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<article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns:ali="http://www.niso.org/schemas/ali/1.0/" article-type="research-article" dtd-version="1.2" xml:lang="en"><front><journal-meta><journal-id journal-id-type="publisher-id">Morphology</journal-id><journal-title-group><journal-title xml:lang="en">Morphology</journal-title><trans-title-group xml:lang="ru"><trans-title>Морфология</trans-title></trans-title-group></journal-title-group><issn publication-format="print">1026-3543</issn><issn publication-format="electronic">2949-2556</issn><publisher><publisher-name xml:lang="en">Eco-Vector</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="publisher-id">687507</article-id><article-id pub-id-type="doi">10.17816/morph.687507</article-id><article-id pub-id-type="edn">IJECVC</article-id><article-categories><subj-group subj-group-type="toc-heading" xml:lang="en"><subject>Original Study Articles</subject></subj-group><subj-group subj-group-type="toc-heading" xml:lang="ru"><subject>Оригинальные исследования</subject></subj-group><subj-group subj-group-type="article-type"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title xml:lang="en">Detection of brain macrophages in rats using different anti-CD68/macrosialin antibodies</article-title><trans-title-group xml:lang="ru"><trans-title>Выявление макрофагов головного мозга у крыс с использованием различных антител к CD68/макросиалину</trans-title></trans-title-group><trans-title-group xml:lang="zh"><trans-title>利用多种抗CD68/巨噬细胞唾液酸蛋白抗体检测大鼠脑巨噬细胞</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0009-0002-8659-733X</contrib-id><contrib-id contrib-id-type="spin">6780-2677</contrib-id><name-alternatives><name xml:lang="en"><surname>Beketova</surname><given-names>Anastasiia A.</given-names></name><name xml:lang="ru"><surname>Бекетова</surname><given-names>Анастасия Алексеевна</given-names></name><name xml:lang="zh"><surname>Beketova</surname><given-names>Anastasiia A.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><email>beketova.anastasiya@yandex.ru</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-3997-2232</contrib-id><contrib-id contrib-id-type="spin">8877-8902</contrib-id><name-alternatives><name xml:lang="en"><surname>Razenkova</surname><given-names>Valeria A.</given-names></name><name xml:lang="ru"><surname>Разенкова</surname><given-names>Валерия Алексеевна</given-names></name><name xml:lang="zh"><surname>Razenkova</surname><given-names>Valeria A.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Biology)</p></bio><bio xml:lang="ru"><p>канд. биол. наук</p></bio><bio xml:lang="zh"><p>Cand. Sci. (Biology)</p></bio><email>valeriya.raz@yandex.ru</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0001-6113-3948</contrib-id><contrib-id contrib-id-type="spin">5725-8742</contrib-id><name-alternatives><name xml:lang="en"><surname>Kirik</surname><given-names>Olga V.</given-names></name><name xml:lang="ru"><surname>Кирик</surname><given-names>Ольга Викторовна</given-names></name><name xml:lang="zh"><surname>Kirik</surname><given-names>Olga V.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Biology)</p></bio><bio xml:lang="ru"><p>канд. биол. наук</p></bio><bio xml:lang="zh"><p>Cand. Sci. (Biology)</p></bio><email>olga_kirik@mail.ru</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-2456-8165</contrib-id><contrib-id contrib-id-type="spin">3252-3029</contrib-id><name-alternatives><name xml:lang="en"><surname>Korzhevskii</surname><given-names>Dmitrii E.</given-names></name><name xml:lang="ru"><surname>Коржевский</surname><given-names>Дмитрий Эдуардович</given-names></name><name xml:lang="zh"><surname>Korzhevskii</surname><given-names>Dmitrii E.