Duct Formation Research Articles

A Cadaveric study on Branching Patterns of the Right Intrahepatic Bile Ducts at the Hepatic Hilum

Objective: Comprehensive anatomical knowledge in branching patterns of intrahepatic bile ducts at the hepatic hilum is useful in liver resection and transplantation. This study was aimed on the anatomical relation of the right anterior and posterior segmental branches to the formation of the right hepatic duct and on the variants of their drainage. Methods: From the year 2022 to 2023, 25 fresh liver specimens were dissected. The extrahepatic bile duct was identified initially. Then the right hepatic duct identified followed by the right anterior and posterior segmental branches were delineated via liver parenchymal dissection. Results: In our study, 19/25 (76%) of liver specimens had the right posterior duct ran posterior to the right anterior duct and fused it from a left (medial) sided approach to form the right hepatic duct. An ectopic drainage of the right anterior segmental duct into the common hepatic duct was noted in 3/25 (12%). A variant of “triple confluence”, was encountered in 1/25 (4%). Drainage of the right posterior hepatic duct into the left hepatic duct before its confluence with the right anterior duct was presented in 1/25 (4%). The direct drainage of the right posterior duct into the common hepatic duct was present in 1/25 (4%). There was no ectopic drainage of the right anterior duct into the left hepatic ductal system, and an ectopic drainage of the right posterior duct into the cystic duct. Conclusion: Variations of the right hepatic duct confluence were less. However, two most common variations were the ectopic drainage of the right anterior duct into the common hepatic duct and the triple confluence.

Generation of human iPSC-derived 3D bile duct within liver organoid by incorporating human iPSC-derived blood vessel

In fetal development, tissue interaction such as the interplay between blood vessel (BV) and epithelial tissue is crucial for organogenesis. Here we recapitulate the spatial arrangement between liver epithelial tissue and the portal vein to observe the formation of intrahepatic bile ducts (BDs) from human induced pluripotent stem cells (hiPSC). We co-culture hiPSC-liver progenitors on the artificial BV consisting of immature smooth muscle cells and endothelial cells, both derived from hiPSCs. After 3 weeks, liver progenitors within hiPSC-BV-incorporated liver organoids (BVLO) differentiate to cholangiocytes and acquire epithelial characteristics, including intercellular junctions, microvilli on the apical membrane, and secretory functions. Furthermore, liver surface transplanted-BVLO temporarily attenuates cholestatic injury symptoms. Single cell RNA sequence analysis suggests that BD interact with the BV in BVLO through TGFβ and Notch pathways. Knocking out JAG1 in hiPSC-BV significantly attenuates bile duct formation, highlighting BVLO potential as a model for Alagille syndrome, a congenital biliary disease. Overall, we develop a novel 3D co-culture method that successfully establishes functional human BDs by emulating liver epithelial-BV interaction.

Exploring LGR5 as a prognostic marker of extrahepatic cholangiocarcinoma: insights from expression analysis and clinical correlations.

Leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5) is a cancer stem cell (CSC) marker of colorectal cancer and may be a CSC marker of other cancer types. Few studies have been conducted on LGR5 expression in extrahepatic cholangiocarcinoma (ECC). We analyzed LGR5 expression using RNAscope, a highly sensitive RNA in situ hybridization technique. Fifty-three ECCs were selected from the medical archives at Shinshu University Hospital and analyzed using a tissue microarray. LGR5 expression levels were divided into expression and no expression groups. LGR5 expression and clinicopathological characteristics were analyzed. Among 25 cases, no LGR5-positive dots were identified. Among 28 cases, some LGR5-positive dots were observed in carcinoma cells, together with a wide range of LGR5-positive cells. LGR5 expression was conspicuous in glandular duct formations. Well- to moderately differentiated types showed significantly higher LGR5 expression than the poorly differentiated type (p = 0.0268). LGR5 expression was associated with good overall survival (p = 0.0219) and good disease-free survival (DFS) (p = 0.0228). High LGR5 expression was associated with well- to moderately-differentiated types, indicating a favorable prognosis. In terms of DFS, multivariate analysis showed that high LGR5 expression was an independent favorable prognostic factor (p = 0.0397). These findings suggest that LGR5 is a promising, novel prognostic marker.

