scholarly journals Shedding new light on early sex determination in zebrafish

2020 ◽  
Vol 94 (12) ◽  
pp. 4143-4158
Author(s):  
Alex C. King ◽  
Michelle Gut ◽  
Armin K. Zenker

Abstract In contrast to established zebrafish gene annotations, the question of sex determination has still not been conclusively clarified for developing zebrafish, Danio rerio, larvae, 28 dpf or earlier. Recent studies indicate polygenic sex determination (PSD), with the genes being distributed throughout the genome. Early genetic markers of sex in zebrafish help unravel co-founding sex-related differences to apply to human health and environmental toxicity studies. A qPCR-based method was developed for six genes: cytochrome P450, family 17, subfamily A, polypeptide 1 (cyp17a1); cytochrome P450, family 19, subfamily A, polypeptide 1a (cyp19a1a); cytochrome P450, family 19, subfamily A, polypeptides 1b (cyp19a1b); vitellogenin 1 (vtg1); nuclear receptor subfamily 0, group B, member 1 (nr0b1), sry (sex-determining region Y)-box 9b (sox9b) and actin, beta 1 (actb1), the reference gene. Sry-box 9a (Sox9a), insulin-like growth factor 3 (igf3) and double sex and mab-3 related transcription factor 1 (dmrt1), which are also known to be associated with sex determination, were used in gene expression tests. Additionally, Next-Generation-Sequencing (NGS) sequenced the genome of two adult female and male and two juveniles. PCR analysis of adult zebrafish revealed sex-specific expression of cyp17a1, cyp19a1a, vtg1, igf3 and dmrt1, the first four strongly expressed in female zebrafish and the last one highly expressed in male conspecifics. From NGS, nine female and four male-fated genes were selected as novel for assessing zebrafish sex, 28 dpf. Differences in transcriptomes allowed allocation of sex-specific genes also expressed in juvenile zebrafish.

2007 ◽  
Vol 194 (1) ◽  
pp. 223-241 ◽  
Author(s):  
Zhihao Liu ◽  
Fengrui Wu ◽  
Baowei Jiao ◽  
Xiuyue Zhang ◽  
Chongjiang Hu ◽  
...  

To address the roles of doublesex and mab-3-related transcription factor 1 (Dmrt1), forkhead transcription factor gene 2 (Foxl2), and aromatase in sex differentiation of Southern catfish, the cDNA sequences of these genes were isolated from the gonads. Dmrt1a and Dmrt1b were found to be expressed in the gonads, being higher in the testis. A low expression level of Dmrt1b was also detected in the intestine and kidney of the male. Foxl2 was found to be expressed extensively in the brain (B), pituitary (P), gill and gonads (G), with the highest level in the ovary, indicating the possible involvement of Foxl2 in the B–P–G axis. Cytochrome P450 (Cyp)19b was found to be expressed in the brain, spleen, and gonads, while Cyp19a was only expressed in the gonads and spleen. All-female Southern catfish fry were treated with fadrozole (F), tamoxifen (TAM), and 17β-estradiol (E2) respectively, from 5 to 25 days after hatching (dah). The expression levels of these genes were measured at 65 dah. In the F-, TAM-, and FTAM-treated groups, Dmrt1a and Dmrt1b were up-regulated in the gonad, whereas Foxl2 and Cyp19a were down-regulated, while the expression of Cyp19b in the gonad remained unchanged. Furthermore, down-regulation of Foxl2 and Cyp19b was also detected in the brain. In the E2-treated group, Dmrt1a and Dmrt1b were down-regulated to an undetectable level in the gonad, whereas Foxl2 and Cyp19b were up-regulated in the brain. Consistent with the observed changes in the expressions of these genes, 56, 70, and 80% sex-reversed male individuals were obtained in the F-, TAM-, and F + TAM-treated groups respectively. These results indicate the significant roles of Dmrt1, Foxl2, and Cyp19 in the sex differentiation of Southern catfish.


