Spatiotemporal Expression Patterns Research Articles

Comparative Transcriptomic Analysis Provides Insight into Spatiotemporal Expression Patterns of Pivotal Genes During Critical Growth Stages in Min Pig Breed

The growth and development of animals are dynamic processes characterized by fluctuations. Min pigs, a local breed renowned for their superior meat quality, present an intriguing yet poorly understood relationship between this quality and their growth and development patterns. To elucidate this relationship, we employed a multi-faceted approach that included comparative transcriptomics, quantitative real-time PCR (qRT-PCR), selection pressure analysis of key genes, and three-dimensional protein structure simulations. Our findings revealed that 150 days (150 d) of age represented a pivotal turning point in the growth and development of Min pigs. Thirteen key genes exhibiting significant differential expression between early and late growth stages were identified. Notably, the CDK2 gene demonstrated specific high expression in the hind limb muscles and adipose tissues during the later growth stages. Comparative analysis with the African warthog revealed that while the CDK2 protein structure remained conserved, base mutations in upstream and downstream non-coding regions resulted in strong positive selection pressure on the CDK2 gene. These results suggest that CDK2 plays a crucial role in defining the spatiotemporal characteristics of meat development during the domestication of Min pigs. This study provides critical insights into the growth and development patterns of domestic pigs and offers a robust scientific foundation for improving meat quality traits through domestication.

The expression of transcription factors in the human fetal subthalamic nucleus suggests its origin from the first hypothalamic prosomere.

In this study, we analyzed the spatio-temporal pattern of expression of specific transcription factors (PITX2, FOXA1, BARHL1, FOXP1, FOXP2) in the human fetal subthalamic nucleus and its neighboring structures from 11 postconceptional weeks (PCW) to 3 postnatal months. We found that all analyzed transcription factors are expressed already during the early fetal period (at 11 PCW). Both FOXP1- and FOXP2-immunoreactive cells were found in the subthalamic nucleus as well as in the striatum, thalamus, reticular nucleus, but not in the zona incerta. FOXP2-ir cells were also found in the lateral hypothalamic-supramamillary area (LHA-SMA) and internal pallidal segment.On the other hand, PITX2, FOXA1 and BARHL1 were expressed exclusively in the subthalamic nucleus and LHA-SMA, from 11 PCW until the birth, the only exception being gradual loss of BARHL1 expression in the LHA-SMA during the late fetal period.Our findings present the first evidence in the human fetal brain that neurons of the subthalamic nucleus do not originate in the diencephalon, as was proposed by classical histological studies, but instead share a common hypothalamic (hp1 prosomere) origin with neurons of the LHA-SMA group, as proposed by the prosomeric model of brain development.

Neuroanatomical and functional substrates of the short video addiction and its association with brain transcriptomic and cellular architecture.

Short video addiction (SVA) has emerged as a growing behavioral and social issue, driven by the widespread use of digital platforms that provide highly engaging, personalized, and brief video content. We investigated the neuroanatomical and functional substrates of SVA symptoms, alongside brain transcriptomic and cellular characteristics, using Inter-Subject Representational Similarity Analysis (IS-RSA) and transcriptomic approaches. Behaviorally, we found that dispositional envy was associated with SVA. Structurally, SVA was positively correlated with increased morphological volumes in the orbitofrontal cortex (OFC) and bilateral cerebellum. Functionally, the dorsolateral prefrontal cortex (DLPFC), posterior cingulate cortex (PCC), cerebellum, and temporal pole (TP) exhibited heightened spontaneous activity, which was positively correlated with SVA severity. Transcriptomic and cellular analyses also showed specific genes linked to gray matter volume (GMV) associated with SVA, with predominant expression in excitatory and inhibitory neurons. These genes showed distinct spatiotemporal expression patterns in the cerebellum during adolescence. This study offers a comprehensive framework integrating structural, functional, and neurochemical evidence to highlight the neural-transcriptomic underpinnings of SVA symptoms in a non-clinical population.