Spatiotemporal Analysis of Sonar Detection Range in Luzon Strait

Sonar serves as a critical submarine detection apparatus for naval vessels, with its detection range forming the foundation of its overall performance in underwater surveillance. The Luzon Strait, in the eastern part of the South China Sea, presents a complex hydrographic setting that profoundly influences sonar performance, necessitating mastery of the detection range variation for enhanced anti-submarine operational efficiency. This study employs the Bellhop acoustic propagation model to estimate the transmission loss. Subsequently, a detection probability integration approach is applied to determine the sonar detection range in the Luzon Strait from 2019 to 2023, which is then subjected to statistical analysis. The findings indicate the following. (1) During the summer and autumn, the shallow mixed layer fails to generate a surface duct, resulting in shorter detection ranges that are primarily dependent on the water depth. In the Shallow Water Zone (<150 m), frequent interactions between sound waves and the sea boundaries lead to considerable acoustic energy attenuation, maintaining a short detection range. In the Intermediate Depth Zone (150–2500 m), sound rays retain adequate energy post-seabed reflection, extending the sonar detection to 5–8 km. Beyond 2500 m, the diminishing reflective energy restricts the range to 2–5 km. (2) Conversely, in the winter and spring, the formation of a surface duct becomes the predominant determinant of the detection range, capable of exceeding 10 km, overshadowing the influence of the water depth.

Sox9 links biliary maturation to branching morphogenesis

During liver development, bipotential epithelial progenitor cells, termed hepatoblasts, give rise to hepatocytes and biliary epithelial cells (BECs). BECs line the intrahepatic bile ducts which are essential for liver function. The adult intrahepatic bile duct (IHBD) network is hierarchical in structure with smaller, peripheral “ductules” draining into larger “ducts” that eventually exit the liver. Despite proposed models from histological and genetic studies, mechanisms differentiating local “duct” or “ductule” hierarchies and driving the global IHBD architecture are poorly understood. Previous studies of IHBD development have focused on mechanisms of BEC specification/ de novo bile duct formation by tubulogenesis. However, the contribution of pre-existing BECs to IHBD expansion and morphogenesis is unclear. Previous studies on liver specific Sox9 knockout (Sox9cKO) mice showed that Sox9 is important for timing of BEC specification during development and prevents cystogenesis in aged mice. We hypothesize that SOX9 is an important regulator of IHBD local and global architecture during development by promoting IHBD branching morphogenesis. Using whole-lobe 3D imaging and iDISCO+, we find that adult, embryonic, and postnatal Sox9cKO mice demonstrate ductal paucity with bias towards ductule, rather than duct, loss. Bulk RNA-sequencing reveals that Sox9cKO BECs are transcriptionally “immature” and have gene expression signatures consistent with a failure to fully acquire BEC identity. scRNA-sequencing identifies ductule-like cells in a subset of Sox9cKO BECs correlating with elevated Inhba, the gene precursor of Activin A. Sox9cKO biliary organoids exhibit non-cystic morphology that is reproduced in Activin A treated WT organoids. In vitro and in vivo, Activin A signaling inhibition experiments partially rescue BEC morphology. Together, our data demonstrate that SOX9 drives BEC maturation and promotes ductule formation, in part through inhibiting Activin A. Our data support a model for IHBD development involving distinct mechanisms governing Sox9-independent large duct tubulogenesis versus Sox9-dependent small ductule branching morphogenesis. F31DK134199; PI: Hannah Hrncir T34GM149511; PI: Anita Corbett, PhD; R01DK132653; PI: Adam Gracz, PhD. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Dual Deletion of Keap1 and Rbpjκ Genes in Liver Leads to Hepatomegaly and Hypercholesterolemia.

