Otoconia are formed embryonically and are instrumental in detecting linear acceleration and gravity. Degeneration and fragmentation of otoconia in elderly patients leads to imbalance resulting in higher frequency of falls that are positively correlated with the incidence of bone fractures and death. In this work we investigate the roles otoconial proteins Otolin-1 and Otoconin 90 (OC90) perform in the formation of otoconia. We demonstrate by rotary shadowing and atomic force microscopy (AFM) experiments that Otolin-1 forms homomeric protein complexes and self-assembled networks supporting the hypothesis that Otolin-1 serves as a scaffold protein of otoconia. Our calcium carbonate crystal growth data demonstrate that Otolin-1 and OC90 modulate in vitro calcite crystal morphology but neither protein is sufficient to produce the shape of otoconia. Coadministration of these proteins produces synergistic effects on crystal morphology that contribute to morphology resembling otoconia.
References
[1]
Weiner S, Wagner HD (1998) THE MATERIAL BONE: Structure-Mechanical Function Relations. Annual Review of Materials Science 28: 271–298. doi: 10.1146/annurev.matsci.28.1.271
[2]
Ross M, Peacor D, Johnsson L, Allard L (1976) Observations on normal and degenerating human otoconia. Ann Otol Rhinol Laryngol 85: 310–326.
[3]
Lim DJ (1984) Otoconia in health and disease. A review. Ann Otol Rhinol Laryngol Suppl 112: 17–24.
[4]
Anniko M, Ylikoski J, Wroblewski R (1984) Microprobe analysis of human otoconia. Acta Otolaryngol 97: 283–289. doi: 10.3109/00016488409130990
[5]
Agrawal Y, Carey JP, Della Santina CC, Schubert MC, Minor LB (2009) Disorders of balance and vestibular function in US adults: data from the National Health and Nutrition Examination Survey, 2001–2004. Arch Intern Med 169: 938–944. doi: 10.1001/archinternmed.2009.66
[6]
Sattin RW (1992) Falls among older persons: a public health perspective. Annu Rev Public Health 13: 489–508. doi: 10.1146/annurev.pu.13.050192.002421
[7]
Baloh RW, Honrubia V, Jacobson K (1987) Benign positional vertigo: clinical and oculographic features in 240 cases. Neurology 37: 371–378. doi: 10.1212/wnl.37.3.371
[8]
Welgampola MS, Bradshaw A, Halmagyi GM (2011) Practical neurology–4: Dizziness on head movement. Med J Aust 195: 518–522. doi: 10.5694/mja11.11001
[9]
Vibert D, Kompis M, Hausler R (2003) Benign paroxysmal positional vertigo in older women may be related to osteoporosis and osteopenia. Ann Otol Rhinol Laryngol 112: 885–889.
[10]
Jeong SH, Choi SH, Kim JY, Koo JW, Kim HJ, et al. (2009) Osteopenia and osteoporosis in idiopathic benign positional vertigo. Neurology 72: 1069–1076. doi: 10.1212/01.wnl.0000345016.33983.e0
[11]
Lu W, Zhou D, Freeman JJ, Thalmann I, Ornitz DM, et al. (2010) In vitro effects of recombinant otoconin 90 upon calcite crystal growth. Significance of tertiary structure. Hearing Research 268: 172–183. doi: 10.1016/j.heares.2010.05.019
[12]
Thalmann I, Hughes I, Tong BD, Ornitz DM, Thalmann R (2006) Microscale analysis of proteins in inner ear tissues and fluids with emphasis on endolymphatic sac, otoconia, and organ of Corti. Electrophoresis 27: 1598–1608. doi: 10.1002/elps.200500768
[13]
Murayama E, Takagi Y, Ohira T, Davis JG, Greene MI, et al. (2002) Fish otolith contains a unique structural protein, otolin-1. Eur J Biochem 269: 688–696. doi: 10.1046/j.0014-2956.2001.02701.x
[14]
Deans MR, Peterson JM, Wong GW (2010) Mammalian Otolin: a multimeric glycoprotein specific to the inner ear that interacts with otoconial matrix protein Otoconin-90 and Cerebellin-1. PLoS One 5: e12765. doi: 10.1371/journal.pone.0012765
[15]
Zhao X, Yang H, Yamoah E, Lundberg Y (2007) Gene targeting reveals the role of Oc90 as the essential organizer of the otoconial organic matrix. Dev Biol 304: 508–524. doi: 10.1016/j.ydbio.2007.01.013
[16]
Yang H, Zhao X, Xu Y, Wang L, He Q, et al. (2011) Matrix recruitment and calcium sequestration for spatial specific otoconia development. PLoS One 6: e20498. doi: 10.1371/journal.pone.0020498
[17]
Albeck S, Aizenberg J, Addadi L, Weiner S (1993) Interactions of various skeletal intracrystalline components with calcite crystals. Journal of the American Chemical Society 115: 11691–11697. doi: 10.1021/ja00078a005
[18]
Parkhurst DL, Appelo CAJ (1999) User guide to PHREEQC (version 2) - A Computer Program for Speciation, Batch Reaction, One Dimensional Transport, and Inverse Geochemical Calculations. US Geological Survey Water-Resources Investigation Report.
