Key Developmental Stages Research Articles

Developmental dynamics of sex reprogramming by high incubation temperatures in a dragon lizard

BackgroundIn some vertebrate species, gene-environment interactions can determine sex, driving bipotential gonads to differentiate into either ovaries or testes. In the central bearded dragon (Pogona vitticeps), the genetic influence of sex chromosomes (ZZ/ZW) can be overridden by high incubation temperatures, causing ZZ male to female sex reversal. Previous research showed ovotestes, a rare gonadal phenotype with traits of both sexes, develop during sex reversal, leading to the hypothesis that sex reversal relies on high temperature feminisation to outcompete the male genetic cue. To test this, we conducted temperature switching experiments at key developmental stages, and analysed the effect on gonadal phenotypes using histology and transcriptomics.ResultsWe found sexual fate is more strongly influenced by the ZZ genotype than temperature. Any exposure to low temperatures (28 °C) caused testes differentiation, whereas sex reversal required longer exposure to high temperatures. We revealed ovotestes exist along a spectrum of femaleness to male-ness at the transcriptional level. We found inter-individual variation in gene expression changes following temperature switches, suggesting both genetic sensitivity to, and the timing and duration of the temperature cue influences sex reversal.ConclusionsThese findings bring new insights to the mechanisms underlying sex reversal, improving our understanding of thermosensitive sex systems in vertebrates.

Effects of endocrine disrupting chemicals on gonad development: Mechanistic insights from fish and mammals.

Over the past century, evidence has emerged that endocrine disrupting chemicals (EDCs) have an impact on reproductive health. An increased frequency of reproductive disorders has been observed worldwide in both wildlife and humans that is correlated with accidental exposures to EDCs and their increased production. Epidemiological and experimental studies have highlighted the consequences of early exposures and the existence of key windows of sensitivity during development. Such early in life exposures can have an immediate impact on gonadal and reproductive tract development, as well as on long-term reproductive health in both males and females. Traditionally, EDCs were thought to exert their effects by modifying the endocrine pathways controlling reproduction. Advances in knowledge of the mechanisms regulating sex determination, differentiation and gonadal development in fish and rodents have led to a better understanding of the molecular mechanisms underlying the effects of early exposure to EDCs on reproduction. In this manuscript, we review the key developmental stages sensitive to EDCs and the state of knowledge on the mechanisms by which model EDCs affect these processes, based on the roadmap of gonad development specific to fish and mammals.

Methodological and theoretical advancements in social impacts of tourism research

This lead article introduces the double special issue dedicated to methodological and theoretical advancements in social impacts of tourism research. We begin by providing an overview of five key developmental stages of research within this area: Definitions, typologies, and conceptual model development; the advent of case study-based, atheoretical empirical inquiry; scale design, development, and testing; further scale development/refinement and theoretical application; and theoretical model development and testing. Brief evolutionary histories of the methodological and theoretical advancements of research dedicated to social impacts of tourism are then discussed. This includes a review of the most pertinent predictor variables (along with a visual display of each and key studies) in explaining residents’ perceptions of social impacts of tourism and a thorough review of most frequently used theoretical frameworks. Following this, brief synopses of the articles are provided along with key themes (e.g. resident-tourist relationships, social impacts and residents’ attitudes, residents’ empowerment, overtourism, and methodologies) and salient points of each work. In closing, we suggest numerous lines of inquiry that will continue to advance research into social impacts of tourism. Though these studies were undertaken prior to the COVID-19 outbreak, we emphasize that future work should be designed with the pandemic in mind. Supplemental data for this article is available online at http://dx.doi.org/10.1080/09669582.2022.2046011

Chromosome-level genome of Schistosoma haematobium underpins genome-wide explorations of molecular variation.

