Single nucleus/cell RNA-seq of the chicken hypothalamicpituitaryovarian axis offers new insights into the molecular regulatory mechanisms of ovarian development

The hypothalamic-pituitary-ovarian(HPO)axis represents a central neuroendocrine network essential for reproductive function.Despite its critical role,the intrinsic heterogeneity within the HPO axis across vertebrates and the complex intercellular interactions remain poorly defined.This study provides the first comprehensive,unbiased,cell type-specific molecular profiling of all three components of the HPO axis in adult Lohmann layers and Liangshan Yanying chickens.Within the hypothalamus,pituitary,and ovary,seven,12,and 13 distinct cell types were identified,respectively.Results indicated that the pituitary adenylate cyclase activating polypeptide(PACAP),follicle-stimulating hormone(FSH),and prolactin(PRL)signaling pathways may modulate the synthesis and secretion of gonadotropin-releasing hormone(GnRH),FSH,and luteinizing hormone(LH)within the hypothalamus and pituitary.In the ovary,interactions between granulosa cells and oocytes involved the KIT,CD99,LIFR,FN1,and ANGPTL signaling pathways,which collectively regulate follicular maturation.The SEMA4 signaling pathway emerged as a critical mediator across all three tissues of the HPO axis.Additionally,gene expression analysis revealed that relaxin 3(RLN3),gastrin-releasing peptide(GRP),and cocaine-and amphetamine regulated transcripts(CART,also known as CARTPT)may function as novel endocrine hormones,influencing the HPO axis through autocrine,paracrine,and endocrine pathways.Comparative analyses between Lohmann layers and Liangshan Yanying chickens demonstrated higher expression levels of GRP,RLN3,CARTPT,LHCGR,FSHR,and GRPR in the ovaries of Lohmann layers,potentially contributing to their superior reproductive performance.In conclusion,this study provides a detailed molecular characterization of the HPO axis,offering novel insights into the regulatory mechanisms underlying reproductive biology.

精氨酸功能化羟基磷灰石纳米颗粒与人脐静脉内皮细胞的相互作用及机制

选用铽掺杂的精氨酸修饰的羟基磷灰石纳米颗粒(Arg@HAP^(Tb)NPs),探讨该纳米颗粒与细胞的相互作用以及Arg@HAP^(Tb)NPs在血管内皮细胞中的内吞机制。选取人脐静脉内皮细胞(HUVEC)作为研究的模型细胞,通过激光扫描共聚焦显微镜监测Arg@HAP^(Tb)NPs在细胞中的摄取情况,并通过细胞流速仪检测不同浓度下的细胞摄取后的平均荧光强度。结果表明,HUVEC吸收Arg@HAP^(Tb)NPs后主要定位在细胞质中,并主要分布在细胞核周围。Arg@HAP^(Tb)NPs的入胞过程具有时间和浓度依赖性,作用时间以4 h为宜,纳米颗粒的浓度以50 mg/L为最佳。Arg@HAP^(Tb)NPs入胞量明显强于未经精氨酸修饰的HAP^(Tb)NPs。Arg@HAP^(Tb)NPs进入HUVECs是一个能量依赖的主动运输过程,通过小凹蛋白介导的内吞作用发生。