The hepatic deletion of Rbpjκ (RbpjF/F::AlbCre) in the mouse leads to exhibition of the Alagille syndrome phenotype during early postnatal liver development with hyperlipidemia and cholestasis due to attenuated disruption of NOTCH signaling. Given the roles of NRF2 signaling in the regulation of lipid metabolism and bile ductal formation, it was anticipated that these symptoms could be alleviated by enhancing NRF2 signaling in the RbpjF/F::AlbCre mouse by hepatic deletion of Keap1 in compound Keap1F/F::RbpjF/F::AlbCre mice. Unexpectedly, these mice developed higher hepatic and plasma cholesterol levels with more severe cholestatic liver damage during the pre-weaning period than in the RbpjF/F::AlbCre mice. In addition, hypercholesterolemia and hepatic damage were sustained throughout the growth period unlike in the RbpjF/F::AlbCre mouse. These enhanced abnormalities in lipid metabolism appear to be due to NRF2-dependent changes in gene expression related to cholesterol synthetic and subsequent bile acid production pathways. Notably, the hepatic expression of Cyp1A7 and Abcb11 genes involved in bile acid homeostasis was significantly reduced in Keap1F/F::RbpjF/F::AlbCre compared to RbpjF/F::AlbCre mice. The accumulation of liver cholesterol and the weakened capacity for bile excretion during the 3 pre-weaning weeks in the Keap1F/F::RbpjF/F::AlbCre mice may aggravate hepatocellular damage level caused by both excessive cholesterol and residual bile acid toxicity in hepatocytes. These results indicate that a tuned balance of NOTCH and NRF2 signaling is of biological importance for early liver development after birth.

Van Gogh-like 2 is essential for the architectural patterning of the mammalian biliary tree

In the developing liver, bipotent epithelial progenitor cells undergo lineage segregation to form hepatocytes, which constitute the bulk of the liver parenchyma, and biliary epithelial cells (cholangiocytes), which comprise the bile duct (a complex tubular network that is critical for normal liver function). Notch and TGFβ signalling promote the formation of a sheet of biliary epithelial cells, the ductal plate, that organises into discontinuous tubular structures. How these structures elongate and connect to form a continuous duct remains undefined. We aimed to define the mechanisms by which the ductal plate transitions from a simple sheet of epithelial cells into a complex and connected bile duct. By combining single-cell RNA sequencing of embryonic mouse livers with genetic tools and organoid models we functionally dissected the role of planar cell polarity in duct patterning. We show that the planar cell polarity protein VANGL2 is expressed late in intrahepatic bile duct development and patterns the formation of cell-cell contacts between biliary cells. The patterning of these cell contacts regulates the normal polarisation of the actin cytoskeleton within biliary cells and loss of Vangl2 function results in the abnormal distribution of cortical actin remodelling, leading to the failure of bile duct formation. Planar cell polarity is a critical step in the post-specification sculpture of the bile duct and is essential for establishing normal tissue architecture. Like other branched tissues, such as the lung and kidney, the bile ducts use planar cell polarity signalling to coordinate cell movements; however, how these biochemical signals are linked to ductular patterning remains unclear. Here we show that the core planar cell polarity protein VANGL2 patterns how cell-cell contacts form in the mammalian bile duct and how ductular cells transmit confluent mechanical changes along the length of a duct. This work sheds light on how biological tubes are patterned across mammalian tissues (including within the liver) and will be important in how we promote ductular growth in patients where the duct is mis-patterned or poorly formed.

Subsurface acoustic ducts in the Northern California current system.

This study investigates the subsurface sound channel or acoustic duct that appears seasonally along the U.S. Pacific Northwest coast below the surface mixed layer. The duct has a significant impact on sound propagation at mid-frequencies by trapping sound energy and reducing transmission loss within the channel. A survey of the sound-speed profiles obtained from archived mooring and glider observations reveals that the duct is more prevalent in summer to fall than in winter to spring and offshore of the shelf break than over the shelf. The occurrence of the subsurface duct is typically associated with the presence of a strong halocline and a reduced thermocline or temperature inversion. Furthermore, the duct observed over the shelf slope corresponds to a vertically sheared along-slope velocity profile, characterized by equatorward near-surface flow overlaying poleward subsurface flow. Two potential duct formation mechanisms are examined in this study, which are seasonal surface heat exchange and baroclinic advection of distinct water masses. The former mechanism regulates the formation of a downward-refracting sound-speed gradient that caps the duct near the sea surface, while the latter contributes to the formation of an upward-refracting sound-speed gradient that defines the duct's lower boundary.

Rhodomyrtus tomentosa as a new anticancer molecular strategy in breast histology via Her2, IL33, EGFR, and MUC1.

