Noggin is required for normal lobe patterning and ductal budding in the mouse prostate

Mesenchymal expression of the BMP antagonist NOGGIN during prostate development plays a critical role in pre-natal ventral prostate development and opposes BMP4-mediated inhibition of cell proliferation during postnatal ductal development. Morphologic examination of newborn Noggin^−/− male fetuses revealed genitourinary anomalies including cryptorchidism, incomplete separation of the hindgut from the urogenital sinus (UGS), absence of the ventral mesenchymal pad, and a complete loss of ventral prostate (VP) budding. Examination of lobe-specific marker expression in the E14 Noggin^−/− UGS rescued by transplantation under the renal capsule of a male nude mouse confirmed a complete loss of VP determination. More modest effects were observed in the other lobes, including decreased number of ductal buds in the dorsal and lateral prostates of newborn Noggin^−/− males. BMP4 and BMP7 have been shown to inhibit ductal budding and outgrowth by negatively regulating epithelial cell proliferation. We show here that NOGGIN can neutralize budding inhibition by BMP4 and rescues branching morphogenesis of BMP4-exposed UGS in organ culture and show that the effects of BMP4 and NOGGIN activities converge on P63⁺ epithelial cells located at nascent duct tips. Together, these studies show that the BMP-NOGGIN axis regulates patterning of the ventral prostate, regulates ductal budding, and controls proliferation of P63⁺ epithelial cells in the nascent ducts of developing mouse prostate.     

Morphogenesis and ductal development of the prostatic complex of the guinea pig

The morphogenesis of glandular architecture of the three lobes of prostate gland of the guinea pig, lateral, dorsal, and coagulating gland was studied from 35 days gestation to 90 postnatal days. Epithelial ductal tubules of various lobes of the gland were microdissected after treatment by collagenase and displayed two dimensionally. The number of ductal tips was counted, and the volume of the ductal network was quantified using a graphic tablet. The results show that the growth and ductal morphogenesis fall into two phases: prenatal and postnatal. The first outgrowth of prostatic buds begins at 35 days gestation (gestational length is 65 days). Ductal growth and branching continues over the next 15–20 days and by 55 days gestation, approximately 60%, 79%, and 71% of the adult number of ductal tips of the lateral and dorsal lobes and coagulating gland respectively, are formed. The figures increase to 89%, 84%, and 106%, respectively, by birth. There is little increase in number of ductal tips thereafter. Postnatal growth is accomplished mainly by elongation of existing ductal network with a little additional branching but with an increase in size (volume) of the tubules. Canalization of ductal tubules occurs prenatally in all lobes but postnatal functional cytodifferentiation takes a slightly different pace among them. Ductal morphogenesis of the guinea pig prostate gland differs significantly in time-course from that of the mouse in which ductal development occurs mainly postnatally. © 1993 Wiley-Liss, Inc.     

Morphogenesis and histochemistry of the developing mouse kidney

A morphological and histochemical investigation was conducted on the pronephros and mesonephros of the mouse embryo from 8.5 through 16.5 days. The pronephros appeared between days 8.5 and 9.5 as a thickening of the somatic layer of the intermediate cell mass. It consisted of three small clusters of cells on either side of the midline dorsally between the somite and the coelom, at the level of somites 8 and 9. The mesonephros arose during day 9 and persisted until day 16. In the male the anterior three tubules were incorporated into the testis at 15.5–16.5 days. The mesonephros consisted of approximately 11 tubules located between somites 10–17. The tubules possessed lumina and connected with the Wolffian duct. Indications of internal and external glomeruli were noted on day 11. The Wolffian duct reached the cloaca at ten days. Strong alkaline phosphatase activity was noted in the differentiating tubules. Cytoplasmic and luminal enzyme activity was observed between 9.3 and 12.5 days indicating possible function at this time. Acid phosphatase was demonstrable in the tubules and duct only on day 11. Ribonucleic acid was observed in the nuclei and cytoplasm of the mesodermal cells as they differentiated into tubules and duct. A decrease in RNA was noted after differentiation was complete. Periodic acid-Schiff material (diastase-stable) was localized in the basement membrane of the tubule and duct cells. A faint positive reaction was also found at the luminal border of the tubules. The strongest reaction was noted in the luminal border at 11.5–12.5 days. Those tubules being incorporated into the genital system in the male were also PAS positive. Morphological and histochemical evidence suggested that the mouse mesonephros, though quasi vestigial, may function for a short time.     

