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Animals : An Open Access Journal From... Feb 2024Zygote arrest-1 (Zar1) and Wilms' tumor 1 (Wt1) play an important role in oogenesis, with the latter also involved in testicular development and gender differentiation....
Zygote arrest-1 (Zar1) and Wilms' tumor 1 (Wt1) play an important role in oogenesis, with the latter also involved in testicular development and gender differentiation. Here, and were identified in Asian seabass (), a hermaphrodite fish, as the valuable model for studying sex differentiation. The cloned cDNA fragments of were 1192 bp, encoding 336 amino acids, and contained a zinc-binding domain, while those of cDNA were 1521 bp, encoding a peptide of 423 amino acids with a Zn finger domain belonging to Wt1b family. RT-qPCR analysis showed that mRNA was exclusively expressed in the ovary, while mRNA was majorly expressed in the gonads in a higher amount in the testis than in the ovary. In situ hybridization results showed that mRNA was mainly concentrated in oogonia and oocytes at early stages in the ovary, but were undetectable in the testis. mRNA was localized not only in gonadal somatic cells (the testis and ovary), but also in female and male germ cells in the early developmental stages, such as those of previtellogenic oocytes, spermatogonia, spermatocytes and spermatids. These results indicated that and possibly play roles in gonadal development. Therefore, the findings of this study will provide a basis for clarifying the mechanism of and in regulating germ cell development and the sex reversal of Asian seabass and even other hermaphroditic species.
PubMed: 38338151
DOI: 10.3390/ani14030508 -
Animals : An Open Access Journal From... Feb 2024is one of the earliest expressed genes in the oocyte during ovarian development. In this study, was characterized in . , one of the main aquaculture species in China,...
is one of the earliest expressed genes in the oocyte during ovarian development. In this study, was characterized in . , one of the main aquaculture species in China, and designated as . The length of cDNA was 1303 bp, encoding 197 amino acids that contained a conserved bHLH domain. revealed a female-biased expression patterns in the gonads of adult fish, and expression was far higher in ovaries than that in testes at all gonadal development stages, especially at 60~180 days post-fertilization (dpf). Furthermore, a noteworthy inverse relationship was observed between expression and the methylation of its promoter in the adult gonads. Gonads at 90 dpf were used for in situ hybridization (ISH), and transcripts were mainly concentrated in oogonia and the primary oocytes in ovaries, but undetectable in the testes. These results indicated that Figla would play vital roles in the ovarian development in . Additionally, the frame-shift mutations of were successfully constructed through the CRISPR/Cas9 system, which established a positive foundation for further investigation on the role of in the ovarian development of . Our study provides valuable clues for exploring the regulatory mechanism of in the fish ovarian development and maintenance, which would be useful for the sex control and reproduction of fish in aquaculture.
PubMed: 38338134
DOI: 10.3390/ani14030491 -
Scientific Reports Feb 2024Spermatogenesis is one of the most dramatic changes in cell differentiation. Remarkable chromatin condensation of the nucleus is observed in animal, plant, and algal...
Spermatogenesis is one of the most dramatic changes in cell differentiation. Remarkable chromatin condensation of the nucleus is observed in animal, plant, and algal sperm. Sperm nuclear basic proteins (SNBPs), such as protamine and sperm-specific histone, are involved in chromatin condensation of the sperm nucleus. Among brown algae, sperm of the oogamous Fucales algae have a condensed nucleus. However, the existence of sperm-specific SNBPs in Fucales algae was unclear. Here, we identified linker histone (histone H1) proteins in the sperm and analyzed changes in their gene expression pattern during spermatogenesis in Sargassum horneri. A search of transcriptomic data for histone H1 genes in showed six histone H1 genes, which we named ShH1.1a, ShH1b, ShH1.2, ShH1.3, ShH1.4, and ShH1.5. Analysis of SNBPs using SDS-PAGE and LC-MS/MS showed that sperm nuclei contain histone ShH1.2, ShH1.3, and ShH1.4 in addition to core histones. Both ShH1.2 and ShH1.3 genes were expressed in the vegetative thallus and the male and female receptacles (the organs producing antheridium or oogonium). Meanwhile, the ShH1.4 gene was expressed in the male receptacle but not in the vegetative thallus and female receptacles. From these results, ShH1.4 may be a sperm-specific histone H1 of S. horneri.
