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The Journal of Reproduction and... May 2024Intracytoplasmic sperm injection (ICSI) is clinically used to treat obstructive/nonobstructive azoospermia. This study compared the efficacy of ICSI with cauda...
Intracytoplasmic sperm injection (ICSI) is clinically used to treat obstructive/nonobstructive azoospermia. This study compared the efficacy of ICSI with cauda epididymal and testicular sperm in Wistar (WI) and Brown-Norway (BN) rats. The transfer of ICSI oocytes with cryopreserved epididymal and testicular WI sperm resulted in offspring production of 26.2% and 3.7%-4.7%, respectively (P < 0.05). Treatments for artificial oocyte activation (AOA) and acrosome removal improved pronuclear formation in BN-ICSI oocytes; however, only AOA treatment was effective in producing offspring (3.7%-6.5%). In the case of ICSI with testicular sperm (TESE-ICSI), one offspring (0.6%) was derived from the BN-TESE-ICSI oocytes. The application of AOA or a hypo-osmotic sperm suspension did not improve the production of TESE-ICSI offspring. Thus, outbred WI rat offspring can be produced by using ICSI and less efficiently by using TESE-ICSI. Challenges in producing offspring by using ICSI/TESE-ICSI in inbred BN strains require further investigation.
PubMed: 38735740
DOI: 10.1262/jrd.2024-030 -
Stem Cell Reports Jun 2024Removal of somatic histone H3 lysine 9 trimethylation (H3K9me3) from the embryonic genome can improve the efficiency of mammalian cloning using somatic cell nuclear...
Removal of somatic histone H3 lysine 9 trimethylation (H3K9me3) from the embryonic genome can improve the efficiency of mammalian cloning using somatic cell nuclear transfer (SCNT). However, this strategy involves the injection of histone demethylase mRNA into embryos, which is limiting because of its invasive and labor-consuming nature. Here, we report that treatment with an inhibitor of G9a (G9ai), the major histone methyltransferase that introduces H3K9me1/2 in mammals, greatly improved the development of mouse SCNT embryos. Intriguingly, G9ai caused an immediate reduction of H3K9me1/2, a secondary loss of H3K9me3 in SCNT embryos, and increased the birth rate of cloned pups about 5-fold (up to 3.9%). G9ai combined with the histone deacetylase inhibitor trichostatin A further improved this rate to 14.5%. Mechanistically, G9ai and TSA synergistically enhanced H3K9me3 demethylation and boosted zygotic genome activation. Thus, we established an easy, highly effective SCNT protocol that would enhance future cloning research and applications.
Topics: Animals; Histones; Nuclear Transfer Techniques; Mice; Histone-Lysine N-Methyltransferase; Methylation; Cloning, Organism; Embryo, Mammalian; Embryonic Development; Hydroxamic Acids; Female; Histone Deacetylase Inhibitors
PubMed: 38729154
DOI: 10.1016/j.stemcr.2024.04.003 -
Marine Environmental Research Jun 2024Marine macroalgal forests are facing unprecedented challenges worldwide due to the accelerating impacts of climate change. These ecosystems play a crucial role in...