Urogenital schistosomiasis is caused by the blood fluke Schistosoma haematobium and is one of the most neglected tropical diseases worldwide, afflicting > 100 million people. It is characterised by granulomata, fibrosis and calcification in urogenital tissues, and can lead to increased susceptibility to HIV/AIDS and squamous cell carcinoma of the bladder. To complement available treatment programs and break the transmission of disease, sound knowledge and understanding of the biology and ecology of S. haematobium is required. Hybridisation/introgression events and molecular variation among members of the S. haematobium-group might effect important biological and/or disease traits as well as the morbidity of disease and the effectiveness of control programs including mass drug administration. Here we report the first chromosome-contiguous genome for a well-defined laboratory line of this blood fluke. An exploration of this genome using transcriptomic data for all key developmental stages allowed us to refine gene models (including non-coding elements) and annotations, discover 'new' genes and transcription profiles for these stages, likely linked to development and/or pathogenesis. Molecular variation within S. haematobium among some geographical locations in Africa revealed unique genomic 'signatures' that matched species other than S. haematobium, indicating the occurrence of introgression events. The present reference genome (designated Shae.V3) and the findings from this study solidly underpin future functional genomic and molecular investigations of S. haematobium and accelerate systematic, large-scale population genomics investigations, with a focus on improved and sustained control of urogenital schistosomiasis.

The Relative Influence of Family and Neighborhood Factors on Child Maltreatment at Critical Stages of Child Development.

This study examines the impact of family and neighborhood factors on physical and psychological abuse across three developmental stages of children: early childhood (age 3), young school age (age 5), and middle childhood (age 9). Data from the Fragile Families and Child Wellbeing Study, a longitudinal national cohort study of children from 20 urban U.S. cities, are used. Path analysis is employed to investigate the longitudinal relationships between family and neighborhood context variables and abuse risk, as well as the importance of different factors at key developmental stages. Economic hardship, maternal substance use, intimate partner violence, and exposure to community violence are found to be related to child abuse risk regardless of developmental stage, while maternal depression and neighborhood informal social control are found to have impacts only within certain child development stages. Findings suggest the need for early intervention and prevention strategies that specifically target economic hardship, poverty, intimate partner violence, and exposure to community violence.

Corn Nitrogen Nutrition Index Prediction Improved by Integrating Genetic, Environmental, and Management Factors with Active Canopy Sensing Using Machine Learning

Accurate nitrogen (N) diagnosis early in the growing season across diverse soil, weather, and management conditions is challenging. Strategies using multi-source data are hypothesized to perform significantly better than approaches using crop sensing information alone. The objective of this study was to evaluate, across diverse environments, the potential for integrating genetic (e.g., comparative relative maturity and growing degree units to key developmental growth stages), environmental (e.g., soil and weather), and management (e.g., seeding rate, irrigation, previous crop, and preplant N rate) information with active canopy sensor data for improved corn N nutrition index (NNI) prediction using machine learning methods. Thirteen site-year corn (Zea mays L.) N rate experiments involving eight N treatments conducted in four US Midwest states in 2015 and 2016 were used for this study. A proximal RapidSCAN CS-45 active canopy sensor was used to collect corn canopy reflectance data around the V9 developmental growth stage. The utility of vegetation indices and ancillary data for predicting corn aboveground biomass, plant N concentration, plant N uptake, and NNI was evaluated using singular variable regression and machine learning methods. The results indicated that when the genetic, environmental, and management data were used together with the active canopy sensor data, corn N status indicators could be more reliably predicted either using support vector regression (R2 = 0.74–0.90 for prediction) or random forest regression models (R2 = 0.84–0.93 for prediction), as compared with using the best-performing single vegetation index or using a normalized difference vegetation index (NDVI) and normalized difference red edge (NDRE) together (R2 < 0.30). The N diagnostic accuracy based on the NNI was 87% using the data fusion approach with random forest regression (kappa statistic = 0.75), which was better than the result of a support vector regression model using the same inputs. The NDRE index was consistently ranked as the most important variable for predicting all the four corn N status indicators, followed by the preplant N rate. It is concluded that incorporating genetic, environmental, and management information with canopy sensing data can significantly improve in-season corn N status prediction and diagnosis across diverse soil and weather conditions.

Identification of High Nitrogen Use Efficiency Phenotype in Rice (Oryza sativa L.) Through Entire Growth Duration by Unmanned Aerial Vehicle Multispectral Imagery.

