The orphan G protein coupled receptor GPR101, which is implicated in X-linked acrogigantism (X-LAG), a rare pituitary disorder characterized by rapid growth a few years after birth, has received significant attention for its expression pattern in adult vertebrate tissues. However, the characterization of GPR101 expression during early embryonic development is poorly characterized. In this study, we investigated the spatiotemporal expression patterns of Gpr101 during early embryonic mouse development (E7.5-E15.5) using a global Gpr101 knock-in mouse model with an inserted LacZ reporter, Gpr101. Similar to previously published studies using adult tissues, we found that LacZ reporter expression was largely restricted to regions of the central nervous system. Expression was not detected until E10.5 in a region near the telencephalic vesicle. In contrast to what has been reported in adult tissues, Gpr101 expression was absent in the hypothalamus and pituitary gland during the developmental timepoints assessed. These novel observations provide a more comprehensive characterization of GPR101's expression and may offer insights into its role in growth and development across species.

The regeneration of amputated body parts depends on the activation and inactivation of signaling pathways that regulate various cellular processes, such as proliferation, migration, differentiation, apoptosis, etc. In the case of annelids, gene expression of several members of the canonical Wnt pathway has been detected during regeneration of the primary body axis and it has been determined that alteration of its activity results in deficiencies in blastema and central nervous system formation, as well as in body segmentation. However, most studies have been carried out at very specific times of regeneration, in a few species and mainly in anterior regeneration. In this work, we analyzed the expression of β-catenin throughout anterior and posterior regeneration in Lumbriculus variegatus, using it as a marker for the potential activation of the pathway. We also carried out pharmacological inhibition and overactivation assays of the pathway during blastema formation and growth. We determined that the expression of β-catenin changes in anterior and posterior regeneration in a highly dynamic manner, both in its intensity and in the tissue in which it is expressed as the process progresses. We also described its potential role in chaeta regeneration. In addition, we showed that inhibition and overactivation of the pathway negatively affect blastema formation and do so differently in the anterior and posterior sections. These findings suggest that the canonical Wnt pathway participates in several cellular processes within annelid regeneration and that its activity must be finely regulated to allow the transition between one process and another.

Shroom3 is an actin binding protein integral to apical constriction and apical-basal elongation during mammalian morphogenesis. Shroom3 function has been demonstrated in the development of the heart, neural tube, gut tube, eye, thyroid bud, and kidneys, with mutations linked to human congenital defects including anencephaly, spina bifida, cleft lip and palate, and ventricular septal defects. Genome-wide association studies implicate Shroom3 in human conditions such as chronic kidney disease and heterotaxy. While this suggests that Shroom3 expression continues postnatally, the extent of adult expression patterns remain unexplored. To address this, we first harvested organs from adult mice heterozygous for a LacZ reporter inserted into the Shroom3 allele (Shroom3). Organs were stained in wholemount with X-gal and cleared in glycerol to identify which tissues expressed Shroom3 and to localize the expression within the tissue. Wildtype organs were then harvested and fluorescently stained to localize Shroom3 protein in tissue sub-structures. From this, we have characterized previously unknown Shroom3 expression in many adult organs including the bladder and reproductive systems, as well as established postnatal Shroom3 expression in organs thought to only have developmental expression including the eye and whisker pad. Our results also demonstrate organs which did not show Shroom3 expression, including the liver and pancreas. This data suggests that Shroom3 has roles in a wider number of tissues than previously thought and provides a foundation for future hypotheses regarding the biological and pathological significance of Shroom3.

