Bloom F, Battenberg E, Rossier J, Ling N, Guillemin R
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1591-5. doi: 10.1073/pnas.75.3.1591.
Well-characterized antisera to porcine beta-endorphin were used to localize immunoreactive sites in cryostat sections of formaldehyde-fixed rat brain by indirect immunohistochemistry. Specificity was established by absorption of immune sera with synthetic peptide fragments. Specific immunoreactivity was localized to neuronal perikarya in the basal tuberal hypothalamus, and to varicose nerve fibers which were distributed to midline nuclear areas throughout the diencephalon and anterior pons. These patterns of reactivity were unaffected by preabsorption of the immune sera with millimolar concentrations of Met5- or Leu5-enkephalin or alpha-endorphin. The beta-endorphin immunoreactive structures were morphologically separate from those cells and fibers reported to react with antisera to the enkephalins. One anti-beta-endorphin serum gave additional immunoreactivity with myelinated axons in limbic cortical zones; when absorbed with purified rat myelin basic protein, only the specific patterns of immunoreactivity remained. Thus, discrete beta-endorphin-containing neuronal circuits exist in rat brain and are anatomically distinguishable from enkephalin-containing nerve cell and fiber pathways.
利用对猪β-内啡肽特性充分了解的抗血清，通过间接免疫组织化学方法在甲醛固定的大鼠脑低温切片中定位免疫反应位点。通过用合成肽片段吸收免疫血清来确定特异性。特异性免疫反应定位于下丘脑基部结节的神经元胞体，以及分布于整个间脑和脑桥前部中线核区域的曲张神经纤维。这些反应模式不受用毫摩尔浓度的甲硫氨酸脑啡肽或亮氨酸脑啡肽或α-内啡肽对免疫血清进行预吸收的影响。β-内啡肽免疫反应结构在形态上与那些据报道与脑啡肽抗血清发生反应的细胞和纤维是分开的。一种抗β-内啡肽血清在边缘皮质区的有髓轴突上产生额外的免疫反应；当用纯化的大鼠髓鞘碱性蛋白吸收后，仅保留特异性免疫反应模式。因此，大鼠脑中存在离散的含β-内啡肽的神经回路，并且在解剖学上可与含脑啡肽的神经细胞和纤维通路区分开来。