Identification of high Nitrogen Use Efficiency (NUE) phenotypes has been a long-standing challenge in breeding rice and sustainable agriculture to reduce the costs of nitrogen (N) fertilizers. There are two main challenges: (1) high NUE genetic sources are biologically scarce and (2) on the technical side, few easy, non-destructive, and reliable methodologies are available to evaluate plant N variations through the entire growth duration (GD). To overcome the challenges, we captured a unique higher NUE phenotype in rice as a dynamic time-series N variation curve through the entire GD analysis by canopy reflectance data collected by Unmanned Aerial Vehicle Remote Sensing Platform (UAV-RSP) for the first time. LY9348 was a high NUE rice variety with high Nitrogen Uptake Efficiency (NUpE) and high Nitrogen Utilization Efficiency (NUtE) shown in nitrogen dosage field analysis. Its canopy nitrogen content (CNC) was analyzed by the high-throughput UAV-RSP to screen two mixed categories (51 versus 42 varieties) selected from representative higher NUE indica rice collections. Five Vegetation Indices (VIs) were compared, and the Normalized Difference Red Edge Index (NDRE) showed the highest correlation with CNC (r = 0.80). Six key developmental stages of rice varieties were compared from transplantation to maturation, and the high NUE phenotype of LY9348 was shown as a dynamic N accumulation curve, where it was moderately high during the vegetative developmental stages but considerably higher in the reproductive developmental stages with a slower reduction rate. CNC curves of different rice varieties were analyzed to construct two non-linear regression models between N% or N% × leaf area index (LAI) with NDRE separately. Both models could determine the specific phenotype with the coefficient of determination (R2) above 0.61 (Model I) and 0.86 (Model II). Parameters influencing the correlation accuracy between NDRE and N% were found to be better by removing the tillering stage data, separating the short and long GD varieties for the analysis and adding canopy structures, such as LAI, into consideration. The high NUE phenotype of LY9348 could be traced and reidentified across different years, locations, and genetic germplasm groups. Therefore, an effective and reliable high-throughput method was proposed for assisting the selection of the high NUE breeding phenotype.

Transcriptome comparative analysis of ovarian follicles reveals the key genes and signaling pathways implicated in hen egg production

BackgroundOvarian follicle development plays an important role in determination of poultry egg production. The follicles at the various developmental stages possess their own distinct molecular genetic characteristics and have different biological roles in chicken ovary development and function. In the each stage, several genes of follicle-specific expression and biological pathways are involved in the vary-sized follicular development and physiological events. Identification of the pivotal genes and signaling pathways that control the follicular development is helpful for understanding their exact regulatory functions and molecular mechanisms underlying egg-laying traits of laying hens.ResultsThe comparative mRNA transcriptomic analysis of ovarian follicles at three key developmental stages including slow growing white follicles (GWF), small yellow follicles (SYF) of recruitment into the hierarchy, and differentiated large yellow follicles (LYF), was accomplished in the layers with lower and higher egg production. Totally, 137, 447, and 229 of up-regulated differentially expressed genes (DEGs), and 99, 97, and 157 of down-regulated DEGs in the GWF, SYF and LYF follicles, including VIPR1, VIPR2, ADRB2, and HSD17B1 were identified, respectively. Moreover, NDUFAB1 and GABRA1 genes, two most promising candidates potentially associated with egg-laying performance were screened out from the 13 co-expressed DEGs in the GWF, SYF and LYF samples. We further investigated the biological effects of NDUFAB1 and GABRA1 on ovarian follicular development and found that NDUFAB1 promotes follicle development by stimulating granulosa cell (GC) proliferation and decreasing cell apoptosis, increases the expression of CCND1 and BCL-2 but attenuates the expression of caspase-3, and facilitates steroidogenesis by enhancing the expression of STAR and CYP11A1. In contrast, GABRA1 inhibits GC proliferation and stimulates cell apoptosis, decreases the expression of CCND1, BCL-2, STAR, and CYP11A1 but elevates the expression of caspase-3. Furthermore, the three crucial signaling pathways such as PPAR signaling pathway, cAMP signaling pathway and neuroactive ligand-receptor interaction were significantly enriched, which may play essential roles in ovarian follicle growth, differentiation, follicle selection, and maturation.ConclusionsThe current study provided new molecular data for insight into the regulatory mechanism underlying ovarian follicle development associated with egg production in chicken.

