Methodology of adipose tissue type detection in mammals
https://doi.org/10.21323/2414-438X-2023-8-1-43-50
Abstract
Nowadays, an interest in studying the composition, properties and functions of adipose tissue (AT) is growing among researchers, which is conditioned by its important role in the normal functioning of the body. Due to different types of adipose tissue (AT) in mammals (white, beige, brown and pink) and different physiological tasks performed by each type of AT, rapid, correct and effective detection of an AT type is highly topical. Methods used today are labor consuming and in the case of NMR and CT expensive, which limits possibilities of scientists. In this connection, the aim of this research was to develop a methodological approach allowing rapid and effective detection of an adipose tissue type. A methodology was formed based on the concept, formalized requirements for the method, step-wise structure of investigations and interpretation of results. The concept is based on differences in the structure of the adipose cell (adipocyte) of different AT types. The method is based on extraction of heme containing proteins. To this end, solvents and parameters of extraction that facilitate their better extraction have been chosen. An AT type has been determined by the total content of iron contained in the cytochrome fragment. Our own modification was selected. This modification includes preliminary mincing of a sample with the ice-cold TES buffer (pH 8.5) in a ratio of 1:5 (g: mL), homogenization at 9,000 rpm for 2 min with the following centrifugation at 10,000 g and 4 °C for 15 min. Effectiveness of the proposed method was confirmed by the histological and electrophoretic analyses. Therefore, the new methodology of identification and differentiation of adipocytes was proposed for rapid and effective detection of an adipose tissue type.
Keywords
white and brown adipose tissue,
heme containing proteins,
adipocytes,
SDS-PAGE,
histology,
extraction
Supporting agencies: The work was performed within the framework of the NIR theme No. FNEN-2019–0008 of the state assignment for the V. M. Gorbatov Federal Research Center of RAS (in part of approbation and design of the method) and within the framework of the grant of the Russian Science Foundation No. 21–76–20032 (in part of acquisition and investigation of research objects).
About the Authors
I. M. Chernukha
V. M. Gorbatov Federal Research Center for Food Systems
Russian Federation
Russian Federation
Irina M. Chernukha, Doctor of Technical Sciences, Professor, Academician of Russian Academy of Sciences, Principal Researcher, Experimental Clinic — Research Laboratory of Biologically Active Substances of an Animal Origin
109316, Moscow, Talalikhina str., 26
Tel: +7–495–676–63–21
N. V. Kupaeva
V. M. Gorbatov Federal Research Center for Food Systems
Russian Federation
Russian Federation
Nadezhda V. Kupaeva, Junior Researcher, Experimental Clinic — Research Laboratory of Biologically Active Substances of an Animal Origin
109316, Moscow, Talalikhina str., 26
Tel: +7–495–676–95–11 (209)
J. A. Smirnova
V. M. Gorbatov Federal Research Center for Food Systems; D. Mendeleev University of Chemical Technology of Russia
Russian Federation
Russian Federation
Jutta A. Smirnova, Senior Laboratory Assistant, Experimental Clinic — Research Laboratory of Biologically Active Substances of an Animal Origin
109316, Moscow, Talalikhina str., 26
Tel: +7–495–676–95–11 (209)
A. G. Akhremko
V. M. Gorbatov Federal Research Center for Food Systems
Russian Federation
Russian Federation
Anastasiya G. Akhremko, Candidate of Technical Sciences, Junior Researcher, Experimental clinic — research laboratory of biologically active substances of an animal origin
109316, Moscow, Talalikhina str., 26
Tel.: +7–495–676–95–11 (207)
V. A. Pchelkina
V. M. Gorbatov Federal Research Center for Food Systems
Russian Federation
Russian Federation