Morphogenesis of tight junctions in the peritoneal mesothelium of the mouse embryo

The peritoneal mesothelium of mouse embryos (12 to 18 day of gestation) was studied by freeze-fracture and in sections in order to reveal the initial formation of the tight junctions. Freeze-fracture observations showed three types of tight junctions. Type I consists of belt-like meshworks of elevations on the P face and of shallow grooves on the E face. No tight junctional particle can be seen either on the elevations or in the grooves. Type II shows rows of discontinuous particles on the elevations on the P face. Type III consists of strands forming ridges on the P face. On the E face, the grooves of Type II and III appear to be narrower and sharper than those of Type I. Quantitatively, Type I junctions are most numerous during the early stages (day 12-13) of embryonic development, while Type III junctions become more common in the later stages, and are the only type seen by day 18. Observations on sections, however, fail to distinguish between the three types. The results suggest that an initial sign of tight junction formation is close apposition of the two cell membranes in the junctional domain, without tight junctional particles. Later, the particles appear to be incorporated in the tight junctions and the strands form by fusion of the particles.     

Morphogenesis and ultrastructure of the peripolar cells in the mouse kidney.

Peripolar cells are located in the outer layer of the Bowman's capsule. They surround the vascular pole of the renal corpuscle and project into the urinary space. Morphologically they are characterized by the presence of secretory granules within their cytoplasm. In order to study their embryological development, we used 60 C57bl mice embryos (15th to 19th gestational day), 10 newborn mice (2 hours to 6 days old), 10 preadult mice (8-30 days old) and 4 adults (4 months old). Some granular cells, dispersed at the outer and inner layer of the Bowman's capsule, appear on the 17th gestational day. Later, these cells are found around the vascular pole of the renal corpuscle, located exclusively at the outer layer of the Bowman's capsule. Their granules are spherical and variously dense, they are surrounded by a membrane and their number increases progressively with time and reaches a maximum on the 4th postnatal day. Following that, there is a diminution and then their population stabilizes. By the end of the first month, there are only a few such cells (mean number 1 to 2). They become smaller and they always project into the urinary space.     

Biopsy of the mouse prostate

Many transgenic and knockout mouse models of prostate cancer have become available over the past decade. In this paper we describe a simple biopsy technique of the murine prostate. This technique allows sequential follow-up of the prostate in an individual mouse. Its use could also reduce the number of mice used in studies of the prostate gland.     

Morphogenesis and cytochemistry of the postacrosomal dense lamina during mouse spermiogenesis

During the elongation phase of spermiogenesis in the mouse, a layer of electron-dense material appears just below the posterior portion of the acrosomal zonule. Subsequently this material accumulates on the outer side of the nuclear envelope immediately subjacent to the caudal tip of the acrosomal zonule--the anlage of the future postnuclear band--as well as on the inner side of the plasma membrane vis-à-vis to this region--the anlage of the future postacrosomal dense lamina (PADL). Corresponding to further development the postacrosomal region of the nucleus becomes directly enveloped by the plasma membrane, and the PADL, situated on its inner side, grows adequately. The postnuclear band, however, staying the same size as in the preceding elongation phase, gets shifted to the caudal end of the PADL, where it closes the perinuclear space. Since the anlagen and the mature PADL and postnuclear band show the same cytochemical reactions as the dense basal plaque of the acrosomal zonule and the thin layer on the nuclear envelope vis-à-vis to it, a relationship between these structures can be assumed. Furthermore, the demonstration of ribonucleoproteins in all these structures is discussed in connection with a possible nucleolar genesis.     