The SH2B family, which includes SH2B1, SH2B2, and SH2B3, consists of adaptor proteins that possess conserved Src homology 2 (SH2) and pleckstrin homology (PH) domains, playing essential roles as signaling mediators. However, the gene expression patterns of this family during embryonic development are still mostly unclear. In this study, we first investigated the evolutionary conservation of SH2B across multiple species using phylogenetic analysis, which revealed high sequence homology between zebrafish Sh2b and its orthologs in other vertebrates. Subsequently, we examined the expression patterns of sh2b during zebrafish embryogenesis through whole mount in situ hybridization. The findings revealed that all sh2b genes are expressed during the early developmental stages of zebrafish embryos, with a significant concentration in the brain, eyes, and spinal cord. Additionally, sh2b1 was found to be expressed in the lateral line neuromast (lln) support cells. In conclusion, our results suggest that the sh2b family is mainly localized in the brain and eyes, with sh2b1 specifically expressed in the lln support cells. This study offers important insights into the role of the sh2b family in zebrafish embryonic development.

CWC27 is a member of cyclophilin-type peptidyl-prolyl cis-trans isomerase family and takes part in the pre-mRNA splicing. The mutation of its gene, Cwc27, has been verified to be related to cancer, craniofacial and skeletal anomalies, retinal degeneration, and hearing loss. However, expression pattern of Cwc27 in the development of inner ear has not been fully understood. In this study, we studied the spatiotemporal expression of Cwc27 mRNA and CWC27 protein in the mouse inner ear from embryonic day (E) 9 to postnatal 0-day by in situ hybridization and immunohistochemistry staining, respectively. The expression level of Cwc27 was analyzed by quantitative reverse transcription polymerase chain reaction. We found that Cwc27 was expressed early in the otocyst at E9. At E13, Cwc27 was expressed in the cochlear duct, spiral ganglion area, and vestibular organ. The expression level of Cwc27 mRNA reached its maximum level at E13. From E14 onward, Cwc27 was expressed in the sensory epithelium in the cochlear duct and spiral ganglion area. From E18 onward, the distribution of CWC27 protein became more limited to the organ of Corti. Our study revealed Cwc27 might play an important part on promoting the development of developing inner ear.

This correspondence addresses several inconsistencies identified in the article "Kurdish Handwritten Character Recognition Using Deep Learning Techniques," published in Gene Expression Patterns. We commend the authors for their contribution to Kurdish handwriting recognition using deep learning methods. However, critical discrepancies are evident in the model architecture description, class labeling, and model summary. This letter outlines these concerns in detail and suggests revisions to enhance transparency and reproducibility.

Deer antlers exhibit rapid growth during the velvet phase. As a critical endogenous growth factor in animals, midkine (MDK) is likely closely associated with the growth of antlers. However, the spatio-temporal expression pattern of MDK during the velvet phase was unclear. This study explored the physiological role of MDK by analyzing its molecular characterization and spatio-temporal expression dynamics during the growth of sika deer antlers. The study cloned the coding sequences (CDS) of MDK, which spanned 429 bp and encoded 142 amino acids. The results of bioinformatics prediction analysis showed that MDK was an extracellular hydrophilic secreted protein, which was mainly composed of random coil. MDK protein was relatively conserved in evolution and MDK protein of sika deer had the closest relatives to ruminants and the furthest relatives to Aves. The tip tissues (dermis, mesenchyme, precartilage, cartilage) of antlers were collected from three important growth and development nodes (early period, EP. middle period, MP. late period, LP), and quantitative real-time polymerase chain reaction (qRT-PCR) was chosen to detect the spatio-temporal expression of the MDK. The results showed that MDK was expressed in all tissue sites of antler tip in EP, MP, LP. MDK had a consistent expression pattern under all growth periods and was strongly expressed in dermis and cartilage. The expression of MDK was consistently up-regulated in precartilage, whereas it was first up-regulated and then down-regulated in other tissues, and it was highly significant in MP compared to EP and LP (P < 0.01). This study suggested that MDK may regulate the growth of dermis and cartilage tissues mainly by participating in the process of angiogenesis and bone formation, thus promoting the rapid growth of antlers.