The prevalence of breast cancer among patients in Indonesia is significant. Indonesian individuals maintain the belief that cancer cannot be cured alone by pharmaceuticals and treatment; herbal remedies must be used in conjunction. Rhodomyrtus tomentosa, also known as Haramonting, is an indigenous Indonesian medicinal plant renowned for its copious antioxidant properties. The objective of study was to assess the impact of haramonting on breast cancer by examining the expression of various biomarker proteins associated with breast cancer. Haramonting was administered to breast cancer model mice at different doses over a period of 30days. Subsequently, blood and breast samples were obtained for immunohistochemistry and enzyme-linked immunosorbent assays (ELISA). Authors have discovered that there has been a notable rise in the proliferation of epithelial cells in the duct lobes, resulting in the formation of ducts and lobules. Additionally, the researchers discovered that the breasts exhibited distinct clinical and histological alterations. Haramonting possesses the capacity to restore the concentrations of malondialdehyde (MDA) and superoxide dismutase (SOD) to normal levels in the blood serum of rats afflicted with cancer. The histopathological analysis of the breast tissue revealed elevated levels of Her2, IL33, EGFR, and MUC1. The authors also discovered a notable increase in the growth of epithelial cells, with two or more layers of cells reaching towards the centre of the duct. The size of the epithelial cells exhibits variability; however, this state ameliorates with the administration of a dosage of 300mg/kgBW of this botanical specimen. This study proposes that Haramonting may be effective in treating breast cancer.

Salivary Duct Carcinoma Arising in the Submandibular Gland in a Patient with Neurofibromatosis Type 1.

A 71-year-old man with neurofibromatosis type 1 (NF1) presented to our department with a 1-week history of a painful mass in the left submandibular area. Computed tomography (CT) and magnetic resonance imaging revealed an irregular-shaped tumor with a diameter of 2.0 cm in the left submandibular gland and a metastatic lymph node with a diameter of 1.0 cm adjacent to the tumor. Fluorodeoxyglucose-positron emission tomography/CT revealed increased uptake in the tumor. Fine-needle aspiration cytology revealed atypical cells, suggesting salivary duct carcinoma (SDC). Left neck dissection with resection of the tumor and submandibular gland was performed under general anesthesia. Histologic examination revealed ductal formation with a solid, cystic, cribriform, and papillary structure with intraductal comedonecrosis, diagnosing as SDC originating in the submandibular gland (pT3N1M0 pStage III). Mutational analysis of 160 cancer-related genes by next-generation sequencing (NGS) revealed a germline and frameshift mutation in the NF1 gene (p.R2408Kfs*14) and a somatic and frameshift mutation in the TP53 gene (p.C176Wfs*22). The patient received postoperative radiotherapy to the left neck area at 66 Gy. No evidence of recurrence or metastasis has been observed as of 10 months postoperatively. This is the first reported case of SDC in the submandibular gland in a patient with NF1. The mutational data by NGS may contribute to a better understanding of the oncogenesis of SDC in patients with NF1.

Constitutive resin ducts formation in Pinus sylvestris responds to water availability and temperature in southern Siberia

Constitutive resin ducts (RDs) are a distinctive anatomical trait in the xylem of most conifers, playing a clear defensive role against biotic and abiotic factors. Although resin duct dynamics are well documented in Mediterranean environments, less attention has been dedicated to continental climate. In this study, we evaluated the occurrence and climate response of resin ducts in Pinus sylvestris L. trees in five sites along a 3422 km transect of increasing continentality in the forest-steppe of southern Siberia over the 1966–2016 period. Our results suggest a strong climatic dependence of tree growth and resin duct production under the Siberian continental climate conditions. Tree-ring width and RDs abundance were positively affected by summer precipitation and negatively influenced by temperature and solar radiation, suggesting a link between RDs formation and the water deficit period that limits RW. However, the RDs climate signal was generally weaker, with the strongest climate response occurring later in the season, showing independent signals when the RW effect was removed. The site played a key role as an explanatory factor of the climate response variability. Although no clear pattern was found along the transect, potential differences are due to phenotypic variations between the P. sylvestris populations. Warming-induced drought conditions during the growing season, associated to ongoing climate warming, might affect tree growth and resin ducts production with an increase of this critical defensive structure abundance in conifers growing in the forest-steppe in southern Siberia.

Vasculogenic mimicry regulates immune infiltration and mutational status of the tumor microenvironment in breast cancer to influence tumor prognosis.