Expression signature of the mouse prostate

Genetically engineered mice are being used increasingly for delineating the molecular mechanisms of prostate cancer development. Epithelium-stroma interactions play a critical role in prostate development and tumorigenesis. To better understand gene expression patterns in the normal sexually mature mouse prostate, epithelium and stroma were laser-capture microdissected from ventral, dorsolateral, and anterior prostate lobes. Genome-wide expression was measured by DNA microarrays. Our analysis indicated that the gene expression pattern in the mouse dorsolateral lobe was closest to that of the human prostate peripheral zone, supporting the hypothesis that these prostate compartments are functionally equivalent. Stroma from a given lobe had closer gene expression patterns with stroma from other lobes than epithelium from the same lobe. Stroma appeared to have higher expression complexity than epithelium. Specifically, stromal cells had higher expression levels of genes implicated in cell adhesion, muscle development, and contraction, in structural constituents of cytoskeleton and actin binding, and in components such as sarcomere and extracellular matrix collagen. Among the genes that were enriched in the epithelium were secretory proteins, including seminal vesicle protein secretion 2 and 5. Surprisingly, prostate stroma expressed many osteogenic molecules, as confirmed by immunohistochemistry. A "bone-like" environment in the prostate may predispose prostate cells for survival in the bone. Chemokine Cxcl12 but not its receptor, Cxcr4, was expressed in normal prostate. In prostate tumors, interestingly, Cxcl12 was up-regulated in epithelial cells with a concomitant expression of Cxcr4. Expression of both the receptor and ligand may provide an autocrine mechanism for tumor cell migration and invasion.     

Morphogenesis of the lower incisor in the mouse from the bud to early bell stage

The development of the lower incisor in the mouse was investigated from histological sections using computer-aided 3D reconstructions. At ED 13.0, the incisor was still at the bud stage. At ED 13.5, the initial cap was delimited by a short cervical loop, the development of which proceeded on the labial side, but was largely retarded on the medial side. This difference was maintained up to ED 15.0. From ED 16.0, the bell stage was achieved. Metaphases had a ubiquitous distribution both in the enamel organ and in the dental papilla from the bud to early bell stage. Apoptosis gradually increased in the mesenchyme posteriorly to the labial cervical loop from ED 13.5 to 14.0 and then disappeared; this apoptosis was not related to the posterior growth of the incisor. From ED 13.5, a high apoptotic activity was observed in the stalk. A focal area of apoptosis was observed at ED 13.5 in the enamel organ, approaching the epithelio-mesenchymal junction at the future tip of the incisor. There, the inner dental epithelium formed a bulbous protrusion towards dental papilla, reminiscent of the secondary enamel knot of mouse molars. This epithelial protrusion was still maintained at the bell stage. The enamel knot in the incisor demonstrated specific features, different from those characterizing the enamel knot in the molar: the concentric arrangement of epithelial cells was much less prominent and the occurrence of apoptosis was very transitory in the incisor at ED 13.5. The disappearance of the enamel knot despite a low apoptotic activity and the maintenance of the protrusion suggested a histological reorganization specific for rodent incisor.     

Roles for Hedgehog signaling in androgen production and prostate ductal morphogenesis

Previous studies have demonstrated that the Hedgehog (Hh) signaling pathway plays a critical role in the development and patterning of many endodermally derived tissues. We have investigated the role of Sonic hedgehog (Shh) in formation of the prostate gland by examining the urogenital phenotype of Shh mutant fetuses. Consistent with earlier work reporting an essential role for Shh in prostate induction, we have found that Shh mutant fetuses display abnormal urogenital development and fail to form prostate buds. Unexpectedly, however, we have discovered that this prostate defect could be rescued by three different methods: renal grafting, explant culture in the presence of androgens, and administration of dihydrotestosterone (DHT) to pregnant mice, indicating that the prostate defect in Shh mutants is due to insufficient levels of androgens. Furthermore, we find that the inhibition of Hh pathway signaling by treatment with cyclopamine does not block prostate formation in explant culture, but instead produces morphological defects consistent with a role for Hh signaling in ductal patterning. Taken together, our studies indicate that the initial organogenesis of the prostate proceeds independently of Shh, but that Shh or other Hh ligands may play a role in subsequent events that pattern the prostate.     