Humans have limited capacity to regenerate lost tissues post injury. The ability to modulate regenerative repair of tissues offers possibilities for restoring loss of tissue (organ) structure and function. Zebrafish (Danio rerio) larvae fin fold regeneration model is a simple system to study the process of regeneration and associated cellular mechanisms. Berberine, a plant alkaloid which is known to have wound healing properties shows potential to modulate regeneration. The present study aimed to explore the modulating influence of berberine on the signaling pathways involved in zebrafish larvae transected tail fin fold regeneration. Tail fin fold transection was performed on 3 dpf (days post fertilization) zebrafish larvae treated with Berberine (0.01%) and untreated control (System water (SW)). The larvae were observed under a microscope at 0, 1, 2, 3, 4, 5, hours post transection (hpt). RNA was extracted from Berberine treated and untreated (control) tail fin transected larvae at 4 hpt to perform RNA-seq analysis. PPI (protein-protein interaction) network, Shiny GO functional enrichment and topology analysis of DEGs (differentially expressed genes) was performed. Berberine treated larvae showed an accelerated regeneration growth in their transected tail fin by 4 hpt. Berberine induced accelerated regeneration is associated with the involvement of Insulin, IGF, stress response, jak-stat, cytokine, and cellular reprogramming signaling pathways as per RNA-seq analysis and String PPI network, and Shiny GO functional enrichment analysis of DEGs. Topological analysis using Cytohubba revealed tnfa, stat3, jak2b, igf1, jak1, hsp90aa1.1, stat1a, stat1b, bag3, hsp70, and fosl1a as the key Hub genes in the PPI network. The present study identifies the pathways and the Hub proteins involved in berberine induced accelerated regeneration process in zebrafish larvae.

The impact of gestational stress on reproductive outcomes is well-established, but the effects of pre-gestational stress remain inconclusive. Using female Wistar rats, we demonstrated that pre-gestational stress negatively affects fertility and pregnancy outcomes. The rats were subjected to restraint stress (RS) for 15 days, with 3 h of stress each day, before mating. The RS group exhibited higher levels of corticosterone and prolactin, along with lower levels of adrenocorticotropic hormone (ACTH), indicating a successful stress model. Stressed rats showed reduced fertility and fecundity indices, longer conception times, and decreased levels of ovarian steroids, such as progesterone, testosterone, and estradiol. Additionally, the ovaries of the RS group had fewer antral follicles and more ovarian cysts. Elevated protein levels of cytochrome P450 side-chain cleavage enzyme (CYP11A1) and aromatase (CYP19A1), along with decreased levels of 17β-hydroxysteroid dehydrogenase (17β-HSD), indicating impaired ovarian steroidogenesis in stress exposed rats. In the RS group, there was a significant increase in proteins associated with folliculogenesis, specifically octamer-binding transcription factor 4 (OCT 4) and growth differentiation factor 9 (GDF 9). Additionally, proteins linked to ovulation, such as the prolactin receptor (PRLR), peroxisome proliferator-activated receptor-γ (PPAR-γ), and cyclooxygenase 2 (COX 2), were elevated. The increased levels of PRLR, progesterone receptor (PR), androgen receptor (AR), and estrogen receptor (ER) combined with heightened oxidative stress in the uteri of the RS group, suggest a potential disruption in uterine function. Overall, this research indicates that pre-gestational stress can significantly impact reproductive health by altering gonadotrophin and ovarian steroid dynamics in the female reproductive tract.

All pancreatic lineages originate from a transitory structure known as the multipotent progenitor epithelium (MPE), which is an endodermal placode formed via epithelial stratification. Cells within the MPE undergo de novo lumenogenesis to give rise to an epithelial plexus, which serves as a progenitor niche for subsequent development of endocrine, ductal and acinar cell types. Recent evidence suggests that Hippo signaling is required for pancreatic cell differentiation, but little is known about the function of Hippo signaling in the development of the MPE. Here, we characterize the expression of YAP1, TAZ, and the Hippo regulators LATS1/2 kinases and MERLIN in early murine pancreatic epithelium, during epithelial stratification, plexus development and emergence of endocrine cells. We find that YAP1 expression is relatively low in the pancreas bud during stratification but increases by E11.5. Intriguingly, we find differing patterns of TAZ and YAP1 immunoreactivty throughout pancreatic development. We further find that MERLIN and LATS1/2 kinases are expressed during the period of rapid stratification and become markedly apical at nascent lumens. To gain a better understanding of how Hippo signaling and lumen formation are connected, we analyzed the subcellular localization of Hippo signaling components during varying stages of lumen formation and found that they are dynamically localized during lumenogenesis. Together, our results point to a previously unsuspected relationship between Hippo signaling and lumen formation during pancreatic development.