Topics: Animals; Male; Histones; Sargassum; Chromatography, Liquid; Semen; Tandem Mass Spectrometry; Cell Nucleus; Chromatin; Spermatozoa
PubMed: 38336896
DOI: 10.1038/s41598-024-53729-2 -
International Journal of Molecular... Jan 2024is gonochoristic and sexually dimorphic. The male prawn grows faster and usually has a larger size than the female. Therefore, a higher male proportion in stock usually...
is gonochoristic and sexually dimorphic. The male prawn grows faster and usually has a larger size than the female. Therefore, a higher male proportion in stock usually results in higher yield. To investigate the impact of temperature on sexual differentiation in , two temperature treatments (26 °C and 31 °C) were conducted. The results showed that compared to the 31 °C treatment (3.20 ± 0.12), the 26 °C treatment displayed a lower female/male ratio (2.20 ± 0.11), which implied that a lower temperature could induce masculinization in The temperature-sensitive sex differentiation phase was 25-35 days post hatching (DPH) at 26 °C while 15-20 DPH at 31 °C. Transcriptome and qPCR analysis revealed that a lower temperature up-regulated the expression of genes related to androgen secretion, and down-regulated the expressions of genes related to oogonia differentiation. Thirty-one temperature-regulated sex-differentiation genes were identified and the molecular mechanism of temperature-regulated sex differentiation was suggested. The finding of this study indicates that temperature regulation can be proposed as an innovative strategy for improving the culture yield of .
Topics: Animals; Male; Female; Palaemonidae; Sex Differentiation; Temperature; Transcriptome; Penaeidae; Arthropod Proteins
PubMed: 38279207
DOI: 10.3390/ijms25021207 -
Plant Disease Jan 2024Peanut ( L.) has long been cultivated worldwide as an important crop for oil and protein production. Among the various diseases in peanut plants, wilt diseases caused by...
Peanut ( L.) has long been cultivated worldwide as an important crop for oil and protein production. Among the various diseases in peanut plants, wilt diseases caused by soil-borne pathogens such as and are especially destructive and substantially diminish both quantity and quality in peanut production (Kokalis-Burelle et al., 1997; Thiessen et al., 2012). In July 2022, wilt symptoms were observed in 1 to 3% of the area of peanut fields in Yeoju-si, Korea (37°23´04.0˝N; 127°33´43.0˝E). The xylem in the stems of the wilted plants was dark brown at the soil-surface, which is a representative symptom of vascular wilt pathogens (Yadeta et al. 2013). To isolate the causative pathogens, the stems exhibiting dark lesions were disinfected with 1% NaOCl for 1 min, rinsed with sterile distilled water, and placed on potato dextrose agar medium. The plates were incubated at 25℃ for 2 days, and white hyphae that grew out from the tissues were subcultured twice on V8 juice agar (V8A) medium. Among the 3 isolates, morphological characteristics of the representative strain YJ1-2 were observed under a microscope. The sporangia were terminal intercalary, filamentous, inflated lobulate, and ranging from 37.4 to 73.6 μm in diameter. The antheridia were diclinous, with clavate, elongate, and crook-necked shapes. The oogonia were mostly globose, with an average of 27.1 μm (range from 20.2 to 35.2 μm, n = 50) in diameter, and mated with one to several antheridia. Both plerotic or aplerotic oospores were observed. Overall, the morphological characteristics of the sporangia, antheridia, oogonia, and oospores indicated that YJ1-2 belongs to the genus . To genetically characterize YJ1-2, genomic DNA was extracted using cetyltrimethylammonium bromide buffer, and the internal transcribed spacer (ITS) region and cytochrome c oxidase subunit I () gene were amplified by PCR using primer sets ITS4/ITS5 and OomCoxI-Levlo/ OomCoxI-Levup, respectively (White et al., 1990; Robideau et al. 2011), sequenced, and identified using BLASTN (NCBI, National Center for Biotechnology Information). The ITS sequence (NCBI Acc. No. OR125595) of YJ1-2 has 99% similarity with that of isolate PY39 (NCBI Acc. No. KX671096). A neighbor-joining phylogenetic tree was constructed from aligned sequence (NCBI Acc. No. OR224334) of the 10 species strains including YJ1-2 by CLUSTALW method was used as an outgroup. The YJ1-2 was most closely related to isolate PM30 (NCBI Acc. No. MT823167). To substantiate the pathogenicity of YJ1-2, the crown roots of peanut plants grown in pots for 4 weeks were wounded using a sterile tweezer, and the mycelial plugs of YJ1-2 cultured for 5 days on V8A were inoculated on the wounds. The inoculated plants were cultivated in a growth chamber at 30℃ and 70% relative humidity with a 12-h photoperiod. The infected peanut plants exhibited wilt symptoms 11 days after inoculation, consistent with the initial observation, while uninoculated plants remained healthy. To satisfy Koch's postulates, white mycelia were re-isolated from the stems of inoculated plants and axenically cultured in V8A. The morphologies and ITS sequences of the re-isolates were consistent with those of YJ1-2. has been reported as a causal pathogen of peanut pod rot in the United States and China. However, to the best of our knowledge, this is the first report of wilt disease in peanut plants caused by in Korea. To prevent the incidence of wilt disease, we will continue our investigations to develop control strategies, including the selection of appropriate agrochemicals.
