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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="ru"><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">696674</article-id><article-id pub-id-type="doi">10.17816/morph.696674</article-id><article-id pub-id-type="edn">KRTZPD</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">Age-dependent changes in cholinergic neurons of the rat small and large intestines</article-title><trans-title-group xml:lang="ru"><trans-title>Возрастные изменения холинергических нейронов тонкой и толстой кишки у крыс</trans-title></trans-title-group><trans-title-group xml:lang="zh"><trans-title>大鼠小肠和大肠胆碱能神经元的年龄相关变化</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-3333-5865</contrib-id><contrib-id contrib-id-type="spin">3691-4817</contrib-id><name-alternatives><name xml:lang="en"><surname>Budnik</surname><given-names>Antonina F.</given-names></name><name xml:lang="ru"><surname>Будник</surname><given-names>Антонина Францевна</given-names></name><name xml:lang="zh"><surname>Budnik</surname><given-names>Antonina F.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>MD, Cand. Sci. (Medicine), Assistant Professor</p></bio><bio xml:lang="ru"><p>канд. мед. наук, доцент</p></bio><bio xml:lang="zh"><p>MD, Cand. Sci. (Medicine), Assistant Professor</p></bio><email>budnik74@mail.ru</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-6230-5024</contrib-id><contrib-id contrib-id-type="spin">7676-0849</contrib-id><name-alternatives><name xml:lang="en"><surname>Masliukov</surname><given-names>Petr M.</given-names></name><name xml:lang="ru"><surname>Маслюков</surname><given-names>Петр Михайлович</given-names></name><name xml:lang="zh"><surname>Masliukov</surname><given-names>Petr M.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>MD, Dr. Sci. (Medicine), Professor</p></bio><bio xml:lang="ru"><p>д-р мед. наук, профессор</p></bio><bio xml:lang="zh"><p>MD, Dr. Sci. (Medicine), Professor</p></bio><email>mpm@ysmu.ru</email><xref ref-type="aff" rid="aff2"/></contrib></contrib-group><aff-alternatives id="aff1"><aff><institution xml:lang="en">Kabardino-Balkarian State University</institution></aff><aff><institution xml:lang="ru">Кабардино-Балкарский государственный университет им. Х.М. Бербекова</institution></aff><aff><institution xml:lang="zh">Kabardino-Balkarian State University</institution></aff></aff-alternatives><aff-alternatives id="aff2"><aff><institution xml:lang="en">Yaroslavl State Medical University</institution></aff><aff><institution xml:lang="ru">Ярославский государственный медицинский университет</institution></aff><aff><institution xml:lang="zh">Yaroslavl State Medical University</institution></aff></aff-alternatives><content-language>ru</content-language><pub-date date-type="preprint" publication-format="electronic" iso-8601-date="2026-04-19"><day>19</day><month>04</month><year>2026</year></pub-date><pub-date date-type="pub" publication-format="electronic" iso-8601-date="2026-08-07"><day>07</day><month>08</month><year>2026</year></pub-date><pub-date date-type="collection"><year>2026</year></pub-date><volume>164</volume><issue>3</issue><issue-title xml:lang="en"/><issue-title xml:lang="ru"/><issue-title xml:lang="zh"/><fpage>404</fpage><lpage>414</lpage><history><date date-type="received" iso-8601-date="2025-11-20"><day>20</day><month>11</month><year>2025</year></date><date date-type="accepted" iso-8601-date="2025-12-24"><day>24</day><month>12</month><year>2025</year></date></history><pub-history><event event-type="received"><event-desc>Received:<date