Bioinformatic analyses of zona pellucida genes in vertebrates and their expression in Nile tilapia
Zona pellucida (ZP) genes encode ZP glycoproteins which constitute the coat surrounding oocytes and early embryos. Genome-wide identification of ZP genes is still lacking in vertebrates, especially in fish species. Herein, we conducted bioinformatic analyses of the ZP genes of the Nile tilapia and other vertebrates. Totally 16, 9, 17, 27, 21, 20, 26, 19, 14,11, 24, 17, 9, 18, 8, 11, 9, 8, 5, and 4 ZP genes belonging to 5 subfamilies (ZPA, ZPB, ZPC, ZPD, and ZPAX) were found in the sea lamprey, elephant shark, coelacanth, spotted gar, zebrafish, medaka, stickleback, Nile tilapia, Amazon molly, platyfish, seahorse, Northern snakehead, cavefish, tetraodon, clawed frog, turtle, chicken, platypus, kangaroo rat, and human genomes, respectively. The expansion of ZP genes in basal vertebrates was mainly achieved by gene duplication of ZPB, ZPC, and ZPAX subfamilies, while the shrink of ZP gene number in viviparous mammals was achieved by keeping only one copy of the ZP genes in each subfamily or even secondary loss of some subfamilies. The number of ZP gene is related to the environment where the eggs are fertilized and the embryos develop in vertebrates. Transcriptomic analysis showed that 14 ZP genes were expressed in the ovary of Nile tilapia, while two (ZPB2b and ZPC2) were highly expressed in the liver. On the other hand, ZPB1a and ZPB2c were not found to be expressed in any tissue or at any developmental stage of the gonads examined. In the ovary, the expression of ZP genes started from 30 dah (days after hatching), significantly upregulated at 90 dah and maintained this level at 180 dah. The expression of ZPC2 in the liver and ZPC5-2 and ZPAX1 in the ovary was confirmed by in situ hybridization. The ovary- and liver-expressed ZP genes are expressed coordinately with oocyte growth in tilapia.
KeywordsZona pellucida genes Isolation Expression profile Nile tilapia Adaptive evolution
zona pellucida glycoprotein
consensus furin cleavage site
days after hatching
reads per kb per million reads
Tianli Wu contributed to the preparation of the manuscript and analysis of the ZP genes. Yunying Cheng and Wenjing Tao carried out the sequence identification, contributed to the phylogenetic and transcriptomic analysis, and manually annotated the ZP genes. Zhilong Liu and Shuqing Zheng conducted experimental validation of expression data. Deshou Wang coordinated the study and was responsible for the final drafting of the manuscript. All authors read and approved the manuscript.
This work was supported by grants 31630082, 91331119, and 31502147 from the National Natural Science Foundation of China and grant cstc2015jcyjB80001 from the Natural Science Foundation Project of Chongqing, Chongqing Science and Technology Commission.
Compliance with ethical standards
All animal experiments were undertaken in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals, with the approval of the Scientific Investigation Board of Southwest University, Chongqing.
The authors declare that they have no competing interests.
- Okumura H, Kohno Y, Iwata Y, Mori H, Aoki N, Sato C, Kitajima K, Nadano D, Matsuda T (2004) A newly identified zona pellucida glycoprotein, ZPD, and dimeric ZP1 of chicken egg envelope are involved in sperm activation on sperm-egg interaction. Biochem J 384:191–199CrossRefPubMedPubMedCentralGoogle Scholar
- Wang AM (1994) Electron microscopic observations on the formation of chorion (egg envelope) of Oreochromis Mossambicus. Oceanologia Et Limnologia Sinica 25:385–389Google Scholar
- Wyrick RE, Nishihara T, Hedrick JL (1974) Fertilization envelope formation by glycoprotein agglutination and block to polyspermy in Xenopus laevis. Fed Proc 33:1453Google Scholar
- Xu J, Li JT, Jiang Y, Peng W et al (2016) Genomic basis of adaptive evolution: the survival of Amur ide (Leuciscuswaleckii) in an extremely alkaline environment. Mol Biol Evol 20:msw230Google Scholar