MZ, Alazami AM, Abdelbasit OB, Al Hussein K, Miqdad AM, Abu-Sa'da O, Mustafa T, Bahjat S and Alkuraya FS: Report of a case of Raine syndrome and literature review. Am J Med Genet A. 167A:2394–2398. 2015. View Article : Google Scholar : PubMed/NCBI
20	Raine J, Winter RM, Davey A and Tucker SM: Unknown syndrome: Microcephaly, hypoplastic nose, exophthalmos, gum hyperplasia, cleft palate, low set ears, and osteosclerosis. J Med Genet. 26:786–788. 1989. View Article : Google Scholar : PubMed/NCBI
21	Wang X, Hao J, Xie Y, Sun Y, Hernandez B, Yamoah AK, Prasad M, Zhu Q, Feng JQ and Qin C: Expression of FAM20C in the osteogenesis and odontogenesis of mouse. J Histochem Cytochem. 58:957–967. 2010. View Article : Google Scholar : PubMed/NCBI
22	Du EX, Wang XF, Yang WC, Kaback D, Yee SP, Qin CL, George A and Hao JJ: Characterization of Fam20C expression in odontogenesis and osteogenesis using transgenic mice. Int J Oral Sci. 7:89–94. 2015. View Article : Google Scholar
23	Wang X, Wang S, Lu Y, Gibson MP, Liu Y, Yuan B, Feng JQ and Qin C: FAM20C plays an essential role in the formation of murine teeth. J Biol Chem. 287:35934–35942. 2012. View Article : Google Scholar : PubMed/NCBI
24	Vogel P, Hansen GM, Read RW, Vance RB, Thiel M, Liu J, Wronski TJ, Smith DD, Jeter-Jones S and Brommage R: Amelogenesis imperfecta and other biomineralization defects in Fam20a and Fam20c null mice. Vet Pathol. 49:998–1017. 2012. View Article : Google Scholar : PubMed/NCBI
25	Tibaldi E, Arrigoni G, Brunati AM, James P and Pinna LA: Analysis of a sub-proteome which co-purifies with and is phosphorylated by the Golgi casein kinase. Cell Mol Life Sci. 63:378–389. 2006. View Article : Google Scholar : PubMed/NCBI
26	Jernvall J and Thesleff I: Reiterative signaling and patterning during mammalian tooth morphogenesis. Mech Dev. 92:19–29. 2000. View Article : Google Scholar : PubMed/NCBI
27	Wagner KU, McAllister K, Ward T, Davis B, Wiseman R and Hennighausen L: Spatial and temporal expression of the Cre gene under the control of the MMTV-LTR in different lines of transgenic mice. Transgenic Res. 10:545–553. 2001. View Article : Google Scholar
28	Ewald D, Li M, Efrat S, Auer G, Wall RJ, Furth PA and Hennighausen L: Time-sensitive reversal of hyperplasia in transgenic mice expressing SV40 T antigen. Science. 273:1384–1386. 1996. View Article : Google Scholar : PubMed/NCBI
29	Wagner KU, Wall RJ, St-Onge L, Gruss P, Wynshaw-Boris A, Garrett L, Li M, Furth PA and Hennighausen L: Cre-mediated gene deletion in the mammary gland. Nucleic Acids Res. 25:4323–4330. 1997. View Article : Google Scholar : PubMed/NCBI
30	Wang X, Wang S, Li C, Gao T, Liu Y, Rangiani A, Sun Y, Hao J, George A, Lu Y, et al: Inactivation of a novel FGF23 regulator, FAM20C, leads to hypophosphatemic rickets in mice. PLoS Genet. 8:e10027082012. View Article : Google Scholar : PubMed/NCBI
31	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
32	Romanenko VG, Nakamoto T, Srivastava A, Begenisich T and Melvin JE: Regulation of membrane potential and fluid secretion by Ca²⁺-activated K⁺ channels in mouse submandibular glands. J Physiol. 581:801–817. 2007. View Article : Google Scholar : PubMed/NCBI
33	Nalbant D, Youn H, Nalbant SI, Sharma S, Cobos E, Beale EG, Du Y and Williams SC: FAM20: An evolutionarily conserved family of secreted proteins expressed in hematopoietic cells. BMC Genomics. 6:112005. View Article : Google Scholar : PubMed/NCBI
34	Gresik EW: The granular convoluted tubule (GCT) cell of rodent submandibular glands. Microsc Res Tech. 27:1–24. 1994. View Article : Google Scholar : PubMed/NCBI
35	Ma T, Song Y, Gillespie A, Carlson EJ, Epstein CJ and Verkman AS: Defective secretion of saliva in transgenic mice lacking aquaporin-5 water channels. J Biol Chem. 274:20071–20074. 1999. View Article : Google Scholar : PubMed/NCBI
