scholarly journals Transcriptome profiling of laser-captured germ cells and functional characterization of zbtb40 during 17alpha-methyltestosterone-induced spermatogenesis in orange-spotted grouper (Epinephelus coioides)

BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Xi Wu ◽  
Yang Yang ◽  
Chaoyue Zhong ◽  
Yin Guo ◽  
Shuisheng Li ◽  
...  
Keyword(s):  
Germ Cells ◽  
Functional Characterization ◽  
Transcriptome Profiling ◽  
Epinephelus Coioides

scholarly journals Transcriptome profiling of laser-captured germ cells and functional characterization of zbtb40 during MT-induced spermatogenesis in orange-spotted grouper (Epinephelus coioides)

2019 ◽  
Author(s):  
Xiaochun Liu ◽  
Xi Wu ◽  
Yang Yang ◽  
Chaoyue Zhong ◽  
Yin Guo ◽  
...  
Keyword(s):  
Germ Cells ◽  
Cellular Localization ◽  
Molecular Mechanisms ◽  
Expression Patterns ◽  
Functional Characterization ◽  
Transcriptome Profiling ◽  
Sex Reversal ◽  
Epinephelus Coioides ◽  
Isolated Cells ◽  
Male Germ Cells

Abstract Background: Spermatogenesis is an intricate process regulated by a finely organized network. The orange-spotted grouper (Epinephelus coioides) is a protogynous hermaphroditic fish, but the process of its spermatogenesis is not well-understood. In the present study, transcriptome sequencing of the male germ cells from orange-spotted grouper was performed to explore the molecular mechanisms underlying spermatogenesis. Results: In this study, the orange-spotted grouper was induced to change sex from female to male by 17alpha-methyltestosterone implantation. During the artificial spermatogenesis, different cell types from cysts containing spermatogonia, spermatocytes, spermatids, and spermatozoa were isolated by laser capture microdissection. Subsequently, transcriptomic analysis for the isolated cells were performed. A series of genes was used to verify and investigate the expression patterns in spermatogenesis. Furthermore, we also analyzed the expression of the same set of genes involved with steroid metabolism and sex throughout spermatogenesis (early-mid, late, and maturing stages) in the orange-spotted grouper. Several generally female-related genes took significantly changes in sex reversal hinted that the female-related genes in previously recognized may also play vital roles in spermatogenesis and sex reversal. In the transcriptomic data, we focused on zbtb family genes, which may be related to the process of spermatogenesis. Their expression patterns and cellular localization were examined, and the location of Eczbtb40 in different gonadal stages was investigated. We found that Eczbtb40 was expressed throughout spermatogenesis. These preliminary findings suggest that Eczbtb40 is highly conserved during vertebrate evolution and plays roles in spermatogenesis. Besides, the expression of Eczbtb40 and Eccyp17a1a overlapped in male germ cells, especially spermatogonium and spermatocyte, which suggested that Eczbtb40 might interact with Eccyp17a1a participant in spermatogenesis and sex reversal. Conclusions: The present study first depicted RNA sequencing of the male germ cells from orange-spotted grouper, and identified many important functional genes and pathways involved in spermatogenesis. The Eczbtb40 gene was subjected to molecular characterization and expression pattern analysis. These results will contribute to future studies of the molecular mechanism of spermatogenesis and sex reversal.

scholarly journals Transcriptome profiling of laser-captured germ cells and functional characterization of zbtb40 during MT-induced spermatogenesis in orange-spotted grouper (Epinephelus coioides)

2019 ◽  
Author(s):  
Xiaochun Liu ◽  
Xi Wu ◽  
Yang Yang ◽  
Chaoyue Zhong ◽  
Yin Guo ◽  
...  
Keyword(s):  
Germ Cells ◽  
Cellular Localization ◽  
Molecular Mechanisms ◽  
Expression Patterns ◽  
Functional Characterization ◽  
Transcriptome Profiling ◽  
Sex Reversal ◽  
Epinephelus Coioides ◽  
Isolated Cells ◽  
Male Germ Cells