PubMed: 38173269
DOI: 10.1094/PDIS-11-23-2311-PDN -
Journal of Fungi (Basel, Switzerland) Nov 2023In recent years, a new root rot disease in barley, which is caused by an species, was found in field surveys in Southern Sweden and Denmark. Its symptoms occurred at...
In recent years, a new root rot disease in barley, which is caused by an species, was found in field surveys in Southern Sweden and Denmark. Its symptoms occurred at the early tillering stage, around the BBCH 21 growth stage, and included the yellowing of leaves, brown coleoptiles, and the discolouration of roots. Prolonged soil wetness after rainfall favoured disease development, which sometimes advanced the yellowing patches to entire fields, resulting in lower yields. Oospores were found in the fine roots of diseased plants, and isolates were obtained from these roots, as well as from the roots of barley plants grown in the greenhouse in soil samples from infected fields. Based on morphological analysis, we found that the new isolates were similar to those already obtained from barley and spinach roots in the 1990s in the same growing area. The morphological and molecular analyses performed in this study clearly separated and distinguished these barley isolates from other known , and hereby sp. nov. is proposed as a new plant pathogenic species. It has larger oogonia and oospores than , , and , with one up to eight diclinous antheridia per oogonium. The phylogenetic analysis of the ITS rDNA region sequences grouped these new isolates in a monophyletic clade, which was clearly distinguished from other plant pathogenic species. The further pathogenicity of on other plants is currently under investigation, but it is clear that it can at least infect barley, spinach, and sugar beet, indicating a wide host range for this species. The widespread presence and presumably broad host range of this new pathogenic species must be considered in crop rotations.
PubMed: 38132745
DOI: 10.3390/jof9121144 -
PeerJ 2023Thyroid-associated orbitopathy (TAO) is a disease associated with autoimmune thyroid disorders and it can lead to proptosis, diplopia, and vision-threatening compressive...
BACKGROUND
Thyroid-associated orbitopathy (TAO) is a disease associated with autoimmune thyroid disorders and it can lead to proptosis, diplopia, and vision-threatening compressive optic neuropathy. To comprehensively understand the molecular mechanisms underlying orbital adipogenesis in TAO, we characterize the intrinsic molecular properties of orbital adipose/connective tissue from patients with TAO and control individuals.
METHODS
RNA sequencing analysis (RNA-seq) was performed to measure the gene expression of orbital adipose/connective tissues of TAO patients. Differentially expressed genes (DEGs) were detected and analyzed through Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and Gene Set Enrichment Analysis (GSEA). The protein-protein interaction (PPI) network was constructed using the STRING database, and hub genes were identified by the Cytoscape plug-in, cytoHubba. We validated several top DEGs through quantitative real-time polymerase chain reaction (qRT-PCR).
RESULTS
We identified 183 DEGs in adipose tissue between TAO patients ( = 3) and control patients ( = 3) through RNA sequencing, including 114 upregulated genes and 69 downregulated genes. The PPI network of these DEGs had 202 nodes and 743 edges. PCR-based validation results of orbital adipose tissue showed multiple top-ranked genes in TAO patients ( = 4) are immune and inflammatory response genes compared with the control individual ( = 4). They include ceruloplasmin isoform x3 (CP), alkaline tissue-nonspecific isozyme isoform x1 (ALPL), and angiotensinogen (AGT), which were overrepresented by 2.27- to 6.40-fold. Meanwhile, protein mab-21-like 1 (MAB21L1), phosphoinositide 3-kinase gamma-subunit (PIK3C2G), and clavesin-2 (CLVS2) decreased by 2.6% to 32.8%. R-spondin 1 (RSPO1), which is related to oogonia differentiation and developmental angiogenesis, was significantly downregulated in the orbital muscle tissues of patients with TAO compared with the control groups ( = 0.024).
CONCLUSIONS
Our results suggest that there are genetic differences in orbital adipose-connective tissues derived from TAO patients. The upregulation of the inflammatory response in orbital fat of TAO may be consistent with the clinical phenotype like eyelid edema, exophthalmos, and excess tearing. Downregulation of MAB21L1, PIK3C2G, and CLVS2 in TAO tissue demonstrates dysregulation of differentiation, oxidative stress, and developmental pathways.
Topics: Humans; Graves Ophthalmopathy; Phosphatidylinositol 3-Kinases; Connective Tissue; Real-Time Polymerase Chain Reaction; Protein Isoforms; Homeodomain Proteins
PubMed: 38130930
DOI: 10.7717/peerj.16569 -
Cells Nov 2023(1) Fshβ and Lhβ showed stronger signals and higher transcript levels from 590 to 1050 dph than at earlier stages, implying their active involvement during primary...