The Multidrug and toxin compound extrusion gene GhTT12 promotes the accumulation of both proanthocyanidins and anthocyanins in Gossypium hirsutum.

The pigments present in the fibers of naturally colored cotton provide excellent antibacterial and environmentally friendly properties, making these colored fibers increasingly favored by the textile industry and consumers. Proanthocyanidins (PAs), the critical pigments responsible for the color of brown cotton fiber, are produced on the endoplasmic reticulum and subsequently transported to the vacuole for polymerization and/or storage. Previous studies have identified GhTT12 as a potential transmembrane transporter of PAs in Gossypium hirsutum, with GhTT12 being a homolog of Arabidopsis Transparent Testa 12 (TT12). Here, we analyzed the spatiotemporal expression pattern of GhTT12, silenced and transiently overexpressed GhTT12 in cotton to confirm its biological function. The GhTT12 protein contains two Multidrug and toxic compound extrusion (MATE) domains and 12 transmembrane helices, and the GhTT12 gene displayed predominant expressions in flowers and fibers of cotton that had higher contents of PAs, particularly in brown cotton, suggesting that GhTT12 may play a role in the transport of PAs in cotton. Silencing or transient overexpression of GhTT12 in cotton resulted in decreased or increased accumulation levels of PAs and anthocyanins (Ans), respectively, accompanied by correspondingly down- or up-regulation of genes involved in PAs synthesis (GhANR) and oxidative polymerization (GhTT10). These findings indicate that GhTT12 may also participate in the biosynthesis of PAs and Ans. Moreover, the silencing of GhTT12 led to a lightening of the color of brown cotton fibers, probably due to the reductions in both PAs content and PAs oxidation. Overall, this study, along with previous research, provides compelling evidence to support the hypothesis that GhTT12 transports PAs and Ans while also regulating their biosynthesis and oxidative polymerization, thereby promoting the accumulation of PAs and Ans in cotton and ultimately affecting the fiber coloration.

FKBP5 as a key regulator of metabolic processes in birds: Insights from chicken pectoral muscle.

FK506-binding protein 5 (FKBP5) is a negative regulator of the glucocorticoid response and may play an important role in regulating metabolic homeostasis in birds. However, limited information is available regarding its role in avian species. This study aimed to clarify the spatiotemporal characteristics of chicken FKBP5 and investigate the effects of exogenous stimuli on its expression. Real-time quantitative PCR (RT-qPCR) was utilized to analyze the spatiotemporal expression patterns of FKBP5 in chickens. Additionally, the impact of exogenous stimuli, including fasting, energy restriction, and insulin/pyruvate/glucose injections, on FKBP5 expression in the pectoral muscle was investigated. The results showed that FKBP5 is broadly expressed in various bird tissues, with dominant expression in insulin-sensitive tissues. The FKBP5 levels in birds are dramatically modulated during development in insulin-sensitive tissues, with the highest expression observed in striated muscle and liver at embryonic day 19 (E19) (P < 0.01). The basal level of FKBP5 in the pectoral muscle of broilers is higher than that in Silky chickens. Insulin administration leads to a rapid drop in blood glucose levels in both breeds, with a slower recovery in AA broilers (P < 0.01). However, FKBP5 expression in the pectoral muscle initially shows a slight drop, followed by a sharp increase over time after insulin administration (P < 0.01). Additionally, breed heterogeneity in FKBP5 expression was observed in the pectoral muscle upon insulin stimulation, with broilers showing stronger insulin sensitivity compared to Silky chickens. A 24-hour fasting period downregulated blood glucose levels (P < 0.01) and FKBP5 expression in the pectoral muscle of AA broilers (P < 0.01). A 30% energy restriction over three weeks slightly reduced blood glucose levels (P = 0.075) and significantly decreased FKBP5 levels in the pectoralis muscle of broilers. In addition, pyruvate increased blood glucose and pectoralis FKBP5 transcription levels at 60 min (P < 0.01), whereas glucose increased blood glucose levels but had no effect on FKBP5 expression. Overall analysis showed a negative correlation between FKBP5 expression in the pectoral muscle and blood glucose levels (ρ = -0.405, P < 0.001). In conclusion, FKBP5 is a key player in the metabolic regulation of birds, with its expression being highly dynamic and tissue-specific. Insulin regulates FKBP5 expression in pectoral muscles in a time-dependent manner, while pyruvate increases FKBP5 levels and fasting/energy restriction reduces FKBP5 mRNA expression in the pectoral muscles.

