Inflammasomes bridge signaling between pathogen identification and the immune response.

Microbial organisms express pathogen-associated molecular patterns (PAMPs) that can stimulate expression of proinflammatory mediators following ligation of pathogen recognition receptors. However, both commensal organisms and pathogens can express PAMPs. The immune system can distinguish between commensals and pathogens in part through secretion of the key inflammatory cytokines interleukin (IL)-1beta and IL-18. A PAMP such as lipopolysaccharide can induce production of intracellular pro-IL-1beta and pro-IL-18, but not their secretion. A second "danger signal", derived from host-cell molecules that are released from stressed or infected cells, or detected as a PAMP that is present in the cytosol, can stimulate assembly of an inflammasome that activates the protease caspase-1. Caspase-1, in turn, is responsible for processing and secretion of the mature IL-1beta and IL-18. Many diverse ligands leading to inflammasome activation have been identified, but the cell signaling pathways initiated by the ligands tend to converge on a small set of common mechanisms.

Regulation of MAP kinase by the BMP-4/TAK1 pathway in Xenopus ectoderm.

Bone morphogenetic protein-4 (BMP-4) induces epidermis and represses neural fate in Xenopus ectoderm. Our previous findings implicate p42 Erk MAP kinase (MAPK) in the response to neural induction. We have examined the effects of BMP-4 on MAPK activity in gastrula ectoderm. Expression of a dominant negative BMP-4 receptor resulted in a 4.5-fold elevation in MAPK activity in midgastrula ectoderm. MAPK activity was reduced in ectoderm expressing a constitutively active BMP-4 receptor, or ectoderm treated with BMP-4 protein in the presence or absence of cycloheximide. Overexpression of TAK1 led to a reduction in MAPK activity in early gastrula ectoderm. The inhibitory effects of TAK1 could be reversed by 1 microM SB 203580, a p38 inhibitor. Treatment of isolated ectoderm with SB 203580 led to expression of otx2, NCAM, and noggin. Western blot analyses indicated that the BMP-4 pathway does not activate JNKs in ectoderm. Our findings indicate that BMP-4 inhibits ectodermal MAPK activity through a TAK1/p38-type pathway. MAPK has been shown to inactivate Smad1. Thus, our results suggest that BMP-4 and MAPK pathways are mutually antagonistic in Xenopus ectoderm, and that interactions between these pathways may govern the choice between epidermal and neural fate.

Male size, sperm transfer, and colony fitness in the western harvester ant, Pogonomyrmex occidentalis.

Mating success in the western harvester ant, Pogonomyrmex occidentalis, increases with male size. We tested the hypothesis that increased mating success increases male fitness and the fitness of colonies that make large males by comparing the sperm content of males prior to and at the conclusion of the mating swarm. The number of sperm a male initially possesses is a function of male size, and large males transfer a greater proportion of their sperm than do small males. For colonies, the payoff per unit of investment is an increasing function of male size, and investment in large males is not equivalent to investing in a larger number of small males. Allocation ratios in species that show size variation in reproductives may need to be modified by the individual fitness functions.

Mitogen-activated protein kinase and neural specification in Xenopus.

We have investigated the activity and function of mitogen-activated protein kinase (MAPK) during neural specification in Xenopus. Ectodermal MAPK activity increased between late blastula and midgastrula stages. At midgastrula, MAPK activity in both newly induced neural ectoderm and ectoderm overexpressing the anterior neural inducer noggin was 5-fold higher than in uninduced ectoderm. Overexpression of MAPK phosphatase-1 (MKP-1) in ectoderm inhibited MAPK activity and prevented neurectoderm-specific gene expression when the ectoderm was recombined with dorsal mesoderm or treated with fibroblast growth factor (FGF). Neurectoderm-specific gene expression was observed, however, in ectoderm overexpressing both noggin and MKP-1. To evaluate the role of MAPK in posterior regionalization, ectodermal isolates were treated with increasing concentrations of FGF and assayed for MAPK activity and neurectoderm-specific gene expression. Although induction of posterior neural ectoderm by FGF was accompanied by an elevation of MAPK activity, relative MAPK activity associated with posterior neural fate was no higher than that of ectoderm specified to adopt an anterior neural fate. Thus, increasingly posterior neural fates are not correlated with quantitative increases in MAPK activity. Because MAPK has been shown to down-regulate Smad1, MAPK may disrupt bone morphogenetic protein 4 (BMP-4) signaling during neural specification. Our results suggest that MAPK plays an essential role in the establishment of neural fate in vivo.

