{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:30:58Z","timestamp":1787027458706,"version":"build-2736575974"},"reference-count":79,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2025,8,4]],"date-time":"2025-08-04T00:00:00Z","timestamp":1754265600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Inner Mongolia Open Competition Project","award":["2022JBGS0025"],"award-info":[{"award-number":["2022JBGS0025"]}]},{"name":"Inner Mongolia Open Competition Project","award":["BR251026"],"award-info":[{"award-number":["BR251026"]}]},{"name":"Inner Mongolia Open Competition Project","award":["BZX202201"],"award-info":[{"award-number":["BZX202201"]}]},{"name":"Inner Mongolia Open Competition Project","award":["CG202410"],"award-info":[{"award-number":["CG202410"]}]},{"name":"Inner Mongolia Open Competition Project","award":["CG202423"],"award-info":[{"award-number":["CG202423"]}]},{"name":"Inner Mongolia Open Competition Project","award":["NDYB2022-1"],"award-info":[{"award-number":["NDYB2022-1"]}]},{"name":"Fundamental Research Founds for Inner Mongolia Autonomous Region Direct Affiliated Universities","award":["2022JBGS0025"],"award-info":[{"award-number":["2022JBGS0025"]}]},{"name":"Fundamental Research Founds for Inner Mongolia Autonomous Region Direct Affiliated Universities","award":["BR251026"],"award-info":[{"award-number":["BR251026"]}]},{"name":"Fundamental Research Founds for Inner Mongolia Autonomous Region Direct Affiliated Universities","award":["BZX202201"],"award-info":[{"award-number":["BZX202201"]}]},{"name":"Fundamental Research Founds for Inner Mongolia Autonomous Region Direct Affiliated Universities","award":["CG202410"],"award-info":[{"award-number":["CG202410"]}]},{"name":"Fundamental Research Founds for Inner Mongolia Autonomous Region Direct Affiliated Universities","award":["CG202423"],"award-info":[{"award-number":["CG202423"]}]},{"name":"Fundamental Research Founds for Inner Mongolia Autonomous Region Direct Affiliated Universities","award":["NDYB2022-1"],"award-info":[{"award-number":["NDYB2022-1"]}]},{"name":"Double First-Class Construction Funds for Inner Mongolia Agricultural University","award":["2022JBGS0025"],"award-info":[{"award-number":["2022JBGS0025"]}]},{"name":"Double First-Class Construction Funds for Inner Mongolia Agricultural University","award":["BR251026"],"award-info":[{"award-number":["BR251026"]}]},{"name":"Double First-Class Construction Funds for Inner Mongolia Agricultural University","award":["BZX202201"],"award-info":[{"award-number":["BZX202201"]}]},{"name":"Double First-Class Construction Funds for Inner Mongolia Agricultural University","award":["CG202410"],"award-info":[{"award-number":["CG202410"]}]},{"name":"Double First-Class Construction Funds for Inner Mongolia Agricultural University","award":["CG202423"],"award-info":[{"award-number":["CG202423"]}]},{"name":"Double First-Class Construction Funds for Inner Mongolia Agricultural University","award":["NDYB2022-1"],"award-info":[{"award-number":["NDYB2022-1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The hypothalamus\u2013pituitary\u2013ovarian (HPO) axis orchestrates reproductive functions through intricate neuroendocrine crosstalk. Here, we integrated single-nucleus RNA sequencing (snRNA-seq) and spatial transcriptomics (ST) to decode the cellular heterogeneity and intercellular communication networks in the reproductive systems of pregnant Mongolian cattle. We retained a total of 6161 high-quality nuclei from the hypothalamus, 14,715 nuclei from the pituitary, and 26,072 nuclei from the ovary, providing a comprehensive cellular atlas across the HPO axis. In the hypothalamus, neurons exhibited synaptic and neuroendocrine specialization, with glutamatergic subtype Glut4 serving as a TGF\u03b2 signaling hub to regulate pituitary feedback, while GABAergic GABA1 dominated PRL signaling, likely adapting maternal behavior. Pituitary stem cells dynamically replenished endocrine populations via TGF\u03b2, and lactotrophs formed a PRL\u2013PRLR paracrine network with stem cells, synergizing mammary development. Ovarian luteal cells exhibited steroidogenic specialization and microenvironmental synergy: endothelial cells coregulated TGF\u03b2-driven angiogenesis and immune tolerance, while luteal\u2013stromal PRL\u2013PRLR interactions amplified progesterone synthesis and nutrient support. Granulosa cells (GCs) displayed spatial-functional stratification, with steroidogenic GCs persisting across pseudotime as luteinization precursors, while atretic GCs underwent apoptosis. Spatial mapping revealed GCs\u2019 annular follicular distribution, mediating oocyte\u2013somatic crosstalk, and luteal\u2013endothelial colocalization supporting vascularization. This study unveils pregnancy-specific HPO axis regulation, emphasizing multi-organ crosstalk through TGF\u03b2\/PRL pathways and stem cell-driven plasticity, offering insights into reproductive homeostasis and pathologies.<\/jats:p>","DOI":"10.3390\/ani15152277","type":"journal-article","created":{"date-parts":[[2025,8,4]],"date-time":"2025-08-04T15:30:06Z","timestamp":1754321406000},"page":"2277","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Single-Nucleus RNA Sequencing and Spatial Transcriptomics Reveal Cellular Heterogeneity and Intercellular Communication Networks in the Hypothalamus\u2013Pituitary\u2013Ovarian Axis of Pregnant Mongolian Cattle"],"prefix":"10.3390","volume":"15","author":[{"given":"Yanchun","family":"Bao","sequence":"first","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"},{"name":"Inner Mongolia Engineering Research Center 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China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3913-2944","authenticated-orcid":false,"given":"Hongxia","family":"Jia","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"},{"name":"Inner Mongolia Engineering Research Center of Genomic Big Data for Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunhan","family":"Li","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"},{"name":"Inner Mongolia Engineering Research Center of Genomic Big Data for Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyi","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"},{"name":"Inner Mongolia Engineering Research Center of Genomic Big Data for Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiajing","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"},{"name":"Inner Mongolia Engineering Research Center of Genomic Big Data for Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengbo","family":"Gu","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"},{"name":"Inner Mongolia Engineering Research Center of Genomic Big Data for Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caixia","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"},{"name":"Inner Mongolia Engineering Research Center of Genomic Big Data for Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8244-9519","authenticated-orcid":false,"given":"Mingjuan","family":"Gu","sequence":"additional","affiliation":[{"name":"College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China"},{"name":"Inner Mongolia Engineering Research Center of Genomic Big Data for Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Zhu","sequence":"additional","affiliation":[{"name":"College of Animal Science, 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