Multiple allelic configurations govern long-range Shh enhancer-promoter communication in the embryonic forebrain.

Developmental gene regulation requires input from enhancers spread over large genomic distances. Our understanding of long-range enhancer-promoter (E-P) communication, characterized as loops, remains incomplete without addressing the role of intervening chromatin. Here, we examine the topology of the entire Sonic hedgehog (Shh) regulatory domain in individual alleles from the mouse embryonic forebrain. Through sequential Oligopaint labeling and super-resolution microscopy, we find that the Shh locus maintains a compact structure that adopts several diverse configurations independent of Shh expression. The most frequent configuration contained distal E-P contacts at the expense of those more proximal to Shh, consistent with an interconnected loop. Genetic perturbations demonstrate that this long-range E-P communication operates by Shh-expression-independent and dependent mechanisms, involving CTCF binding sites and active enhancers, respectively. We propose a model whereby gene regulatory elements secure long-range E-P interactions amid an inherent architectural framework to coordinate spatiotemporal patterns of gene expression.

CD2AP deficiency aggravates Alzheimer’s disease phenotypes and pathology through p38 MAPK activation

BackgroundAlzheimer’s disease (AD) is the most common form of neurodegenerative disorder, which is characterized by a decline in cognitive abilities. Genome-wide association and clinicopathological studies have demonstrated that the CD2-associated protein (CD2AP) gene is one of the most important genetic risk factors for AD. However, the precise mechanisms by which CD2AP is linked to AD pathogenesis remain unclear.MethodsThe spatiotemporal expression pattern of CD2AP was determined. Then, we generated and characterized an APP/PS1 mouse model with neuron-specific Cd2ap deletion, using immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay, electrophysiology and behavioral tests. Additionally, we established a stable CD2AP-knockdown SH-SY5Y cell line to further elucidate the specific molecular mechanisms by which CD2AP contributes to AD pathogenesis. Finally, the APP/PS1 mice with neuron-specific Cd2ap deletion were treated with an inhibitor targeting the pathway identified above to further validate our findings.ResultsCD2AP is widely expressed in various regions of the mouse brain, with predominant expression in neurons and vascular endothelial cells. In APP/PS1 mice, neuronal knockout of Cd2ap significantly aggravated tau pathology, synaptic impairments and cognitive deficits. Mechanistically, the knockout of Cd2ap activated p38 mitogen-activated protein kinase (MAPK) signaling, which contributed to increased tau phosphorylation, synaptic injury, neuronal apoptosis and cognitive impairment. Furthermore, the phenotypes of neuronal Cd2ap knockout were ameliorated by a p38 MAPK inhibitor.ConclusionOur study presents the first in vivo evidence that CD2AP deficiency exacerbates the phenotypes and pathology of AD through the p38 MAPK pathway, identifying CD2AP/p38 MAPK as promising therapeutic targets for AD.

The lawc gene emerged de novo from conserved genomic elements and acquired a broad expression pattern in Drosophila.