Abstract Background: Spermatogenesis is an intricate process regulated by a finely organized network. The orange-spotted grouper (Epinephelus coioides) is a protogynous hermaphroditic fish, but the process of its spermatogenesis is not well-understood. In the present study, transcriptome sequencing of the male germ cells from orange-spotted grouper was performed to explore the molecular mechanisms underlying spermatogenesis. Results: In this study, the orange-spotted grouper was induced to change sex from female to male by 17alpha-methyltestosterone implantation. During the artificial spermatogenesis, different cell types from cysts containing spermatogonia, spermatocytes, spermatids, and spermatozoa were isolated by laser capture microdissection. Subsequently, transcriptomic analysis for the isolated cells were performed. A series of genes was used to verify and investigate the expression patterns in spermatogenesis. Furthermore, we also analyzed the expression of the same set of genes involved with steroid metabolism and sex throughout spermatogenesis (early-mid, late, and maturing stages) in the orange-spotted grouper. Several generally female-related genes took significantly changes in sex reversal hinted that the female-related genes in previously recognized may also play vital roles in spermatogenesis and sex reversal. In the transcriptomic data, we focused on zbtb family genes, which may be related to the process of spermatogenesis. Their expression patterns and cellular localization were examined, and the location of Eczbtb40 in different gonadal stages was investigated. We found that Eczbtb40 was expressed throughout spermatogenesis. These preliminary findings suggest that Eczbtb40 is highly conserved during vertebrate evolution and plays roles in spermatogenesis. Besides, the expression of Eczbtb40 and Eccyp17a1a overlapped in male germ cells, especially spermatogonium and spermatocyte, which suggested that Eczbtb40 might interact with Eccyp17a1a participant in spermatogenesis and sex reversal. Conclusions: The present study first depicted RNA sequencing of the male germ cells from orange-spotted grouper, and identified many important functional genes and pathways involved in spermatogenesis. The Eczbtb40 gene was subjected to molecular characterization and expression pattern analysis. These results will contribute to future studies of the molecular mechanism of spermatogenesis and sex reversal.

Identification and functional characterization of two Secretogranin II genes in orange-spotted grouper (Epinephelus coioides)

2018 ◽  
Vol 261 ◽  
pp. 115-126 ◽  
Author(s):  
Hu Shu ◽  
Lidong Yang ◽  
Yong Zhang ◽  
Xiaochun Liu ◽  
Haoran Lin ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Epinephelus Coioides ◽  
Secretogranin Ii

scholarly journals Characterization of novel regulators for heat stress tolerance in tomato from Indian sub-continent

2019 ◽  
Author(s):  
Sonia Balyan ◽  
Sombir Rao ◽  
Sarita Jha ◽  
Chandni Bansal ◽  
Jaishri Rubina Das ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Heat Stress ◽  
Functional Characterization ◽  
Transcriptome Profiling ◽  
Negative Regulator ◽  
Comparative Transcriptome ◽  
Regulatory Pathways ◽  
Cool Region ◽  
Promoter Swapping

AbstractThe footprint of tomato cultivation, a cool region crop that exhibits heat stress (HS) sensitivity, is increasing in the tropics/sub-tropics. Knowledge of novel regulatory hot-spots from varieties growing in the Indian sub-continent climatic zones could be vital for developing HS-resilient crops. Comparative transcriptome-wide signatures of a tolerant (CLN1621L) and sensitive (CA4) cultivar-pair short-listed from a pool of varieties exhibiting variable thermo-sensitivity using physiological, survival and yield-related traits revealed redundant to cultivar-specific HS-regulation with more up-regulated genes for CLN1621L than CA4. The anatgonisiticly-expressing genes include enzymes; have roles in plant defense and response to different abiotic stresses. Functional characterization of three antagonistic genes by overexpression and TRV-VIGS silencing established Solyc09g014280 (Acylsugar acyltransferase) and Solyc07g056570 (Notabilis), that are up-regulated in tolerant cultivar, as positive regulators of HS-tolerance and Solyc03g020030 (Pin-II proteinase inhibitor), that is down-regulated in CLN1621L, as negative regulator of thermotolerance. Transcriptional assessment of promoters of these genes by SNPs in stress-responsive cis-elements and promoter swapping experiments in opposite cultivar background showed inherent cultivar-specific orchestration of transcription factors in regulating transcription. Moreover, overexpression of three ethylene response transcription factors (ERF.C1/F4/F5) also improved HS-tolerance in tomato. This study identifies several novel HS-tolerance genes and provides proof of their utility in tomato-thermotolerance.HighlightNovel heat stress regulatory pathways uncovered by comparative transcriptome profiling between contrasting tomato cultivars from Indian sub-continent for improving thermotolerance. (20/30)

Functional characterization of Cystatin C in orange-spotted grouper, Epinephelus coioides