Chronological Changes in Gonadotropin-Releasing Hormone 1, Gonadotropins, and Sex Steroid Hormones along the Brain-Pituitary-Gonadal Axis during Gonadal Sex Differentiation and Development in the Longtooth Grouper, .
(1) Fshβ and Lhβ showed stronger signals and higher transcript levels from 590 to 1050 dph than at earlier stages, implying their active involvement during primary oocyte development. (2) Fshβ and Lhβ at lower levels were detected during the phases of ovarian differentiation and oogonial proliferation. (3) E concentrations increased significantly at 174, 333, and 1435 dph, while T concentrations exhibited significant increases at 174 and 333 dph. These findings suggest potential correlations between serum E concentrations and the phases of oogonial proliferation and pre-vitellogenesis.
Topics: Female; Animals; Bass; Sex Differentiation; Gonadotropin-Releasing Hormone; Gonadal Steroid Hormones; Follicle Stimulating Hormone, beta Subunit; Luteinizing Hormone, beta Subunit; Brain
PubMed: 37998369
DOI: 10.3390/cells12222634 -
Brazilian Journal of Biology = Revista... 2023Specimens of Cnemidocarpa amphora were collected monthly from the Arabian Gulf from September 2017 to August 2018. Parts of their gonads were prepared for histological...
Gonadal proliferation and reproductive cycle of the exotic sea squirt Cnemidocarpa amphora () (Pleurogona, Styelidae) sampled for the first time from the northern coast of Arabian Gulf in Saudi Arabia.
Specimens of Cnemidocarpa amphora were collected monthly from the Arabian Gulf from September 2017 to August 2018. Parts of their gonads were prepared for histological testing. The gonads' diameters varied by month. Each gonad contained many ovarian follicles with different morphologies and was surrounded by several testicular follicles. The ovarian and testicular follicles were separate, although the latter were always present near the former. Repeated measures ANOVA tests were conducted to investigate monthly changes in oocyte stages. In squirts measuring 12-13 cm in length, the gonads measured 30-50 mm from July to August; 20-25 mm from September to October; 15-20 mm from November to February; and 25-40 mm from March to June. Oogonia budded from the germinal epithelium with diameters of 20-30 µm. Previtellogenic oocytes measuring 70-120 µm were characterized by the deposition of small granules of protein around the nucleus, a continuous layer of follicular cuboidal epithelium, and scattered vacuoles in the ooplasm. The measurement of gonads and oocyte diameters were performed by image analysis (Image scope 2.3, Image Line, Inc.) and stage micrometer. The vitellogenic oocytes measured 130-220 µm and the follicular epithelium consisted of flattened and cuboidal layers. Beneath the vitelline membrane, scattered test cells appeared in the ooplasm and different granules of protein and MPS were deposited in the ooplasm. In the later phase, lipid droplets began to appear in the ooplasm. Yolk bodies formed after the impregnation of various granules together and the oocyte was ready to be shed. Before spawning, a yolk membrane appeared above the ooplasm. Post-vitellogenic oocytes, in which the homogeneity of ooplasm was restored, underwent gradual lysis and entered the atretic phase. Different stages of sperm development were present year-round in different follicles of the same squirt; hence, the testes were always mature.
Topics: Animals; Female; Male; Urochordata; Saudi Arabia; Semen; Oocytes; Ovary; Cell Proliferation
PubMed: 37970899
DOI: 10.1590/1519-6984.273666 -
Animals : An Open Access Journal From... Oct 2023This study sought to examine the ovarian cellular and stromal components of the zebrafish () throughout the spawning season using light and electron microscopic tools....
This study sought to examine the ovarian cellular and stromal components of the zebrafish () throughout the spawning season using light and electron microscopic tools. The ovaries of zebrafish showed oocytes in all stages of follicular development and degeneration (atresia). Six stages of oogenesis were demonstrated: oogonia, early oocytes, late oocytes, vacuolated follicles, the yolk globule stage (vitellogenesis), and mature follicles. The SOX9 protein was expressed in the ooplasm of the primary and previtellogenic oocytes and the theca cell layer of the mature follicles. Myostatin was expressed in the granulosa and theca cells. Many stem cells in the ovarian stroma expressed myostatin and SOX9. During the spawning season, the EM results indicated that the zona radiata increased in thickness and was crossed perpendicularly by pore canals that contained processes from both oocytes and zona granulosa. The granulosa cells contained many mitochondria, rER, sER, and vesicles. Meanwhile, the thecal layer consisted of fibroblast-like cells. Atretic follicles could be demonstrated that involved both oocytes and their follicular walls. Several types of cells were distinguished in the ovarian stroma, including mast cells, telocytes, lymphocytes, fibroblasts, endocrine cells, macrophages, adipocytes, dendritic cells, and steroidogenic (stromal) cells. The ovary of the zebrafish serves as a model to investigate follicular development.
PubMed: 37958117
DOI: 10.3390/ani13213362