Development of the nervous system in mouse liver.
Development of the nervous system in mouse liver.
复制标题
DOI：
10.4254/wjh.v14.i2.386
复制
发表时间：
2022-02-27
影响因子：
2.4
通讯作者：
Taniguchi H
中科院分区：
文献类型：
--
作者：
Koike N;Tadokoro T;Ueno Y;Okamoto S;Kobayashi T;Murata S;Taniguchi H
肝脏神经系统在肝脏发育中的作用尚不清楚。我们之前通过将人肝内皮细胞与内皮细胞和间充质细胞共同培养，在小鼠身上创造了具有功能的人微肝组织。然而，它们缺乏Glisson鞘[门静脉束(PT)]。PT由肝动脉(HA)、门静脉和肝内胆管(IHBD)的分支组成，统称为门静脉三联体，以及自主神经。用免疫组织化学方法评价小鼠肝神经网络在PT中的发育情况。取C57BL/6J小鼠胚胎10.5天至出生后56天不同发育时期的肝脏标本。分别用蛋白基因产物9.5(PGP9.5)和细胞角蛋白19(CK19)抗体、神经纤维标记物(NFs)和胆管上皮细胞(BECs)对肝门表面薄片进行检测。分别以肝门周围(中心)和肝脏周边(周边)为研究对象，比较各发育阶段中心和周边的平均值。对PT中的核因子-IHBD和核因子-HA接触者进行计数，并量化它们之间的关系。BEC标志物SRY相关高迁移率族盒基因9(SOX9)、肝细胞标志物HNF4α(HNF4α)和Notch配体Jagge1也进行了免疫组织化学染色。胚胎10.5胎肝组织中HNF4α呈阳性表达，Jagge1呈弥漫性阳性表达，而胎肝中则呈阴性表达。E12.5时SOX9阳性细胞散在分布于肝门周围区域。Jagge-1主要表达于门静脉周围组织，SOX9阳性细胞数在E16.5时增加。SOX9阳性细胞构成导管板和主要位于中心的原始IHBD，SOX-9阳性IHBD在同一时期部分获得CK19阳性。PGP9.5阳性小体最早发现于胎龄16.5天，阳性小体最早发现于胎龄0天的中央门静脉周围组织。因此，原始的PT结构首先在中心的P0处构建。随着门静脉周围组织的重塑，CK19阳性的IHBD和PGP9.5阳性的神经纤维数量逐渐增加，直到P5，门静脉周围也形成PTS。从E16.5到P5，中心区的神经纤维网和IHBD数显著高于周边区。出生后28天，神经纤维束和IHBD的数目达到成体水平，中央和外周的差异缩小。在出生后早期，在PTS中，与HAS相关的NSF比与IHBD相关的频率更高，此后，NFIHBD接触的数量逐渐增加。小鼠肝脏的神经纤维在出生前最早出现在中心，并向外围延伸。在PT结构的形态发生过程中，NBS与IHBDS或HAS之间的相互作用起着重要作用。
The role of the hepatic nervous system in liver development remains unclear. We previously created functional human micro-hepatic tissue in mice by co-culturing human hepatic endodermal cells with endothelial and mesenchymal cells. However, they lacked Glisson’s sheath [the portal tract (PT)]. The PT consists of branches of the hepatic artery (HA), portal vein, and intrahepatic bile duct (IHBD), collectively called the portal triad, together with autonomic nerves. To evaluate the development of the mouse hepatic nervous network in the PT using immunohistochemistry. Liver samples from C57BL/6J mice were harvested at different developmental time periods, from embryonic day (E) 10.5 to postnatal day (P) 56. Thin sections of the surface cut through the hepatic hilus were examined using protein gene product 9.5 (PGP9.5) and cytokeratin 19 (CK19) antibodies, markers of nerve fibers (NFs), and biliary epithelial cells (BECs), respectively. The numbers of NFs and IHBDs were separately counted in a PT around the hepatic hilus (center) and the peripheral area (periphery) of the liver, comparing the average values between the center and the periphery at each developmental stage. NF-IHBD and NF-HA contacts in a PT were counted, and their relationship was quantified. SRY-related high mobility group-box gene 9 (SOX9), another BEC marker; hepatocyte nuclear factor 4α (HNF4α), a marker of hepatocytes; and Jagged-1, a Notch ligand, were also immunostained to observe the PT development. HNF4α was expressed in the nucleus, and Jagged-1 was diffusely positive in the primitive liver at E10.5; however, the PGP9.5 and CK19 were negative in the fetal liver. SOX9-positive cells were scattered in the periportal area in the liver at E12.5. The Jagged-1 was mainly expressed in the periportal tissue, and the number of SOX9-positive cells increased at E16.5. SOX9-positive cells constructed the ductal plate and primitive IHBDs mainly at the center, and SOX-9-positive IHBDs partly acquired CK19 positivity at the same period. PGP9.5-positive bodies were first found at E16.5 and HAs were first found at P0 in the periportal tissue of the center. Therefore, primitive PT structures were first constructed at P0 in the center. Along with remodeling of the periportal tissue, the number of CK19-positive IHBDs and PGP9.5-positive NFs gradually increased, and PTs were also formed in the periphery until P5. The numbers of NFs and IHBDs were significantly higher in the center than in the periphery from E16.5 to P5. The numbers of NFs and IHBDs reached the adult level at P28, with decreased differences between the center and periphery. NFs associated more frequently with HAs than IHBDs in PTs at the early phase after birth, after which the number of NF-IHBD contacts gradually increased. Mouse hepatic NFs first emerge at the center just before birth and extend toward the periphery. The interaction between NFs and IHBDs or HAs plays important roles in the morphogenesis of PT structure.
登录
查看更多内容
影响因子：
64.5
作者：
Mukouyama, Y;Shin, D;Anderson, DJ
通讯作者：
Anderson, DJ
影响因子：
2.2
作者：
Ikeda, Osamu;Ozaki, Michitaka;Ohkohchi, Nobuhiro
通讯作者：
Ohkohchi, Nobuhiro
影响因子：
1.9
作者：
ELSALHY, M;STENLING, R;GRIMELIUS, L
通讯作者：
GRIMELIUS, L
影响因子：
2.2
作者：
Hamada, Takayuki;Eguchi, Susumu;Kanematsu, Takashi
通讯作者：
Kanematsu, Takashi
影响因子：
2.4
作者：
Shiojiri, Nobuyoshi;Kametani, Harunobu;Kawakami, Hayato
通讯作者：
Kawakami, Hayato