The role of intracellular alkalinization in the establishment of anterior neural fate in Xenopus.

Our previous work demonstrated that Xenopus ectoderm cells undergo an alkalinization in response to planar inductive signals during neural induction in explants. We have examined the role of intracellular alkalinization in the establishment of anterior neural fate. First, RT-PCR was used to examine neural-specific gene expression in planar explants in which the alkalinization is prevented by treatment with 4,4'-dihydrodiisothiocyanatostilbene-2,2'-disulfonate (H2DIDS). In explants cultured in the presence of H2DIDS, expression of NCAM and the anterior neural gene otx2 is greatly reduced or absent. Second, neural-specific gene expression was examined in isolates of uninduced animal cap ectoderm cultured in the presence of either methylamine or ammonium chloride. NCAM, otx2, and the anterior neural inducer noggin were expressed in alkalinized ectoderm, while the more posterior neural markers krox-20 and Hox B9 were undetectable. Expression of NCAM, otx2, and noggin was observed at stage 11 in both alkalinized ectoderm and the newly induced neural plate, suggesting that intracellular alkalinization could contribute to propagation of noggin signaling through the dorsal ectoderm. Alkalinization of uninduced ectoderm at stage 10.5 led to an upregulation of otx2 within 15 min. Activation of NCAM expression in alkalinized dissociated cells was identical to that observed in intact animal caps, indicating that alkalinization-mediated changes in gene expression do not require cell-cell contact. Finally, the effects of intracellular alkalinization on protein tyrosine phosphorylation were investigated using 2D gel electrophoresis and immunoblots probed with an antiphosphotyrosine antibody. Several phosphorylated protein detected in induced and alkalinized ectoderm were greatly reduced or absent in uninduced ectoderm, indicating that alkalinization elicits alterations in tyrosine phosphorylation similar to some of those observed during neural induction in vivo. Our results indicate that intracellular alkalinization plays a critical role in the activation of anterior neural-specific gene expression and that alkalinization may act by regulating the activity of a tyrosine kinase or phosphatase.

Inhibition of morphogenetic movement during Xenopus gastrulation by injected sulfatase: implications for anteroposterior and dorsoventral axis formation.

In order to explore the role of morphogenetic movement in the establishment of anteroposterior and dorsoventral axes, we sought to identify novel in vivo inhibitors of gastrulation movements in Xenopus laevis. Injection of hydrolytic sulfatase into the blastocoels of gastrula stage embryos resulted in severe anteroposterior truncation, without a corresponding truncation of the dorsoventral axis. Confocal microscopy of whole embryos revealed that gastrulation movements are severely disrupted by sulfatase; in addition, sulfatase dramatically inhibited chordomesodermal cell elongation and convergent extension movements in planar dorsal marginal zone explants. The phenotype of anteroposterior reduction elicited by sulfatase is distinctly different from commonly generated dorsoanterior phenotypes (e.g., ultraviolet irradiation of the vegetal cortex prior to cortical rotation or suramin injection), and the two varieties of phenotype appear to result from inhibition of distinct, separable components of the axis-generating machinery.

USF in the Lytechinus sea urchin embryo may act as a transcriptional repressor in non-aboral ectoderm cells for the cell lineage-specific expression of the LpS1 genes.

Expression of the aboral ectoderm-specific LpS1 gene in Lytechinus was used to study lineage-specific transcriptional regulation during sea urchin development. Band shift assays using anti-USF antibody showed that a USF-like protein bound the USF core sequence 5'-CACGTG-3' in the promoter of the LpS1 gene. DNA constructs consisting of a wild-type LpS1 promoter and the same LpS1 promoter with a mutated USF binding site fused to the bacterial chloramphenicol acetyltransferase reporter gene were tested. The mutation in the USF binding site caused an increase in chloramphenicol acetyltransferse activity. We selected a clone that encodes USF, LvUSF, from a gastrula-stage cDNA library representing Lytechinus variegatus. Transactivation experiments, in which LvUSF RNA or a DNA construct consisting of the LvUSF cDNA clone fused to the Lytechinus pictus metallothionein promoter coinjected with the wild-type or mutated LpS1 promoter-chloramphenicol acetyltransferase gene construct, showed that chloramphenicol acetyltransferase activity from the wild-type construct was repressed, while the construct mutated at the USF binding site was active. The same wild-type and mutated LpS1 promoter DNA fragments ligated to the green fluorescent protein reporter gene were used to examine spatial expression. The reporter gene constructs containing the mutated USF binding site were expressed inappropriately in all cell types including the gut and oral ectoderm in gastrula and larva stage embryos, while the wild-type constructs were expressed primarily in the aboral ectoderm. USF was expressed in all cells of the early embryo and in all tissues except the aboral ectoderm in later embryos. The data are consistent with a model depicting Lytechinus USF, as a temporal and spatial regulator by repressing LpS1 gene transcription in non-aboral ectoderm cells.