The critical importance and remodeling capacity of the blood vasculature within the ovary have been extensively analyzed, while the lymphatic vasculature has received limited attention, and its characteristics were reported by only a few studies.

Chchd10 protein is crucial for sustaining mitochondrial dynamics, physiology and functions, and has been reported to be most abundantly in myocardial cells and skeletal muscle. However, nothing is known for the expression pattern of Chchd10 in gonadal development. Here, we characterized the expression patterns of Chchd10 gene during embryonic gonad development and postnatal testis development in mice, as well as the expression pattern of Chchd10 gene in human puberty testis and young adult testis using publicly available datasets. Besides, we investigated the expression and distribution of Chchd10 in mice testis by RT-qPCR and immunofluorescence and analyzed the possible role and mechanism of Chchd10 in the testis. We noticed that Chchd10 showed abundant expression in embryonic testis compared to ovaries and dynamically expressed during embryonic and postnatal testis development in mice. In addition, Chchd10 was highly abundant within testicular Sertoli cells populations both in embryonic and postnatal mice and mainly located in the mitochondria of Sertoli cells in mice. Furthermore, CHCHD10 was not only enriched in Sertoli cells, but also highly expressed in tMΦ of human puberty testis and adult testis. CHCHD10 may participate in testicular development by regulating multiple biological processes of Sertoli cells. Taken together, our data indicated that Chchd10 appears to be important during testicular development, particularly in the functional modulation of Sertoli cells. Our study revealed the expression profile of Chchd10 gene during testicular development for the first time and will provide new ideas for further studying the function and molecular mechanism of Chchd10.

Recently, transcriptomic analysis has been key in identifying therapeutic targets in cardiovascular regeneration. The postnatal loss of cardiomyocyte proliferative capacity has been linked to the transition from glycolysis to fatty acid oxidation in rodent models of acute myocardial infarction (AMI). However, the transcriptomic profile of these processes in large mammals more similar to humans is still unknown. The aim of this study was to examine the transcriptomic profile, from the proliferative fetal stage to the non-regenerative infarcted adult stage, in an ovine AMI model.

The lateral plate mesoderm of vertebrates, which borders the other mesodermal territories, develops during embryogenesis into a variety of tissues and organs such as blood, heart, vasculature, kidney and smooth muscles. This mesoderm compartment, as well as the unsegmented pharyngeal mesoderm which gives rise to head muscles and part of the heart, have been proposed as vertebrate innovations. Indeed, in the two other chordate clades, the tunicates and the cephalochordates, no such mesoderm regions are formed during development. However, in ascidians, the most studied tunicate group, some cells in the larva which participate to siphon muscles and heart formation are thought to be homologous to the cardiopharyngeal field of vertebrates. Moreover, in the cephalochordate amphioxus, lateral plate and pharyngeal mesoderm marker genes were shown to be expressed in different regions of the fully segmented paraxial mesoderm. In this work, we decided to look at the embryonic expression in amphioxus of several of these mesoderm marker genes, that could give new insights into the putative homology between cephalochordate somite regions and vertebrates' mesoderm compartments. Here, we describe the expression pattern of Erg/Fli1a, Erg/Fli1b, Lmo2, Mesp, Npas4/4l, Osr1/2a, Osr1/2b, Tcf21/Msc and Tcf21/Mscb. Our results highlight the presence of a putative hematopoietic field in the first somite pair as previously proposed, and suggest that some genes were probably specifically recruited during vertebrate evolution for the development of pharyngeal or lateral plate mesoderm derivatives.

