Bao Yanchun, Ma Fengying, Huo Chenxi, Jia Hongxia, Li Yunhan, Yang Xiaoyi, Liu Jiajing, Gu Pengbo, Shi Caixia, Gu Mingjuan, Zhu Lin, Wang Yu, Liu Bin, Na Risu, Zhang Wenguang
College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
Inner Mongolia Engineering Research Center of Genomic Big Data for Agriculture, Inner Mongolia Agricultural University, Hohhot 010018, China.
Animals (Basel). 2025 Aug 4;15(15):2277. doi: 10.3390/ani15152277.
The hypothalamus-pituitary-ovarian (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 TGFβ signaling hub to regulate pituitary feedback, while GABAergic GABA1 dominated PRL signaling, likely adapting maternal behavior. Pituitary stem cells dynamically replenished endocrine populations via TGFβ, and lactotrophs formed a - paracrine network with stem cells, synergizing mammary development. Ovarian luteal cells exhibited steroidogenic specialization and microenvironmental synergy: endothelial cells coregulated TGFβ-driven angiogenesis and immune tolerance, while luteal-stromal - 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' annular follicular distribution, mediating oocyte-somatic crosstalk, and luteal-endothelial colocalization supporting vascularization. This study unveils pregnancy-specific HPO axis regulation, emphasizing multi-organ crosstalk through TGFβ/PRL pathways and stem cell-driven plasticity, offering insights into reproductive homeostasis and pathologies.
下丘脑-垂体-卵巢（HPO）轴通过复杂的神经内分泌相互作用协调生殖功能。在此，我们整合了单核RNA测序（snRNA-seq）和空间转录组学（ST），以解码怀孕蒙古牛生殖系统中的细胞异质性和细胞间通讯网络。我们总共保留了来自下丘脑的6161个高质量细胞核、来自垂体的14715个细胞核和来自卵巢的26072个细胞核，提供了一个跨越HPO轴的全面细胞图谱。在下丘脑中，神经元表现出突触和神经内分泌特化，谷氨酸能亚型Glut4作为TGFβ信号枢纽来调节垂体反馈，而γ-氨基丁酸能的GABA1主导催乳素信号，可能调节母体行为。垂体干细胞通过TGFβ动态补充内分泌细胞群，而催乳素细胞与干细胞形成旁分泌网络，协同促进乳腺发育。卵巢黄体细胞表现出类固醇生成特化和微环境协同作用：内皮细胞共同调节TGFβ驱动的血管生成和免疫耐受，而黄体-基质相互作用增强孕酮合成和营养支持。颗粒细胞（GCs）表现出空间功能分层，类固醇生成的GCs在整个伪时间内作为黄体化前体持续存在，而闭锁的GCs则发生凋亡。空间映射揭示了GCs的环形卵泡分布，介导卵母细胞-体细胞相互作用，以及黄体-内皮细胞共定位支持血管化。本研究揭示了妊娠特异性的HPO轴调节，强调了通过TGFβ/PRL途径和干细胞驱动的可塑性进行的多器官相互作用，为生殖稳态和病理学提供了见解。
