Muslimov I A, Santi E, Homel P, Perini S, Higgins D, Tiedge H
Department of Pharmacology, State University of New York, Health Science Center at Brooklyn, Brooklyn, New York 11203, USA.
J Neurosci. 1997 Jun 15;17(12):4722-33. doi: 10.1523/JNEUROSCI.17-12-04722.1997.
In nerve cells, a select group of RNAs has been localized to dendritic domains. Here we have examined dendritic RNA transport in sympathetic neurons in primary culture, using a microinjection protocol with neuronal BC1 RNA and with BC1-derived sequence segments. After cytoplasmic microinjection, full-length BC1 RNA was selectively transported to dendrites; in contrast, control RNAs such as nuclear RNAs and random-sequence irrelevant RNAs remained restricted to cytoplasmic areas proximal to the injection sites. Chimeric RNAs were constructed that contained the full-length BC1 sequence inserted upstream or downstream of the coding regions of nondendritic mRNAs. After microinjection, such chimeric RNAs were specifically targeted to dendrites; microinjected corresponding nonchimeric mRNAs were not. Dendritic transport of BC1 RNA was rapid: the average dendritic delivery rate within the first hour after microinjection was 242 +/- 25 microm/hr. Whereas a 5'-BC1 segment of 62 nucleotides was transported to dendrites to extents and at levels similar to full-length BC1 RNA, a 3'-BC1 segment of 60 nucleotides did not exit injected somata to any significant degree. A cis-acting dendritic targeting element is thus contained in the 5' part of neuronal BC1 RNA. These results demonstrate that mechanisms exist in neurons for fast and specific transport of selected RNAs to dendrites.
在神经细胞中,一小部分特定的RNA已被定位到树突区域。在此,我们利用一种显微注射方案,使用神经元BC1 RNA和BC1衍生的序列片段,研究了原代培养的交感神经元中的树突RNA转运。细胞质显微注射后,全长BC1 RNA被选择性地转运到树突;相比之下,对照RNA,如核RNA和随机序列无关RNA,仍局限于注射部位近端的细胞质区域。构建了嵌合RNA,其包含插入非树突mRNA编码区域上游或下游的全长BC1序列。显微注射后,此类嵌合RNA被特异性地靶向树突;而显微注射的相应非嵌合mRNA则没有。BC1 RNA的树突转运很快:显微注射后第一小时内的平均树突递送速率为242±25微米/小时。虽然62个核苷酸的5'-BC1片段被转运到树突的程度和水平与全长BC1 RNA相似,但60个核苷酸的3'-BC1片段在很大程度上并未离开注射的胞体。因此,神经元BC1 RNA的5'部分包含一个顺式作用的树突靶向元件。这些结果表明,神经元中存在将选定RNA快速且特异性地转运到树突的机制。