Modulation by cocaine of dopamine receptors through miRNA-133b in zebrafish embryos.
Modulation by cocaine of dopamine receptors through miRNA-133b in zebrafish embryos.
复制标题
DOI：
10.1371/journal.pone.0052701
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Rodríguez RE
中科院分区：
文献类型：
--
作者：
Barreto-Valer K;López-Bellido R;Macho Sánchez-Simón F;Rodríguez RE
怀孕期间使用可卡因会影响母亲，并可能间接改变胚胎/胎儿的发育。因此，在目前的工作中，我们的目标是在体内（斑马鱼胚胎）研究可卡因对多巴胺受体和miR-133 b表达的影响。这些胚胎在受精后5小时（hpf）暴露于盐酸可卡因（HCl），然后在8、16、24、48和72 hpf收集以通过定量真实的时间PCR（qPCR）和原位杂交（ISH，仅在24 hpf）研究多巴胺受体drd 1、drd 2a、drd 2b和drd 3的表达。我们的研究结果表明，可卡因改变了所研究的基因的表达，这取决于发育胚胎的阶段和多巴胺受体的类型。我们通过qPCR发现可卡因降低了中枢神经系统（CNS）和外周中24和48 hpf的miR-133 b表达，并且miR-133 b的空间分布主要见于体节中，这一发现表明miR-133 b参与骨骼肌的发育。相比之下，在CNS水平，miR-133 b在24和48 hpf具有弱和中等表达。我们还通过显微注射Pitx 3 - 3 'UTR序列分析了miR-133 b与Pitx 3和Pitx 3靶基因drd 2a和drd 2b、酪氨酸羟化酶（th）和多巴胺转运蛋白（dat）的相互作用。显微注射Pitx 3 - 3 'UTR影响了pitx 3，drd 2a，drd 2b，th和dat的表达。总之，在目前的工作中，我们描述了一种可能的机制来解释可卡因活性通过控制miR-133 b在斑马鱼中的转录。通过miR-133 b，可卡因可以调节pitx 3的表达，随后调节多巴胺受体dat和th的表达。这些结果表明，miRNAs在胚胎发育和药物成瘾中发挥重要作用。
The use of cocaine during pregnancy can affect the mother and indirectly might alter the development of the embryo/foetus. Accordingly, in the present work our aim was to study in vivo (in zebrafish embryos) the effects of cocaine on the expression of dopamine receptors and on miR-133b. These embryos were exposed to cocaine hydrochloride (HCl) at 5 hours post-fertilization (hpf) and were then collected at 8, 16, 24, 48 and 72 hpf to study the expression of dopamine receptors, drd1, drd2a, drd2b and drd3, by quantitative real time PCR (qPCR) and in situ hybridization (ISH, only at 24 hpf). Our results indicate that cocaine alters the expression of the genes studied, depending on the stage of the developing embryo and the type of dopamine receptor. We found that cocaine reduced the expression of miR-133b at 24 and 48 hpf in the central nervous system (CNS) and at the periphery by qPCR and also that the spatial distribution of miR-133b was mainly seen in somites, a finding that suggests the involvement of miR-133b in the development of the skeletal muscle. In contrast, at the level of the CNS miR-133b had a weak and moderate expression at 24 and 48 hpf. We also analysed the interaction of miR-133b with the Pitx3 and Pitx3 target genes drd2a and drd2b, tyrosine hydroxylase (th) and dopamine transporter (dat) by microinjection of the Pitx3-3'UTR sequence. Microinjection of Pitx3-3'UTR affected the expression of pitx3, drd2a, drd2b, th and dat. In conclusion, in the present work we describe a possible mechanism to account for cocaine activity by controlling miR-133b transcription in zebrafish. Via miR-133b cocaine would modulate the expression of pitx3 and subsequently of dopamine receptors, dat and th. These results indicate that miRNAs can play an important role during embryogenesis and in drug addiction.
登录
查看更多内容
DOI：
10.1523/jneurosci.2419-10.2010
发表时间：
2010-07-28
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
He Y;Yang C;Kirkmire CM;Wang ZJ
通讯作者：
Wang ZJ
DOI：
10.1006/bbrc.1998.8496
发表时间：
1998-04-17
影响因子：
3.1
作者：
Barrallo, A;González-Sarmiento, R;Rodríguez, RE
通讯作者：
Rodríguez, RE
影响因子：
4.1
作者：
Dienes-Nagy, A;Rivier, L;Mangin, P
通讯作者：
Mangin, P
影响因子：
2.7
作者：
Callier, S;Snapyan, M;Vernier, P
通讯作者：
Vernier, P
DOI：
10.1073/pnas.191380698
发表时间：
2001-09-25
影响因子：
11.1
作者：
Darland, T;Dowling, JE
通讯作者：
Dowling, JE