Axon guidance signaling pathways, including the Eph/ephrin, Semaphorin, and Slit/Robo pathways, have been found to play crucial roles in cardiac development. Netrin signaling is another well-studied signaling pathway important for axon guidance, but its role in the developing heart has not been investigated. Here, we describe the novel expression pattern of Netrin-1 in the developing murine heart. Transcriptomic analysis of embryonic mouse hearts shows dynamic Netrin-1 expression from E8.5 through E14.5, where Netrin-1 expression preferentially co-localizes with developing trabecular cardiomyocytes. We further demonstrate the spatiotemporal expression pattern of Netrin-1 using a combination of RNA in situ hybridization and Netrin-1 reporter mice. Netrin-1 is expressed in the developing cardiomyocytes with the highest degree of expression within the left ventricular trabecular myocardium, which has not been previously recognized. Additionally, Netrin-1 expression is observed at lower levels in the cardiomyocytes of the right ventricle and atria. This expression pattern supports a role for Netrin signaling in the developing murine myocardium requiring further functional characterization.

Septins are a group of cytoskeletal GTP binding proteins which are involved in different cellular processes, like cell division, exocytosis and axon growth. Their function, especially in the nervous system, is not clear. In zebrafish 16 different septins are described and for some of them the expression in the brain is described. Interestingly, the expression pattern of several of them is highly specific. Here we describe the expression of sept10 and sept12 in the developing zebrafish brain and found that these show a very defined expression pattern. Interestingly, they show an overlap with a group, but not all proliferating PCNA positive cells in nervous tissue.

Deafness is a common genetic disorder, where mutations,in the OTOF gene can disrupt the normal functionof the Otoferlin protein, leading to impaired neurotransmitter release in the inner ear and subsequent deafness. Despite the complexity of the pathogenic mechanism,it is not fully understood. Zebrafish are an excellent model for studying genetically-induced deafness,but there have been no previous reports on the pathogenesis of OTOF in zebrafish.This study successfully established a zebrafish model with mutated OTOF genes using CRISPR/Cas9 gene editing technology to investigate the molecular basis of OTOF-induced deafness. Compared to AB wild type zebrafish, those with low otof expression showed injury and apoptosis of hair cells in the posterior lateral neuromasts along with significant increase in the number of macrophages and apoptotic cells in this region. Additionally, these mutants exhibited a reduction in body length. To further elucidate differences at 5dpf (days post-fertilization) between mutant and wild type zebrafish embryos, RNA-seq analysis was conducted to examine differentially expressed genes (DEGs).A total of 334 up-regulated DEGs and 111 down-regulated DEGs were identified in mutants compared to wild types.KEGG and GO enrichment analyses were performed on these DEGs to identify key signaling pathways and hub DEGs. The findings revealedan increased expression of several genes involved in the HSP70 oxidative stress system, suggesting that OTOF may protect cochlear hair cell from apoptosis induced by oxidative stress through regulation of MAPK signal and HSP70 expression.In summary, the establishment of a zebrafish model with OTOF knockout provides a valuable tool for investigating the function of Otoferlin and understanding the role of the OTOF gene in deafness. These potential molecular insights offer significant contributions towards understanding the pathogenesis of deafness experimental models and serves as a foundation for comprehending the involvement of the OTOF gene.

The Hippo pathway is a critical regulator of animal development. Activation of the Hippo pathway causes a cascade of phosphorylation events that culminate in the phosphorylation of the transcriptional co-factors YAP and TAZ, which limits their entry into the nucleus. When the Hippo pathway is 'off', however, YAP and TAZ can enter the nucleus, where they interact with the transcription factors of the TEA Domain (TEAD) family to regulate transcriptional activity. Despite the importance of the Hippo pathway for development, including within the nervous system, the expression of the TEAD family remains poorly defined in mammals. Here, we mapped the expression of TEAD1 in the developing mouse brain. We find that TEAD1 expression is confined to progenitor cells during embryonic development, namely radial glia and intermediate progenitor cells. TEAD1 expression is not evident in post-mitotic neurons of the cortical plate. We also identify expression of TEAD1 in developing and mature ependymal cells of the lateral and third ventricle, including within the subcommissural organ, as well as by cells within the choroid plexuses and the forebrain neurogenic niches. Finally, we find that adult mice conditionally heterozygous for Tead1 in the central nervous system exhibit a significantly smaller brain. Collectively, these findings reveal a specific pattern of expression for TEAD1 during telencephalic development and implicate this factor in regulating neural progenitor cell proliferation.