Characterization of phytohormone and transcriptome profiles during protocorm-like bodies development of Paphiopedilum

BackgroundPaphiopedilum, commonly known as slipper orchid, is an important genus of orchid family with prominent horticultural value. Compared with conventional methods such as tillers and in vitro shoots multiplication, induction and regeneration of protocorm-like bodies (PLBs) is an effective micropropagation method in Paphiopedilum. The PLB initiation efficiency varies among species, hybrids and varieties, which leads to only a few Paphiopedilum species can be large-scale propagated through PLBs. So far, little is known about the mechanisms behind the initiation and maintenance of PLB in Paphiopedilum.ResultsA protocol to induce PLB development from seed-derived protocorms of Paphiopedilum SCBG Huihuang90 (P. SCBG Prince × P. SCBG Miracle) was established. The morphological characterization of four key PLB developmental stages showed that significant polarity and cell size gradients were observed within each PLB. The endogenous hormone level was evaluated. The increase in the levels of indoleacetic acid (IAA) and jasmonic acid (JA) accompanying the PLBs differentiation, suggesting auxin and JA levels were correlated with PLB development. Gibberellic acid (GA) decreased to a very low level, indicated that GA inactivation may be necessary for shoot apical meristem (SAM) development.Comparative transcriptomic profiles of four different developmental stages of P. SCBG Huihuang90 PLBs explore key genes involved in PLB development. The numbers of differentially expressed genes (DEGs) in three pairwise comparisons (A vs B, B vs C, C vs D) were 1455, 349, and 3529, respectively. KEGG enrichment analysis revealed that DEGs were implicated in secondary metabolite metabolism and photosynthesis. DEGs related to hormone metabolism and signaling, somatic embryogenesis, shoot development and photosynthesis were discussed in detail.ConclusionThis study is the first report on PLB development in Paphiopedilum using transcriptome sequencing, which provides useful information to understand the mechanisms of PLB development.

Analysis on characteristics of female gametophyte and functional identification of genes related to inflorescences development of Kentucky bluegrass.

The inflorescence is composed of spikes, and the spike is the carrier of grass seed formation and development, so the development status of inflorescence implies grass seed yield and quality. So far, the systematic analysis of inflorescence development of Kentucky bluegrass has not been reported. The development process of the female gametophyte of wild germplasm materials of Kentucky bluegrass in Gannan, Gansu Province of China (KB-GN), was observed. Based on this, the key developmental stages of inflorescence in KB-GN were divided into premeiosis (GPreM), meiosis (GM), postmeiosis (GPostM), and anthesis (GA), and four stages of inflorescence were selected to analyze the transcriptome expression profile. We found that its sexual reproduction formed a polygonum-type embryo sac. Transcriptome analysis showed that 4256, 1125, 1699, and 3127 genes were highly expressed in GPreM, GM, GPostM, and GA, respectively. And a large number of transcription factors (TFs) such as MADS-box, MYB and NAC, AP2, C2H2, FAR1, B3, bHLH, WRKY, and TCP were highly expressed throughout the inflorescence development stages. KEGG enrichment and MapMan analysis showed that genes involved in plant hormone metabolism were also highly expressed at the entire stages of inflorescence development. However, a few TFs belong to stage-specific genes, such as TRAF proteins with unknown function in plants was screened firstly, which was specifically and highly expressed in the GPreM, indicating that TRAF may regulate the preparatory events of meiosis or be essential for the development of megaspore mother cell (MMC). The expression patterns of 15 MADS-box genes were analyzed by qRT-PCR, and the expression results were consistent with that of the transcriptome. The study on the inflorescence development of KB-GN will be great significant works and contribution to illustrate the basic mechanism of grass seeds formation and development.

