IMMUNOHISTOCHEMICAL DEMONSTRATION OF POLY(ADENOSINE DIPHOSPHATE-RIBOSE) IN NUCLEI OF VARIOUS RAT-TISSUES
IMMUNOHISTOCHEMICAL DEMONSTRATION OF POLY(ADENOSINE DIPHOSPHATE-RIBOSE) IN NUCLEI OF VARIOUS RAT-TISSUES
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DOI：
10.1177/28.7.6993553
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发表时间：
1980-01-01
影响因子：
3.2
通讯作者：
HAYAISHI, O
中科院分区：
文献类型：
--
作者：
IKAI, K;UEDA, K;HAYAISHI, O
用间接免疫荧光技术研究了真核生物中一种新的大分子聚腺苷二磷酸核糖的自然分布。该抗体是在兔体内产生的针对聚(ADP-核糖)的抗体，与链长约为25个ADP-核糖单元的聚合物反应最强，与短低聚物反应弱；与单体完全惰性。用这种抗体进行免疫染色，发现这种聚合物存在于各种大鼠组织中。免疫染色似乎是多聚ADP-核糖的特异性，通过用纯化的多聚ADP-核糖预吸收抗血清或用多聚ADP-核糖降解酶预处理组织切片来判断其消失。通过与NAD预先孵育来增强荧光，NAD是合成聚(ADP-核糖)的底物，也支持这一观点。聚腺苷二磷酸核糖的免疫荧光主要分布于肝(成人、新生儿、再生和肝癌)、脑、心脏、肠、胰腺、肾、脾、睾丸、甲状腺和骨骼肌等组织。例外是血细胞；在外周血白细胞的核中几乎没有检测到荧光。只有在与NAD预先孵育后，淋巴细胞和单核细胞才显示出荧光，而粒细胞则从未显示出荧光。这些细胞似乎代表了真核细胞中的聚(ADP-核糖)合成活性第一次缺失。
Natural distribution of poly(ADP-ribose), a novel macromolecule in eukaryotes, was investigated using an indirect immunofluorescence technique. The antibody, produced in a rabbit toward poly(ADP-ribose), was most reactive with polymers having the chain length of about 25 ADP-ribose units and weakly reactive with short oligomers; it was totally inert with monomers. Immunostaining with this antibody revealed the existence of the polymer in various rat tissues. The immunostaining seems to be specific for poly(ADP-ribose), as judged by its disappearance by preabsorption of the antiserum with purified poly(ADP-ribose) or pretreatment of tissue sections with poly(ADP-ribose)-degrading enzymes. Intensification of the fluorescence by preincubation with NAD, a substrate for poly(ADP-ribose) synthesis, also supported this view. The immunofluorescence of poly(ADP-ribose) was found exclusively in the nucleus of almost all tissues tested, including liver (adult, newborn, regenerating and hepatoma), brain, heart, intestine, pancreas, kidney, spleen, testis, thyroid gland and skeletal muscle. Exceptions were blood cells; little fluorescence was detectable in nuclei of peripheral leukocytes. Only after preincubation with NAD, did lymphocytes and monocytes exhibit fluorescence; granulocytes never did exhibit fluorescence. The cells appeared to represent the 1st instance where poly(ADP-ribose) synthesis activity among eukaryotic cells was missing.