Advertisement

57Fe enrichment in mice for β-thalassaemia studies via Mössbauer spectroscopy of blood samples

  • George CharitouEmail author
  • Charalambos Tsertos
  • Yannis Parpottas
  • Marina Kleanthous
  • Carsten W. Lederer
  • Marios Phylactides
Original Article

Abstract

In this work, wild-type and heterozygous β-thalassaemic mice were enriched with 57Fe via gastrointestinal absorption to characterize in greater detail the iron complexes then identifiable via Mössbauer spectroscopy. The 57Fe enrichment method was validated and Mössbauer spectra were obtained at 80 K from blood samples from wild-type and β-thalassaemic mice at 1, 3, 6, and 9 months of age. As expected, the haemoglobin levels of the thalassaemic mice were lower than from normal mice, indicating anaemia. Furthermore, significant amounts of ferritin-like iron were observed in the thalassaemic mice samples, which decreased with mouse age, reflecting the pattern of reticulocyte count reduction reported in the literature.

Keywords

Mössbauer spectroscopy β-Thalassaemia Mice 57Fe enrichment Ferritin Blood 

Notes

Acknowledgements

We gratefully acknowledge the personnel of the Transgenic Mouse Facility of the Cyprus Institute of Neurology and Genetics for the planning of the mouse breeding and mouse feed preparation guidance.

Compliance with ethical standards

Ethical approval

All applicable international, national and/or institutional guidelines for the care and use of animals were followed.