It has recently become evident that the de novo emergence of genes is widespread and documented for a variety of organisms. De novo genes frequently emerge in proximity to existing genes, forming gene overlaps. Here, we present an analysis of the evolutionary history of a putative de novo gene, lawc, which overlaps with the conserved Trf2 gene, which encodes a general transcription factor in Drosophila melanogaster. We demonstrate that lawc emerged approximately 68 million years ago in the 5'-untranslated region of Trf2 and displays an extensive spatiotemporal expression pattern. One of the most remarkable features of the lawc evolutionary history is that its emergence was facilitated by the engagement of Drosophilidae-specific short highly conserved regions located in Trf2 introns. This represents a unique example of putative de novo gene birth involving conserved DNA regions localized in introns of conserved gene. The observed lawc expression pattern may be due to overlap of lawc with the 5'-untranslated region of Trf2. This study not only enriches our understanding of gene evolution but also highlights the complex interplay between genetic conservation and innovation.

Regional homogeneity patterns reveal the genetic and neurobiological basis of State-Trait Anxiety

ObjectiveState anxiety and trait anxiety are differentially mapped in brain function. However, the genetic and neurobiological basis of anxiety-related functional changes remain largely unknown.MethodsParticipants aged 18–30 from the community underwent resting-state fMRI and were assessed with the State-Trait Anxiety Inventory. Using a general linear regression model, we analyzed the effects of state and trait anxiety, as well as their sum and difference (delta), on regional homogeneity (ReHo) in cortical areas. ReHo patterns denote the spatial distribution of ReHo associated with anxiety scores. We further explored the spatial correlations between ReHo patterns and neuromaps, including gene expression, neurotransmitter receptor density, myelination, and functional connectivity gradients, to elucidate the genetic and molecular substrates of these ReHo patterns.ResultsOur findings demonstrated robust spatial correlations between whole-brain ReHo patterns for state and trait anxiety, with trait anxiety and the delta value exhibiting stronger network correlations, notably in the dorsal attention, salience, visual, and sensorimotor networks. Genes highly correlated with ReHo patterns exhibited unique spatiotemporal expression patterns, involvement in oxidative stress, metabolism, and response to stimuli, and were expressed in specific cell types. Furthermore, ReHo patterns significantly correlated with neuromaps of neurotransmitter receptor density, myelination, and functional connectivity gradients.ConclusionsThe ReHo patterns associated with anxiety may be driven by genetic and neurobiological traits. Our findings contribute to a deeper understanding of the pathogenesis of anxiety from a genetic and molecular perspective.

A Single-Cell Atlas of Crab Ovary Provides New Insights Into Oogenesis in Crustaceans.

Oogenesis is crucial for sexual reproduction and provides the material basis for population continuation. Nonetheless, the identity of the cells involved, the nature of transformation, and underlying regulators of oogenesis in crustaceans remain elusive. Here, an atlas of the ovary is plotted via single-nuclei RNA sequencing (snRNA-seq) in the mud crab Scylla paramamosain, resulting in five cell types, including germ cells, somatic cells, and three follicle cell types identified, which in turn provides abundant candidate markers for them. Moreover, profiles of ligand-receptor in different cells of the crab ovary indicate the roles of cell communication in oogenesis. Dozens of transcription factors in the trajectory from oogonia to oocytes as well as the key molecules/pathways in somatic cells and follicle cells relevant to oogenesis are screened, which is evolutionarily conserved and its underlying regulatory mechanism is subject to some modification across various phyla. The spatiotemporal expression patterns of seven markers are further verified and the RNAi confirms the essential roles of piwi and VgR in oogenesis. These data help to elucidate the mechanism underlying gametogenesis and the evolution of reproductive strategy in invertebrates.

