Expression pattern of the zona pellucida 3 (ZP3) gene during ovarian development and the location of ZP3 protein in oocytes in a natural, wild triploid crucian carp mutant, Carassius auratus var. Pingxiangnensis

Abstract

ABSTRACT. Carassius auratus var. Pingxiangnensis (designated CaP), distributed in the Pingxiang region of Jiangxi Province, China, is a natural, wild triploid crucian carp mutant that has two reproductive development modes: gynogenesis and bisexual reproduction. Little information is available about the expression pattern of the zona pellucida 3 (ZP3) gene during ovarian development and the location of the ZP3 protein in oocytes of this fish. In this study, we obtained the full-length cDNA of ZP3 (CaP_ ZP3). CaP_ZP3 contains an open reading frame of 1305 bp that encodes 435 amino acid residues. Real-time polymerase chain reaction (PCR) was used to determine the CaP_ZP3 mRNA expression levels in the ovary at different stages of maturation. Results revealed high levels of CaP_ZP3 expression in 4-to 8-month-old ovaries (stage II-stage III), with a significant decline in 9-to 12-month-old ovaries (stages IV-stage V). The high levels of CaP_ ZP3 transcripts during the early growth period suggest an important role for CaP_ZP3 in early oocyte development. In addition, a polyclonal antibody was prepared against CaP_ZP3, and the immunofluorescence localization was determined. CaP_ZP3 protein was detected close to the oocyte plasma membrane. The results also showed that no fluorescent signal was detected in stage I and II oocytes. CaP_ZP3 protein is primarily detected in stage III oocytes, and the protein accumulates as oocytes develop into stage IV oocytes. These results suggested that the transcription and translation of the CaP_ZP3 gene is asynchronous and that the transcription of the CaP_ZP3 protein occurs prior to its translation in this triploid fish.

Large-scale tissue-specific and temporal gene expression profiles in Pengze crucian carp

“…Pcc-zp2 was highly significant in brain, hepatopancreas, kidney, ovary, and spleen tissues, and these results were the same as those reported in the rare minnow. Ovarian-specific zp2 was found in zebrafish, crucian carp, and goldfish (Mold et al, 2001;Shi et al, 2013), and zp was found in both the hepatopancreas and ovary tissues of medaka and pufferfish (Kanamori et al, 2003). The results of the present study suggested that Pcc-zp2 is associated with the ovarian/hepatic-shared expression pattern.…”

“…Gene expression profiles in different genders (Hale et al, 2011), tissues, and developmental stages (Ijiri et al, 2008;Shi et al, 2013) may have dual functions in sex determination and differentiation. The present study detected all four gene catalogues associated with tissue-specific and temporal distributions in Pcc.…”

Comparative omics and feeding manipulations in chicken indicate a shift of the endocrine role of visceral fat towards reproduction

“…The specificity of this expression to the female red junglefowl demonstrates the relevance of this expression to female reproduction. In other oviparous vertebrates, these proteins are found in the liver either in addition or instead of the ovary [ 37 , 38 ]. In birds, extra gonadal expression of ZP1 was reported in the liver of quail [ 31 ], where it was also shown that intravenously injected ZP1 incorporates into the perivitelline membrane [ 39 ].…”

De novotranscriptome sequencing to identify the sex-determination genes inHyriopsis schlegelii

“…Total RNA was treated with RNase-free DNase I (Promega, USA) to eliminate genomic DNA contamination. 23,24) cDNA was synthesized using the Transcriptor First Strand cDNA Synthesis Kit (Promega, USA). RTqPCR was performed using a real-time PCR SYBR Green detection system (Eppendorf).…”