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Dr. Sci. (Medicine)</p></bio><bio xml:lang="ru"><p>д-р мед. наук</p></bio><bio xml:lang="zh"><p>Dr. Sci. (Medicine)</p></bio><email>dek2@yandex.ru</email><xref ref-type="aff" rid="aff1"/></contrib></contrib-group><aff-alternatives id="aff1"><aff><institution xml:lang="en">Institute of Experimental Medicine</institution></aff><aff><institution xml:lang="ru">Институт экспериментальной медицины</institution></aff><aff><institution xml:lang="zh">Institute of Experimental Medicine</institution></aff></aff-alternatives><pub-date date-type="preprint" iso-8601-date="2025-11-24" publication-format="electronic"><day>24</day><month>11</month><year>2025</year></pub-date><pub-date date-type="pub" iso-8601-date="2026-03-23" publication-format="electronic"><day>23</day><month>03</month><year>2026</year></pub-date><volume>164</volume><issue>2</issue><issue-title xml:lang="en"/><issue-title xml:lang="ru"/><fpage>235</fpage><lpage>244</lpage><history><date date-type="received" iso-8601-date="2025-07-14"><day>14</day><month>07</month><year>2025</year></date><date date-type="accepted" iso-8601-date="2025-07-29"><day>29</day><month>07</month><year>2025</year></date></history><permissions><copyright-statement xml:lang="en">Copyright ©; 2026, Eco-Vector</copyright-statement><copyright-statement xml:lang="ru">Copyright ©; 2026, Эко-Вектор</copyright-statement><copyright-statement xml:lang="zh">Copyright ©; 2026,</copyright-statement><copyright-year>2026</copyright-year><copyright-holder xml:lang="en">Eco-Vector</copyright-holder><copyright-holder xml:lang="ru">Эко-Вектор</copyright-holder><ali:free_to_read xmlns:ali="http://www.niso.org/schemas/ali/1.0/" start_date="2029-03-23"/><license><ali:license_ref xmlns:ali="http://www.niso.org/schemas/ali/1.0/">https://creativecommons.org/licenses/by-nc-nd/4.0/</ali:license_ref></license></permissions><self-uri xlink:href="https://j-morphology.com/1026-3543/article/view/687507">https://j-morphology.com/1026-3543/article/view/687507</self-uri><abstract xml:lang="en"><p><bold>BACKGROUND:</bold> Macrophages represent a heterogeneous cell population with diverse origins, phenotypic characteristics, and functional properties. In the brain, resident macrophages are associated with barrier structures, including the meninges, blood vessels, and choroid plexus. Anti-CD68/macrosialin antibodies are commonly used to identify macrophages. Published data and experimental experience indicate a low degree of conservation of this protein, necessitating the use of species-specific antibodies and careful selection of primary reagents for macrophage detection in laboratory rodent tissues.</p> <p><bold>AIM:</bold> To perform a comparative analysis of macrophage detection in rats using rabbit polyclonal anti-CD68 antibodies and mouse monoclonal antibodies of the ED1 clone.</p> <p><bold>METHODS:</bold> Brain sections obtained from Wistar rats (<italic>n</italic> = 6) were examined; heart and liver sections from the same strain served as controls. CD68/macrosialin-positive macrophages were identified using mouse monoclonal antibodies (clone ED1) and rabbit polyclonal antibodies in various immunohistochemical staining protocols.</p> <p><bold>RESULTS:</bold> Both antibody types specifically detected brain macrophages in three locations: the pia mater, vascular walls, and the choroid plexus. Rabbit polyclonal antibodies demonstrated higher intensity of specific staining compared with mouse monoclonal antibodies, with no nonspecific binding observed. The use of mouse monoclonal antibodies required heat-induced antigen retrieval and blocking reagents within the staining protocol.</p> <p><bold>CONCLUSION:</bold> Comparative analysis of macrophage detection in rat brain sections using antibodies of different origins and clonality demonstrated that rabbit polyclonal antibodies provide reliable macrophage labeling using a simpler immunohistochemical protocol compared with mouse monoclonal antibodies. Therefore, rabbit polyclonal antibodies may be recommended as a useful alternative for CD68/macrosialin staining in rat tissue sections.</p></abstract><trans-abstract xml:lang="ru"><p><bold>Обоснование. </bold>Макрофаги — это разнородная по происхождению, фенотипическим и функциональным особенностям популяция клеток. В головном мозге резидентные макрофаги ассоциированы с барьерными органами: мозговыми оболочками, сосудами и сосудистым сплетением. Для выявления макрофагов обычно используют антитела к CD68/ макросиалину. Данные литературы и опыт исследователей свидетельствуют о низкой степени консервативности этого белка, что обусловливает необходимость использования видоспецифических антител и тщательного подбора первичных реагентов для выявления макрофагов в тканях лабораторных грызунов.