2021 ◽  
Vol 905 (1) ◽  
pp. 012148
Author(s):  
S Prastowo ◽  
A Ratriyanto

Abstract Sex determination in dimorphic animal, such as chicken (G. Gallus), is controlled by the expression of doublesex and mab-3 related transcription factor 1 (DMRT1) gene. This gene act as sex determination switch by critically needed for testis differentiation and as antagonist of ovarian development. miRNA, is belongs to short noncoding RNA which modulate gene expression in specific or board targeted genes. This study was aimed to predict miRNA(s) candidate targeted to DMRT1 expression in chicken. In silico method was employed to mining miRNA targeted to DMRT1 using three online databases namely miRDB, TargetScan, and microT-CDS. Following prediction, clustering was performed to select common miRNA(s) in minimal two databases for gene ontology (GO) analysis. Totally 78 miRNAs were targeted to DMRT1 3’UTR, and eight miRNA(s) were found in minimal two databases. The GO analysis found seven distinct biological functions in membrane, cytoplasm, protein binding, nucleus, integral component of membrane and molecular function, and all are related to the cell growth namely cell proliferation. According to the result, it shows the possibility to use selected candidate of miRNA(s) targeted to DMRT1 to reveal the sex determination mechanism at early stage of chicken development.


2019 ◽  
Vol 102 (1) ◽  
pp. 27-37
Author(s):  
Jinxiu Dong ◽  
Lei Xiong ◽  
Hengwu Ding ◽  
Hui Jiang ◽  
Jiawei Zan ◽  
...  

Abstract A number of genes relevant for sex determination have been found in species with temperature-dependent sex determination. Epigenetics play a key role in sex determination, but characterization of deoxyribonucleic acid methylation of sex-related genes on temperature-dependent sex determination remains unclear. Mauremys reevesii is a typical species with temperature-dependent sex determination. In this study, we analyzed the Cytosine Guanine (CpG) methylation status of the proximal promoters, the messenger ribonucleic acid expression patterns and the correlation between methylation and expression levels of Aromatase, Forkhead box protein L2, Doublesex and mab3-related transcription factor 1, sex-determining region on Y chromosome-box 9, and anti-Müllerian hormone, which are key genes in sex determination in other species. We also analyzed the expression level of genes that encode enzymes involved in methylation and demethylation. The expression levels of Aromatase and Forkhead box protein L2 at the female producing temperature were higher than those at the male producing temperature; the expression levels of Doublesex and mab3-related transcription factor 1, sex-determining region on Y chromosome-box 9, and anti-Müllerian hormone were higher at MPT. The expression of some genes involved in methylation and demethylation is significantly different between male producing temperature and female producing temperature. The expression of messenger ribonucleic acid of genes involved in deoxyribonucleic acid methylation and demethylation affected by temperature, together with other factors, may change the methylation level of the regulatory regions of sex-related genes, which may further lead to temperature-specific expression of sex-related genes, and eventually affect the differentiation of the gonads.


2020 ◽  
Vol 49 (1) ◽  
pp. 97-104
Author(s):  
Wei Zheng ◽  
Xueming Dong ◽  
Qiuying Zhang ◽  
Xuefei Yu ◽  
Wenlan Li ◽  
...  

The MYB gene family is one of the most widespread plant transcription factor (TF) families, and MYB TFs play key roles in plant development, hormone signal transduction, disease resistance, abiotic stress tolerance and secondary metabolism. Recently, many MYBs have been characterized in various plants. However, little is known about the MYBs involved in secondary metabolite biosynthesis in Dimocarpus longan Lour. (D. longan). Based on transcriptome data profiling (Accession number: SRP155595), 35 MYBs from D. longan (DlMYBs) were identified. On the basis of their physicochemical properties, phylogenetic relationships, conserved motifs, and tissue-specific expression profiles these Dimocarpus longan MYBs (DlMYBs) were analyzed. Fifteen motifs in DlMYBs using MEME were found and a phylogenetic tree analysis showed that the DlMYBs identified here were divided into three groups. Group A contained the greatest number (25) of DlMYBs, followed by group B (6) and group C (4). Quantitative real time PCR (qRT-PCR) analysis demonstrated that, of the 35 MYBs studied DlMYB-12 and DlMYB-22 showed large differences in tissue-specific expression, with both MYBs showing very high expression in leaf tissue. These results lay the foundation for further studies of the biosynthesis of secondary metabolites in D. longan and further highlight the importance of MYB TFs in plants.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yanshan Zhong ◽  
Xiaodan Lu ◽  
Zhiwei Deng ◽  
Ziqing Lu ◽  
Minghui Fu

