Faculty of Life Science and Bioengineering, Ningbo University, 315211, Zhejiang, People's Republic of China.
Mol Biol Rep. 2012 May;39(5):5589-98. doi: 10.1007/s11033-011-1363-4. Epub 2011 Dec 20.
KIF3B is known for maintaining and assembling cilia and flagellum. To date, the function of KIF3B and its relationship with KIF3A during spermiogenesis in the cephalopod Octopus tankahkeei remains unknown. In the present study, we characterized a gene encoding a homologue of rat KIF3B in the O. tankahkeei testis and examined its temporal and spatial expression pattern during spermiogenesis. The cDNA of KIF3B was obtained with degenerate and RACE PCR and the distribution pattern of ot-kif3b were observed with RT-PCR. The morphological development during spermiogenesis was illustrated by histological and transmission electron microscopy and mRNA expression of ot-kif3b was observed by in situ hybridization. The 2,365 nucleotides cDNA consisted of a 102 bp 5' untranslated region (UTR), a 2,208 bp open reading frame (ORF) encoding a protein of 736 amino acids, and a 55 bp 3' UTR. Multiple alignments revealed that the putative Ot-KIF3B shared 68, 68, 69, 68, and 67% identity with that of Homo sapiens, Mus musculus, Gallus gallus, Danio rerio, and Xenopus laevis, respectively, along with high identities with Ot-KIF3A in fundamental structures. Ot-kif3b transcripts appeared gradually in early spermatids, increased in intermediate spermatids and maximized in drastically remodeled and final spermatids. The kif3b gene is identified and its expression pattern is demonstrated for the first time in O. tankahkeei. Compared to ot-kif3a reported by our laboratory before, our data suggested that the putative heterodimeric motor proteins Ot-KIF3A/B may be involved in intraspermatic transport and might contribute to structural changes during spermiogenesis.
KIF3B 已知用于维持和组装纤毛和鞭毛。迄今为止,KIF3B 的功能及其在头足类章鱼(Octopus tankahkeei)精子发生过程中与 KIF3A 的关系尚不清楚。在本研究中,我们鉴定了章鱼睾丸中编码大鼠 KIF3B 同源物的基因,并研究了其在精子发生过程中的时空表达模式。使用简并和 RACE PCR 获得了 KIF3B 的 cDNA,并通过 RT-PCR 观察了 ot-kif3b 的分布模式。通过组织学和透射电子显微镜观察精子发生过程中的形态发育,并通过原位杂交观察 ot-kif3b 的 mRNA 表达。2365 个核苷酸的 cDNA 由 102 bp 5'非翻译区(UTR)、2208 bp 开放阅读框(ORF)组成,编码 736 个氨基酸的蛋白质,以及 55 bp 3'UTR。多重比对显示,推定的 Ot-KIF3B 与 Homo sapiens、Mus musculus、Gallus gallus、Danio rerio 和 Xenopus laevis 的同一性分别为 68%、68%、69%、68%和 67%,与 Ot-KIF3A 的基本结构高度一致。Ot-kif3b 转录本在早期精母细胞中逐渐出现,在中间精母细胞中增加,并在剧烈重塑和最终精母细胞中达到最大值。本研究首次在章鱼中鉴定了 kif3b 基因并展示了其表达模式。与我们实验室之前报道的 ot-kif3a 相比,我们的数据表明,假定的异二聚体马达蛋白 Ot-KIF3A/B 可能参与精子内运输,并可能有助于精子发生过程中的结构变化。