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Development & Reproduction Sep 2023Effects of changes in photoperiod on the reproductive events in fish are suggested to be mediated mainly via the action of melatonin (MEL). Changing levels of plasma MEL...
Effects of changes in photoperiod on the reproductive events in fish are suggested to be mediated mainly via the action of melatonin (MEL). Changing levels of plasma MEL throughout the day and year are suggested to influence the hypothalamus-pituitary-gonadal axis in fish. Therefore, in this study, we aimed to investigate the effects of MEL on oocyte maturation and germinal vesicle breakdown (GVBD) in the marine fish, , . Oocytes at three different stages (pre-, mid-, and late-vitellogenesis) were incubated with (a) only MEL (5, 10, 50, 100, 500, and 1,000 pg/mL) and (b) 50 pg/mL of 17α,20β-dihydroxy-4-pregnen-3-one (17α20βP), maturation-inducing hormone (MIH) of this species, and MEL (4-h incubation before addition of MIH). Any single MEL treatment did not significantly induce GVBD. However, treatment with 50 pg/mL MEL or MIH significantly induced GVBD. These results suggest that preincubation with MEL accelerates the effect of MIH on longchin goby oocyte maturation.
PubMed: 38074459
DOI: 10.12717/DR.2023.27.3.127 -
Environmental Pollution (Barking, Essex... Feb 2024In this study, the disrupting effects of glyphosate (GLY), aminomethylphosphonic acid (AMPA), and three glyphosate-based herbicides (GBHs) on vitellogenesis in a...
In this study, the disrupting effects of glyphosate (GLY), aminomethylphosphonic acid (AMPA), and three glyphosate-based herbicides (GBHs) on vitellogenesis in a non-concentration-dependent manner are reported for the first time in 120 h of acute exposure of zebrafish at environmentally relevant concentrations. GBHs are commonly used worldwide in weed control management. Due to their extensive application, they frequently occur in aquatic ecosystems and may affect various organisms. The active substance GLY and its major by-product, AMPA, are the most thoroughly studied chemicals; however, the adverse effects of the complex formulas of GBHs with diverse and unknown content of co-formulants are still not sufficiently researched. This study focused on the embryotoxicity, sublethal malformations, and estrogenic potency of GLY, AMPA, and four commonly used GBHs on zebrafish embryos using a wild type and an estrogen-sensitive, transgenic zebrafish line (Tg(vtg1:mCherry)). After 120 h of exposition, AMPA did not cause acute toxicity, while the LC of GLY was 160 mg/L. The GBHs were more toxic with LC values ranging from 31 to 111 GLY active equivalent (a.e.) mg/L. Exposure to 0.35-2.8 mg/L GBHs led to sublethal abnormalities: typical symptoms were structural deformation of the lower jaw and anomalies in the olfactory region. Deformity rates were 10-30% in the treated groups. In vivo, fluorescently expressed vtg1 mCherry protein in embryonic liver was detected by a non-invasive microscopic method indicating estrogenic action through vitellogenin production by GLY, AMPA, and GBHs. To confirm the in vivo findings, RT-qPCR method was performed to determine the levels of the estrogenicity-related vtg1 mRNA. After 120 h of exposure to GLY, AMPA, and three GBHs at a concentration of 0.35 mg/L, the expression of vtg1 gene was significantly up-regulated. Our results highlight the risk that short-term GLY and GBH exposure can cause developmental malformations and disrupt the hormonal balance in zebrafish embryos.
Topics: Animals; Glyphosate; Zebrafish; alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; Glycine; Ecosystem; Herbicides; Animals, Genetically Modified; Estrone; Organophosphonates
PubMed: 38072021
DOI: 10.1016/j.envpol.2023.123113 -
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 -
Open Biology Nov 2023The halophilic zooplankton brine shrimp has been used as an experimental animal in multidisciplinary studies. However, the reproductive patterns and its regulatory...