Transciptome Analysis Molecular Mechanism of Starch Synthesis During Tuber Development in Chinese Yam (Dioscorea opposita)

Yam (Dioscorea opposita) is a kind of vegetables with important nutritional, medicinal and economic value. To reveal the relationship between starch synthesis and gene expression in yam tubers at gene transcription level, transcriptome profiling was conducted by RNA-Seq in Bikeqi yam (Dioscorea opposita Thunb.) tubers at five key developmental stages (105, 120, 135, 150, and 165 days after sowing, DAS). Based on transcriptome sequencing data, a total of 45,867 unigenes were obtained. The results showed that 135 days after sowing are the key period of starch accumulation. During yam tuber development, 1,941 candidate differentially expressed genes (DEGs) were successfully classified into three GO categories, respectively, and there were 292, 267 and 478 unigenes in cellular component, molecular function and biological process. There were 767, 90 and 73 DEGs enriched in metabolic, plant hormone signal transduction and Plant-pathogen interaction pathway by Kyoto Encyclopedia of Genes and Genomes (KEGG), individually. Especially 72 DEGs were enriched in starch and sucrose metabolism pathway. In this pathway, the metabolic process was mainly positive regulated by genes encoding sucrose synthase, glucose-1-phosphate adenylyltransferase, alpha-trehalase, and so on. There was negative regulated by genes encoding beta-glucosidase. 10 DEGs involved in starch synthesis were selected to prove the accuracy of the RNA-Seq data by qPCR, 85% (34/40) of the results were consistent. The results lay a theoretical foundation be used for further understanding the starch synthesis mechanism of yam tubers development and accelerating breeding progress.

Transcriptome Reveals Allele Contribution to Heterosis in Maize.

Heterosis, which has greatly increased maize yields, is associated with gene expression patterns during key developmental stages that enhance hybrid phenotypes relative to parental phenotypes. Before heterosis can be more effectively used for crop improvement, hybrid maize developmental gene expression patterns must be better understood. Here, six maize hybrids, including the popular hybrid Zhengdan958 (ZC) from China, were studied. Maize hybrids created in-house were generated using an incomplete diallel cross (NCII)-based strategy from four elite inbred parental lines. Differential gene expression (DEG) profiles corresponding to three developmental stages revealed that hybrid partial expression patterns exhibited complementarity of expression of certain parental genes, with parental allelic expression patterns varying both qualitatively and quantitatively in hybrids. Single-parent expression (SPE) and parent-specific expression (PSE) types of qualitative variation were most prevalent, 43.73 and 41.07% of variation, respectively. Meanwhile, negative super-dominance (NSD) and positive super-dominance (PSD) types of quantitative variation were most prevalent, 31.06 and 24.30% of variation, respectively. During the early reproductive growth stage, the gene expression pattern differed markedly from other developmental stage patterns, with allelic expression patterns during seed development skewed toward low-value parental alleles in hybrid seeds exhibiting significant quantitative variation-associated superiority. Comparisons of qualitative gene expression variation rates between ZC and other hybrids revealed proportions of SPE-DEGs (41.36%) in ZC seed DEGs that significantly exceeded the average proportion of SPE-DEGs found in seeds of other hybrids (28.36%). Importantly, quantitative gene expression variation rate comparisons between ZC and hybrids, except for transgressive expression, revealed that the ZC rate exceeded the average rate for other hybrids, highlighting the importance of partial gene expression in heterosis. Moreover, enriched ZC DEGs exhibiting distinct tissue-specific expression patterns belonged to four biological pathways, including photosynthesis, plant hormone signal transduction, biology metabolism and biosynthesis. These results provide valuable technical insights for creating hybrids exhibiting strong heterosis.

