Vigh S, Arimura A, Gottschall P E, Kitada C, Somogyvári-Vigh A, Childs G V
U.S.-Japan Biomedical Research Laboratories, Tulane University Hebert Center, Belle Chasse, LA 70137.
Peptides. 1993 Jan-Feb;14(1):59-65. doi: 10.1016/0196-9781(93)90011-5.
Two novel peptides, named PACAP (pituitary adenylate cyclase activating polypeptide) containing 38 (PACAP38) and 27 residues (PACAP27) were recently isolated from ovine hypothalami. In order to investigate the pituitary cell type(s) that bear a receptor for PACAP, PACAP38 was biotinylated and used for cytochemical examination of binding. The cells were also identified by immunocytochemical methods using the antisera against each of the rat anterior pituitary hormones or an antiserum against S-100 protein, a marker for pituitary folliculo-stellate (FS) cells. Biotinylated PACAP38 (biot-PACAP) exhibited adenylate cyclase stimulating activity (ACSA) comparable to PACAP38 in rat pituitary cell cultures, and displaced the bound 125I-PACAP27 to the rat pituitary membrane preparation to the same extent as PACAP38. After 2-4 days of culture, dispersed rat pituitary cells were incubated with varying concentrations of biot-PACAP at room temperature or 4 degrees C. The bound biot-PACAP38 was visualized by avidin-biotin-peroxidase complex (ABC) method with nickel intensification. Biot-PACAP-positive and pituitary hormone or S-100-positive cells were counted. More than 90% of S-100-positive cells bound biot-PACAP38. A considerable number of GH and PRL cells and a lesser number of ACTH cells also bound biot-PACAP38, whereas only a few identified LH, FSH, or TSH cells bound biot-PACAP38. These results suggest that FS cells are a major target cell type for PACAP. A recent study from our laboratory demonstrated that PACAP stimulated the release of interleukin (IL)-6 in rat pituitary cell cultures. FS cells are known to produce IL-6.
最近从绵羊下丘脑分离出两种新的肽，分别为含38个氨基酸的垂体腺苷酸环化酶激活多肽（PACAP38）和含27个氨基酸的（PACAP27）。为了研究表达PACAP受体的垂体细胞类型，对PACAP38进行生物素化，并用于结合的细胞化学检测。还通过免疫细胞化学方法，使用针对每种大鼠垂体前叶激素的抗血清或针对S-100蛋白（垂体滤泡星状细胞（FS）的标志物）的抗血清对细胞进行鉴定。在大鼠垂体细胞培养物中，生物素化的PACAP38（生物素-PACAP）表现出与PACAP38相当的腺苷酸环化酶刺激活性（ACSA），并且与PACAP38一样，能将结合到大鼠垂体膜制剂上的125I-PACAP27置换到相同程度。培养2-4天后，将分散的大鼠垂体细胞在室温或4℃下与不同浓度的生物素-PACAP孵育。通过抗生物素蛋白-生物素-过氧化物酶复合物（ABC）法并经镍强化来观察结合的生物素-PACAP38。对生物素-PACAP阳性以及垂体激素或S-100阳性细胞进行计数。超过90%的S-100阳性细胞结合生物素-PACAP38。相当数量的生长激素（GH）和催乳素（PRL）细胞以及较少数量的促肾上腺皮质激素（ACTH）细胞也结合生物素-PACAP38，而仅少数已鉴定的促黄体生成素（LH）、促卵泡生成素（FSH）或促甲状腺激素（TSH）细胞结合生物素-PACAP38。这些结果表明，FS细胞是PACAP的主要靶细胞类型。我们实验室最近的一项研究表明，PACAP可刺激大鼠垂体细胞培养物中白细胞介素（IL）-6的释放。已知FS细胞可产生IL-6。
