角膜中神经肽的纯化、表征及定位

Purification, characterization, and localization of neuropeptides in the cornea.

作者信息

Tinsley P W, Fridland G H, Killmar J T, Desiderio D M

机构信息

Charles B. Stout Neuroscience Mass Spectrometry Laboratory, University of Tennessee, Memphis 38163.

出版信息

Peptides. 1988 Nov-Dec;9(6):1373-9. doi: 10.1016/0196-9781(88)90205-7.

DOI:10.1016/0196-9781(88)90205-7

PMID:2470068

Abstract

The immunologically detected neuropeptides methionine enkephalin (ME), substance P (SP), beta-endorphin (beta-End), and alpha-melanocyte stimulating hormone (alpha-MSH) were purified from bovine corneal extracts by gradient, followed by isocratic, reversed phase-high performance liquid chromatography (RP-HPLC) and characterized, after both chromatographic steps, by radioimmunoassay (RIA). Immunologically detected ME and SP were purified from canine corneal extracts by gradient RP-HPLC and characterized by RIA. An anatomical study of the bovine cornea separated the cornea into an epithelium-enriched and a stroma-enriched portion. After gradient RP-HPLC, RIA demonstrated that all the ME-like immunoreactivity was located in the corneal epithelium, whereas the SP-like immunoreactivity was distributed between the stroma and epithelium in an approximate two-to-one ratio.

摘要

通过梯度洗脱，随后进行等度洗脱、反相高效液相色谱法（RP-HPLC），从牛角膜提取物中纯化出经免疫检测的神经肽甲硫氨酸脑啡肽（ME）、P物质（SP）、β-内啡肽（β-End）和α-黑素细胞刺激素（α-MSH），并在两个色谱步骤之后通过放射免疫测定法（RIA）对其进行表征。通过梯度RP-HPLC从犬角膜提取物中纯化出经免疫检测的ME和SP，并通过RIA对其进行表征。一项对牛角膜的解剖学研究将角膜分为富含上皮的部分和富含基质的部分。梯度RP-HPLC后，RIA显示所有ME样免疫反应性都位于角膜上皮中，而SP样免疫反应性则以大约二比一的比例分布在基质和上皮之间。