Repercussions of castration and vasectomy on the ductal system of the rat ventral prostate

Diseases, such as cancer and benign prostatic hyperplasia, are related to disruption of the mechanism regulating the balance between cell proliferation and apoptosis in prostatic cells. Since castration and vasectomy might alter that balance, this study evaluates the cell proliferation, apoptosis and height of the secretory epithelium of the ventral-prostate ductal system post-castration and vasectomy. Immunohistochemical (PCNA and Ki67), cytochemical (Fuelgen reaction) and morphometric investigation have been carried out. Cell proliferation indices decreased significantly in both regions of the ventral-prostate ductal system after castration compared to the sham-operated group. The apoptotic index increased significantly after 48 h, declining 7 days post-castration. The cell proliferation indices did not differ after 48 h significantly; however, they increased 7 days post-vasectomy in both regions. The apoptotic index did not differ significantly in either time post-vasectomy. Castration caused an imbalance in favor of apoptosis, whereas vasectomy caused an imbalance in favor of cell proliferation.     

Analysis of dynamic molecular networks for pancreatic ductal adenocarcinoma progression

Background

Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest solid tumors. The rapid progression of PDAC results in an advanced stage of patients when diagnosed. However, the dynamic molecular mechanism underlying PDAC progression remains far from clear.

Methods

The microarray GSE62165 containing PDAC staging samples was obtained from Gene Expression Omnibus and the differentially expressed genes (DEGs) between normal tissue and PDAC of different stages were profiled using R software, respectively. The software program Short Time-series Expression Miner was applied to cluster, compare, and visualize gene expression differences between PDAC stages. Then, function annotation and pathway enrichment of DEGs were conducted by Database for Annotation Visualization and Integrated Discovery. Further, the Cytoscape plugin DyNetViewer was applied to construct the dynamic protein–protein interaction networks and to analyze different topological variation of nodes and clusters over time. The phosphosite markers of stage-specific protein kinases were predicted by PhosphoSitePlus database. Moreover, survival analysis of candidate genes and pathways was performed by Kaplan–Meier plotter. Finally, candidate genes were validated by immunohistochemistry in PDAC tissues.

Results

Compared with normal tissues, the total DEGs number for each PDAC stage were 994 (stage I), 967 (stage IIa), 965 (stage IIb), 1027 (stage III), 925 (stage IV), respectively. The stage-course gene expression analysis showed that 30 distinct expressional models were clustered. Kyoto Encyclopedia of Genes and Genomes analysis indicated that the up-regulated DEGs were commonly enriched in five fundamental pathways throughout five stages, including pathways in cancer, small cell lung cancer, ECM-receptor interaction, amoebiasis, focal adhesion. Except for amoebiasis, these pathways were associated with poor PDAC overall survival. Meanwhile, LAMA3, LAMB3, LAMC2, COL4A1 and FN1 were commonly shared by these five pathways and were unfavorable factors for prognosis. Furthermore, by constructing the stage-course dynamic protein interaction network, 45 functional molecular modules and 19 nodes were identified as featured regulators for all PDAC stages, among which the collagen family and integrins were considered as two main regulators for facilitating aggressive progression. Additionally, the clinical relevance analysis suggested that the stage IV featured nodes MLF1IP and ITGB4 were significantly correlated with shorter overall survival. Moreover, 15 stage-specific protein kinases were identified from the dynamic network and CHEK1 was particularly activated at stage IV. Experimental validation showed that MLF1IP, LAMA3 and LAMB3 were progressively increased from tumor initiation to progression.

Conclusions

Our study provided a view for a better understanding of the dynamic landscape of molecular interaction networks during PDAC progression and offered potential targets for therapeutic intervention.

     

Morphogenesis and secretory activity of mouse mammary cultures on EHS biomatrix

Summary The nature of the substratum profoundly influences the growth and function of epithelia in tissue culture. Mammospheres, hollow spherical structures, develop when epithelial clusters are plated on a biomatrix derived from the Engelbreth-Holm-Swarm murine tumour (EHS matrix). Morphologic examination of mammosphere development demonstrates that morphogenesis is a two stage process. Over the first 48 h the cells aggregate into spheres, drawing the matrix up and over themselves to become buried within the material. Changes in matrix morphology emphasize the importance of the quasi-fluid nature of the substratum on which the cells are plated. Lumen formation ensues over the next 2 to 5 days as the cells, polarized by basal contact with the matrix, differentiate. They form tight junctions at their apical borders and synthesize milk proteins, secreting caseins into the enlarging interior cavity and transferrin from their basal surfaces into the medium. These experiments demonstrate that the physical properties of the EHS matrix allow epithelial cells to develop the cuboidal shape necessary for secretory activity.Abbreviations EHS Engelbreth-Holm-Swarm biomatrix     