Vasculogenic mimicry (VM) refers to the direct formation of microcirculatory ducts by invasive malignant tumors via cellular phenotypic transformation. However, there is a lack of VM-based biomarkers for breast cancer. We obtained transcriptomic expression data, single cell sequencing data, and clinical data of patients from The Cancer Genome Atlas Program (TCGA) database and GEO database, performed single cell analysis to obtain specific type annotations of breast cancer cells and analyzed their spatial expression analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene ontology (GO) analyses as well as Gene Set Enrichment Analysis (GSEA) analyses were performed to clarify the biological pathways and tumor functional enrichment relationships of the major expressed genes of VM in the breast cancer bulk data specimens. VM biomarkers were constructed. Meanwhile, the relationship between VM scores and tumor immune infiltration in breast cancer was analyzed using MCPcounter and ssGSEA methods. In addition, we assessed the specific relationship between NDRG1, a key VM gene in breast cancer, and tumor colonization, adhesion and invasion by biological experiments in breast cancer cell lines. The main cell types of breast cancer (BRCA) samples were annotated by single cell transcriptome analysis. Most of the VM-high group was present in epithelial cells, whereas the VM-low group was present in immune and stromal cells. Multiple tumor pathways such as TGFβ p53 and MAPK were closely associated with VM-mediated breast cancer infiltration and invasion. A prognostic model of breast cancer based on VM key genes was constituted. Prognostic stratification of breast cancer was successfully achieved for the TCGA-BRCA and GSE58812 datasets. Through immune infiltration analysis, we found that differential expression of VM markers was associated with multiple immune cell regulation. In MDA-MB-231 and MDA-MB-453 cell lines, we found that the NDRG1 gene significantly promoted colony formation of breast cancer cells. Our constructed VM-related gene-based model of breast cancer biology holds promise for prognostic prediction and patient stratification of breast cancer. This may provide a potentially clinically valuable aid in promoting a deeper understanding of the biological regulation of VM in breast cancer and exploring the specific mechanisms of tumor angiogenesis and breast cancer development.

Gamete-exporting organs of vertebrates: dazed and confused.

Mature gametes are transported externally for fertilization. In vertebrates, the gonads are located within the coelom. Consequently, each species has specific organs for export, which often vary according to sex. In most vertebrates, sperm ducts and oviducts develop from the Wolffian and Müllerian ducts, respectively. However, exceptions exist. Both sexes of cyclostomes, as well as females of basal teleosts, lack genital ducts but possess genital pores. In teleosts of both sexes, genital ducts are formed through the posterior extensions of gonads. These structures appear to be independent of both Wolffian and Müllerian ducts. Furthermore, the development of Wolffian and Müllerian ducts differs significantly among various vertebrates. Are these gamete-exporting organs homologous or not? A question extensively debated around the turn of the 20th century but now largely overlooked. Recent research has revealed the indispensable role of Wnt4a in genital duct development in both sexes of teleosts: zebrafish and medaka. wnt4a is an ortholog of mammalian Wnt4, which has functions in Müllerian duct formation. These results suggest a potential homology between the mammalian Müllerian ducts and genital ducts in teleosts. To investigate the homology of gamete-exporting organs in vertebrates, more detailed descriptions of their development across vertebrates, using modern cellular and genetic tools, are needed. Therefore, this review summarizes existing knowledge and unresolved questions on the structure and development of gamete-exporting organs in diverse vertebrate groups. This also underscores the need for comprehensive studies, particularly on cyclostomes, cartilaginous fishes, basal ray-finned fishes, and teleosts.

SMAD2/3 signaling regulates initiation of mouse Wolffian ducts and proximal differentiation in Müllerian ducts.

Male and female reproductive tracts develop from anterior intermediate mesoderm with similar differentiation processes. The anterior intermediate mesoderm develops into the mesonephros, and the Wolffian duct initiates by epithelialization in the mesonephros. The Müllerian duct invaginates from the coelomic epithelium of the cranial mesonephros for ductal formation and is then regionalized into proximal to caudal female reproductive tracts. In this study, we focused on the epithelialization of the Wolffian duct, initiation of the Müllerian duct, and the regionalization step of the Müllerian ducts as a continuous process. By using intermediate mesodermal cells from mouse pluripotent stem cells, we identified that inhibition of SMAD2/3 signaling might be involved in the differentiation into mesenchymal cells, after which mesonephric cells might be then epithelialized during differentiation of the Wolffian duct. Aggregation of coelomic epithelial cells might be related to initiation of the Müllerian duct. Transcriptomic analysis predicted that consensus sequences of SMAD3/4 were enriched among highly expressed genes in the proximal Müllerian duct. SMAD2/3 signaling to regulate differentiation of the Wolffian duct was continuously activated in the proximal Müllerian duct and was involved in proximal and oviductal regionalization. Therefore, SMAD2/3 signaling may be finely tuned to regulate differentiation from initiation to regionalization steps.

