Molecular cloning and expression patterns of a sex-biased transcriptional factor Foxl2 in the giant freshwater prawn (Macrobrachium rosenbergii)

Zhu, Yuankui; Chen, Honglu; Zhang, Qingfei; Tan, Kianann; Liao, Shouli; Wang, Weimin

doi:10.1007/s11033-022-07526-7

Cited by 2 publications

(4 citation statements)

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“…In this study, no positive signal was detected in follicular cells, and the positive signal was stronger in the testis, indicating that the levels of Foxl2 mRNA in cells vary among invertebrates. The results of the in situ hybridization revealed that the Foxl2 transcript was found in the spermatocyte, oocyte, and vasosecretory epithelial cells of Macrobrachium rosenbergii [26]. The in situ hybridization analysis found that the Foxl2 signal was strong during the early development of the gonads in the Macrobrachium nipponense [39], which is similar to the results of this study.…”

Section: Discussionsupporting

confidence: 88%

“…The results showed that the PcFoxl2 CDS sequence was 1614bp long and encoded 537 amino acids, and it had the closest homologous relationship with C. quadricarinatus and E. sinensis. The Foxl2 gene of M. rosenbergii was found to be most closely related to the Foxl2 gene of E. sinensis; the cDNA of the Foxl2 gene of M. rosenbergii encodes a protein with 489 amino acids and contains a highly conserved forkhead (FH) domain [26]. Similar to other Foxl2 proteins, PcFoxl2 contains a highly conserved forkhead (FH) domain, suggesting that PcFoxl2 is conserved during evolution.…”

Section: Discussionmentioning

confidence: 99%

“…PrimeSTAR Max DNA Polymerase (high-fidelity enzyme) was added at a volume of 5 µL, and dd H 2 O was added to give 20.0 µL. The amplification procedure was as follows: pre-denaturation was performed at 98 • C for 1 min; then, 35 cycles were performed at 98 • C for 10 s, 58 • C for 30 s, and 72 • C for 40 s. The amplification was extended for 5 min at 72 • C and terminated at 4 • C [26]. The PCR product was gel-purified and cloned into competent cell Top10 (TaKaRa, Osaka City, Osaka Prefecture, Japan) for sequencing.…”

Section: Pcfoxl2 Gene Cloningmentioning

confidence: 99%

“…The Foxl2 gene is highly expressed in the testis, vas deferens, and ovaries. During ovarian development, Foxl2 expression is highest during stage I. Foxl2 may play a role in gonadal development in both female and male M. rosenbergii [26]. Significant progress has been made in the study of Foxl2 gene function in animals, but no relevant studies have been reported on the function and silencing effect of the Foxl2 gene in P. clarkii.…”

Section: Introductionmentioning

confidence: 99%

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Cloning, Identification, and Functional Analysis of the Foxl2 Gene in Procambarus clarkii

Huang,

Zhu,

Peng

et al. 2023

Genes

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Procambarus clarkii is the most widely distributed freshwater shrimp in China, with important economic value and great potential for development. The forkheadboxL2 (Foxl2) gene has been found to be involved in the reproductive development of many crustaceans. To understand the role of the Foxl2 gene in the gonad development of P. clarkii, we designed CDS-specific primers for the P. clarkii Foxl2 (PcFoxl2) gene and cloned its CDS sequence using RT-PCR. The nucleotide and protein sequence information was then analyzed through bioinformatics analysis. The expression and subcellular localization of PcFoxl2 in various tissues were detected using qRT-PCR and in situ hybridization. The effects of PcFoxl2 knockdown on gonad development were investigated using RNA interference. The results showed that the CDS length of the PcFoxl2 gene was 1614 bp and encoded 537 amino acids. Protein sequence comparison and phylogenetic analysis showed that PcFoxl2 was the closest relative to Crayfish. qRT-PCR analysis indicated that the expression level of PcFoxl2 in the testis was significantly higher (>40 fold) than that in the ovary (p < 0.01). The in situ hybridization results showed that PcFoxl2 was expressed in both the cytoplasm and the nucleus of egg cells, and that the expression was strongest in egg cells at the early stage of yolk synthesis, while weak in the secondary oocytes. The positive signal was strongest in the spermatocyte nucleolus, while only a trace signal was observed in the cytoplasm. After interfering with the PcFoxl2 gene using dsRNA, the expression of PcFoxl2 in the RNA interference group was significantly lower than that in the control group, and this interference effect lasted for one week. Moreover, the gonad index of the experimental group was significantly lower than that of the control group (p < 0.05) after 10 days of P. clarkii cultivation following PcFoxl2 knockdown. The expression levels of the nanos and S3a genes, which are related to gonad development, decreased significantly after PcFoxl2 gene interference. The results suggest that the Foxl2 gene is involved in the growth and development of gonads, particularly in the development of testis, and is related to the early development of oocytes. This study provides a theoretical basis for the artificial breeding of P. clarkii.

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Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 99%

Section: Pcfoxl2 Gene Cloningmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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