date-type="received" iso-8601-date="2025-11-20"><day>20</day><month>11</month><year>2025</year></date></event-desc></event><event event-type="accepted"><event-desc>Accepted:<date date-type="accepted" iso-8601-date="2025-12-24"><day>24</day><month>12</month><year>2025</year></date></event-desc></event><event event-type="pub"><event-desc>Published:<pub-date date-type="pub" publication-format="electronic" iso-8601-date="2026-08-07"><day>07</day><month>08</month><year>2026</year></pub-date></event-desc></event></pub-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="2029-08-07"/><license><ali:license_ref xmlns:ali="http://www.niso.org/schemas/ali/1.0/">https://eco-vector.com/for_authors.php#07</ali:license_ref></license></permissions><self-uri xlink:href="https://j-morphology.com/1026-3543/article/view/696674">https://j-morphology.com/1026-3543/article/view/696674</self-uri><abstract xml:lang="en"><p><bold>BACKGROUND:</bold> Neurons containing the acetylcholine-synthesizing enzyme, choline acetyltransferase (ChAT), constitute a significant proportion of cells in the myenteric (MP) and submucosal (SP) plexuses of the small and large intestines. However, current studies lack data on age-related changes in this group of neurons during postnatal ontogenesis.</p> <p><bold>AIM: </bold>To determine postnatal changes in ChAT-containing neurons in the intramural ganglia of the rat intestines.</p> <p><bold>METHODS:</bold> The study was conducted on rats of the following age groups: newborns; animals aged 10, 20, 30, and 60 days; and animals aged 12 and 24 months. Changes in the percentage and average cross-sectional area of ChAT-immunoreactive (IR) neurons in the MP and SP of the small and large intestines of rats during postnatal ontogenesis were analyzed using double-labeling immunohistochemistry with antibodies to ChAT and PGP9.5.</p> <p><bold>RESULTS: </bold>ChAT was detected in the majority of neurons in the MP and SP of the small and large intestines across all age groups. In the MP of the small intestine, the percentage of ChAT-IR neurons increased from the neonatal period to day 20 of life and then gradually decreased until old age. In the MP of the large intestine, the proportion of ChAT-IR neurons increased from the neonatal period to day 10 of life and then also decreased starting from day 20 until old age. In the SP of the small intestine, the percentage of ChAT-IR neurons was higher in newborns compared with 10-day-old rats, and in newborns and day 20 rats compared with aged animals. In the SP of the large intestine, the percentage of ChAT-IR neurons increased during the first 10 days of life, decreased between days 10 and 20, and then remained unchanged at older ages.</p> <p>No differences were found in the average cross-sectional areas of ChAT-IR and ChAT-negative neurons in the small and large intestines. The average cross-sectional area of ChAT-IR neurons increased from birth to day 60 in both the MP and SP of the small and large intestines.</p> <p><bold>CONCLUSION: </bold>These data indicate age-related changes in the neurotransmitter composition of the intestinal intramural ganglia. During postnatal ontogenesis, the percentage of ChAT-IR neurons in the nerve plexuses of the small and large intestines changes: it increases in the early postnatal period and decreases in old age.</p></abstract><trans-abstract xml:lang="ru"><p><bold>Обоснование. </bold>Нейроны, содержащие фермент синтеза ацетилхолина — холинацетилтрансферазу (ХАТ), составляют значительную долю клеток в межмышечном (МС) и подслизистом (ПС) сплетениях тонкой и толстой кишки. Однако в современной литературе отсутствуют сведения о возрастных изменениях этой группы нейронов в постнатальном онтогенезе.