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垂体腺苷酸环化酶激活多肽及其前体编码信使核糖核酸在人和大鼠组织中的分布与分子特征

Distribution, molecular characterization of pituitary adenylate cyclase-activating polypeptide and its precursor encoding messenger RNA in human and rat tissues.

作者信息

Ghatei M A, Takahashi K, Suzuki Y, Gardiner J, Jones P M, Bloom S R

机构信息

Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, U.K.

出版信息

J Endocrinol. 1993 Jan;136(1):159-66. doi: 10.1677/joe.0.1360159.

Abstract

The distribution of a novel neuropeptide, pituitary adenylate cyclase-activating polypeptide (PACAP), was studied in the brain of the rat and man and a variety of other rat tissues using Northern blot hybridization and two radioimmunoassays for PACAP 1-38 and PACAP 1-27. The assay, using PACAP 1-38 as standard and an antibody to PACAP 21-38 and radiolabelled tracer, revealed immunoreactive PACAP in all brain regions examined, with the highest concentrations in the rat being in the hypothalamus, nucleus accumbens and substantia nigra (380 +/- 34, 310 +/- 37 and 346 +/- 30 pmol/g wet tissue, means +/- S.E.M., n = 5 respectively), whilst in man the highest concentrations were found in the pituitary gland (15.8 +/- 4.7 pmol/g). Immunoreactive PACAP 1-38 was also detected in the rat gastrointestinal tract, adrenal gland and testis. The assay using PACAP 1-27 as standard and label and an antibody to PACAP 1-27 detected immunoreactive PACAP only in the rat hypothalamus (12.6 +/- 1.8 pmol/g wet tissue, n = 5). PACAP mRNA of approximately 2.7 kb in size was detectable in all brain regions of both rat and man, and its distribution paralleled that of the immunoreactive peptide. Gel permeation chromatography of different regions of human and rat hypothalamus, and also rat spinal cord and small intestine, showed a broad immunoreactive peak corresponding to PACAP 1-38. Fast protein liquid chromatography (FPLC) resolved this peak into two immunoreactive peaks, the majority eluting in the position of synthetic PACAP 1-38.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

利用Northern印迹杂交以及针对垂体腺苷酸环化酶激活多肽(PACAP)1 - 38和PACAP 1 - 27的两种放射免疫分析法,研究了一种新型神经肽——垂体腺苷酸环化酶激活多肽(PACAP)在大鼠和人类大脑以及多种其他大鼠组织中的分布情况。以PACAP 1 - 38为标准品,使用针对PACAP 21 - 38的抗体和放射性标记示踪剂的分析方法显示,在所检测的所有脑区均存在免疫反应性PACAP,大鼠中浓度最高的区域为下丘脑、伏隔核和黑质(分别为380±34、310±37和346±30 pmol/g湿组织,均值±标准误,n = 5),而在人类中浓度最高的区域是垂体(15.8±4.7 pmol/g)。在大鼠胃肠道、肾上腺和睾丸中也检测到了免疫反应性PACAP 1 - 38。以PACAP 1 - 27为标准品和标记物,使用针对PACAP 1 - 27的抗体的分析方法仅在大鼠下丘脑检测到了免疫反应性PACAP(12.6±1.8 pmol/g湿组织,n = 5)。在大鼠和人类的所有脑区均可检测到大小约为2.7 kb的PACAP mRNA,其分布与免疫反应性肽平行。对人类和大鼠下丘脑的不同区域以及大鼠脊髓和小肠进行凝胶渗透色谱分析,显示出一个与PACAP 1 - 38相对应的宽免疫反应峰。快速蛋白质液相色谱(FPLC)将该峰分离为两个免疫反应峰,大部分在合成PACAP 1 - 38的位置洗脱。(摘要截短于250字)

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