Nozaki M, Gorbman A
Gen Comp Endocrinol. 1984 Mar;53(3):335-52. doi: 10.1016/0016-6480(84)90261-2.
The occurrence and localization of molecular components of pro-opiocortin in the pituitary and brain of two species of adult lamprey, Petromyzon marinus and Entosphenus tridentatus, were studied immunocytochemically using antisera generated against human pro-gamma-MSH (N-terminal fragment 1Trp to 71Gly of pro-opiocortin), porcine ACTH, alpha-MSH, human beta-endorphin, gamma-endorphin, and methionine enkephalin. (1) In both species of lamprey most cells of the rostral pars distalis and some cells of the caudal pars distalis contained Met-enkephalin-like immunoreactivity. Some of these cells also contained gamma-endorphin-like immunoreactivity. After preabsorbing the antisera with their corresponding antigens or related peptides, the Met-enkephalin/gamma-endorphin-like material was found to be related to Met-enkephalin, but not identical with either Met-enkephalin or gamma-endorphin. However, results of anti-pro-gamma-MSH, anti-ACTH, anti-alpha-MSH, or anti-beta-endorphin were consistently negative in the pars distalis of both lamprey species. (2) Immunoreaction to anti-Met-enkephalin was found in some cells of the pars intermedia in both species of lamprey. Although the positive reaction had been eliminated by preabsorption with synthetic Met-enkephalin, the diffuseness of the positive stain in the pars intermedia cells resembled an artifactual cross-reaction rather than a specific reaction. In P. marinus, but not in E. tridentatus, similar inconsistent and questionable immunoreactions corresponding to ACTH and alpha-MSH also occurred in some pars intermedia cells. Results of other antisera (anti-pro-gamma-MSH, anti-beta-endorphin, or anti-gamma-endorphin) were consistently negative in the pars intermedia of both lamprey species. (3) In both species of lamprey beta-endorphin-like material was found in the hypothalamus. In E. tridentatus only Met-enkephalin-like material was observed in the hypothalamus, and these two substances were distributed in different neuronal elements. After application of anti-pro-gamma-MSH, anti-ACTH, anti-alpha-MSH, or anti-gamma-endorphin, no positive reaction was found in the brain of either species of lamprey. These findings suggest that if a pro-opiocortin-related prohormone exists in the lamprey, it may be chemically different from those of more advanced vertebrates, and it clearly differs in distribution between the brain and parts of the pituitary gland.
利用针对人促γ-促黑激素(促阿片皮质素的N端片段1Trp至71Gly)、猪促肾上腺皮质激素、α-促黑激素、人β-内啡肽、γ-内啡肽和甲硫氨酸脑啡肽产生的抗血清,通过免疫细胞化学方法研究了两种成年七鳃鳗(海七鳃鳗和三齿七鳃鳗)垂体和脑内促阿片皮质素分子成分的发生和定位。(1)在两种七鳃鳗中,吻部远侧部的大多数细胞和尾侧部远侧部的一些细胞含有甲硫氨酸脑啡肽样免疫反应性。其中一些细胞还含有γ-内啡肽样免疫反应性。用相应抗原或相关肽预吸收抗血清后,发现甲硫氨酸脑啡肽/γ-内啡肽样物质与甲硫氨酸脑啡肽有关,但与甲硫氨酸脑啡肽或γ-内啡肽均不相同。然而,抗促γ-促黑激素、抗促肾上腺皮质激素、抗α-促黑激素或抗β-内啡肽的结果在两种七鳃鳗的远侧部均始终为阴性。(2)在两种七鳃鳗的中间部一些细胞中发现了抗甲硫氨酸脑啡肽的免疫反应。虽然用合成甲硫氨酸脑啡肽预吸收已消除了阳性反应,但中间部细胞中阳性染色的弥散性更像是一种人为交叉反应而非特异性反应。在海七鳃鳗中,而不是在三齿七鳃鳗中,中间部一些细胞中也出现了与促肾上腺皮质激素和α-促黑激素相应的类似不一致且有疑问的免疫反应。其他抗血清(抗促γ-促黑激素、抗β-内啡肽或抗γ-内啡肽)的结果在两种七鳃鳗的中间部均始终为阴性。(3)在两种七鳃鳗的下丘脑均发现了β-内啡肽样物质。在三齿七鳃鳗的下丘脑中仅观察到甲硫氨酸脑啡肽样物质,且这两种物质分布于不同的神经元成分中。应用抗促γ-促黑激素、抗促肾上腺皮质激素、抗α-促黑激素或抗γ-内啡肽后,在两种七鳃鳗的脑中均未发现阳性反应。这些发现表明,如果七鳃鳗中存在与促阿片皮质素相关的前体激素,其化学性质可能与更高级脊椎动物的不同,并且在脑和垂体部分之间的分布也明显不同。