Mettl3-Mediated m6A Methylation Controls Pancreatic Bipotent Progenitor Fate and Islet Formation.

The important role of m6A RNA modification on β cell function has been established, yet how it regulates pancreas development and endocrine differentiation remains unknown. Here, we generated transgenic mice lacking RNA methyltransferase-like 3 (Mettl3) specifically in Pdx1+ pancreatic progenitor cells and found the mutant mice developed hyperglycemia and hypo-insulinemia at 2 weeks of age, with atrophic pancreas, reduced islet mass and abnormal increase in duct formation. At E15.5, Mettl3 deletion caused a significant loss of Ngn3+ endocrine progenitor cells, which was accompanied by increased Sox9+ duct precursor cells. We identified histone deacetylase 1(Hdac1) as the critical direct m6A target in bipotent progenitor, whose degeneration caused abnormal activation of Wnt/Notch signaling pathway and blocked endocrine differentiation. This transformation could be manipulated in embryonic pancreas culture in vitro through the regulation of the axis of Mettl3-Hdac1-Wnt/Notch signaling. Our finding that Mettl3 determines endocrine lineage through modulating Hdac1 activity during the transition of bipotent progenitor might help in the development of targeted endocrine cell protocols for diabetes treatment.

Cholangiocellular adenoma in Franciscana dolphins (Pontoporia blainvillei)

AbstractPontoporia blainvillei, commonly known as the Franciscana dolphin, inhabits the coastal waters of South America and is classified as vulnerable and as undergoing significant population declines. This species is generally found in turbid coastal waters, although it is also found in clear waters, as well as marine areas associated with islands and rocky shores. Tumours originating from cholangiocytes have been rarely described in cetaceans, with descriptions limited to cholangiocellular carcinomas. Cholangiocellular adenomas are characterised by the formation of bile ducts and are rarer than cholangiocellular carcinoma. This study described the first report of a tumour originating from cholangiocytes in P. blainvillei, raising concerns on this endangered cetacean's health status and conservation. The underlying causes of this tumour in Franciscana dolphins remain unknown, highlighting the need for further research.

Cutaneous Metastasis as a Diagnostic Prelude in a 48-year-old Female

Abstract Cutaneous metastasis (CM) describes the spread of a distant primary tumor into the skin. The overall incidence of CM ranges from 5% to 10% with breast cancer having the highest rate in women. CM of breast carcinoma origin may manifest as erysipelas-like erythema on the chest, having distinct raised borders and edema due to lymphatic obstruction termed as carcinoma erysipeloides. In most cases, CM is recognized after the initial diagnosis of primary internal malignancy. However, in 0.6–1% of cases, CM served as the first presenting sign of malignancy. A 48-year-old female presented with multiple, erythematous patches, and plaques with clear-cut raised margins, some topped with violaceous pinpoint papules and nodules on the chest, abdomen, and back. No palpable breast mass was appreciated. There was noted nipple retraction and axillary lymphadenopathy. A 4-mm skin punch biopsy revealed nests of large pleomorphic cells on the papillary dermis admixed with mitotic figures and attempts of ductal formation. CK7 and CEA were positive. Results of ultrasonography and mammogram were highly suspicious of malignancy. Core needle biopsy of the breast mass revealed an invasive ductal carcinoma. In the context of an eczematous presentation on the chest area without palpable nodules or mass on breast examination, a diagnostic challenge is expected. Interestingly, our patient represents a small group of CM having cutaneous lesions as their primary manifestation. A high index of suspicion supplemented with proper clinicopathologic and radiologic correlation is crucial for the diagnosis of CM. A multidisciplinary referral is required for adequate management and overall survival rate.

Wnt4a Is Indispensable for Genital Duct Elongation but Not for Gonadal Sex Differentiation in the Medaka, Oryzias latipes.

