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A vasa gene from green mud crab Scylla paramamosain and its expression during gonadal development and gametogenesis

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Abstract

VASA is one of the important regulatory factors that determine the development of the reproductive system. However, no information on vasa gene from Pleocyemata Brachyura is available. By using Race, we obtained a full-length cDNA of Sp-vasa of the green mud crab Scylla paramamosain. The full-length (2,851 bp) cDNA of vasa encodes a peptide of 631 amino acids. Real-time PCR results indicated that the expression level of Sp-vasa in the growth stage of ovary was higher than in the maturation stage, and in stage I and II of testis, the expression level of Sp-vasa were higher than in stage III. By using in situ hybridization, Sp-vasa RNAs were detected in the large part of oocyte plasm in stage I, nucleus zone in stage III and perinuclear zone in stage V. As the size of oocytes increases during oogenesis, the signals change from strong to weak. In addition, in stage I and II of testis, the expression levels of Sp-vasa were higher than in stage III, and the hybridization intensity of Sp-vasa gene gradually increased during spermatogenesis from spermatogonia to spermatids. However, no hybridization signal was detected in spermatozoon. Real-time PCR and in situ hybridization were consistent. These findings suggest that Sp-vasa is likely to serve as a useful and specific marker for germ cell development of S. paramamosain.

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Acknowledgments

The work was support by Natural Science Foundation of China (No.30571430, No. 31072200), Natural Science Foundation of Fujian Province (No.2009J01179) and Innovation Team Foundation of Jimei University (No.2010A001). We thank Mr. Ion Beldorth, Department of Chemistry and Biochemistry, Texas State University for his proof reading of the manuscript.

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  1. Ziping Zhang

    Present address: Department of Chemistry and Biochemistry, Texas State University, San Marcos, TX, 78666, USA

Authors and Affiliations

  1. The Key Laboratory of Science and Technology for Aquaculture and Food Safety, Fisheries College, Jimei University, Xiamen, 361021, China

    Yilei Wang, Yudong Chen, Kunhuang Han, Zhihua Zou & Ziping Zhang

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Additional information

GenBank accession number: ACZ92304.

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11033_2011_1220_MOESM1_ESM.tif

Fig. S1. The nucleotide and deduced amino acid sequences of Sp–vasa gene. The nucleotide sequence is displayed in the 5′ → 3′ directions and numbered to the right and left. Nine conserved domains are underlined with bold letter. GG Rich region is highlighted in grey color. Two CCHC structures of VASA are in bold. RGG repeat region is double underlined. The predicted phosphorylation sites are in boxes. (TIFF 157 kb)

Supplementary material 2 (TIFF 21 kb)

11033_2011_1220_MOESM3_ESM.tif

Fig. S2. Multiple alignment of the VASA amino acid sequence between S. paramamosain and other species. The abbreviations were used for species with GenBank Accession no. as follows: Scylla paramamosain, ACZ92304; Litopenaeus vannamei, AAY89069; Fenneropenaeus chinensis, ABQ00071; Parhyale hawaiensis, ACH92926; Daphnia magna, BAD99522; Crassostrea gigas, AAR37337; Chlamys farreri, ABE27759; Haliotis asinina, ACT35657; Strongylocentrotus purpuratus, NP_001139665; Ciona savignyi, BAB12217; Platynereis dumerilii, CAJ38803; Danio rerio, AAL89410; Gallus gallus, BAB12337; Homo sapiens, AAF72705; Apis mellifera, ABC41341; Drosophila melanogaster, NP_723899; Bombyx mori, NP_001037347. (TIFF 635 kb)

Supplementary material 4 (TIFF 873 kb)

Supplementary material 5 (TIFF 950 kb)

Supplementary material 6 (TIFF 576 kb)

11033_2011_1220_MOESM7_ESM.tif

Fig. S3. Phylogenetic tree of the VASA amino acid sequence between S. paramamosain and other species. Mus. musculus Ddx5 protein was used as the out-group. Numbers indicate the bootstrap confidence values of 1000 replicates. The accession numbers of the selected VASA sequences see Fig. S2. (TIFF 193 kb)

Supplementary material 8 (PDF 168 kb)

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Wang, Y., Chen, Y., Han, K. et al. A vasa gene from green mud crab Scylla paramamosain and its expression during gonadal development and gametogenesis. Mol Biol Rep 39, 4327–4335 (2012). https://doi.org/10.1007/s11033-011-1220-5

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