Translation. Region: Russian Federal

Source: Novosibirsk State University – Novosibirsk State University –

What processes occur in the embryo at the molecular level after freezing? Can a slowdown in metabolism indicate risks for the future organism? A unique study in which a master’s student is participating helps to find answers to these questions Faculty of Physics Anastasia Omelchenko of Novosibirsk State University. A team of scientists from the Laboratory of Condensed Matter Spectroscopy of the Institute of Automation and Electrometry of the Siberian Branch of the Russian Academy of Sciences (where Anastasia works) and the Cryopreservation and Reproductive Technologies Sector of the Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, for the first time in the world, used the method of Raman scattering of deuterated labels to study the metabolism of embryos.

— Raman scattering is a contactless and non-destructive method of optical spectroscopy that allows us to understand the chemical composition, structure and phase state of a substance. We use it to look inside living cells and see what molecules are formed as a result of their metabolism, — explained Anastasia Omelchenko.

Scientists are studying the metabolism of early-stage mouse embryos. To do this, they fed the embryos specially labeled (deuterated) molecules — such as amino acids, glucose, stearic acid — and used Raman spectroscopy to track how these substances were transformed inside the cells.

— When we want to track how one compound is converted into another during metabolism, we must separate these compounds from other organic molecules present in the cells. This is a fairly complex task and is similar to finding one person in a crowd. To simplify it, you can give this person a “flag”, that is, mark him – this will allow you to identify him against the background of the rest of the crowd. In Raman spectroscopy, it is convenient to use deuterated labels, that is, molecules in which some hydrogen atoms are replaced by heavier deuterium. Due to the isotopic shift, such labels have a spectrum that is different from other molecules, which allows you to track the number and nature of deuterated molecules in the sample, — the researcher explained.

It turned out that amino acids are steadily converted into proteins at all stages of development, and as the embryo grows, synthesis increases. Contrary to expectations, glucose is not so much broken down to obtain energy as it is stored as a glycogen polymer. Fatty acids, such as stearic acid, accumulate in lipid granules (the cell’s energy storage facilities).

But what was particularly interesting was the effect of cryopreservation on metabolism – a technology used to freeze and store embryos at ultra-low temperatures. After freezing and thawing, the scientists analyzed how the metabolism of the embryos changed and found that at later stages of development, fatty acids were processed worse – it was as if the cell “decided” to accumulate them rather than use them.

— The method allows us to see which processes are disrupted as a result of cryopreservation, — noted Anastasia Omelchenko. — This is especially important in conditions when more and more biomaterial — both in medicine and in agriculture — is frozen for storage or transportation. Our approach can help us understand how to preserve the viability of such cells.

In addition to cryopreservation, the study looked at how another unique condition, diapause, affects embryo metabolism. This is a natural stop in embryo development in response to unfavorable conditions, which occurs in a number of mammals (about 130 species). The scientists worked with mouse embryos with induced diapause and found that their protein synthesis decreases by about 23% compared to normal ones.

— This is consistent with other data: in embryos that enter the implantation stage, the activity of key metabolic processes increases. And our method allows us to measure this quantitatively, quickly and without harm to the object itself, — the researcher emphasizes.

The research is only just entering the stage of systemic application, but it is already clear that optical non-invasive methods of metabolic analysis may well become an effective diagnostic tool in reproductive medicine and biotechnology.

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