A tissue-specific repressor in the sea urchin embryo of Lytechinus pictus binds the distal G-string element in the LpS1-beta promoter.

LpS1 RNA transcripts and proteins are expressed exclusively in the aboral ectoderm of the embryo in the sea urchin Lytechinus pictus. We have characterized the LpS1-beta promoter to identify the cis-acting elements that may be involved in the aboral ectoderm-specific expression of the LpS1-beta gene. The distal G-string site, composed of six contiguous guanine deoxynucleotides located at -721 to -726, was analyzed. A mutation at the distal G-string caused over a two-fold increase in reporter chloramphenicol acetyltransferase gene activity and inappropriate expression of reporter green fluorescent protein in nonaboral ectoderm cells in L. pictus embryos. These results suggest that the proteins that bind the distal G-string act as a spatial repressor in the nonaboral ectoderm cells of the developing embryo.

An increase in intracellular pH during neural induction in Xenopus.

In this paper, we show that an intracellular alkalinization of the dorsal ectoderm cells is among the earliest responses to neural induction in Xenopus. Planar explants of the dorsal marginal zone were prepared from embryos that had been microinjected during cleavage stages with the fluorescent pH indicator bis-carboxyethyl-carboxyfluorescein-dextran (BCECF-dextran), and intracellular pH (pHi) was monitored continuously by emission ratio microfluorimetry. During stage 10.5, the dorsal ectoderm cells undergo a sustained intracellular alkalinization of approximately 0.1 pH units in response to neural induction; in the absence of the inductive signal, the pH of the dorsal ectoderm cells decreases slightly. Ectoderm cells within planar explants of the ventral marginal zone show little change in pH during a similar period. This increase in intracellular pH is inhibited by 4, 4'-dihydrodiisothiocyanatostilbene-2, 2'-disulfonate (H2DIDS) or a low Na+/high Cl- medium, treatments that presumably affect anion transport. Under these conditions, expression of the anterior neural-specific homeobox gene engrailed is not detected, while the notochord-specific epitope recognized by the Tor-70 antibody is expressed in the presence of H2DIDS. This characteristic alkalinization is not evoked by pharmacological agents that reportedly alter ectodermal developmental pathways in Xenopus embryos, such as NH4Cl, phorbol esters, or cAMP-dependent protein kinase agonists. Our results suggest that an ionic regulatory event may participate in the regulation of gene expression in response to neural induction.

Induction of neuronal differentiation by planar signals in Xenopus embryos.

The induction of the central nervous system in amphibian embryos is mediated both by early planar signals produced by mesoderm at the dorsal lip and later vertical signals emanating from the dorsal mesoderm after involution. We have examined the role and spatial extent of planar signals in the induction of neuronal differentiation. Planar explants that included only the deep layer of the dorsal marginal zone, comprising both the dorsal mesoderm and the contiguous dorsal ectoderm, were isolated at the beginning of gastrulation. After removal of the epithelial layer, explants were maintained in modified Danilchik's medium until mid-neurula stages, when they were transferred to modified Danilchik's medium + 0.1% bovine serum albumin and cultured on laminin. Neurite outgrowth occurred in 90% of these planar explants. In contrast, little or no neuronal differentiation occurred in either ventral planar explants or explants of ectoderm alone. Video analysis of cell movements shows that large-scale cell mixing does not occur between mesoderm cells and ectoderm cells in planar explants. Retrograde labelling of neuronal cell bodies indicates that cells throughout the ectoderm undergo neuronal differentiation; neurons also differentiate in cultures of distal ectoderm isolated at early neurula stages from planar explants prepared at the beginning of gastrulation. These observations indicate that planar signals act over an extended range to induce neuronal differentiation. The inductive capacity of vertical signals was examined by recombining animal caps from ultra-violet (UV) irradiated embryos with involuted mesoderm from normal midgastrula embryos. Differentiation of either neurons or anterior neural structures occurred in 73% of vertical recombinates. Our results demonstrate that planar signals from the dorsal lip of the blastopore are capable of inducing neuronal differentiation over a considerable distance in the absence of epithelial confinement, convergence and extension, and mixing between the mesoderm and ectoderm.