Induction of mouse pancreatic ductal differentiation, an in vitro assay

Despite recent technical advances for studying lineage tracing and gene functions, our knowledge of pancreatic duct progenitor cells and mechanisms involved in their differentiation remains a huge void in our understanding of pancreatic development. A deeper insight into ductal differentiation is needed because ductal cells may harbor pancreatic stem/progenitor cells that could give rise to new islets. Also, since the most common pancreatic tumors form structures expressing ductal cell-specific markers, studies of ductal development may provide better markers for pancreatic tumor classification. One major longstanding problem in the study of pancreatic ductal differentiation has been the lack of an effective in vitro model. We thus wished to develop an in vitro system for the study of pancreatic duct development. In doing so, we have developed a specific culture condition to promote ductal differentiation of E11.5 pancreatic rudiments. Normally, pancreatic explants cultured in vitro develop to form endocrine, acinar, as well as ductal cells. Here, we report that addition of a combination of EGF, fibroblast growth factor-10, and platelet-derived growth factor-AA to the explant cultures promotes ductal differentiation, while preventing endocrine and acinar differentiation. This culture system for differentiation and enrichment of pancreatic ductal cells may allow identification of gene(s) involved in ductal development.     

SHARPIN regulates collagen architecture and ductal outgrowth in the developing mouse mammary gland

SHARPIN is a widely expressed multifunctional protein implicated in cancer, inflammation, linear ubiquitination and integrin activity inhibition; however, its contribution to epithelial homeostasis remains poorly understood. Here, we examined the role of SHARPIN in mammary gland development, a process strongly regulated by epithelial–stromal interactions. Mice lacking SHARPIN expression in all cells (Sharpin^cpdm), and mice with a stromal (S100a4‐Cre) deletion of Sharpin, have reduced mammary ductal outgrowth during puberty. In contrast, Sharpin^cpdm mammary epithelial cells transplanted in vivo into wild‐type stroma, fully repopulate the mammary gland fat pad, undergo unperturbed ductal outgrowth and terminal differentiation. Thus, SHARPIN is required in mammary gland stroma during development. Accordingly, stroma adjacent to invading mammary ducts of Sharpin^cpdm mice displayed reduced collagen arrangement and extracellular matrix (ECM) stiffness. Moreover, Sharpin^cpdm mammary gland stromal fibroblasts demonstrated defects in collagen fibre assembly, collagen contraction and degradation in vitro. Together, these data imply that SHARPIN regulates the normal invasive mammary gland branching morphogenesis in an epithelial cell extrinsic manner by controlling the organisation of the stromal ECM.     

FKBP52对小鼠前列腺发育的影响

目的通过FKBP52基因敲除小鼠模型探索FKBP52在小鼠前列腺发育过程中的作用。方法分别对胚胎第17．5天、新生的和出生后3周的野生型和FKBP52基因敲除小鼠的前列腺进行切片HE染色,观察不同发育时期里野生型和FKBP52基因敲除小鼠前列腺发育的异同。结果（1）小鼠前列腺发育的起始不依赖于FKBP52基因的参与;（2）随着胚胎的发育,FKBP52在雄鼠前列腺发育中的作用逐渐显现出来,即FKBP52的缺失会导致前列腺叶发育受阻,最终不能形成成熟的前列腺。结论FKBP52在小鼠前列腺的发育过程中具有重要作用,它不参与前列腺的发育起始过程,但其缺失会导致前列腺发育受阻,即不能形成成熟的前列腺。     

Morphogenesis of the Acanthocephala

Postzygotic development of acanthocephalans is described. The major conclusion that can be drawn is that there has been a great deal of speculation, and that there are still many questions left unanswered. For example, exactly when does the acanthor become a complete syncytium? Is the formulation of the central nuclear mass really homologous to gastrulation, and does this mass really represent endoderm? What is endoderm in an animal with no trace of a digestive system? What is the significance, if any, of the 90 degree change in polarity from the acanthor to the adult? How do a few giant nuclei determine the fate and function of a massive synctial body? These and other questions continue to intrigue and perplex the investigator. Perhaps by the time of ICOPA-VII we will have some of the answers.