The halophilic zooplankton brine shrimp has been used as an experimental animal in multidisciplinary studies. However, the reproductive patterns and its regulatory mechanisms in remain unclear. In this study, the ovarian development process of parthenogenetic () was divided into five stages, and oogenesis or egg formation was identified in six phases. The oogenesis mode was assumed to be polytrophic. We also traced the dynamic translocation of candidate germline stem cells (cGSCs) using EdU labelling and elucidated several key cytological events in oogenesis through haematoxylin and eosin staining and fluorescence imaging. Distinguished from the ovary structure of insects and crustaceans, germarium originated from ovariole buds and are located at the base of the ovarioles. RNA-seq based on five stages of ovarian development identified 2657 upregulated genes related to reproduction by pair-to-pair comparison. , , , , , and genes associated with cGSCs recognition and reproductive development were screened and verified using qPCR. Silencing of the gene in () at ovarian development Stage II led to a low level of gene expression (less than 10%) within 5 days, which resulted in variations in oogenesis-related gene expression and significantly inhibited vitellogenesis, impeded oocyte maturation, and eventually decreased the number of offspring. In conclusion, we have illustrated the patterns of ovarian development, outlined the key spatio-temporal features of oogenesis and identified the negative impacts of VgR gene knockdown on oogenesis using as an experimental animal. The findings of this study also lay a foundation for the further study of reproductive biology of invertebrates.
Topics: Animals; Female; Artemia; Oogenesis; Ovary; Gene Silencing; Fertility
PubMed: 37963545
DOI: 10.1098/rsob.230172 -
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 -
Biology Sep 2023Coleoptera is the largest taxa of animals by far. The robust reproductive capacity is one of the main reasons for such domination. Successful female reproduction...
Coleoptera is the largest taxa of animals by far. The robust reproductive capacity is one of the main reasons for such domination. Successful female reproduction partially relies on effective vitellogenesis. However, the hormone regulation of vitellogenesis remains to be explored. In the present paper, in vitro culture of 1-day-old adult fat bodies in the 20E-contained median did not activate juvenile hormone production and insulin-like peptide pathways, but significantly stimulated the expression of two genes, in a cycloheximide-dependent pattern. In vivo RNA interference (RNAi) of either ecdysone receptor () or ultraspiracle () by injection of corresponding dsRNA into 1-day-old female adults inhibited oocyte development, dramatically repressed the transcription of genes in fat bodies and of in ovaries; application of JH into the or RNAi females did not restore the oocyte development, partially rescued the decreased mRNA levels but over-compensated expression levels. The same RNAi experiments were performed in another Coleoptera species, . Little yolk substances were seen in the misshapen oocytes in the or RNAi ovaries, in contrast to larger amounts of yolk granules in the normal oocytes. Correspondingly, the transcript levels of in the fat bodies and ovaries decreased significantly in the and RNAi samples. Our results here show that 20E signaling is indispensable in the activation of vitellogenesis in the developing oocytes of the two beetle species.
PubMed: 37886994
DOI: 10.3390/biology12101284 -
Frontiers in Endocrinology 2023Bursicon, a neuropeptide hormone comprising two subunits-bursicon (burs) and partner of burs (pburs), belongs to the cystine-knot protein family. Bursicon heterodimers...
Bursicon, a neuropeptide hormone comprising two subunits-bursicon (burs) and partner of burs (pburs), belongs to the cystine-knot protein family. Bursicon heterodimers and homodimers bind to the lucine-rich G-protein coupled receptor (LGR) encoded by s to regulate multiple physiological processes in arthropods. Notably, these processes encompass the regulation of female reproduction, a recent revelation in . In this study we investigated the role of burs/pburs/rickets in mediating female vitellogenesis and reproduction in a hemipteran insect, the whitefly, . Our investigation unveiled a synchronized expression of and , with their transcripts persisting detectable in the days following eclosion. RNAi-mediated knockdown of , or significantly suppressed the transcript levels of () and in the female whiteflies. These effects also impaired ovarian maturation and female fecundity, as evidenced by a reduction in the number of eggs laid per female, a decrease in egg size and a decline in egg hatching rate. Furthermore, knockdown of , or led to diminished juvenile hormone (JH) titers and reduced transcript level of . However, this impact did not extend to genes in the insulin pathway or target of rapamycin pathway, deviating from the results observed in . Taken together, we conclude that burs/pburs/rickets regulates the vitellogenesis and reproduction in the whiteflies by coordinating with the JH signaling pathway.