Abstract Background Glutamine synthetase (GS) acts as a key enzyme in plant nitrogen (N) metabolism. It is important to understand the regulation of GS expression in plant. Promoters can initiate the transcription of its downstream gene. Eichhornia crassipes is a most prominent aquatic invasive plant, which has negative effects on environment and economic development. It also can be used in the bioremediation of pollutants present in water and the production of feeding and energy fuel. So identification and characterization of GS promoter in E. crassipes can help to elucidate its regulation mechanism of GS expression and further to control its N metabolism. Results A 1232 bp genomic fragment upstream of EcGS1b sequence from E. crassipes (EcGS1b-P) has been cloned, analyzed and functionally characterized. TSSP-TCM software and PlantCARE analysis showed a TATA-box core element, a CAAT-box, root specific expression element, light regulation elements including chs-CMA1a, Box I, and Sp1 and other cis-acting elements in the sequence. Three 5′-deletion fragments of EcGS1b upstream sequence with 400 bp, 600 bp and 900 bp length and the 1232 bp fragment were used to drive the expression of β-glucuronidase (GUS) in tobacco. The quantitative test revealed that GUS activity decreased with the decreasing of the promoter length, which indicated that there were no negative regulated elements in the EcGS1-P. The GUS expressions of EcGS1b-P in roots were significantly higher than those in leaves and stems, indicating EcGS1b-P to be a root-preferential promoter. Real-time Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) analysis of EcGS1b gene also showed higher expression in the roots of E.crassipes than in stems and leaves. Conclusions EcGS1b-P is a root-preferential promoter sequence. It can specifically drive the transcription of its downstream gene in root. This study will help to elucidate the regulatory mechanisms of EcGS1b tissue-specific expression and further study its other regulatory mechanisms in order to utilize E.crassipes in remediation of eutrophic water and control its overgrowth from the point of nutrient metabolism.


2021 ◽  
Vol 22 (6) ◽  
pp. 3233
Author(s):  
Christopher Kapitza ◽  
Rittika Chunder ◽  
Anja Scheller ◽  
Katherine S. Given ◽  
Wendy B. Macklin ◽  
...  

Multiple sclerosis (MS) has been considered to specifically affect the central nervous system (CNS) for a long time. As autonomic dysfunction including dysphagia can occur as accompanying phenomena in patients, the enteric nervous system has been attracting increasing attention over the past years. The aim of this study was to identify glial and myelin markers as potential target structures for autoimmune processes in the esophagus. RT-PCR analysis revealed glial fibrillary acidic protein (GFAP), proteolipid protein (PLP), and myelin basic protein (MBP) expression, but an absence of myelin oligodendrocyte glycoprotein (MOG) in the murine esophagus. Selected immunohistochemistry for GFAP, PLP, and MBP including transgenic mice with cell-type specific expression of PLP and GFAP supported these results by detection of (1) GFAP, PLP, and MBP in Schwann cells in skeletal muscle and esophagus; (2) GFAP, PLP, but no MBP in perisynaptic Schwann cells of skeletal and esophageal motor endplates; (3) GFAP and PLP, but no MBP in glial cells surrounding esophageal myenteric neurons; and (4) PLP, but no GFAP and MBP in enteric glial cells forming a network in the esophagus. Our results pave the way for further investigations regarding the involvement of esophageal glial cells in the pathogenesis of dysphagia in MS.


Author(s):  
Hyuk Soo Eun ◽  
Sang Yeon Cho ◽  
Byung Seok Lee ◽  
In‑Ock Seong ◽  
Kyung‑Hee Kim

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