36	Krane CM, Melvin JE, Nguyen HV, Richardson L, Towne JE, Doetschman T and Menon AG: Salivary acinar cells from aquaporin 5-deficient mice have decreased membrane water permeability and altered cell volume regulation. J Biol Chem. 276:23413–23420. 2001. View Article : Google Scholar : PubMed/NCBI
37	Penschow JD, Drinkwater CC, Haralambidis J and Coghlan JP: Sites of expression and induction of glandular kallikrein gene expression in mice. Mol Cell Endocrinol. 81:135–146. 1991. View Article : Google Scholar : PubMed/NCBI
38	Smith FJ, Porter RM, Corden LD, Lunny DP, Lane EB and McLean WH: Cloning of human, murine, and marsupial keratin 7 and a survey of K7 expression in the mouse. Biochem Biophys Res Commun. 297:818–827. 2002. View Article : Google Scholar : PubMed/NCBI
39	Melnick M and Jaskoll T: Mouse submandibular gland morphogenesis: A paradigm for embryonic signal processing. Crit Rev Oral Biol Med. 11:199–215. 2000. View Article : Google Scholar
40	Jaskoll T, Chen H, Min Zhou Y, Wu D and Melnick M: Developmental expression of survivin during embryonic submandibular salivary gland development. BMC Dev Biol. 1:52001. View Article : Google Scholar : PubMed/NCBI
41	Fiaschi M, Kolterud A, Nilsson M, Toftgård R and Rozell B: Targeted expression of GLI1 in the salivary glands results in an altered differentiation program and hyperplasia. Am J Pathol. 179:2569–2579. 2011. View Article : Google Scholar : PubMed/NCBI
42	Barka T: Biologically active polypeptides in submandibular glands. J Histochem Cytochem. 28:836–859. 1980. View Article : Google Scholar : PubMed/NCBI
43	Yoshida S, Ohbo K, Takakura A, Takebayashi H, Okada T, Abe K and Nabeshima Y: Sgn1, a basic helix-loop-helix transcription factor delineates the salivary gland duct cell lineage in mice. Dev Biol. 240:517–530. 2001. View Article : Google Scholar
44	Yamagishi R, Wakayama T, Nakata H, Adthapanyawanich K, Kumchantuek T, Yamamoto M and Iseki S: Expression and localization of alpha-amylase in the submandibular and sublingual glands of mice. Acta Histochem Cytochem. 47:95–102. 2014. View Article : Google Scholar : PubMed/NCBI
45	Marchetti L, Gabrielli MG, Materazzi G and Menghi G: Cellular compartmentation of lysozyme and alpha-amylase in the mouse salivary glands. Immunogold approaches at light and electron microscopy level. Histol Histopathol. 15:337–346. 2000.PubMed/NCBI
46	Menghi G, Marchetti L, Bondi AM, Accili D, Sabbieti MG and Materazzi G: Double-sided staining with a gold probe and silver enhancement to detect alpha-amylase and sugar moieties in the mouse salivary glands. Histol Histopathol. 14:687–695. 1999.PubMed/NCBI
47	Smith RJ and Frommer J: Effects of prepubertal castration on development of granular tubules and amylase activity in the male mouse submandibular gland. Arch Oral Biol. 17:1561–1571. 1972. View Article : Google Scholar : PubMed/NCBI
48	Gresik EW: The postnatal development of the sexually dimorphic duct system and of amylase activity in the submandibular glands of mice. Cell Tissue Res. 157:411–422. 1975. View Article : Google Scholar : PubMed/NCBI
49	Morgan-Bathke M, Lin HH, Chibly AM, Zhang W, Sun X, Chen CH, Flodby P, Borok Z, Wu R, Arnett D, et al: Deletion of ATG5 shows a role of autophagy in salivary homeostatic control. J Dent Res. 92:911–917. 2013. View Article : Google Scholar : PubMed/NCBI
50	Deretic V, Jiang S and Dupont N: Autophagy intersections with conventional and unconventional secretion in tissue development, remodeling and inflammation. Trends Cell Biol. 22:397–406. 2012. View Article : Google Scholar : PubMed/NCBI
51	Dikeakos JD and Reudelhuber TL: Sending proteins to dense core secretory granules: Still a lot to sort out. J Cell Biol. 177:191–196. 2007. View Article : Google Scholar : PubMed/NCBI
52	Musselmann K, Green JA, Sone K, Hsu JC, Bothwell IR, Johnson SA, Harunaga JS, Wei Z and Yamada KM: Salivary gland gene expression atlas identifies a new regulator of branching morphogenesis. J Dent Res. 90:1078–1084. 2011. View Article : Google Scholar : PubMed/NCBI