Marine macroalgal forests are facing unprecedented challenges worldwide due to the accelerating impacts of climate change. These ecosystems play a crucial role in supporting biodiversity, coastal ecosystem functions and services, and are indeed object of several conservation and restoration measures. The Mediterranean Sea is warming faster than the oceans and thermal anomalies are occurring with increasing intensity, frequency and duration. Along the Mediterranean coasts, Cystoseira sensu lato species are the main representatives of macroalgal forests and their decline has been widely documented. Some relevant achievements in the implementation of ecological restoration have been obtained, but rising temperatures and the occurrence of thermal anomalies increasingly threaten the success of these restoration attempts. In the summer of 2022, ex-situ restoration actions of Ericaria amentacea were carried out by collecting fertile material from three donor sites of the Italian coasts along a latitudinal gradient, during the period of sexual maturity (June/July). Noteworthy during the summer of 2022, anomalous thermal conditions were recorded at the donor sites, with sea surface temperatures exceeding the climatological mean up to 4.3 °C and heatwaves lasting up to 78 days. Our results suggest that these thermal anomalies may have affected the culture of the embryos in both the pre- and post-zygotic phases, resulting in significantly low culture efficiency at the three donor sites. The reproductive structures showed some abnormalities, fertilization of eggs was lower and embryo growth was slower, resulting in lower percent cover of seedlings on the tiles and lower survival rate. The observations underscore the vulnerability of Mediterranean algal forests to global change and highlight additional challenges for their restoration due to the increasing frequency and severity of thermal anomalies, emphasizing the need for adaptive strategies and a comprehensive understanding of the species in a changing climate. Marine forest restoration requires long lasting projects, to allow for long-term monitoring and better understanding the biology of the species and for mitigating stochastic events that can cause the temporary failure of efforts.
Topics: Climate Change; Mediterranean Sea; Temperature; Ecosystem; Forests; Conservation of Natural Resources; Seaweed; Ericaceae; Environmental Restoration and Remediation; Italy
PubMed: 38728798
DOI: 10.1016/j.marenvres.2024.106537 -
Frontiers in Allergy 2024Progesterone is an endogenous hormone, produced by the adrenal cortex, the gonads and in women, its source is the corpus luteum. Progesterone is produced in the late... (Review)
Review
Progesterone is an endogenous hormone, produced by the adrenal cortex, the gonads and in women, its source is the corpus luteum. Progesterone is produced in the late phase of the menstrual cycle, when implantation of the zygote does not occur, the corpus luteum involutes and the release of progesterone is suppressed, thus initiating menstruation. Progestogen Hypersensitivity were initially identified as hormone allergy and were related to endogenous reactions to hormones and alteration of ovarian function. Skin manifestations such as dermatitis or urticaria were initially reported and described as progesterone autoimmune dermatitis, although the immune-mediated mechanism was not clear. Currently there is no standardization for or tests for Progestogen Hypersensitivity diagnosis. In this review, we will address the different diagnostic methods of this disease.
PubMed: 38720769
DOI: 10.3389/falgy.2024.1384140 -
Nature Neuroscience Jun 2024Cortical malformations such as focal cortical dysplasia type II (FCDII) are associated with pediatric drug-resistant epilepsy that necessitates neurosurgery. FCDII...
Cortical malformations such as focal cortical dysplasia type II (FCDII) are associated with pediatric drug-resistant epilepsy that necessitates neurosurgery. FCDII results from somatic mosaicism due to post-zygotic mutations in genes of the PI3K-AKT-mTOR pathway, which produce a subset of dysmorphic cells clustered within healthy brain tissue. Here we show a correlation between epileptiform activity in acute cortical slices obtained from human surgical FCDII brain tissues and the density of dysmorphic neurons. We uncovered multiple signatures of cellular senescence in these pathological cells, including p53/p16 expression, SASP expression and senescence-associated β-galactosidase activity. We also show that administration of senolytic drugs (dasatinib/quercetin) decreases the load of senescent cells and reduces seizure frequency in an Mtor FCDII preclinical mouse model, providing proof of concept that senotherapy may be a useful approach to control seizures. These findings pave the way for therapeutic strategies selectively targeting mutated senescent cells in FCDII brain tissue.
Topics: Animals; TOR Serine-Threonine Kinases; Mice; Humans; Seizures; Senotherapeutics; Cellular Senescence; Dasatinib; Epilepsy; Male; Malformations of Cortical Development; Neurons; Female
PubMed: 38710875
DOI: 10.1038/s41593-024-01634-2 -
Open Biology May 2024The nucleolus is the most prominent liquid droplet-like membrane-less organelle in mammalian cells. Unlike the nucleolus in terminally differentiated somatic cells,...
