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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 -
Pesticide Biochemistry and Physiology Nov 2023The serine/threonine kinase Akt is an important component of the insulin signalling pathway (ISP) in regulating insect metabolism, growth, and reproduction. The psocid...
The serine/threonine kinase Akt is an important component of the insulin signalling pathway (ISP) in regulating insect metabolism, growth, and reproduction. The psocid Liposcelis entomophila (Enderlein) is a distasteful stored products pest for its fecundity. However, the molecular mechanism of Akt that controls vitellogenesis and oviposition in L. entomophila remains obscure. In this study, the function of the Akt gene in the female reproduction of L. entomophila (designated as LeAkt) was characterized and investigated. LeAkt contains a 1587 bp open reading frame encoding a 529 amino acid protein that possesses a conserved Pleckstrin Homology domain (PH) and a Ser/Thr-type protein kinase (S_TKc) domain. The mRNA expression of LeAkt was the highest in female adult stages and peaked for 7-day female adults. In female adult tissues, LeAkt was highly expressed in the head and the ovary, indicating that LeAkt was closely correlated with female ovarian development. LeAkt transcription level was significantly suppressed by oral feeding on artificial diets mixed with dsRNA-LeAkt. RNAi-mediated silencing of LeAkt led to a severe inhibition of vitellogenein (Vg) expression and ovarian development, together with lower fecundity and hatchability compared to that of the normal feeding group, suggesting a critical role for LeAkt in L. entomophila reproduction. Further studies revealed that LeAkt silencing significantly decreased the mRNA levels of several signalling and biosynthetic genes in the juvenile hormone (JH) signalling pathway, such as methoprene-tolerant (LeMet), krüppel homolog 1 (LeKr-h1) and JH methyltransferase (LeJHAMT), leading to a severe inhibition of JH biosynthesis in L. entomophila female adults. These results suggested that LeAkt was affecting JH synthesis, thereby influencing Vg synthesis and ultimately L. entomophila reproduction.
Topics: Animals; Proto-Oncogene Proteins c-akt; Juvenile Hormones; Fertility; RNA, Messenger; Serine; Insect Proteins
PubMed: 37945269
DOI: 10.1016/j.pestbp.2023.105583 -
International Journal of Biological... Jan 2024Ribosomal proteins (RPs) have mang extraribosomal functions including regulation of ovarian development in some organisms. In order to solve the problem of rapid ovarian...
Ribosomal proteins (RPs) have mang extraribosomal functions including regulation of ovarian development in some organisms. In order to solve the problem of rapid ovarian maturation in Macrobrachium nipponense aquaculture, this study identified a RPS24 (MnRPS24) gene from M. nipponense, which encodes a protein of ββαβαααα folding structure type. MnRPS24 exhibited the greatest expressions in the female adult stage among the six growth stages, in the ovary among the nine tissues, and in the stage I ovary among the six ovarian development stages. The MnRPS24 protein located in the cytoplasm of oogonia, previtellogenic and early-vitellogenic oocytes, and the follicular cells surrounding the oocytes. The expression of the vitellogenin (MnVg), vitellogenin receptor (MnVgr), cell cycle protein B (MnCyclin B) and cell division cyclin 2 (MnCdc2) genes were increased by recombinant MnRPS24 protein incubation. Conversely, the expression of the Wee1 kinase (MnWee1) gene was decreased. MnRPS24 gene silencing downregulated the expression for MnVg, MnVgr, MnCyclin B and MnCdc2 and upregulated the expression for MnWee1. Furthermore, MnRPS24 gene silencing delayed the vitellogenesis of oocytes, halting the progression of ovarian development. The findings of this research demonstrate that MnRPS24 could potentially function as a stimulator in promoting the development of ovaries in M. nipponense.
Topics: Animals; Female; Palaemonidae; Oocytes; Ovary; Ribosomes
PubMed: 37939777
DOI: 10.1016/j.ijbiomac.2023.127934 -
General and Comparative Endocrinology Jan 2024Estrogen receptors (ERs) play a critical role in vitellogenesis (Vtgs). However, the contribution of each ER for the regulation of vtgs expression was not analyzed...