Late gestation hyperthermia: epigenetic programming of daughter's mammary development and function

Exposure to stressors during early developmental windows, such as prenatally (i.e., in utero), can have life-long implications for an animal's health and productivity. The mammary gland starts developing in utero and, like other developing tissues and organs, may undergo fetal programming. Previous research has implicated factors, such as prenatal exposure to endocrine disruptors or alterations in maternal diet (e.g., maternal over or undernutrition), that can influence the developmental trajectory of the offspring mammary gland in postnatal life. However, the direct links between prenatal insults and future productive outcomes are less documented in livestock species. Research on in utero hyperthermia effects on early-life mammary development is scarce. This review will provide an overview of key developmental milestones taking place in the bovine mammary gland during the pre- and postnatal stages. We will showcase how intrauterine hyperthermia, experienced by the developing fetus during the last trimester of gestation, derails postnatal mammary gland development and impairs its synthetic capacity later in life. We will provide insights into the underlying histological, cellular, and molecular mechanisms taking place at key postnatal developmental life stages, including birth, weaning and the first lactation, that might explain permanent detriments in productivity long after the initial exposure to hyperthermia. Collectively, our studies indicate that prenatal hyperthermia jeopardizes the normal developmental trajectory of the mammary gland from fetal development to lactation. Further, in utero hyperthermia epigenetically programs the udder, and possibly other organs critical to lactation, yielding a less resilient and less productive cow for multiple lactations.

An Experimental Model of Bronchopulmonary Dysplasia Features Long-Term Retinal and Pulmonary Defects but Not Sustained Lung Inflammation.

Bronchopulmonary dysplasia (BPD) is a severe lung disease that affects preterm infants receiving oxygen therapy. No standardized, clinically-relevant BPD model exists, hampering efforts to understand and treat this disease. This study aimed to evaluate and confirm a candidate model of acute and chronic BPD, based on exposure of neonatal mice to a high oxygen environment during key lung developmental stages affected in preterm infants with BPD. Neonatal C57BL/6 mouse pups were exposed to 75% oxygen from postnatal day (PN)-1 for 5, 8, or 14 days, and their lungs were examined at PN14 and PN40. While all mice showed some degree of lung damage, mice exposed to hyperoxia for 8 or 14 days exhibited the greatest septal wall thickening and airspace enlargement. Furthermore, when assessed at PN40, mice exposed for 8 or 14 days to supplemental oxygen exhibited augmented septal wall thickness and emphysema, with the severity increased with the longer exposure, which translated into a decline in respiratory function at PN80 in the 14-day model. In addition to this, mice exposed to hyperoxia for 8 days showed significant expansion of alveolar epithelial type II cells as well as the greatest fibrosis when assessed at PN40 suggesting a healing response, which was not seen in mice exposed to high oxygen for a longer period. While evidence of lung inflammation was apparent at PN14, chronic inflammation was absent from all three models. Finally, exposure to high oxygen for 14 days also induced concurrent outer retinal degeneration. This study shows that early postnatal exposure to high oxygen generates hallmark acute and chronic pathologies in mice that highlights its use as a translational model of BPD.

P–206 Does oocyte vitrifcation affect morphokinetics of subsequent embryo development?