Viktoriya A. Pchelkina, Candidate of Technical Sciences, Leading Researcher, Experimental Clinic-Laboratory of Biologically Active Substances of Animal Origin
109316, Moscow, Talalikhina str., 26
Tel.: +7–495–676–95–11 (242)
E. A. Kotenkova
V. M. Gorbatov Federal Research Center for Food Systems
Russian Federation
Russian Federation
Elena A. Kotenkova, Candidate of Technical Sciences, Senior Researcher, Experimental Clinic-Laboratory of Biologically Active Substances of Animal Origin
109316, Moscow, Talalikhina str., 26
Tel.: +7–495–676–95–11 (129)
References
1. Rosen, E. D., Spiegelman, B. M. (2014). What we talk about when we talk about fat. Cell, 156(1–2), 20–44. https://doi.org/10.1016/j.cell.2013.12.012
2. Fantuzzi, G. (2005). Adipose tissue, adipokines, and inflammation. Journal of Allergy and Clinical Immunology, 115(5), 911–919. https://doi.org/10.1016/j.jaci.2005.02.023
3. Choe, S. S., Huh, J. Y., Hwang, I. J., Kim, J. I., Kim, J. B. (2016). Adipose tissue remodeling: Its role in energy metabolism and metabolic disorders. Frontiers in Endocrinology, 7, Article 30. https://doi.org/10.3389/fendo.2016.00030
4. Scheja, L., Heeren, J. (2016). Metabolic interplay between white, beige, brown adipocytes and the liver. Journal of Hepatology, 64(5), 1176–1186. https://doi.org/10.1016/j.jhep.2016.01.025
5. Richard, A. J., White, U., Elks, C. M., Stephens, J. M. (2020). Adipose tissue: physiology to metabolic dysfunction. Endotext [Internet]. Retrieved from https://www.ncbi.nlm.nih.gov/books/NBK555602/ Accessed January 20, 2022
6. Fonseca-Alaniz, M. H., Takada, J., Alonso-Vale, M. I. C., Lima, F. B. (2007). Adipose tissue as an endocrine organ: from theory to practice. Jornal de Pediatria, 83, S192-S203. https://doi.org/10.2223/JPED.1709
7. Alkhouli, N., Mansfield, J., Green, E., Bell, J., Knight, B., Liversedge, N. et al. (2013). The mechanical properties of human adipose tissues and their relationships to the structure and composition of the extracellular matrix. American Journal of Physiology-Endocrinology and Metabolism, 305(12), E1427-E1435. https://doi.org/10.1152/ajpendo.00111.2013
8. Kershaw, E. E., Flier, J. S. (2004). Adipose tissue as an endocrine organ. The Journal of Clinical Endocrinology and Metabolism, 89(6), 2548–2556. https://doi.org/10.1210/jc.2004–0395
9. Trayhurn, P. (2017). Origins and early development of the concept that brown adipose tissue thermogenesis is linked to energy balance and obesity. Biochimie, 134, 62–70. https://doi.org/10.1016/j.biochi.2016.09.007
10. Villarroya, J., Cereijo, R., Villarroya, F. (2013). An endocrine role for brown adipose tissue? American Iournal of PhysiologyEndocrinology and Metabolism, 305(5), E567-E572. https://doi.org/10.1152/ajpendo.00250.2013
11. Oelkrug, R., Polymeropoulos, E. T., Jastroch, M. (2015). Brown adipose tissue: physiological function and evolutionary significance. Journal of Comparative Physiology B, 185, 587–606. https://doi.org/10.1007/s00360–015–0907–7
12. Gonzalez-Hurtado, E., Lee, J., Choi, J., Wolfgang, M. J. (2018). Fatty acid oxidation is required for active and quiescent brown adipose tissue maintenance and thermogenic programing. Molecular Metabolism, 7, 45–56. https://doi.org/10.1016/j.mol-met.2017.11.004
13. Lee, J., Ellis, J. M., Wolfgang, M. J. (2015). Adipose fatty acid oxidation is required for thermogenesis and potentiates oxidative stress-induced inflammation. Cell Reports, 10(2), 266–279. https://doi.org/10.1016/j.celrep.2014.12.023
14. Giralt, M., Villarroya, F. (2013). White, brown, beige/brite: different adipose cells for different functions? Endocrinology, 154(9), 2992–3000. https://doi.org/10.1210/en.2013–1403
15. Wang, H., Willershäuser, M., Karlas, A., Gorpas, D., Reber, J., Ntziachristos, V. et al. (2019). A dual Ucp1 reporter mouse model for imaging and quantitation of brown and brite fat recruitment. Molecular Metabolism, 20, 14–27. https://doi.org/10.1016/j.molmet.2018.11.009