The nucleolus is the most prominent liquid droplet-like membrane-less organelle in mammalian cells. Unlike the nucleolus in terminally differentiated somatic cells, those in totipotent cells, such as murine zygotes or two-cell embryos, have a unique nucleolar structure known as nucleolus precursor bodies (NPBs). Previously, it was widely accepted that NPBs in zygotes are simply passive repositories of materials that will be gradually used to construct a fully functional nucleolus after zygotic genome activation (ZGA). However, recent research studies have challenged this simplistic view and demonstrated that functions of the NPBs go beyond ribosome biogenesis. In this review, we provide a snapshot of the functions of NPBs in zygotes and early two-cell embryos in mice. We propose that these membrane-less organelles function as a regulatory hub for chromatin organization. On the one hand, NPBs provide the structural platform for centric and pericentric chromatin remodelling. On the other hand, the dynamic changes in nucleolar structure control the release of the pioneer factors (i.e. double homeobox (Dux)). It appears that during transition from totipotency to pluripotency, decline of totipotency and initiation of fully functional nucleolus formation are not independent events but are interconnected. Consequently, it is reasonable to hypothesize that dissecting more unknown functions of NPBs may shed more light on the enigmas of early embryonic development and may ultimately provide novel approaches to improve reprogramming efficiency.
Topics: Animals; Humans; Mice; Cell Nucleolus; Chromatin; Chromatin Assembly and Disassembly; Embryonic Development; Gene Expression Regulation, Developmental; Zygote
PubMed: 38689555
DOI: 10.1098/rsob.230358 -
Molecular Biology and Evolution May 2024Maternal genes have a pivotal role in regulating metazoan early development. As such their functions have been extensively studied since the dawn of developmental... (Comparative Study)
Comparative Study
Maternal genes have a pivotal role in regulating metazoan early development. As such their functions have been extensively studied since the dawn of developmental biology. The temporal and spatial dynamics of their transcripts have been thoroughly described in model organisms and their functions have been undergoing heavy investigations. Yet, less is known about the evolutionary changes shaping their presence within diverse oocytes. Due to their unique maternal inheritance pattern, a high degree is predicted to be present when it comes to their expression. Insofar only limited and conflicting results have emerged around it. Here, we set out to elucidate which evolutionary changes could be detected in the maternal gene expression patterns using phylogenetic comparative methods on RNAseq data from 43 species. Using normalized gene expression values and fold change information throughout early development we set out to find the best-fitting evolutionary model. Through modeling, we find evidence supporting both the high degree of divergence and constraint on gene expression values, together with their temporal dynamics. Furthermore, we find that maternal gene expression alone can be used to explain the reproductive modes of different species. Together, these results suggest a highly dynamic evolutionary landscape of maternal gene expression. We also propose a possible functional dichotomy of maternal genes which is influenced by the reproductive strategy undertaken by examined species.
Topics: Animals; Reproduction; Biological Evolution; Female; Phylogeny; Maternal Inheritance; Evolution, Molecular
PubMed: 38679468
DOI: 10.1093/molbev/msae081 -
Cancer Genetics Jun 2024Germline heterozygous TP53 pathogenic variants (PVs) cause Li Fraumeni Syndrome (LFS, OMIM#151623). TP53 PVs at lower-than-expected variant allele frequencies (VAF) may...
BACKGROUND
Germline heterozygous TP53 pathogenic variants (PVs) cause Li Fraumeni Syndrome (LFS, OMIM#151623). TP53 PVs at lower-than-expected variant allele frequencies (VAF) may reflect postzygotic mosaicism (PZM) or clonal hematopoiesis (CH); however, no guidelines exist for workup and clinical management.
PATIENTS AND METHODS
Retrospective analysis of probands who presented to an academic cancer genetics program with a TP53 PV result on germline genetic testing.
