Characterization and expression pattern of p53 during spermatogenesis in the Chinese mitten crab Eriocheir sinensis
Characterization and expression pattern of p53 during spermatogenesis in the Chinese mitten crab Eriocheir sinensis
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中华绒螯蟹精子发生过程中p53的特征及表达模式。
DOI：
10.1007/s11033-012-2145-3
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发表时间：
2013-02-01
影响因子：
2.8
通讯作者：
Yang, Wan-Xi
中科院分区：
文献类型：
--
作者：
Hou, Cong-Cong;Yang, Wan-Xi
p53作为“基因组的守护者”，在包括睾丸在内的多种组织中，在细胞周期阻滞、凋亡、DNA修复和抑制血管生成等方面发挥着重要作用。p53基因及其蛋白在哺乳动物精子发生中起着重要作用。为探讨p53基因在中华绒螯蟹精子发生中的作用，采用简并引物PCR和快速扩增cDNA末端的方法，从中华绒螯蟹精巢中克隆了p53基因的cDNA（1,218 bp），并进行了序列测定。p53蛋白质序列比对显示其保守的DNA结合结构域、二聚化位点和锌结合位点与预测的结构一致。系统发育分析表明，p53基因与中国囊对虾（Marsupenaeus）和虎足虾（Tigriopus）的亲缘关系较近。组织表达分析表明，p53在附性腺、肌肉、鳃、心脏、肝胰腺和精巢中均有表达。原位杂交结果显示，早期p53 mRNA在细胞核周围有较弱的分布，而在顶体小管内陷处有较强的分布。中期p53 mRNA信号较早期增强，呈点状分布于杯状核表面。顶体帽上的信号比顶体小管上的信号强，尽管顶体小管上的信号也不同。晚期信号仍主要位于顶体帽和顶体小管。杯状核周围可见散在信号，但信号很弱。在成熟精子中，信号显著降低。杯状核和顶体小管上的信号虽然清晰，但比中期弱。因此，我们认为p53可能在精子发生过程中的顶体形成和核形成中起重要作用。sinensis。
p53, as a "Guardian of the Genome", plays an important role in cell cycle arrest, apoptosis, DNA repair and inhibition of angiogenesis in different tissues including testis. p53 gene and its protein perform many essential roles for mammalian spermatogenesis. To explore its functions during spermatogenesis in Eriocheir sinensis, we have cloned and sequenced the cDNA (1,218 bp) of p53 from the testis by degenerating primer PCR and rapid-amplification of cDNA ends. The protein alignment of p53 shows the conserved DNA binding domain, dimerization site and zinc binding site consisted of the predicted structures. Phylogenetic analysis revealed that p53 was more closer to Marsupenaeus japonicus and Tigriopus japonicus than other examined species. Tissue expression analysis of p53 mRNA showed p53 was distinctly expressed in accessory sexual gland, muscle, gill, heart, hepatopancreas and testis. In situ hybridization revealed that the p53 mRNA was weakly distributed around the nucleus, but stronger in the invaginated acrosomal tubule at the early stage. At the middle stage, p53 mRNA signal was increased than the early stage and the signal displayed dot-like pattern on the surface of cup-like nucleus. The signal on acrosomal cap is stronger than on the acrosomal tubule, despite acrosomal tubule signal was also distinct. At the late stage, the signal was still mainly located in acrosomal cap and acrosomal tubule. Sporadic signal were found surrounding the cup-like nucleus, but they were very weak. In the mature sperm, the signal was dramatically decreased. Even though the signal on cup-like nucleus and acrosomal tubule were distinct, they were weaker than those in middle stage. Based on these results, we concluded that p53 may play an important role in formation of acrosome biogenesis and nuclear shaping during spermiogenesis of E. sinensis.