</p> <p><bold>Цель исследования</bold> — провести сравнительный анализ результатов выявления макрофагов у крыс при использовании кроличьих поликлональных антител к CD68 и мышиных моноклональных антител клона ED1.</p> <p><bold>Методы.</bold> Объектом исследования служили срезы головного мозга крыс линии Вистар (<italic>n</italic> = 6), а в качестве контроля использовали срезы сердца и печени крыс той же линии. Макрофаги, положительные по CD68/макросиалину, выявляли с помощью мышиных моноклональных антител клона ED1 и кроличьих поликлональных антител в различных протоколах иммуногистохимической реакции.</p> <p><bold>Результаты.</bold> Оба типа антител позволяют специфично выявлять макрофаги головного мозга трёх локализаций: в мягкой мозговой оболочке, стенках сосудов и сосудистом сплетении. При использовании кроличьих поликлональных антител интенсивность специфической реакции выше по сравнению с мышиными моноклональными антителами, а неспецифическое связывание отсутствует. Установлена необходимость теплового демаскирования антигена и применения блокирующих реагентов в протоколах с использованием мышиных моноклональных антител.</p> <p><bold>Заключение. </bold>Анализ результатов выявления макрофагов на срезах головного мозга крыс при использовании антител разного происхождения и клональности показал, что кроличьи поликлональные антитела хорошо выявляют макрофаги при более простом протоколе иммуногистохимической реакции по сравнению с мышиными моноклональными антителами, а потому могут быть рекомендованы в качестве полезной альтернативы для окрашивания CD68/макросиалина на срезах тканей крысы.</p></trans-abstract><trans-abstract xml:lang="zh"><p><bold>论证。</bold>巨噬细胞是一类在起源、表型及功能特征上均具有异质性的细胞群体。在脑内，常驻巨噬细胞与屏障器官：脑膜、血管和脉络丛密切相关。抗CD68/巨噬细胞唾液酸蛋白抗体常用于检测巨噬细胞。文献资料及研究经验表明，该蛋白保守性较低，因此，在实验室啮齿动物组织中检测巨噬细胞时，需使用物种特异性抗体并审慎选择一抗。</p> <p><bold>目的。</bold>本研究旨在比较使用兔抗CD68多克隆抗体和小鼠单克隆抗体ED1检测大鼠脑巨噬细胞的结果。</p> <p><bold>方法。</bold>以Wistar大鼠（n = 6）的脑组织切片为研究对象，并以同品系大鼠的心、肝组织切片作为对照。采用小鼠单克隆抗体ED1及兔多克隆抗体，在不同的免疫组织化学染色方案中检测CD68/巨噬细胞唾液酸蛋白阳性的巨噬细胞。</p> <p><bold>结果。</bold>两种抗体均可特异性显示巨噬细胞在三个部位的分布：软脑膜、血管壁及脉络丛。与小鼠单克隆抗体相比，使用兔多克隆抗体时特异性反应强度更高，且无非特异性结合。研究表明，使用小鼠单克隆抗体时，必须进行抗原热修复并应用封闭试剂。</p> <p><bold>结论。</bold>通过对比分析不同来源及克隆型抗体在大鼠脑组织切片中检测巨噬细胞的效果，发现相较于小鼠单克隆抗体，兔多克隆抗体能够以更简捷的免疫组织化学反应方案有效显示巨噬细胞，因此可作为大鼠组织切片CD68/巨噬细胞唾液酸蛋白染色的优质替代方案。</p></trans-abstract><kwd-group xml:lang="en"><kwd>CD68</kwd><kwd>macrophages</kwd><kwd>monoclonal antibodies</kwd><kwd>polyclonal antibodies</kwd><kwd>immunohistochemistry</kwd><kwd>rat</kwd></kwd-group><kwd-group xml:lang="ru"><kwd>CD68</kwd><kwd>макрофаги</kwd><kwd>моноклональные антитела</kwd><kwd>поликлональные антитела</kwd><kwd>иммуногистохимия</kwd><kwd>крыса</kwd></kwd-group><kwd-group xml:lang="zh"><kwd>CD68</kwd><kwd>巨噬细胞</kwd><kwd>单克隆抗体</kwd><kwd>多克隆抗体</kwd><kwd>免疫组织化学</kwd><kwd>大鼠</kwd></kwd-group><funding-group><award-group><funding-source><institution-wrap><institution xml:lang="ru">Российский научный фонд</institution></institution-wrap><institution-wrap><institution xml:lang="en">Russian Science Foundation</institution></institution-wrap><institution-wrap><institution xml:lang="zh">Russian Science Foundation</institution></institution-wrap></funding-source><award-id>24-15-00032</award-id></award-group><funding-statement xml:lang="en">The study was supported by the Russian Science Foundation, Project No. 24-15-00032, https://rscf.ru/project/24-15-00032/</funding-statement><funding-statement xml:lang="ru">Исследование выполнено при финансовой поддержке Российского научного фонда, проект № 24-15-00032, https://rscf.ru/project/24-15-00032/</funding-statement><funding-statement xml:lang="zh">The study was supported by the Russian Science Foundation, Project No. 24-15-00032, https://rscf.ru/project/24-15-00032/</funding-statement></funding-group></article-meta></front><body></body><back><ref-list><ref id="B1"><label>1.</label><mixed-citation>Lazarov T, Juarez-Carreño S, Cox N, Geissmann F. 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