Identification and characterization of ATP-binding cassette transporters involved in chlorantraniliprole tolerance of model insect Drosophila melanogaster and agricultural pest Spodoptera frugiperda

ATP-binding cassette (ABC) transporter family is one of the largest transporter families, which plays an important role in insecticide tolerance. In this study, we found that the ABC transporter inhibitor verapamil could significantly enhance the toxicity of chlorantraniliprole (CHL) to the model insect Drosophila melanogaster. Forty-six ABC transporter genes of D. melanogaster were knocked down through the daughterless-GAL4 (Da-GAL4) strain. The subsequent bioassay result showed that D. melanogaster with DmCG5772, DmCG1494, and DmCG5853 genes silencing significantly increased mortality after CHL treatment. Based on the genome of the fall armyworm (FAW), three genes with the best hits were identified, and SfABCA1 (XM_035576510.2) and SfABCG10 (XM_035577893.2) were successfully cloned. Spatiotemporal expression pattern analysis showed that SfABCA1 and SfABCG10 were both highly expressed in adult and pupal stages. Hemolymph was also a tissue with high expression of these two genes. LC10 dose of CHL could induce the expression levels of SfABCA1 and SfABCG10, with SfABCG10 upregulated 8-fold after 48 h of CHL treatment. Furthermore, overexpression of SfABCA1 and SfABCG10 increased the viability of Sf9 cell under CHL treatment. Our findings indicate that SfABCA1 and SfABCG10 might associate with the tolerance of CHL in S. frugiperda. These results are not only helpful in understanding the role of ABC transporters in CHL tolerance of other agricultural pests, but also lay a theoretical foundation for delaying the development of CHL resistance in pest management.

Multi-omics integration analysis identifies pathogenic candidate genes of congenital heart defects with vascular malformation

Abstract Background Congenital heart defects (CHD) with vascular malformation harbors numerous genetic and epigenetic aberrations of DNA copy numbers and methylation. Proteomic deregulation by the aberrations plays key driver roles in heterogeneous progression of CHD. However, it still remains largely unknown about integrative analysis of genetic, epigenetic regulation and proteome in CHD. Purpose We employed a multi-omics approach to explore pathogenic candidate genes in CHD with vascular malformation. Methods Previously, we gathered WES data for 216 CHD patients with vascular malformation and 100 healthy controls. Meanwhile, we carried out DNA methylation analysis from 11 patients and 5 controls. Here, we profile proteomics of vascular tissues from 11 patients and 5 controls. Integration analysis of DNA copy-number-correlated (CNVcor) genes, methylation-correlated (METcor) genes and differentially proteins reveal the innovative and crucial pathogenetic genes. We then detected spatiotemporal expression patterns of candidate genes in zebrafish embryos using in situ hybridization and qPCR, and verified the roles in vascular development by morpholino-mediated gene knockdown. Results 90 differentially expressed proteins (DEPs) were obtained using proteomics and were mainly related to cell adhesion and migration, angiogenesis, amino acid metabolism, and HIF-1 signaling pathway through GO and KEGG pathway enrichment analysis. 15 hub proteins were identified by WGCNA analysis (Fig 1). Additionally, we found that the frequencies of the aberrancies of the CNVcor genes were significantly correlated with the METcor genes (Fig 2 C). We obtained 1924 differentially CNVcor genes and 952 differentially METcor genes. Then, we computed the Pearson’s correlation coefficients of all three-omics by pairwise comparison and found that the distribution of correlations was bimodal (Fig 2 D). Meanwhile, 28 overlapped genes were observed among three-omics. We screened out 11 candidate genes (TIMP1, BGN, LTBP1, AKR1C3, LOXL3, MAOA, MAOB, GNA12, IFI30, CNTNAP1, IGF2BP1) according to the node degree of PPI network analysis between 28 genes and known vascular development related genes. We further figured out LOXL3, MAOA, CNTNAP1 and IGF2BP1 are the novel important genes for vascular malformation, of which CNTNAP1 is the joint gene of three-omics. Through expression patterns and gene knockdown in zebrafish embryos, we revealed that LOXL3, MAOA, CNTNAP1 and IGF2BP1 affect vascular development (Fig 2 E-G). Conclusions We identify novel key candidate genes (LOXL3, MAOA, CNTNAP1 and IGF2BP1) potentially related to CHD with vascular malformation and elucidate the possible molecular pathogenesis of CHD. Our multi-omics integrative analysis provides new insights on the multi-layered etiology of CHD, which might be helpful in developing clinical diagnosis and precaution for CHD.Figure 1Figure 2

