Barnea A, Hajibeigi A, Cho G, Magni P
Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas.
Neuroendocrinology. 1991 Jul;54(1):7-13. doi: 10.1159/000125844.
The aim of this study was to establish a culture system of fetal brain cells that could serve as a model for the study of the developmental regulation of the neuropeptide Y (NPY) neuron. Single cell suspensions were prepared from the hypothalamic-olfactory tubercle region of 18-day-old rat fetuses, and aggregates were formed by incubation in serum-free medium under constant rotation. Aggregate formation was complete within 24-48 h, and cultures were maintained for up to 23 days. The content of immunoreactive (IR) NPY in the medium and in the aggregates increased progressively with time in culture and at each time point, the medium contained 5- to 10-fold more NPY-IR. A 48-hour exposure to forskolin resulted in a 2-fold increase in the accumulation of NPY-IR in the aggregates and in the medium, indicating that both production and secretion of NPY are regulated by the cAMP intracellular pathway. Sephadex gel filtration revealed the presence of proNPY- and NPY-size substances. The ratio of NPY- to proNPY-size substances increased progressively with age of the aggregates as well as in tissues obtained from perinatal rats of comparable age. Thus, production and secretion of NPY-IR in the cultured aggregates are regulated processes and hence, this culture system can serve as a model to study regulatory processes in the developing NPY neuron.
本研究的目的是建立一种胎儿脑细胞培养系统，该系统可作为研究神经肽Y（NPY）神经元发育调节的模型。从18日龄大鼠胎儿的下丘脑 - 嗅结节区域制备单细胞悬液，并在无血清培养基中持续旋转孵育形成聚集体。聚集体在24 - 48小时内形成，培养物维持长达23天。培养基和聚集体中免疫反应性（IR）NPY的含量在培养过程中随时间逐渐增加，并且在每个时间点，培养基中含有的NPY - IR比聚集体多5至10倍。用福司可林处理48小时导致聚集体和培养基中NPY - IR的积累增加2倍，表明NPY的产生和分泌均受细胞内cAMP途径调节。葡聚糖凝胶过滤显示存在前NPY和NPY大小的物质。随着聚集体年龄的增加以及从同龄围产期大鼠获得的组织中，NPY大小物质与前NPY大小物质的比例逐渐增加。因此，培养聚集体中NPY - IR的产生和分泌是受调节的过程，因此，该培养系统可作为研究发育中NPY神经元调节过程的模型。