Estrogen receptors (ERs) play a critical role in vitellogenesis (Vtgs). However, the contribution of each ER for the regulation of vtgs expression was not analyzed clearly in teleosts. In the present study, three ers isoforms (erα, erβ1, and erβ2) were cloned in pompano (Trachinotus ovatus). Real-time PCR and enzyme-linked immunosorbent assay (ELISA) was used to detect the effects of 17β-estradiol (E) on ERs and Vtgs in the liver of pompano. In vivo injection experiments showed that E significantly increased the expressions of ers and vtgs. ER broad spectrum antagonist Fulvestrant significantly attenuated the E- induced up-regulation of ers and vtgs in a dose-dependent manner. ERα antagonist Methyl-piperidino pyrazole (MPP) significantly attenuated the up-regulation of erα, erβ2, vtg-B and vtg-C, and promoted the expressions of erβ1 and vtg-A. ERβ antagonist Cyclofenil significantly inhibited the expressions of erβ1, erβ2, vtg-A and vtg-C, and promoted the expressions of erα and vtg-B. In addition, E significantly increased the protein level of Vtg, while Fulvestrant, MPP and Cyclofenil significantly inhibited the protein level of Vtg in a dose-dependent manner. Our results indicate that E may regulate the expression of each vtg with different subtypes of ERs, and shows a distinct compensatory expression effect on the regulation for ers and vtgs, which provides a theoretical basis for reproductive endocrinology study in pompano.
Topics: Animals; Receptors, Estrogen; Estrogen Receptor alpha; Vitellogenesis; Fulvestrant; Cyclofenil; Estrogen Receptor beta; Cloning, Molecular; Fishes; Estradiol
PubMed: 37923147
DOI: 10.1016/j.ygcen.2023.114403 -
Comparative Biochemistry and... Dec 2023Blue mussels (Mytilus sp.) are an economically important species for European aquaculture. Their importance as a food source is expected to increase in the coming...
Blue mussels (Mytilus sp.) are an economically important species for European aquaculture. Their importance as a food source is expected to increase in the coming net-zero society due to their low environmental footprint; however, their production is affected by anthropogenic stressors and climate change. During reproduction, lipids are key molecules for mussels as they are the main source of energy on which newly hatched embryos depend in the first days of their development. In this work, blue mussels of different origins are analysed, focusing on the differences in lipid composition between the ovary (BMO) and the testis (BMT). The lipidome of blue mussel gonads (BMG) is studied here by combining traditional lipid profiling methods, such as fatty acid and lipid class analysis, with untargeted liquid chromatography-mass spectrometry (LC-MS) lipidomics. The approach used here enabled the identification of 770 lipid molecules from 23 different lipid classes in BMG. BMT, which consists of billions of spermatocytes, had greater amounts of cell membrane and membrane lipid components such as FA18:0, C20 polyunsaturated fatty acids (PUFA), free sterols (ST), ceramide phosphoethanolamines (CerPE), ceramide aminoethylphosphonates (CAEP), cardiolipins (CL), glycerophosphocholines (PC), glycerophosphoethanolamines (PE) and glycerophosphoserines (PS). In BMO, saturated fatty acids (FA14:0 and FA16:0), monounsaturated fatty acids (MUFA) and other storage components such as C18-PUFA accumulated in triradylglycerolipids (TG) and alkyldiacylglycerols (neutral plasmalogens, TG O-), which, together with terpenes, wax esters and cholesterol esters, make up most of oocytes yolk reserves. BMO also had higher levels of ceramides (Cer) and generally alkyl/alkenyl glycerophospholipids (mainly plasmanyl/plasmenyl PC), suggesting a role for these lipids in vitellogenesis. Non-methylene interrupted dienoic fatty acids (NMID FA), typically found in plasmalogens, were the only membrane-forming PUFA predominantly detected in BMO. The results of this study are of great importance for clarifying the lipid composition of BMG and provide an important basis for future studies on the reproductive physiology of these organisms.
Topics: Male; Female; Animals; Mytilus edulis; Mytilus; Lipidomics; Plasmalogens; Sex Characteristics; Fatty Acids; Fatty Acids, Unsaturated; Gonads; Ceramides
PubMed: 37913700
DOI: 10.1016/j.cbd.2023.101150 -
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