</p> <p><bold>Цель исследования </bold>— изучить постнатальные изменения ХАТ-содержащих нейронов в интрамуральных ганглиях кишечника крыс.</p> <p><bold>Методы.</bold> Исследование проведено на крысах следующих возрастных групп: новорождённые; 10 суток после рождения (Р10, postnatal day 10), Р20, Р30 и Р60; 12 и 24 месяца. Анализировили изменения процентного соотношения и средней площади сечения ХАТ-иммунореактивных нейронов в МС и ПС тонкой и толстой кишки крыс в постнатальном онтогенезе. Для этого проводили иммуногистохимическое окрашивание с использованием антител к ХАТ и белку PGP9.5.</p> <p><bold>Результаты. </bold>ХАТ определялась в большинстве нейронов МС и ПС тонкой и толстой кишки во всех возрастных группах. В МС тонкой кишки процент ХАТ<sup>+</sup> нейронов возрастал от периода новорождённости до Р20, а затем постепенно снижался вплоть до старости. В МС толстой кишки доля ХАТ<sup>+</sup> нейронов увеличивалась от периода новорождённости до Р10, после чего также уменьшалась. В ПС тонкой кишки процентное содержание ХАТ<sup>+</sup> нейронов больше у новорождённых по сравнению с крысами в возрасте Р10, а также у новорождённых и Р20 по сравнению со старыми животными. В ПС толстой кишки процентное содержание ХАТ<sup>+</sup> нейронов увеличивалось в первые 10 суток после рождения, уменьшалось между Р10 и Р20 и не изменялось в более старшем возрасте.</p> <p>В тонкой и толстой кишке не выявлено различий между средней площадью сечения ХАТ<sup>+</sup> и ХАТ<sup>−</sup> нейронов. Средняя площадь сечения ХАТ<sup>+ </sup>нейронов увеличивалась с момента рождения до Р60 как в МС, так и в ПС тонкой и толстой кишки.</p> <p><bold>Заключение.</bold> Полученные данные свидетельствуют о возрастных преобразованиях нейротрансмиттерного состава интрамуральных ганглиев кишечника. В постнатальном онтогенезе изменяется доля ХАТ<sup>+</sup> нейронов в нервных сплетениях тонкой и толстой кишки — она увеличивается в раннем постнатальном периоде и снижается в старости.</p></trans-abstract><trans-abstract xml:lang="zh"><p><bold>论证。</bold>含有乙酰胆碱合成酶 — — 胆碱乙酰转移酶 (ChAT) 的神经元在小肠和大肠的肌间神经丛 и 黏膜下神经丛中占有相当大的比例。然而，现代文献中尚无关于这类神经元在出生后发育过程中年龄相关变化的信息。</p> <p><bold>目的。</bold>本研究旨在探讨大鼠肠壁神经节中含 ChAT 神经元的出生后变化。</p> <p><bold>方法。</bold>本研究选取以下年龄组的大鼠：新生大鼠；出生后第10天（P10）、第20天（P20）、第30天（P30）和第60天（P60）；12个月和24个月。分析了出生后发育过程中大鼠小肠和大肠中肌间神经丛 и 黏膜下神经丛区域 ChAT 免疫反应性神经元的百分比和平均横截面积的变化。为此，采用 ChAT 和 PGP9.5 蛋白抗体进行免疫组织化学染色。</p> <p><bold>结果。</bold>在所有年龄组的小肠和大肠的肌间神经丛 и 黏膜下神经丛中，大多数神经元均检测到了 ChAT。在小肠肌间神经丛中，ChAT 阳性神经元的比例从新生儿期到 P20 逐渐增加，之后逐渐下降直至老年。在大肠肌间神经丛中，ChAT 阳性神经元的比例从新生儿期到 P10 逐渐增加，之后也逐渐下降。在小肠黏膜下神经丛中，新生儿的 ChAT 阳性神经元比例高于 P10 的大鼠，新生儿和 P20 的大鼠的 ChAT 阳性神经元比例也高于老年大鼠。在大肠黏膜下神经丛中，ChAT 阳性神经元的比例在出生后前10天增加，在 P10 至 P20 之间下降，之后在更年长的年龄段保持不变。</p> <p>在小肠和大肠中，ChAT 阳性神经元和 ChAT 阴性神经元的平均横截面积无显著差异。在大小肠的肌间神经丛和黏膜下神经丛中，ChAT 阳性神经元的平均横截面积从出生到 P60 均有所增加。</p> <p><bold>结论。</bold>所得数据表明肠壁神经节的神经递质组成存在年龄相关性变化。在出生后的个体发育过程中，小肠和大肠神经丛中 ChAT 阳性神经元的比例发生变化：在出生后早期增加，在老年期减少。</p></trans-abstract><kwd-group xml:lang="en"><kwd>choline acetyltransferase</kwd><kwd>small intestine</kwd><kwd>large intestine</kwd><kwd>myenteric plexus</kwd><kwd>submucous plexus</kwd><kwd>ontogenesis</kwd></kwd-group><kwd-group xml:lang="ru"><kwd>холинацетилтрансфераза</kwd><kwd>тонкая кишка</kwd><kwd>толстая кишка, межмышечное сплетение</kwd><kwd>подслизистое сплетение</kwd><kwd>онтогенез</kwd></kwd-group><kwd-group xml:lang="zh"><kwd>胆碱乙酰转移酶</kwd><kwd>小肠</kwd><kwd>大肠</kwd><kwd>肌间神经丛</kwd><kwd>黏膜下神经丛</kwd><kwd>个体发育</kwd></kwd-group><funding-group/></article-meta></front><body></body><back><ref-list><ref id="B1"><label>1.