Abstract Study question Are the morphokinetic profiles, as assessed using time-lapse technology, of human embryos developed from vitrified oocytes different to those from fresh oocytes. Summary answer Vitrification of oocytes does have an effect on early developmental morphokinetic profiles, but this is normalized by the time the embryo has reached blastocyst. What is known already Vitrification of oocytes is now commonplace, but little is known about the effect this may have on subsequent embryo development. Study design, size, duration This was a retrospective data analysis, from 8 fertility clinics in the UK between 2012 and 2019. Embryos from patients in the vitrified group (n = 557) were matched to fresh patient controls (n = 539). The matching was performed based on the following criteria: type of treatment, patient age, cause of infertility and number of embryos. Participants/materials, setting, methods The embryos in each group were compared for mean morphokinetics of key developmental stages in hours post insemination (hpi). Parameters compared included early cleavage divisions (t2-t8), time to start of compaction (tSC), time to morula (tM), time to start blastulation (tSB), time to full blastocyst (tB) and duration of compaction (tB-tSC). Treatment outcomes were compared between the two groups, including percentage of blastocyst formation, clinical pregnancy rate, implantation rate and live birth rate. Main results and the role of chance The results showed a significant delay across all early cleavage divisions as follows for vitrified and fresh oocytes respectively: 2-cell (28.14 vs 26.10 (p < 0.001)), 3 cell (37.56 vs 35.37 (p < 0.001)), 4 cell (40.58 vs 37.54 (p < 0.001)), 5 cell (50.31 vs 47.14 (p < 0.001)), 6 cell (53.99 vs 50.87 (p < 0.001)), 7 cell (57.08 vs 54.48 (p < 0.001)) and 8 cell (61.26 vs 58.91 (p < 0.01)). In addition, tSC was also significantly delayed in the vitrified group (80.65 vs 76.36 (p < 0.001)). However, the compaction stage was significantly shorter in the vitrified oocytes (19.02 vs 22.45 (p < 0.001)). Therefore, there was no difference in the time that embryos derived from fresh and vitrified oocytes reached the blastocyst stage (108.03 vs 107.78 (p > 0.05)). No difference was found in clinical pregnancy, implantation or live birth rates but significantly fewer blastocyst developed from vitrified oocytes compared to fresh (36.09% vs 42.4% (p < 0.05)). Limitations, reasons for caution Although this was a matched analysis, it was a retrospective in nature therefore is subject to confounders. However, it would be problematic to perform a prospective randomized controlled trial to address this study question given the need to randomize patients to elective freezing of oocytes prior to embryo creation. Wider implications of the findings: Vitrification of oocytes may affect early developmental morphokinetic profiles, but any effect is normalized by the time the embryo has reached blastocyst. However, fewer blastocysts may develop following oocyte vitrification. This may have implications for oocyte donation banks and those patients choosing to cryopreserve oocytes. Trial registration number NA

Dual role for CXCL12 signaling in semilunar valve development.

SummaryCxcl12-null embryos have dysplastic, misaligned, and hyperplastic semilunar valves (SLVs). In this study, we show that CXCL12 signaling via its receptor CXCR4 fulfills distinct roles at different stages of SLV development, acting initially as a guidance cue to pattern cellular distribution within the valve primordia during the endocardial-to-mesenchymal transition (endoMT) phase and later regulating mesenchymal cell proliferation during SLV remodeling. Transient, anteriorly localized puncta of internalized CXCR4 are observed in cells undergoing endoMT. In vitro, CXCR4+ cell orientation in response to CXCL12 requires phosphatidylinositol 3-kinase (PI3K) signaling and is inhibited by suppression of endocytosis. This dynamic intracellular localization of CXCR4 during SLV development is related to CXCL12 availability, potentially enabling activation of divergent downstream signaling pathways at key developmental stages. Importantly, Cxcr7-/- mutants display evidence of excessive CXCL12 signaling, indicating a likely role for atypical chemokine receptor CXCR7 in regulating ligand bioavailability and thus CXCR4 signaling output during SLV morphogenesis.

An integrated transcriptome and metabolome approach reveals the accumulation of taste-related metabolites and gene regulatory networks during watermelon fruit development.

Accumulation patterns and gene regulatory networks of sugars and cucurbitacins and related primary and secondary metabolites during cultivated watermelon 'Cheng Lan' and wild watermelon 'PI 632,751' fruit development were identified. Metabolites are the end products of cellular regulatory processes and play important roles in fruit taste formation. However, comprehensive studies on the accumulation patterns of watermelon fruit metabolites and transcriptional regulatory networks are still scarce. In this study, 451 annotated metabolites were identified at four key fruit developmental stages in wild watermelon 'PI 632,751' and modern cultivated watermelon 'Cheng Lan'. Interestingly, 11 sugars and 25 major primary metabolites were mainly accumulated in 'Cheng Lan' during fruit development, which are considered to be the potential metabolites beneficial to the formation of watermelon taste. Cucurbitacins and the main flavonoids were mainly specifically accumulated in 'PI 632,751', not being considered to be responsible for the taste. Moreover, forty-seven genes involved in carbohydrate metabolism, glycolysis, and TCA cycle were highly expressed in 'Cheng Lan', which was positively correlated with the accumulation of major primary metabolites. Alternatively, seven UDP-glycosyltransferase genes are closely related to the glycosylation of cucurbitacins through co-expression analysis. Our findings established a global map of metabolite accumulation and gene regulation during fruit development in wild and cultivated watermelons and provided valuable information on taste formation in watermelon fruit.