Analysis of rosmarinic acid synthase (RAS) gene family and functional study of SmRAS1/2/4 in Salvia miltiorrhiza

Rosmarinic acid synthase is an essential enzyme involved in the biosynthesis of rosmarinic acid (RA), which facilitates the coupling of phenylpropanoid and tyrosine-derived pathway products. Our study identified six SmRAS genes in Salvia miltiorrhiza, with SmRAS1 being the only one functionally characterized to date. Real-time quantitative PCR was employed to analyze the expression profiles of the SmRAS gene family, revealing that SmRAS1/2/4 are predominantly expressed in the roots, which are the medicinal components of S. miltiorrhiza. SmRAS2 and SmRAS4 exhibited significant responses to abscisic acid (ABA), gibberellin (GA3), and methyl jasmonate (MeJA) stimuli, while SmRAS1 had notable responses to GA3 and MeJA. β-glucuronidase (GUS) staining in transgenic Arabidopsis thaliana confirmed a spatiotemporal expression pattern of SmRAS1/2/4 that was consistent with the qRT-PCR results. SmRAS1/2/4 are primarily localized to the cytoplasm and plasma membrane. Our findings suggested that the overexpressions of SmRAS1 or SmRAS4 led to increased levels of salvianolic acid B (SalB) and RA, with a concomitant decrease in the Danshensu (DSS) content, which served as a substrate. In contrast, RNA interference lines exhibited a downward trend in the content of these substances. Interestingly, no significant changes were detected in the SalB, RA, or DSS contents due to the overexpression of SmRAS2 or RNA interference lines. Collectively, our study demonstrated that SmRAS1 and SmRAS4 are key regulators of RA and SalB biosynthesis in S. miltiorrhiza, while SmRAS2's role appears less impactful, suggesting a complex regulatory network that influences the medicinal properties of this plant.

A spatiotemporal transcriptomic atlas of mouse placentation

The placenta, a temporary but essential organ for gestational support, undergoes intricate morphological and functional transformations throughout gestation. However, the spatiotemporal patterns of gene expression underlying placentation remain poorly understood. Utilizing Stereo-seq, we constructed a Mouse Placentation Spatiotemporal Transcriptomic Atlas (MPSTA) spanning from embryonic day (E) 7.5 to E14.5, which includes the transcriptomes of large trophoblast cells that were not captured in previous single-cell atlases. We defined four distinct strata of the ectoplacental cone, an early heterogeneous trophectoderm structure, and elucidated the spatial trajectory of trophoblast differentiation during early postimplantation stages before E9.5. Focusing on the labyrinth region, the interface of nutrient exchange in the mouse placenta, our spatiotemporal ligand–receptor interaction analysis unveiled pivotal modulators essential for trophoblast development and placental angiogenesis. We also found that paternally expressed genes are exclusively enriched in the placenta rather than in the decidual regions, including a cluster of genes enriched in endothelial cells that may function in placental angiogenesis. At the invasion front, we identified interface-specific transcription factor regulons, such as Atf3, Jun, Junb, Stat6, Mxd1, Maff, Fos, and Irf7, involved in gestational maintenance. Additionally, we revealed that maternal high-fat diet exposure preferentially affects this interface, exacerbating inflammatory responses and disrupting angiogenic homeostasis. Collectively, our findings furnish a comprehensive, spatially resolved atlas that offers valuable insights and benchmarks for future explorations into placental morphogenesis and pathology.

Transcriptomic Analysis of Genes Associated with Stinger Development at Different Life Stages of Apis mellifera.