</label><mixed-citation>Klein J. The central cholinergic synapse: a primer. Int J Mol Sci. 2025;26(19):9670. doi: 10.3390/ijms26199670 EDN: QXNNMM</mixed-citation></ref><ref id="B2"><label>2.</label><mixed-citation>Sharkey KA, Mawe GM. The enteric nervous system. Physiol Rev. 2023;103(2):1487–1564. doi: 10.1152/physrev.00018.2022 EDN: XRQNJM</mixed-citation></ref><ref id="B3"><label>3.</label><mixed-citation>Costa M, Spencer NJ, Brookes SJH. The role of enteric inhibitory neurons in intestinal motility. Auton Neurosci. 2021;235:102854. doi: 10.1016/j.autneu.2021.102854 EDN: HKJGZB</mixed-citation></ref><ref id="B4"><label>4.</label><mixed-citation>Furness JB. Comparative and evolutionary aspects of the digestive system and its enteric nervous system control. Adv Exp Med Biol. 2022;1383:165–177. doi: 10.1007/978-3-031-05843-1_16</mixed-citation></ref><ref id="B5"><label>5.</label><mixed-citation>Fung C, Vanden Berghe P. Functional circuits and signal processing in the enteric nervous system. Cell Mol Life Sci. 2020; 77(22):4505–4522. doi: 10.1007/s00018-020-03543-6 EDN: FJKRTL</mixed-citation></ref><ref id="B6"><label>6.</label><mixed-citation>Spencer NJ, Hu H. Enteric nervous system: sensory transduction, neural circuits and gastrointestinal motility. Nat Rev Gastroenterol Hepatol. 2020;17(6):338–351. doi: 10.1038/s41575-020-0271-2 EDN: COPBEW</mixed-citation></ref><ref id="B7"><label>7.</label><mixed-citation>Furness JB, Stebbing MJ. The first brain: Species comparisons and evolutionary implications for the enteric and central nervous systems. Neurogastroenterol Motil. 2018;30(2):e13234. doi: 10.1111/nmo.13234 EDN: YDLCEX</mixed-citation></ref><ref id="B8"><label>8.</label><mixed-citation>Masliukov PM, Budnik AF, Nozdrachev AD. Neurochemical features of metasympathetic system ganglia in the course of ontogenesis. Advances in Gerontology. 2017;7(4):281–289. doi: 10.1134/S2079057017040087 EDN: XXGXFR</mixed-citation></ref><ref id="B9"><label>9.</label><mixed-citation>Masliukov PM, Nozdratchev AD, Timmermans JP. The age-bound specifics of the stellate ganglion neurons’ transmitter composition. Russian Journal of Physiology. 2006;92(2):214–221. EDN: JSCIHP</mixed-citation></ref><ref id="B10"><label>10.</label><mixed-citation>Scott-Solomon E, Boehm E, Kuruvilla R. The sympathetic nervous system in development and disease. Nat Rev Neurosci. 2021;22(11):685–702. doi: 10.1038/s41583-021-00523-y EDN: KPKSEE</mixed-citation></ref><ref id="B11"><label>11.</label><mixed-citation>Erickson CS, Lee SJ, Barlow-Anacker AJ, et al. Appearance of cholinergic myenteric neurons during enteric nervous system development: comparison of different ChAT fluorescent mouse reporter lines. Neurogastroenterol Motil. 2014;26(6):874–884. doi: 10.1111/nmo.12343</mixed-citation></ref><ref id="B12"><label>12.</label><mixed-citation>Hao MM, Bornstein JC, Young HM. Development of myenteric cholinergic neurons in ChAT-Cre;R26R-YFP mice. J Comp Neurol. 2013;521(14):3358–3370. doi: 10.1002/cne.23354</mixed-citation></ref><ref id="B13"><label>13.</label><mixed-citation>Korzhevskii DE, Grigor’ev IP, Gusel’nikova VV, et al. Immunohistochemical markers for neurobiology. Medical academic journal. 2019;19(4):7–24. doi: 10.17816/MAJ16548 EDN: BQAXWZ</mixed-citation></ref><ref id="B14"><label>14.</label><mixed-citation>Chumasov EI, Petrova ES, Korzhevskii DE. Study of the rat duodenal innervation using neural immunohistochemical markers. Russian Journal of Physiology. 2020;106(7):853–865. doi: 10.31857/S086981392007002X EDN: XGGZHF</mixed-citation></ref><ref id="B15"><label>15.