MONOCLONAL-ANTIBODIES DISTINGUISH PHOSPHORYLATED AND NONPHOSPHORYLATED FORMS OF NEUROFILAMENTS INSITU
MONOCLONAL-ANTIBODIES DISTINGUISH PHOSPHORYLATED AND NONPHOSPHORYLATED FORMS OF NEUROFILAMENTS INSITU
复制标题
DOI：
10.1073/pnas.80.19.6126
复制
发表时间：
1983-01-01
期刊：
影响因子：
--
通讯作者：
STERNBERGER, NH
中科院分区：
文献类型：
--
作者：
STERNBERGER, LA;STERNBERGER, NH
上述37种神经元特异性单抗的免疫细胞化学染色模式分为4组：(1)抗突触相关抗体，(2)抗神经纤维抗体，(3)抗核周-神经纤维抗体，(4)与广泛分布的表位反应的单一抗体，覆盖了组2和组3的模式。组2、组3和组4的抗体被证明是针对神经丝三联体亚基的，尽管组2和组3之间的染色模式几乎没有重叠。用磷酸酶消化后，组2的5个抗体取消了神经丝成分的电印迹染色，组3的3个抗体增强了神经丝成分的电印迹染色，抗体4不影响神经丝成分的免疫细胞化学染色。对于组2的抗体，胰酶消化也不影响染色，但当用磷酸酶消化时，5种抗体中有3种染色减弱。胰酶消化取消了第3组各抗体的所有染色。如果用磷酸酶消化，则再次出现染色，但第3组模式被第2组模式所取代。在用胰酶进行第二次处理时，这种图案的染色再次被取消。经胰酶消化后，第4组抗体失去了第2组和第3组的大部分染色模式。组2的模式再次出现，并在随后的磷酸酶治疗时得到加强，在第二次胰酶治疗后再次减少。组2的5个抗体中有4个的染色被无机磷抑制。这些数据表明，某些神经细胞体、树突和至少近端的轴突含有非磷酸化的神经丝，而长纤维，包括终末轴突，含有磷酸化的神经丝。磷酸化可能是稳定致密形式神经细丝的一个因素，致密结构的异质性可能在单个神经细胞内可能的功能多样性中发挥作用。
The immunocytochemical staining patterns of 37 neuron-specific monoclonal antibodies previously described fell into 4 groups: (1) anti-synapse-associated, (2) anti-neurofibrillar, (3) anti-perikaryonal-neurofibrillar, and (4) a single antibody reactive with a widely distributed epitope that covered the patterns of groups 2 and 3. Antibodies of gropus 2, 3 and 4 were shown to be specific to neurofilament triplet subunits, even though there was little overlap in staining patterns between groups 2 and 3. Nine of these antibodies were examined to distinguish functional states of neurofilaments dependent upon phosphorylation. Upon digestion with phosphatase, electroblot staining of neurofilament components was abolished with the 5 antibodies of group 2, enhanced with the 3 antibodies from group 3, and unaffected with antibody 4. Immunocytochemical staining of Bouin-fixed paraffin sections of rat brain was unaffected by phosphatase pretreatment. With antibodies of group 2, digestion with trypsin also left staining unaffected, but when followed by digestion with phosphatase, staining was diminished with 3 out of 5 antibodies. Digestion with trypsin abolished all staining with each antibody from group 3. If followed by digestion with phosphatase, staining reappeared, but the group 3 pattern was replaced by a group 2 pattern. Staining of this pattern was again abolished upon a 2nd treatment with trypsin. The antibody from group 4 lost most its groups 2 and 3 staining patterns when sections were digested with trypsin. The group 2 pattern reappeared and was enhanced upon a subsequent phosphatase treatment and was reduced again upon a 2nd trypsin treatment. Staining by 4 out of 5 antibodies from group 2 was inhibited by inorganic phosphate. The data indicate that certain nerve cell bodies, their dendrites, and at least proximal axons possess nonphosphorylated neurofilaments and that long fibers, including terminal axons, possess phosphorylated neurofilaments. Phosphorylation may be a factor in stabilizing compacted forms of neurofilaments and that heterogeneity of the compacted structures may play a role in a possible multiplicity of function within individual nerve cells.