[19]
Frischholz S, Beier F, Girkontaite I, Wagner K, Poschl E, et al. (1998) Characterization of Human Type X Procollagen and Its NC-1 Domain Expressed as Recombinant Proteins in HEK293 Cells. J Biol Chem 273: 4547–4555. doi: 10.1074/jbc.273.8.4547
[20]
Kwan AP, Cummings CE, Chapman JA, Grant ME (1991) Macromolecular organization of chicken type X collagen in vitro. J Cell Biol 114: 597–604. doi: 10.1083/jcb.114.3.597
[21]
Narayanan B, Gilmer GH, Tao J, De Yoreo JJ, Ciobanu CV (2014) Self-assembly of collagen on flat surfaces: the interplay of collagen-collagen and collagen-substrate interactions. Langmuir 30: 1343–1350. doi: 10.1021/la4043364
[22]
Lins U, Farina M, Kurc M, Riordan G, Thalmann R, et al. (2000) The otoconia of the guinea pig utricle: internal structure, surface exposure, and interactions with the filament matrix. J Struct Biol 131: 67–78. doi: 10.1006/jsbi.2000.4260
[23]
Andrade LR, Lins U, Farina M, Kachar B, Thalmann R (2012) Immunogold TEM of otoconin 90 and otolin - relevance to mineralization of otoconia, and pathogenesis of benign positional vertigo. Hearing Research 292: 14–25. doi: 10.1016/j.heares.2012.07.003
[24]
Huss D, Dickman JD (2003) Histological preparation of developing vestibular otoconia for scanning electron microscopy. Journal of Neuroscience Methods 125: 129–136. doi: 10.1016/s0165-0270(03)00048-7
[25]
Teng HH, Dove PM, De Yoreo JJ (2000) Kinetics of calcite growth: Surface processes and relationships to macroscopic rate laws. Geochimica et Cosmochimica Acta 64: 2255–2266. doi: 10.1016/s0016-7037(00)00341-0
[26]
Orme CA, Noy A, Wierzbicki A, McBride MT, Grantham M, et al. (2001) Formation of chiral morphologies through selective binding of amino acids to calcite surface steps. Nature 411: 775–779. doi: 10.1038/35081034
[27]
Guo SW, Ward MD, Wesson JA (2002) Direct visualization of calcium oxalate monohydrate crystallization and dissolution with atomic force microscopy and the role of polymeric additives. Langmuir 18: 4284–4291. doi: 10.1021/la011754+
[28]
Qiu SR, Wierzbicki A, Orme CA, Cody AM, Hoyer JR, et al. (2004) Molecular modulation of calcium oxalate crystallization by osteopontin and citrate. Proceedings of the National Academy of Sciences of the United States of America 101: 1811–1815. doi: 10.1073/pnas.0307900100
[29]
Fu G, Qiu SR, Orme CA, Morse DE, De？Yoreo JJ (2005) Acceleration of Calcite Kinetics by Abalone Nacre Proteins. Advanced Materials 17: 2678–2683. doi: 10.1002/adma.200500633
[30]
Elhadj S, De Yoreo JJ, Hoyer JR, Dove PM (2006) Role of molecular charge and hydrophilicity in regulating the kinetics of crystal growth. Proceedings of the National Academy of Sciences of the United States of America 103: 19237–19242. doi: 10.1073/pnas.0605748103
[31]
De Yoreo JJ, Wierzbicki A, Dove PM (2007) New insights into mechanisms of biomolecular control on growth of inorganic crystals. Crystengcomm 9: 1144–1152. doi: 10.1039/b713006f
[32]
Maruyama M, Tsukamoto K, Sazaki G, Nishimura Y, Vekilov PG (2009) Chiral and Achiral Mechanisms of Regulation of Calcite Crystallization. Crystal Growth & Design 9: 127–135. doi: 10.1021/cg701219h
[33]
Friddle RW, Weaver ML, Qiu SR, Wierzbicki A, Casey WH, et al. (2010) Subnanometer atomic force microscopy of peptide-mineral interactions links clustering and competition to acceleration and catastrophe. Proceedings of the National Academy of Sciences of the United States of America 107: 11–15. doi: 10.1073/pnas.0908205107
[34]
De Yoreo JJ, Zepeda-Ruiz LA, Friddle RW, Qiu SR, Wasylenki LE, et al. (2009) Rethinking Classical Crystal Growth Models through Molecular Scale Insights: Consequences of Kink-Limited Kinetics. Crystal Growth & Design 9: 5135–5144. doi: 10.1021/cg900543g
[35]
Chen C-L, Qi J, Zuckermann RN, DeYoreo JJ (2011) Engineered biomimetic polymers as tunable agents for controlling CaCO3 mineralization. Journal of the American Chemical Society 133: 5214–5217. doi: 10.1021/ja200595f
[36]
Cho KR, Salter EA, De Yoreo JJ, Wierzbicki A, Elhadj S, et al. (2013) Growth inhibition of calcium oxalate monohydrate crystal by linear aspartic acid enantiomers investigated by in situ atomic force microscopy. Crystengcomm 15: 54–64. doi: 10.1039/c2ce25936b