Stingers, evolved from ovipositors, are an important defense organ for the Apidae, Vespidae, and Formicidae species. However, the molecular mechanism of stinger development remains unclear. Here, we show that the earliest time point for the appearance of stingers in Apis mellifera is at the 1-day-old worker pupal stage based on morphological observations and anatomy from the pre-pupal to adult stages. To discover the genes related to stinger development, we first comprehensively compared the stinger transcriptome at different stages and screened 1282, 186, and 166 highly expressed genes in the stingers of 1- and 5-day-old worker pupae and newly emerged worker bees (NEBs), respectively, then identified 25 DEGs involved in the early stage of stinger development. We found that Dll was a key candidate gene in the early development of A. mellifera stingers by combining analyses of the protein-protein interaction network and spatiotemporal expression patterns. An RNAi experiment showed that about 20% of individuals exhibited tip bending in the piercing parts of their stingers in the Dll-dsRNA-treated group, with the morphology presenting as side-side or front-back tip bending. This indicates that Dll plays a vital role in the early development of A. mellifera stingers. Together, our study provides insight into the molecular mechanism of Hymenoptera stinger development and an inspiration for the molecular breeding of gentle honeybee species with stinger abnormalities.

A role of Lhx2 in the migration and axonal projection of cortical postmitotic neurons in the cortical upper layer of the mouse neocortex

The transcription factor LHX2 contains a LIM domain and plays an important role in the development of the vertebrate nervous system. Although much research has been conducted on the function of Lhx2 during cerebral development, its role in postmitotic neuron differentiation in the cerebral cortex remains unknown. Therefore, this study was conducted to determine the function of Lhx2 in dynamic and elaborate developmental processes, including neurogenesis. We first created and confirmed an Lhx2-BAC Gfp transgenic model to three-dimensionally confirm the spatiotemporal expression pattern of Lhx2 during brain development. On this basis, we used the bilateral in utero electroporation technique to express the dominant-negative form of LHX2. LHX2 was confirmed to be important for the migration and callosal projection of postmitotic neurons that form the upper layer of the cerebral cortex during neurogenesis. Additionally, transcriptome analysis confirmed that LHX2 affected the genes involved in neuronal migration and axonal projection. We demonstrated that Lhx2 is important for postmitotic neurons in the cerebral cortex, which migrate to normal positions and extend nerve axons. Taken together, our findings can provide important clues to understanding the relationship between human Lhx2 gene mutations and brain developmental diseases.

Genome-wide investigation of glycoside hydrolase 9 (GH9) gene family unveils implications in orchestrating the mastication trait of Citrus sinensis fruits

Mastication trait of citrus significantly influences the fruit’s overall quality and consumer preference. The accumulation of cellulose in fruits significantly impacts the mastication trait of citrus fruits, and the glycoside hydrolase 9 (GH9) family plays a crucial role in cellulose metabolism. In this study, we successfully identified 32 GH9 genes from the Citrus sinensis genome and subsequently conducted detailed bioinformatics analyses of the GH9 family. Additionally, we profiled the spatiotemporal expression patterns of CsGH9 genes across four distinct fruit tissue types and six crucial developmental stages of citrus fruits, leveraging transcriptome data. Parallel to this, we undertook a comparative analysis of transcriptome profiles and cellulose content among diverse fruit tissues spanning six developmental stages. Furthermore, to identify the pivotal genes involved in cellulose metabolism within the GH9 family during fruit maturity, we employed correlation analysis between cellulose content and gene expression in varying tissues across diverse citrus varieties. This analysis highlighted key genes such as CsGH9A2/6 and CsGH9B12/13/14/22. Collectively, this study provides an in-depth analysis of the GH9 gene family in citrus and offers novel molecular insights into the underlying mechanisms governing the mastication trait formation in citrus fruits.

