agronaukaАграрная наука Евро-Северо-ВостокаAgricultural Science Euro-North-East2072-90812500-1396FARC North-East10.30766/2072-9081.2025.26.5.963-974agronauka-2212Research ArticleОБЗОРЫREVIEWSОплодотворяющая способность сперматозоидов: факторы ее обуславливающие и методы определения (обзор)Fertilizing ability of spermatozoa: its conditioning factors and methods of determination (review)https://orcid.org/0009-0003-5630-8987БерелетТ. Ю.BereletT. Y.

Берелет Татьяна Юрьевна, студент,

ул. Черниговская, д. 5, г. Санкт-Петербург, 196084

 

Tatiana Y. Berelet, student,

Chernigovskaya St., 5, Saint Petersburg, 196084

secretary@spbguvm.ruhttps://orcid.org/0000-0002-7011-4594КорочкинаЕ. А.KorochkinaE. A.

Корочкина Елена Александровна, доктор вет. наук, доцент,

ул. Черниговская, д. 5, г. Санкт-Петербург, 196084

secretary@spbguvm.ru

Elena A. Korochkina, DSc in Veterinary Science, Associate Professor,

Chernigovskaya St., 5, Saint Petersburg, 196084

e.kora@mail.ruФГБОУ ВО «Санкт-Петербургский государственный университет ветеринарной медицины»РоссияSaint Petersburg State University of Veterinary MedicineRussian Federation202503112025265963974Copyright © Берелет Т.Ю., Корочкина Е.А., 20252025Берелет Т.Ю., Корочкина Е.А.Berelet T.Y., Korochkina E.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.agronauka-sv.ru/jour/article/view/2212

Оплодотворение – сложный процесс, результатом которого является слияние гамет. Для получения оплодотворяющей способности сперматозоидам необходимо пройти постэякуляционные процессы в репродуктивном тракте самок. К ним относятся капацитация, гиперактивация и акросомальная реакция. Капацитация является сложным процессом, во время которого сперматозоид подвергается различным изменениям метаболизма, внутриклеточных концентраций ионов и других веществ. Основными факторами капацитации являются концентрация внутриклеточных ионов кальция, изменение свойств и структуры плазматической мембраны, pH среды, а также воздействие прогестерона и холестерола на сперматозоид. Возникающая далее гиперактивация приводит к изменению подвижности сперматозоида, вследствие чего он достигает яйцеклетки для дальнейшего проникновения в нее. Затем происходит акросомальная реакция – выброс из акросомы веществ, обеспечивающих проникновение через прозрачную оболочку ооцита. Определить оплодотворяющую способность можно различными методами: HBA-тест основан на связывании сперматозоидов с гиалуроновой кислотой, SpermSlow используется для замедления сперматозоидов в гиалуронсодержащей среде. Целостность акросомы и подвижность сперматозоида можно определить с помощью метода Acrobeads, основанного на образовании у подвижных сперматозоидов комплекса с иммуноглобулинами, покрытыми антителами против белка с внутренней поверхности акросомальной мембраны. Метод SPA позволяет установить функциональную способность сперматозоидов связываться с оболочкой яйцеклетки. Помимо прямых методов, исследующих основные параметры и функции сперматозоида, есть методы, определяющие дополнительные параметры, такие как уровень окислительного стресса и факторов его вызывающих, наличие нарушений в генетическом аппарате мужской гаметы. Уровень окислительного стресса, количество активных форм кислорода можно найти с помощью реакции на тиобарбитуровую кислоту, реакцией с нитросиним тетразолием, путем оценки количества карбонильных производных аминокислотных остатков в белках, хемолюминисцентным анализом. Разрывы в ДНК можно определить тестом TUNEL, метод ДНК-комет используют для идентификации степени повреждения геномной ДНК, метод FISH применяют для анализа хромосомного набора сперматозоида. В шейке матки содержится цервикальная слизь, являющаяся необходимым фактором для оплодотворения. Способность сперматозоидов проникать через нее можно определить с помощью вспомогательных методов, например, swim-up. Его суть заключается в том, что в культуральной среде имитируют естественное перемещение мужских гамет, и отбирают те, которые отвечают требованиям. Концентрацию сперматозоидов и степень их подвижности в цервикальной слизи определяют посткоитальным тестом. Учитывая вышесказанное, и главным образом, доступность использования данных методик, актуальным является разработка отечественных наборов.

Fertilization is a complex process that results in the fusion of gametes. In order to obtain fertilizing ability, spermatozoa need to undergo post-ejaculation processes in the female reproductive tract. These include capacitation, hyperactivation, and acrosomal reaction. Capacitation is a complex process during which the sperm undergoes various changes in metabolism, intracellular concentrations of ions and other substances. The main factors of capacitation are the concentration of intracellular calcium ions, changes in the properties and structure of the plasma membrane, the pH of the medium, as well as the effect of progesterone and cholesterol on the sperm. The resulting hyperactivation leads to a change in the motility of the sperm, as a result of which it reaches the egg for further penetration into it. Then an acrosomal reaction occurs – the release of substances from the acrosome that ensure penetration through the transparent membrane of the oocyte. The fertilizing ability can be determined by various methods: the HBA test is based on the binding of spermatozoa to hyaluronic acid, SpermSlow is used to slow down spermatozoa in a hyaluronic-containing medium. The integrity of the acrosome and the motility of the sperm can be determined using the Acrobeads method, based on the formation of a complex of motile spermatozoa with immunoglobulins coated with antibodies against the protein from the inner surface of the acrosomal membrane. The SPA method allows us to determine the functional ability of spermatozoa to bind to the egg shell. In addition to direct methods that examine the basic parameters and functions of the sperm, there are methods that determine additional parameters, such as the level of oxidative stress and the factors that cause it, and the presence of disorders in the genetic apparatus of the male gamete. The level of oxidative stress and the amount of reactive oxygen species can be determined by reaction to thiobarbituric acid, reaction with nitrosine tetrazolium, by estimating the amount of carbonyl derivatives of amino acid residues in proteins, and by chemoluminescence analysis. DNA breaks can be detected using the TUNEL test, the DNA comet method is used to identify the degree of damage to genomic DNA, and the FISH method is used to analyze the chromosome set of a sperm cell. The cervix contains cervical mucus, which is a necessary factor for fertilization. The ability of spermatozoa to penetrate through it can be determined using auxiliary methods, for example, swim-up. Its essence lies in the fact that in a cultural environment, the natural movement of male gametes is imitated, and those that meet the requirements are selected. The concentration of spermatozoa and the degree of their motility in the cervical mucus are determined by a postcoital test. Taking into account the availability of these techniques from a logistical point of view, the development of domestic kits is relevant.

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The authors declare that there are no conflicts of interest present.