Topics: Animals; Female; Hemiptera; Invertebrate Hormones; Juvenile Hormones; Vitellogenesis; Neuropeptides; Rickets
PubMed: 37854192
DOI: 10.3389/fendo.2023.1277439 -
International Journal of Molecular... Sep 2023In insects, the ecdysteroid hormone regulates development and reproduction. However, its function in the reproduction process of spider mites is still unclear. In this...
In insects, the ecdysteroid hormone regulates development and reproduction. However, its function in the reproduction process of spider mites is still unclear. In this study, we investigated the effect of the Halloween gene on the oviposition of the reproduction process in a spider mite, . The expression patterns of the ecdysteroid biosynthesis and signaling pathway genes, as analyzed by RT-qPCR, showed that the expression pattern of the Halloween genes was similar to the oviposition pattern of the female mite and the expression patterns of the vitellogenesis-related genes and , suggesting that the Halloween genes are involved in the oviposition of spider mites. To investigate the function of the ecdysteroid hormone on the oviposition of the reproduction process, we carried out an RNAi assay against the Halloween gene by injection in female mites. Effective silencing of led to a significant reduction of oviposition. In summary, these results provide an initial study on the effect of Halloween genes on the reproduction in and may be a foundation for a new strategy to control spider mites.
Topics: Animals; Female; Oviposition; Tetranychidae; Ecdysteroids; Reproduction; RNA Interference
PubMed: 37834248
DOI: 10.3390/ijms241914797 -
In vivo direct cell-penetrating peptide mediated protein transduction system in Acyrthosiphon pisum.BMC Research Notes Sep 2023The principal delivery method for CRISPR-based genome editing in insects is now based on microinjection into single cells or embryos. The direct protein transduction...
OBJECTIVE
The principal delivery method for CRISPR-based genome editing in insects is now based on microinjection into single cells or embryos. The direct protein transduction systems cannot be employed in aphids because oogenesis occurs without apparent vitellogenesis. Given the limited timing of injection into the embryonic stage in oviparous aphids, a protein delivery system from the hemolymph to the germline and embryos would be a useful tool for genome editing. This study reports a newly developed direct protein delivery system for aphids using cell-penetrating peptides (CPPs). CPPs are short peptides that translocate across the plasma membrane when bound to cargo proteins.
RESULTS
Penetratin (PEN), a widely conserved CPP among insects, was identified in this study. We used mVenus, a recombinant fluorescent protein, as a visual marker for CPP availability assessments, and fused it with PEN by bacterial protein expression. The mVenus-PEN recombinant proteins were introduced into the hemolymph of adult unwinged Acyrthosiphon pisum females using a nanoinjector. Fluorescence emitted by mVenus-PEN was observed in various tissues, such as the gut, trachea, bacteriocytes, and their progeny. This study shows that PEN can deliver exogenously expressed proteins into tissues in vivo, indicating that CPPs are powerful tools for protein transduction.
Topics: Female; Animals; Cell-Penetrating Peptides; Pisum sativum; Aphids; Bacterial Proteins; Cell Membrane
PubMed: 37749584
DOI: 10.1186/s13104-023-06514-9 -
International Journal of Molecular... Aug 2023is an important plant pest. Comparative feeding experiments showed that the egg production, oviposition duration and survival rate of beetles were significantly higher...
is an important plant pest. Comparative feeding experiments showed that the egg production, oviposition duration and survival rate of beetles were significantly higher when they fed on elm leaves than when they fed on willow or purpus privet leaves. RNA sequencing was used to determine transcriptomic changes associated with oviposition. Comparative transcriptome analysis revealed that the beetles that fed on elm and willow had a total of 171 genes with differential expression. When the beetles fed on elm and purpus privet, 3568 genes had differential expression. The vitellogenesis, ovarian serine protease, odorant-binding proteins, acyl-CoA synthetase and follicle cell proteins were commonly upregulated genes in elm-fed beetles compared with those fed on willow/purpus privet leaves. The involvement of the follicle cell protein 3C gene in the regulation of oviposition was confirmed using RNA interference. The results provide insights into the molecular mechanisms underlying oviposition in feeding on different host plants. This study also describes a method for identifying potentially effective genes for pest control.
Topics: Female; Animals; Transcriptome; Oviposition; Gene Expression Profiling; Coleoptera; Chlorophyceae
PubMed: 37685945
DOI: 10.3390/ijms241713138