</label><mixed-citation>Avtandilov GG. Medical morphometry. Moscow; Meditsina: 1990. (In Russ.)</mixed-citation></ref><ref id="B16"><label>16.</label><mixed-citation>Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9(7):671–675. doi: 10.1038/nmeth.2089</mixed-citation></ref><ref id="B17"><label>17.</label><mixed-citation>Mead R, Gilmour SG, Mead A. Statistical principles for the design of experiments: applications to real experiments. Cambridge: Cambridge University Press; 2012. doi: 10.1017/CBO9781139020879</mixed-citation></ref><ref id="B18"><label>18.</label><mixed-citation>Parathan P, Wang Y, Leembruggen AJ, et al. The enteric nervous system undergoes significant chemical and synaptic maturation during adolescence in mice. Dev Biol. 2020;458(1):75–87. doi: 10.1016/j.ydbio.2019.10.011 EDN: ERMPCV</mixed-citation></ref><ref id="B19"><label>19.</label><mixed-citation>Masliukov PM, Budnik AF. Morphometric characteristics of intramural autonomic nerve ganglia of the myenteric and submucosal plexuses of the small and large intestines in rats during postnatal ontogenesis. Morphology. 2025;163(4):305–315. doi: 10.17816/morph.677905 EDN: GICHGI</mixed-citation></ref><ref id="B20"><label>20.</label><mixed-citation>Budnik AF, Masliukov PM. Postnatal development of the enteric neurons expressing neuronal nitric oxide synthase. Anat Rec (Hoboken). 2023;306(9):2276–2291. doi: 10.1002/ar.24947 EDN: FDQLJQ</mixed-citation></ref><ref id="B21"><label>21.</label><mixed-citation>Masliukov PM, Emanuilov AI, Budnik AF. Sympathetic innervation of the development, maturity, and aging of the gastrointestinal tract. Anat Rec (Hoboken). 2023;306(9):2249–2263. doi: 10.1002/ar.25015 EDN: BVUDEQ</mixed-citation></ref><ref id="B22"><label>22.</label><mixed-citation>Emanuilov AI, Budnik AF, Masliukov PM. Somatostatin-immunoreactive neurons of the rat gut during the development. Histochem Cell Biol. 2024;162(5):385–402. doi: 10.1007/s00418-024-02322-9 EDN: HOGJDV</mixed-citation></ref><ref id="B23"><label>23.</label><mixed-citation>Masliukov PM, Budnik AF, Vishnyakova PA, Pavlov AV. Neurochemical features of the neuropeptide Y-ergic enteric submucous neurons of the small intestine in postnatal ontogenesis. Russian Journal of Physiology. 2021;107(10):1209–1218. doi: 10.31857/S0869813921100083 EDN: SMRRDK</mixed-citation></ref><ref id="B24"><label>24.</label><mixed-citation>Liu S. Neurotrophic factors in enteric physiology and pathophysiology. Neurogastroenterol Motil. 2018;30(10):e13446. doi: 10.1111/nmo.13446</mixed-citation></ref><ref id="B25"><label>25.</label><mixed-citation>Furness JB. The enteric nervous system and neurogastroenterology. Nat Rev Gastroenterol Hepatol. 2012;9(5):286–294. doi: 10.1038/nrgastro.2012.32</mixed-citation></ref><ref id="B26"><label>26.</label><mixed-citation>Stern T, Davis AM. Evaluation and treatment of patients with constipation. JAMA. 2016;315(2):192–193. doi: 10.1001/jama.2015.16995</mixed-citation></ref><ref id="B27"><label>27.</label><mixed-citation>Yeh KM, Johansson O, Le H, et al. Cystic fibrosis transmembrane conductance regulator modulates enteric cholinergic activities and is abnormally expressed in the enteric ganglia of patients with slow transit constipation. J Gastroenterol. 2019;54(11):994–1006. doi: 10.1007/s00535-019-01610-9 EDN: OPIXFB</mixed-citation></ref><ref id="B28"><label>28.</label><mixed-citation>Lu XY, Wen YX, Jiang N, et al. DREADDs-based chemogenetics induced slow transit constipation via inhibition of enteric neurons. J Dig Dis. 2025;26(1–2):62–73. doi: 10.1111/1751-2980.13344 EDN: WRDUKU</mixed-citation></ref></ref-list></back></article>