16. Chu, D.-T., Gawronska-Kozak, B. (2017). Brown and brite adipocytes: Same function, but different origin and response. Biochimie, 138, 102–105. https://doi.org/10.1016/j.bio-chi.2017.04.017
17. Pilkington, A.-C., Paz, H. A., Wankhade, U. D. (2021). Beige adipose tissue identification and marker specificity — Overview. Frontiers in Endocrinology, 12, Article 599134. https://doi.org/10.3389/fendo.2021.599134
18. Harms, M., Seale, P. (2013). Brown and beige fat: development, function and therapeutic potential. Nature Medicine, 19(10), 1252–1263. https://doi.org/10.1038/nm.3361
19. Keipert, S., Jastroch, M. (2014). Brite/beige fat and UCP1 — is it thermogenesis? Biochimica et Biophysica Acta (BBA)-Bioenergetics, 1837(7), 1075–1082. https://doi.org/10.1016/j.bbabio.2014.02.008
20. Bartesaghi, S., Hallen, S., Huang, L., Svensson, P.-A., Momo, R. A., Wallin, S. et al. (2015). Thermogenic activity of UCP1 in human white fat-derived beige adipocytes. Molecular Endocrinology, 29(1), 130–139. https://doi.org/10.1210/me.2014–1295
21. Zinngrebe, J., Debatin, K.-M., Fischer-Posovszky, P. (2020). Adipocytes in hematopoiesis and acute leukemia: friends, enemies, or innocent bystanders? Leukemia, 34(9), 2305–2316. https://doi.org/10.1038/s41375–020–0886-x
22. Price, N. L., Fernández-Hernando, C. (2016). miRNA regulation of white and brown adipose tissue differentiation and function. Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids, 1861(12, Part B), 2104–2110. https://doi.org/10.1016/j.bbalip.2016.02.010
23. Bondarenko, V.M., Pimanov, S.I., Makarenko, E.V. (2017). Ultrasound diagnosing of visceral. Vestnik Vitebskogo Gosudarstvennogo Meditsinskogo Universiteta, 16(1), 71–79. https://doi.org/10.22263/2312–4156.2017.1.71 (In Russian)
24. Kuznetsova T. G., Nasonova V. V., Tuniyeva E. K., Spiridonov K. I. (2020). Influence of histological features on functional properties of animal proteins. Vsyo o Myase, 4, 38–41 https://doi.org/10.21323/2071–2499–2020–4–38–41 (In Russian)
25. Gondret, F., Guével, B., Pere, M. C., Quesnel, H., Billon, Y., Com, E. et al. (2018). Proteomic analysis of adipose tissue during the last weeks of gestation in pure and crossbred Large White or Meishan fetuses gestated by sows of either breed. Journal of Animal Science and Biotechnology, 9, Article 28. https://doi.org/10.1186/s40104–018–0244–2
26. Akhremko, A.G., Vetrova, E.S. (2021). Comparative proteomic study of pig muscle proteins during growth and development of an animal. Theory and Practice of Meat Processing, 6(4), 320–327. https://doi.org/10.21323/2414–438X-2021–6–4–320–327
27. Vasilevskaya, E.R., Aryuzina, M.A., Vetrova, E.S. (2021). Saline extraction as a method of obtaining a mixture of biologically active compounds of protein nature from a porcine pancreas. Food Systems, 4(2), 97–105. https://doi.org/10.21323/2618–9771–2021–4–2–97–105 (In Russian)
28. Romeis B. (1989). Microscopic technique. München: Urban und Schwarzenberg. (In German).
29. UniProt, Retrieved from https://www.uniprot.org/uniprotkb?query=Cytochrome%20rat. Accessed January 20, 2022
30. UniProt, Retrieved from https://www.uniprot.org/uniprotkb/P10715/entry#expression. Accessed January 20, 2022
31. UniProt, Retrieved from https://www.uniprot.org/uniprotkb/P62898/entry#interaction. Accessed January 20, 2022
32. Zhang, F., Hao, G., Shao, M., Nham, K., An, Y., Wang, Q. et al. (2018). An adipose tissue atlas: an image-guided identification of human-like BAT and beige depots in rodents. Cell Metabolism, 27(1), 252–262 https://doi.org/10.1016/j.cmet.2017.12.004
33. Cohen, P., Kajimura, S. (2021). The cellular and functional complexity of thermogenic fat. Nature Reviews Molecular Cell Biology, 22(6), 393–409. https://doi.org/10.1038/s41580–021–00350–0
Review
For citations:
Chernukha I.M., Kupaeva N.V., Smirnova J.A., Akhremko A.G., Pchelkina V.A., Kotenkova E.A. Methodology of adipose tissue type detection in mammals. Theory and practice of meat processing. 2023;8(1):43-50. https://doi.org/10.21323/2414-438X-2023-8-1-43-50
Views:
583
Email this article 