References

  1. Abreu MS, Sanchís ME, Peñalver JA, Kanter F (1989) Application of Mossbauer spectroscopy to the study of hemoglobinopathies. Preliminary experience. Sangre (Barc) 34:325–328Google Scholar
  2. Bauminger ER, Cohen SG, Ofer S, Rachmilewitz EA (1979) Mössbauer studies of ferritin-like iron in red blood cells of thalassemia sickle-cell anemia and hemoglobin Hammersmith. J Phys Colloq 40:C2-502–C2-504.  https://doi.org/10.1051/jphyscol:19792176 Google Scholar
  3. Brissot P, Ropert M, Le Lan C, Loréal O (2012) Non-transferrin bound iron: a key role in iron overload and iron toxicity. Biochim Biophys Acta BBA Gen Subj 1820:403–410.  https://doi.org/10.1016/j.bbagen.2011.07.014 CrossRefGoogle Scholar
  4. Cappellini M, Cohen A, Porter J, et al. (eds) (2014) Guidelines for the management of transfusion dependent thalassaemia (TDT). Thalassaemia International Federation Publications, NicosiaGoogle Scholar
  5. Charitou G, Petousis V, Tsertos C et al (2018) First study on iron complexes in blood and organ samples from thalassaemic and normal laboratory mice using Mössbauer spectroscopy. Eur Biophys J 47:131–138.  https://doi.org/10.1007/s00249-017-1234-6 CrossRefGoogle Scholar
  6. Chua-anusorn W, Webb J, Macey DJ et al (1999) The effect of prolonged iron loading on the chemical form of iron oxide deposits in rat liver and spleen. Biochim Biophys Acta BBA Mol Basis Dis 1454:191–200.  https://doi.org/10.1016/S0925-4439(99)00036-8 CrossRefGoogle Scholar
  7. Darshan D, Vanoaica L, Richman L et al (2009) Conditional deletion of ferritin H in mice induces loss of iron storage and liver damage. Hepatology 50:852–860.  https://doi.org/10.1002/hep.23058 CrossRefGoogle Scholar
  8. Eleftheriou A (2003) About thalassaemia. Thalassaemia International Federation Publications, CyprusGoogle Scholar
  9. Galanello R, Origa R (2010) Beta-thalassemia. Orphanet J Rare Dis 5:11.  https://doi.org/10.1186/1750-1172-5-11 CrossRefGoogle Scholar
  10. Gardenghi S, Marongiu MF, Ramos P et al (2007) Ineffective erythropoiesis in thalassemia is characterized by increased iron absorption mediated by down-regulation of hepcidin and up-regulation of ferroportin. Blood 109:5027–5035.  https://doi.org/10.1182/blood-2006-09-048868 CrossRefGoogle Scholar
  11. Gkouvatsos K, Papanikolaou G, Pantopoulos K (2012) Regulation of iron transport and the role of transferrin. Biochim Biophys Acta BBA Gen Subj 1820:188–202.  https://doi.org/10.1016/j.bbagen.2011.10.013 CrossRefGoogle Scholar
  12. Gütlich P, Bill E, Trautwein A (2011) Mössbauer spectroscopy and transition metal chemistry: fundamentals and application. Springer, BerlinCrossRefGoogle Scholar
  13. Harrison PM, Arosio P (1996) The ferritins: molecular properties, iron storage function and cellular regulation. Biochim Biophys Acta BBA Bioenerg 1275:161–203.  https://doi.org/10.1016/0005-2728(96)00022-9 CrossRefGoogle Scholar
  14. Hoy GR, Cook DC, Berger RL, Friedman FK (1986) Mössbauer spectroscopic studies of hemoglobin and its isolated subunits. Biophys J 49:1009–1015.  https://doi.org/10.1016/S0006-3495(86)83729-8 CrossRefGoogle Scholar
  15. Jiang K, Ma WY, Ortalli I et al (1994) Some comments on the effectiveness of the therapy for Β-thalassemia. Hyperfine Interact 91:859–863.  https://doi.org/10.1007/BF02064619 CrossRefGoogle Scholar
  16. Kaufman KS, Papaefthymiou GC, Frankel RB, Rosenthal A (1980) Nature of iron deposits on the cardiac walls in β-thalassemia by Mössbauer spectroscopy. Biochim Biophys Acta BBA Gen Subj 629:522–529.  https://doi.org/10.1016/0304-4165(80)90157-9 CrossRefGoogle Scholar
  17. Knovich MA, Storey JA, Coffman LG et al (2009) Ferritin for the clinician. Blood Rev 23:95–104.  https://doi.org/10.1016/j.blre.2008.08.001 CrossRefGoogle Scholar
  18. Maeda Y (1979) Mössbauer studies on the iron-ligand binding in hemoproteins and their related compounds. J Phys Colloq 40:C2-514–C2-522.  https://doi.org/10.1051/jphyscol:19792180 Google Scholar
  19. National Research Council (1995) Nutrient requirements of laboratory animals: fourth revised edition, 1995. National Academies Press, Washington, D.C.Google Scholar
  20. Oshtrakh MI, Semionkin VA (1986) Mössbauer spectroscopy of haemoglobins: study of the relationship of Fe2+ electronic and molecular structure of the active site. FEBS Lett 208:331–336.  https://doi.org/10.1016/0014-5793(86)81044-4 CrossRefGoogle Scholar
  21. Oshtrakh MI, Berkovsky AL, Kumar A et al (2010) 57Fe quadrupole splitting and isomer shift in various oxyhemoglobins: study using Mössbauer spectroscopy. Hyperfine Interact 197:301–307.  https://doi.org/10.1007/s10751-010-0232-1 CrossRefGoogle Scholar
  22. Oshtrakh MI, Alenkina IV, Milder OB, Semionkin VA (2011) Mössbauer spectroscopy with a high velocity resolution in the study of iron-containing proteins and model compounds. Spectrochim Acta A Mol Biomol Spectrosc 79:777–783.  https://doi.org/10.1016/j.saa.2010.08.052 CrossRefGoogle Scholar
  23. Patel M, Ramavataram DVSS (2012) Non transferrin bound iron: nature, manifestations and analytical approaches for estimation. Indian J Clin Biochem 27:322–332.  https://doi.org/10.1007/s12291-012-0250-7 CrossRefGoogle Scholar
  24. Piperno A, Taddei MT, Sampietro M et al (1984) Erythrocyte ferritin in thalassemia syndromes. Acta Haematol 71:251–256.  https://doi.org/10.1159/000206596 CrossRefGoogle Scholar
  25. Ponka P, Koury MJ, Sheftel AD (2013) Erythropoiesis, hemoglobin synthesis, and erythroid mitochondrial iron homeostasis. Handbook of Porphyrin Science. World Scientific Publishing Company, Singapore, pp 41–84Google Scholar
  26. Schechter AN (2008) Hemoglobin research and the origins of molecular medicine. Blood 112:3927–3938.  https://doi.org/10.1182/blood-2008-04-078188 CrossRefGoogle Scholar
  27. Xuanhui G, Nanming Z, Xiufang Z et al (1988) Study on Mössbauer spectra of hemoglobin in thalassemia. Hyperfine Interact 42:897–900.  https://doi.org/10.1007/BF02395534 CrossRefGoogle Scholar
  28. Yang B, Kirby S, Lewis J et al (1995) A mouse model for beta 0-thalassemia. Proc Natl Acad Sci USA 92:11608–11612CrossRefGoogle Scholar

Copyright information

© European Biophysical Societies' Association 2019

Authors and Affiliations

  1. 1.Department of PhysicsUniversity of CyprusNicosiaCyprus
  2. 2.School of Engineering and Applied SciencesFrederick UniversityNicosiaCyprus
  3. 3.Molecular Genetics Thalassaemia DepartmentThe Cyprus Institute of Neurology and GeneticsNicosiaCyprus

Personalised recommendations