2019 ◽  
Vol 96 ◽  
pp. 37-46
Author(s):  
Shina Wei ◽  
Jia Cai ◽  
Shaowen Wang ◽  
Yepin Yu ◽  
Jingguang Wei ◽  
...  
Keyword(s):  
Cystatin C ◽  
Functional Characterization ◽  
Epinephelus Coioides

scholarly journals Structural and Functional Characterization of a Testicular Long Non-coding RNA (4930463O16Rik) Identified in the Meiotic Arrest of the Mouse Topaz1–/– Testes

2021 ◽  
Vol 9 ◽  
Author(s):  
Manon Chadourne ◽  
Elodie Poumerol ◽  
Luc Jouneau ◽  
Bruno Passet ◽  
Johan Castille ◽  
...  
Keyword(s):  
Germ Cells ◽  
Developmental Stages ◽  
Functional Characterization ◽  
Sperm Concentration ◽  
Specific Gene ◽  
Meiotic Arrest ◽  
Expression Of Genes ◽  
Non Coding Rna ◽  
Long Non Coding Rna

Spermatogenesis involves coordinated processes, including meiosis, to produce functional gametes. We previously reported Topaz1 as a germ cell-specific gene highly conserved in vertebrates. Topaz1 knockout males are sterile with testes that lack haploid germ cells because of meiotic arrest after prophase I. To better characterize Topaz1–/– testes, we used RNA-sequencing analyses at two different developmental stages (P16 and P18). The absence of TOPAZ1 disturbed the expression of genes involved in microtubule and/or cilium mobility, biological processes required for spermatogenesis. Moreover, a quarter of P18 dysregulated genes are long non-coding RNAs (lncRNAs), and three of them are testis-specific and located in spermatocytes, their expression starting between P11 and P15. The suppression of one of them, 4939463O16Rik, did not alter fertility although sperm parameters were disturbed and sperm concentration fell. The transcriptome of P18-4939463O16Rik–/– testes was altered and the molecular pathways affected included microtubule-based processes, the regulation of cilium movement and spermatogenesis. The absence of TOPAZ1 protein or 4930463O16Rik produced the same enrichment clusters in mutant testes despite a contrasted phenotype on male fertility. In conclusion, although Topaz1 is essential for the meiosis in male germ cells and regulate the expression of numerous lncRNAs, these studies have identified a Topaz1 regulated lncRNA (4930463O16Rik) that is key for both sperm production and motility.

scholarly journals Transcriptome Profiling of Two Ornamental and Medicinal Papaver Herbs

2018 ◽  
Vol 19 (10) ◽  
pp. 3192 ◽  
Author(s):  
Jaehyeon Oh ◽  
Younhee Shin ◽  
In Ha ◽  
Min Lee ◽  
Seok-Geun Lee ◽  
...  
Keyword(s):  
Secondary Metabolite ◽  
Plant Development ◽  
Functional Characterization ◽  
Transcriptome Profiling ◽  
Single Copy ◽  
Isoquinoline Alkaloid ◽  
Metabolite Profiles ◽  
Secondary Metabolite Biosynthesis ◽  
Core Genes

The Papaver spp. (Papaver rhoeas (Corn poppy) and Papaver nudicaule (Iceland poppy)) genera are ornamental and medicinal plants that are used for the isolation of alkaloid drugs. In this study, we generated 700 Mb of transcriptome sequences with the PacBio platform. They were assembled into 120,926 contigs, and 1185 (82.2%) of the benchmarking universal single-copy orthologs (BUSCO) core genes were completely present in our assembled transcriptome. Furthermore, using 128 Gb of Illumina sequences, the transcript expression was assessed at three stages of Papaver plant development (30, 60, and 90 days), from which we identified 137 differentially expressed transcripts. Furthermore, three co-occurrence heat maps are generated from 51 different plant genomes along with the Papaver transcriptome, i.e., secondary metabolite biosynthesis, isoquinoline alkaloid biosynthesis (BIA) pathway, and cytochrome. Sixty-nine transcripts in the BIA pathway along with 22 different alkaloids (quantified with LC-QTOF-MS/MS) were mapped into the BIA KEGG map (map00950). Finally, we identified 39 full-length cytochrome transcripts and compared them with other genomes. Collectively, this transcriptome data, along with the expression and quantitative metabolite profiles, provides an initial recording of secondary metabolites and their expression related to Papaver plant development. Moreover, these profiles could help to further detail the functional characterization of the various secondary metabolite biosynthesis and Papaver plant development associated problems.

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