The cellular basis of the convergence and extension of the Xenopus neural plate.

There is great interest in the patterning and morphogenesis of the vertebrate nervous system, but the morphogenetic movements involved in early neural development and their underlying cellular mechanisms are poorly understood. This paper describes the cellular basis of the early neural morphogenesis of Xenopus laevis. The results have important implications for neural induction. Mapping the fate map of the midneurula (Eagleson and Harris: J. Neurobiol. 21:427-440, 1990) back to the early gastrula with time-lapse video recording demonstrates that the prospective hindbrain and spinal cord are initially very wide and very short, and thus at the beginning of gastrulation all their precursor cells lie within a few cell diameters of the inducing mesoderm. In the midgastrula, the prospective hindbrain and spinal cord undergo very strong convergence and extension movements in two phases: In the first phase they primarily undergo thinning in the radial direction and lengthening (extension) in the animal-vegetal direction, and the second phase is characterized primarily by mediolateral narrowing (convergence) and anterior-posterior lengthening (extension). These movements also occur in sandwich explants of the gastrula, thus demonstrating the local autonomy of the forces producing them. Tracing cell movements with fluorescein dextran-labeled cells in embryos or explants shows that the initial thinning and extension occurs by radial intercalation of deep cells to form fewer layers of greater area, all of which is expressed as increased length. The subsequent convergence and extension occurs by mediolateral intercalation of deep cells to form a longer, narrower array. These results establish that a similar if not identical sequence of radial and mediolateral cell intercalations underlie convergence and extension of the neural and the mesoderm tissues (Wilson and Keller: Development, 112:289-300, 1991). Moreover, these results establish that radial and mediolateral intercalation are the principal neural cell behaviors induced by the planar signals emanating from the dorsal involuting marginal zone (the Spemann organizer) in the early gastrula (Keller et al: Develop. Dynamics, 193: 218-234, 1992). Radial and mediolateral intercalation are induced among the 5 to 7 rows of cells comprising the prospective hindbrain and spinal cord, thus producing the massive convergence and extension movements that narrow and elongate these regions of the nervous system in the late gastrula. A more general significance of these results is that neural induction is best analyzed and understood in terms of the dynamics of the morphogenetic processes involved.

Planar induction of convergence and extension of the neural plate by the organizer of Xenopus.

This paper demonstrates that convergence and extension within the neural plate of Xenopus laevis are regulated by planar inductive interactions with the adjacent Spemann organizer. The companion article (Keller et al.: Developmental Dynamics 193:199-217, 1992) showed that the prospective hindbrain and spinal cord occupy a very short and very wide area just above the Spemann organizer in the early gastrula and that these regions converge and extend greatly during gastrulation and neurulation, using a sequence of radial and mediolateral cell intercalations. In this article, we show that "planar" contact of these regions with the organizer at their vegetal edge until stage 11 is sufficient to induce convergence and extension, after which their convergence and extension become autonomous. Grafts of the organizer in planar contact with uninduced ectodermal tissues induce these ectodermal tissues to converge and extend by a planar inductive signal from the organizer. Labeling of the inducing or responding tissues confirms that only planar interactions occur. Neural convergence and extension are actually hindered in explants deliberately constructed so that vertical interactions occur. These results show unambiguously that the Spemann organizer induces the extraordinary and precocious convergence and extension movements of the Xenopus neural plate by planar interactions acting over short distances.

[Acute urinary retention secondary to cavernous metastases from a prostatic tumor]. / Rétention urinaire aiguë secondaire aux métastases caverneuses d'une tumeur prostatique.

Prostatic carcinoma metastatic to the penis is a rare phenomenon. We report two cases of acute urinary retention due to penile metastases from a prostatic carcinoma. Patients may have a penile mass or diffuse swelling of the penis. Some patients develop priapism and urinary retention can be rarely observed. The most effective treatment seems to be local excision with wide resection and, if necessary, total penectomy. The prognosis remains poor regardless of the type of therapy employed.

The restriction of the heart morphogenetic field in Xenopus laevis.