RESULTS
Twenty-one of 125 unrelated probands (17 %) were found to harbor a TP53 PV with VAF<30 % or a designation of "mosaic". A diagnosis of PZM was made in nine (43 %) due to a clinical phenotype consistent with LFS with (n = 8) or without (n = 1) positive ancillary tissue testing. Twelve patients (57 %) were diagnosed with presumed CH (pCH) due to a diagnosis of a myeloproliferative neoplasm, negative ancillary tissue testing, clinical phenotype not meeting LFS criteria, no cancer, and/or no first cancer age<50. Of the 19 patients with biological offspring, nine had either partial or complete offspring testing, all negative.
CONCLUSIONS
Determining the etiology of low VAF TP53 PVs requires ancillary tissue testing and incorporation of clinical phenotype. Discerning PZM versus CH is important to provide optimal care and follow-up.
Topics: Humans; Genetic Testing; Tumor Suppressor Protein p53; Female; Mosaicism; Germ-Line Mutation; Male; Li-Fraumeni Syndrome; Retrospective Studies; Adult; Middle Aged; Young Adult; Adolescent
PubMed: 38677009
DOI: 10.1016/j.cancergen.2024.04.002 -
Genes Apr 2024Haploinsufficiency of the gene is implicated in a human neuro-ocular syndrome. Although identified as a nuclear protein highly expressed in the embryonic mouse brain,...
Haploinsufficiency of the gene is implicated in a human neuro-ocular syndrome. Although identified as a nuclear protein highly expressed in the embryonic mouse brain, molecular function remains elusive. This study explores the spatio-temporal expression of zebrafish co-orthologs, and , as a first step to elucidate their function. In silico analysis reveals high evolutionary conservation in the DNA-interacting domains for both orthologs, with significant syntenic conservation observed for the locus. In situ hybridization and RT-qPCR analyses on zebrafish embryos and larvae reveal distinct expression patterns: is expressed early in zygotic development, mainly in the central nervous system, while expression initiates during gastrulation, localizing later to dopaminergic telencephalic and diencephalic cell clusters. Both transcripts are enriched in the ganglion cell and inner neural layers of the 72 hpf retina, with widely distributed in the ciliary marginal zone. In the adult brain, and are found in the cerebellum, amygdala and ventral telencephalon, which represent the main areas affected in autistic patients. Overall, this study suggests 's potential involvement in eye and brain development, laying the groundwork for further investigations into -related neurobehavioral disorders.
Topics: Animals; Brain; Gene Expression Regulation, Developmental; Retina; Zebrafish; Zebrafish Proteins; Nerve Tissue Proteins; Membrane Proteins
PubMed: 38674426
DOI: 10.3390/genes15040492 -
Life (Basel, Switzerland) Apr 2024Commencing with sperm-egg fusion, the early stages of metazoan development include the cleavage and formation of blastula and gastrula. These early embryonic events play...
Commencing with sperm-egg fusion, the early stages of metazoan development include the cleavage and formation of blastula and gastrula. These early embryonic events play a crucial role in ontogeny and are accompanied by a dramatic remodeling of the gene network, particularly encompassing the maternal-to-zygotic transition. Nonetheless, the gene expression dynamics governing early embryogenesis remain unclear in most metazoan lineages. We conducted transcriptomic profiling on two types of gametes (oocytes and sperms) and early embryos (ranging from the four-cell to the gastrula stage) of an economically valuable flatfish-the Chinese tongue sole (Pleuronectiformes: Cynoglossidae). Comparative transcriptome analysis revealed that large-scale zygotic genome activation (ZGA) occurs in the blastula stage, aligning with previous findings in zebrafish. Through the comparison of the most abundant transcripts identified in each sample and the functional analysis of co-expression modules, we unveiled distinct functional enrichments across different gametes/developmental stages: actin- and immune-related functions in sperms; mitosis, transcription inhibition, and mitochondrial function in oocytes and in pre-ZGA embryos (four- to 1000-cell stage); and organ development in post-ZGA embryos (blastula and gastrula). These results provide insights into the intricate transcriptional regulation of early embryonic development in Cynoglossidae fish and expand our knowledge of developmental constraints in vertebrates.
PubMed: 38672775
DOI: 10.3390/life14040505