Utilizing In Vivo Postnatal Electroporation to Study Cerebellar Granule Neuron Morphology and Synapse Development.

Neurons undergo dynamic changes in their structure and function during brain development to form appropriate connections with other cells. The rodent cerebellum is an ideal system to track the development and morphogenesis of a single cell type, the cerebellar granule neuron (CGN), across time. Here, in vivo electroporation of granule neuron progenitors in the developing mouse cerebellum was employed to sparsely label cells for subsequent morphological analyses. The efficacy of this technique is demonstrated in its ability to showcase key developmental stages of CGN maturation, with a specific focus on the formation of dendritic claws, which are specialized structures where these cells receive the majority of their synaptic inputs. In addition to providing snapshots of CGN synaptic structuresthroughout cerebellar development, this technique can be adapted to genetically manipulate granule neurons in a cell-autonomous manner to study the role of any gene of interest and its effect on CGN morphology, claw development, and synaptogenesis.

Highest Defoliation Tolerance in Amaranthus cruentus Plants at Panicle Development Is Associated With Sugar Starvation Responses.

Defoliation tolerance (DT) in Amaranthus cruentus is known to reach its apex at the panicle emergence (PE) phase and to decline to minimal levels at flowering (FL). In this study, defoliation-induced changes were recorded in the content of non-structural carbohydrates and raffinose family oligosaccharides (RFOs), and in the expression and/or activity of sugar starvation response-associated genes in plants defoliated at different vegetative and reproductive stages. This strategy identified sugar-starvation-related factors that explained the opposite DT observed at these key developmental stages. Peak DT at PE was associated with increased cytosolic invertase (CI) activity in all organs and with the extensive induction of various class II trehalose-phosphate synthase (TPS) genes. Contrariwise, least DT at FL coincided with a sharp depletion of starch reserves and with sucrose (Suc) accumulation, in leaves and stems, the latter of which was consistent with very low levels of CI and vacuolar invertase activities that were not further modified by defoliation. Increased Suc suggested growth-inhibiting conditions associated with altered cytosolic Suc-to-hexose ratios in plants defoliated at FL. Augmented cell wall invertase activity in leaves and roots, probably acting in a regulatory rather than hydrolytic role, was also associated with minimal DT observed at FL. The widespread contrast in gene expression patterns in panicles also matched the opposite DT observed at PE and FL. These results reinforce the concept that a localized sugar starvation response caused by C partitioning is crucial for DT in grain amaranth.

Metabolome and Transcriptome Integration Reveals Insights Into Flavor Formation of 'Crimson' Watermelon Flesh During Fruit Development.

Metabolites have been reported as the main factor that influences the fruit flavor of watermelon. But the comprehensive study on the dynamics of metabolites during the development of watermelon fruit is not up-to-date. In this study, metabolome and transcriptome datasets of ‘Crimson’ watermelon fruit at four key developmental stages were generated. A total of 517 metabolites were detected by ultrahigh-performance liquid chromatography–electrospray ionization–tandem mass spectrometry and gas chromatography–solid-phase microextraction–mass spectrometry. Meanwhile, by K-means clustering analysis, the total differentially expressed genes were clustered in six classes. Integrating transcriptome and metabolome data revealed similar expression trends of sugars and genes involved in the glycolytic pathway, providing molecular insights into the formation of taste during fruit development. Furthermore, through coexpression analysis, we identified five differentially expressed ADH (alcohol dehydrogenase) genes (Cla97C01G013600, Cla97C05G089700, Cla97C01G001290, Cla97C05G095170, and Cla97C06G118330), which were found to be closely related to C9 alcohols/aldehydes, providing information for the formation of fruit aroma. Our findings establish a metabolic profile during watermelon fruit development and provide insights into flavor formation.