We have examined the spatial restriction of heart-forming potency in Xenopus laevis embryos, using an assay system in which explants or explant recombinates are cultured in hanging drops and scored for the formation of a beating heart. At the end of neurulation at stage 20, the heart morphogenetic field, i.e., the area that is capable of heart formation when cultured in isolation, includes anterior ventral and ventrolateral mesoderm. This area of developmental potency does not extend into more posterior regions. Between postneurula stage 23 and the onset of heart morphogenesis at stage 28, the heart morphogenetic field becomes spatially restricted to the anterior ventral region. The restriction of the heart morphogenetic field during postneurula stages results from a loss of developmental potency in the lateral mesoderm, rather than from ventrally directed morphogenetic movements of the lateral mesoderm. This loss of potency is not due to the inhibition of heart formation by migrating neural crest cells. During postneurula stages, tissue interactions between the lateral mesoderm and the underlying anterior endoderm support the heart-forming potency in the lateral mesoderm. The lateral mesoderm loses the ability to respond to this tissue interaction by stages 27-28. We speculate that either formation of the third pharyngeal pouch during stages 23-27 or lateral inhibition by ventral mesoderm may contribute to the spatial restriction of the heart morphogenetic field.

The role of the dorsal lip in the induction of heart mesoderm in Xenopus laevis.

We have examined the tissue interactions responsible for the expression of heart-forming potency during gastrulation. By comparing the specification of different regions of the marginal zone, we show that heart-forming potency is expressed only in explants containing both the dorsal lip of the blastopore and deep mesoderm between 30 degrees and 45 degrees lateral to the dorsal midline. Embryos from which both of these 30 degrees-45 degrees dorsolateral regions have been removed undergo heart formation in two thirds of cases, as long as the dorsal lip is left intact. If the dorsal lip is removed along with the 30 degrees-45 degrees regions, heart formation does not occur. These results indicate that the dorsolateral deep mesoderm must interact with the dorsal lip in order to express heart-forming potency. Transplantation of the dorsal lip into the ventral marginal zone of host embryos results in the formation of a secondary axis; in over half of cases, this secondary axis includes a heart derived from the host mesoderm. These findings suggest that the establishment of heart mesoderm is initiated by a dorsalizing signal from the dorsal lip of the blastopore.

The specification of heart mesoderm occurs during gastrulation in Xenopus laevis.

The establishment of heart mesoderm during Xenopus development has been examined using an assay for heart differentiation in explants and explant combinations in culture. Previous studies using urodele embryos have shown that the heart mesoderm is induced by the prospective pharyngeal endoderm during neurula and postneurula stages. In this study, we find that the specification of heart mesoderm must begin well before the end of gastrulation in Xenopus embryos. Explants of prospective heart mesoderm isolated from mid- or late neurula stages were capable of heart formation in nearly 100% of cases, indicating that the specification of heart mesoderm is complete by midneurula stages. Moreover, inclusion of pharyngeal endoderm had no statistically significant effect upon either the frequency of heart formation or the timing of the initiation of heartbeat in explants of prospective heart mesoderm isolated after the end of gastrulation. When the superficial pharyngeal endoderm was removed at the beginning of gastrulation, experimental embryos formed hearts, as did explants of prospective heart mesoderm from such embryos. These results indicate that the inductive interactions responsible for the establishment of heart mesoderm occur prior to the end of gastrulation and do not require the participation of the superficial pharyngeal endoderm.

Abscess of corpus cavernosum.

A rare case of abscess of the corpus cavernosum is described. The patient was treated successfully by surgical drainage and antibiotic therapy. A right penile deviation and a relatively poor erection were reported by the patient 6 weeks postoperatively but he did not require further surgery. The possible causes, diagnosis and treatment of this rare disease are discussed.

Features of embryonic induction.

The patterned distribution of different organs in the amphibian embryo begins with the establishment of two domains, the animal and vegetal regions, that differ in developmental potency. Differences amplify as inductive interactions occur across boundaries between areas of different potency. Embryonic induction establishes a temporally and spatially dynamic area of developmental potency - a morphogenetic field. The final arrangement and differentiation of cell types within the field emerge from subsequent interactions occurring primarily within the field. These principles are illustrated in a review of the induction of the lens and the heart. Recent studies show that the induction of the lens of the eye and the induction of the heart begin early in development. Most of lens inductions occurs before the formation of the optic vesicle, and the heart appears to be part of a complex of dorsal structures whose formation is dependent upon the establishment of the dorsoventral axis. Suppressive as well as inductive tissue interactions occur during the determination of both of these organs, affecting their position and time of appearance. The complex processes of induction defined by the past nine decades of experimental work present many challenging questions that can now be addressed, especially in terms of the molecular events, cellular behaviour and regulatory physiology of the responding tissue.