The Gene Expression And Functional Analysis Of Kifc1during The Spermatogenesis Of Sepiella Maindroni

Sepiella maindroni, as an important economical mollusk, has completed mariculturedevelopment artificially in Zhejiang and Fujian province recently. Because of the study onreproduction and development biology has received great attention; the mechanism ofspermatogenesis of this species became especially focused. In this research, we studied thegene sequence and characteristic of kifc1in Sepiella Maindroni through PCR and RACEtechnology, and concentrated on the spatial-temporal expression pattern of kifc1in thedevelopmental testis through in situ hybridization, with aim to figure out what role doeskifc1play in the spermiogenesis of Sepiella Maindroni.1The gene sequence and characteristic of kifc1.Based on multiple protein sequences alignment of KIFC1homologues and using acombined method of degenerate and RACE PCR, a gene from the testis of Sepiellamaindroni is identified, which encode homologue of rat KIFC1and ot-KIFC1. And also itsfull-length cDNA is obtained. The2432bp cDNA include a2109in-frame ORFcorresponding to703continuous amino acids. The encoded polypeptide shares a highestsimilarity with Octopus Tankahkeei. Through the prediction of secondary structure andtertiary structure, the motor domain of KIFC1is conserved at the C-terminal, having theputative ATP-binding and microtubule-binding motifs, while the N-terminal is morespecific to bind different types of cargoes for cellular events. The stalk domain connectingbetween the C-terminal and N-terminal, determines the direction of movement. FromRT-PCR results, kifc1gene is not tissue-specific, commonly detected in different tissues,for example, testis, liver, stomach, muscle, caecus and gills.2The spatial-temporal expression pattern of kifc1in the spermiogenesis of SepiellaMaindroni.Through in situ hybridization research, we can mimic the expression pattern ofKIFC1protein in spermiogenesis of Sepiella maindroni. During the primary stage of thespermatogenesis, the kifc1mRNA signal was hardly detected. At the early spermatids, the signal starts to present. With the development of spermatids, the signals increased a lot. Itpeaks and gathers around the acrosome area when the spermatids elongate into the spindleshape. As the spermatids develope into mature spermatids, the signal was vanished. Insummary, the expression of kfic1at specific stages during spermiogenesis and thedistribution shed light on the potential functions of this motor in major cytologicaltransformations. The KIFC1homologue may provid direct shaping force the nuclear orinfluence the shaping process through indirect regulation.