In most vertebrates, the oviducts and sperm ducts are derived from the Müllerian ducts and Wolffian ducts, respectively. However, in teleosts, the genital ducts are formed by the posterior extension of gonads in both sexes. Whether the genital ducts of teleosts are newly evolved organs or variants of Müllerian ducts is an important question for understanding evolutionary mechanisms of morphogenesis. One of the genes essential for Müllerian duct formation in mice is Wnt4, which is expressed in the mesenchyme and induces invagination of the coelomic epithelium and its posterior elongation. Here, we addressed the above question by examining genital duct development in mutants of two Wnt4 genes in the medaka (wnt4a is orthologous to mouse Wnt4, and wnt4b is paralogous). The wnt4b mutants had a short body but were fertile with normal genital ducts. In contrast, both male and female wnt4a mutants had their posterior elongation of the gonads stopped within or just outside the coelom. The mutants retained the posterior parts of ovarian cavities or sperm duct primordia, which are potential target tissues of Wnt4a. The gonads of female scl mutants (unable to synthesize sex steroids) lacked these tissues and did not develop genital ducts. Medaka wnt4a was expressed in the mesenchyme ventral to the genital ducts in both sexes. Taken together, the data strongly suggest that the mouse Müllerian ducts and the medaka genital ducts share homologous developmental processes. Additionally, the wnt4a or wnt4b single mutants and the double mutants did not show sex-reversal, implying that both genes are dispensable for gonadal sex differentiation in the medaka.

Lymphatic Defects in Zebrafish sox18 Mutants Are Exacerbated by Perturbed VEGFC Signaling, While Masked by Elevated sox7 Expression.

Mutations in the transcription factor-coding gene SOX18, the growth factor-coding gene VEGFC and its receptor-coding gene VEGFR3/FLT4 cause primary lymphedema in humans. In mammals, SOX18, together with COUP-TFII/NR2F2, activates the expression of Prox1, a master regulator in lymphatic identity and development. Knockdown studies have also suggested an involvement of Sox18, Coup-tfII/Nr2f2, and Prox1 in zebrafish lymphatic development. Mutants in the corresponding genes initially failed to recapitulate the lymphatic defects observed in morphants. In this paper, we describe a novel zebrafish sox18 mutant allele, sa12315, which behaves as a null. The formation of the lymphatic thoracic duct is affected in sox18 homozygous mutants, but defects are milder in both zygotic and maternal-zygotic sox18 mutants than in sox18 morphants. Remarkably, in sox18 mutants, the expression of the closely related sox7 gene is elevated where lymphatic precursors arise. Sox7 could thus mask the absence of a functional Sox18 protein and account for the mild lymphatic phenotype in sox18 mutants, as shown in mice. Partial knockdown of vegfc exacerbates lymphatic defects in sox18 mutants, making them visible in heterozygotes. Our data thus reinforce the genetic interaction between Sox18 and Vegfc in lymphatic development, previously suggested by knockdown studies, and highlight the ability of Sox7 to compensate for Sox18 lymphatic dysfunction.

Three-dimensional human bile duct formation from chemically induced human liver progenitor cells.

Background: The intrahepatic bile ducts (BDs) play an important role in the modification and transport of bile, and the integration between the BD and hepatocytes is the basis of the liver function. However, the lack of a source of cholangiocytes limits in vitro research. The aim of the present study was to establish three-dimensional BDs combined with human mature hepatocytes (hMHs) in vitro using chemically induced human liver progenitor cells (hCLiPs) derived from hMHs. Methods: In this study, we formed functional BDs from hCLiPs using hepatocyte growth factor and extracellular matrix. BDs expressed the typical biliary markers CK-7, GGT1, CFTR and EpCAM and were able to transport the bile-like substance rhodamine 123 into the lumen. The established three-dimensional BDs were cocultured with hMHs. These cells were able to bind to the BDs, and the bile acid analog CLF was transported from the culture medium through the hMHs and accumulated in the lumen of the BDs. The BDs generated from the hCLiPs showed a BD function and a physiological system (e.g., the transport of bile within the liver) when they were connected to the hMHs. Conclusion: We present a novel in vitro three-dimensional BD combined with hMHs for study, drug screening and the therapeutic modulation of the cholangiocyte function.