Reproductive development in Trithuria submersa (Hydatellaceae: Nymphaeales): the involvement of AGAMOUS-like genes

Main conclusionIn the early diverging angiosperm Trithuria submersaTsAG1 and TsAG2 are expressed in different flower organs, including bracts, while TsAG3 is more ovule-specific, probably functioning as a D-type gene.Species of Trithuria, the only genus of the family Hydatellaceae, represent ideal candidates to explore the biology and flower evolution of early diverging angiosperms. The life cycle of T. submersa is generally known, and the “reproductive units” are morphologically well described, but the availability of genetic and developmental data of T. submersa is still scarce. To fill this gap, a transcriptome analysis of the reproductive structures was performed and presented in this work. This analysis provided sequences of MADS-box transcription factors, a gene family known to be involved in flower and fruit development. In situ hybridization experiments on floral buds were performed to describe the spatiotemporal expression patterns of the AGAMOUS genes, revealing the existence of three AG genes with different expression domains in flower organs and in developing ovules. Trithuria may offer important clues to the evolution of reproductive function among early angiosperms and Nymphaeales in particular, and this study aims to broaden relevant knowledge regarding key genes of reproductive development in non-model angiosperms, shaping first flower appearance and evolution.

Involvement of extracellular vesicle microRNA clusters in developing healthy and Rett syndrome brain organoids

Rett syndrome (RTT) is a neurodevelopmental disorder caused by de novo mutations in the MECP2 gene. Although miRNAs in extracellular vesicles (EVs) have been suggested to play an essential role in several neurological conditions, no prior study has utilized brain organoids to profile EV-derived miRNAs during normal and RTT-affected neuronal development. Here we report the spatiotemporal expression pattern of EV-derived miRNAs in region-specific forebrain organoids generated from female hiPSCs with a MeCP2:R255X mutation and the corresponding isogenic control. EV miRNA and protein expression profiles were characterized at day 0, day 13, day 40, and day 75. Several members of the hsa-miR-302/367 cluster were identified as having a time-dependent expression profile with RTT-specific alterations at the latest developmental stage. Moreover, the miRNA species of the chromosome 14 miRNA cluster (C14MC) exhibited strong upregulation in RTT forebrain organoids irrespective of their spatiotemporal location. Together, our results suggest essential roles of the C14MC and hsa-miR-302/367 clusters in EVs during normal and RTT-associated neurodevelopment, displaying promising prospects as biomarkers for monitoring RTT progression.Graphical

Systematic Identification and Characterization of O-Methyltransferase Gene Family Members Involved in Flavonoid Biosynthesis in Chrysanthemum indicum L.

Chrysanthemum indicum L. capitulum is an enriched source of flavonoids with broad-ranging biological activities, mainly due to their anti-inflammatory, anti-cancer, immune regulation, anti-microbial activity, hepatoprotective, and neuroprotective effects. The O-methylation of various secondary metabolites has previously been demonstrated to be mainly catalyzed by S-adenosyl-L-methionine-dependent O-methyltransferase (OMT) proteins encoded by the OMT gene family. However, limited comprehensive study was published on the OMT gene family, especially the CCoAOMT subfamily, involved in the O-methylation of flavonoids in Chrysanthemum. Here, we analyzed the spatiotemporal expression patterns of C. indicum OMT genes in leaf and flower at different developmental stages. Transcriptome sequencing and qRT-PCR analysis showed that COMTs were mainly highly expressed in capitulum, especially in full bloom, while CCoAOMTs were mainly highly expressed in leaves. Correlation analysis of OMT gene expression and flavonoids accumulation revealed that four OMTs (CHR00029120, CHR00029783, CHR00077404, and CHR00078333) were putatively involved in most methylated flavonoids biosynthesis in the capitulum. Furthermore, we identified a true CCoAOMT enzyme, CiCCoAOMT1, and found that it catalyzed O-methylation of quercetin and luteolin at the 3'-OH position. In summary, this work provides an important theoretical basis for further research on the biological functions of OMTs in C. indicum.