Smoc1 and Smoc2, members of the SPARC family of genes, encode signaling molecules downstream of growth factors such as the TGF-β, FGF, and PDGF families. Smoc1 has been implicated in playing a crucial role in microphthalmia with limb anomalies in humans and mice, while Smoc2 deficiency causes dental developmental defects. Although developmental cytokines/growth factors including TGF-β superfamily have been shown to play critical roles in postnatal spermatogenesis, there are no reports analyzing the spatial and temporal expression of Smoc1 and Smoc2 in the postnatal testis. In this study, we investigated the mRNA and protein expression of Smoc1 and Smoc2 in neonatal, juvenile, and adult mouse testes by RNA in situ hybridization, immunofluorescence, and single-cell RNA-seq analysis. We show that Smoc1 and Smoc2 have distinct expression patterns in male germ cells: Smoc1 is more highly expressed than Smoc2 in the germline. In contrast, Smoc2 is highly expressed in testicular somatic cells from neonatal to juvenile stages. The Smoc2-expressing cells then switch from somatic cells to germ cells in adults. Thus, although SMOC1 and SMOC2 proteins are structurally very similar, their spatial and temporal expression patterns in the postnatal testis differ significantly, suggesting their distinct roles in reproduction.

The ABC transporter gene family encodes proteins that form transmembrane channels and hydrolyze ATP to pump various substrates into and out of cells. Adult stem cell populations in numerous mammalian tissues express members of this gene family in order to efflux toxins or vital dyes, thereby conferring the "side population" trait, and cancer stem cells exhibit multidrug resistance upon expression of members of this gene family. In this study we investigated the expression of ABC transporters in a leading model organism stem cell niche, the Drosophila testis. We screened enhancer and gene trap lines with insertions near ABC transporter genes and identified six transporters with tissue-specific expression in the testis (ABCB7, MRP, rdog, CG3164, CG31121, and CG9663). Contrary to our expectation, we did not observe mostly stem cell-restricted expression patterns for these genes. We also report the expression patterns of ABC transporter genes in the testis from the published Fly Cell Atlas consortium single-cell sequencing of the testis (Li et al., 2022). Comparison of the expression levels and patterns of the six positive genes from the enhancer/gene trap screen with the FCA data showed only weak correlation between the two gene expression approaches. While there are reasons that the techniques might show different results, our work highlights the need for caution in over-reliance on single techniques to investigate gene expression.

ROBO4 involves in the stabilization of blood vessel and mediates the migration of hematopoietic stem cell and newborn neuron. However, the patterns of expression and regulation are not quite clear. To resolve this, we analyzed the single cell sequence data, and confirmed that Robo4 mainly expresses in various endothelial cells, but also in epithelial cells, pericytes, and stem or progenitor cells of bone marrow, fibroblast cells/mesenchymal stem cell of adipose tissues, muscle cells and neuron. Robo4 expressions in endothelial cells derived from capillary vessel, tip/stalk/activated endothelial cells were higher than that in artery and large vein (matured endothelial cells). On the other hand, via mining the gene expression data deposited in the NCBI Gene Expression Omnibus database as well as National Genomics Data Center (NGDC), we uncovered that the expression of Robo4 were regulated by different stimulus and variable in diseases' condition.Moreover, we constructed enhanced GFP (eGFP) transgene mouse controlled by Robo4 promoter using CRISPR/CAS9 system. We found GFP signals in many cell types from the embryonic section, confirming a widely expression of Robo4. Together, Robo4 widely and dynamically express in multiple cell types, and can be regulated by diverse factors.