Comparison of Pretectal Genoarchitectonic Pattern between Quail and Chicken Embryos

Regionalization of the central nervous system is controlled by local networks of transcription factors that establish and maintain the identities of neuroepithelial progenitor areas and their neuronal derivatives. The conserved cerebral Bauplan of vertebrates must result essentially from conserved patterns of developmentally expressed transcription factors. We have previously produced detailed molecular maps for the alar plate of prosomere 1 (the pretectal region) in chicken (Ferran et al., 2007, 2008, 2009). Here we compare the early molecular signature of the pretectum of two closely related avian species of the family Phasianidae, Coturnix japonica (Japanese quail) and Gallus gallus (chicken), aiming to test conservation of the described pattern at a microevolutionary level. We studied the developmental pretectal expression of Bhlhb4, Dbx1, Ebf1, Gata3, Gbx2, Lim1, Meis1, Meis2, Pax3, Pax6, Six3, Tal2, and Tcf7l2 (Tcf4) mRNA, using in situ hybridization, and PAX7 immunohistochemistry. The genoarchitectonic profile of individual pretectal domains and strata was produced, using comparable section planes. Remarkable conservation of the combinatorial genoarchitectonic code was observed, fundamented in a tripartite anteroposterior subdivision. However, we found that at corresponding developmental stages the pretectal region of G. gallus was approximately 30% larger than that of C. japonica, but seemed relatively less mature. Altogether, our results on a conserved genoarchitectonic pattern highlight the importance of early developmental gene networks that causally underlie the production of homologous derivatives in these two evolutionarily closely related species. The shared patterns probably apply to sauropsids in general, as well as to more distantly related vertebrate species

Ontogenetic Expression of Sonic Hedgehog in the Chicken Subpallium

Sonic hedgehog (SHH) is a secreted signaling factor that is implicated in the molecular patterning of the central nervous system (CNS), somites, and limbs in vertebrates. SHH has a crucial role in the generation of ventral cell types along the entire rostrocaudal axis of the neural tube. It is secreted early in development by the axial mesoderm (prechordal plate and notochord) and the overlying ventral neural tube. Recent studies clarified the impact of SHH signaling mechanisms on dorsoventral patterning of the spinal cord, but the corresponding phenomena in the rostral forebrain are slightly different and more complex. This notably involves separate Shh expression in the preoptic part of the forebrain alar plate, as well as in the hypothalamic floor and basal plates. The present work includes a detailed spatiotemporal description of the singular alar Shh expression pattern in the rostral preoptic forebrain of chick embryos, comparing it with FoxG1, Dlx5, Nkx2.1, and Nkx2.2 mRNA expression at diverse stages of development. As a result of this mapping, we report a subdivision of the preoptic region in dorsal and ventral zones; only the dorsal part shows Shh expression. The positive area impinges as well upon a median septocommissural preoptic domain. Our study strongly suggests tangential migration of Shh-positive cells from the preoptic region into other subpallial domains, particularly into the pallidal mantle and the intermediate septum

Topography of Somatostatin Gene Expression Relative to Molecular Progenitor Domains during Ontogeny of the Mouse Hypothalamus

The hypothalamus comprises alar, basal, and floor plate developmental compartments. Recent molecular data support a rostrocaudal subdivision into rostral (terminal) and caudal (peduncular) halves. In this context, the distribution of neuronal populations expressing somatostatin (Sst) mRNA was analyzed in the developing mouse hypothalamus, comparing with the expression pattern of the genes Orthopedia (Otp), Distal-less 5 (Dlx5), Sonic Hedgehog (Shh), and Nk2 homeobox 1 (Nkx2.1). At embryonic day 10.5 (E10.5), Sst mRNA was first detectable in the anterobasal nucleus, a Nkx2.1-, Shh-, and Otp-positive basal domain. By E13.5, nascent Sst expression was also related to two additional Otp-positive domains within the alar plate and one in the basal plate. In the alar plate, Sst-positive cells were observed in rostral and caudal ventral subdomains of the Otp-positive paraventricular complex. An additional basal Sst-expressing cell group was found within a longitudinal Otp-positive periretromamillary band that separates the retromamillary area from tuberal areas. Apart of subsequent growth of these initial populations, at E13.5 and E15.5 some Sst-positive derivatives migrate tangentially into neighboring regions. A subset of cells produced at the anterobasal nucleus disperses ventralward into the shell of the ventromedial hypothalamic nucleus and the arcuate nucleus. Cells from the rostroventral paraventricular subdomain reach the suboptic nucleus, whereas a caudal contingent migrates radially into lateral paraventricular, perifornical, and entopeduncular nuclei. Our data provide a topologic map of molecularly defined progenitor areas originating a specific neuron type during early hypothalamic development. Identification of four main separate sources helps to understand causally its complex adult organization

The IDENTIFY study: the investigation and detection of urological neoplasia in patients referred with suspected urinary tract cancer - a multicentre observational study

Objective To evaluate the contemporary prevalence of urinary tract cancer (bladder cancer, upper tract urothelial cancer [UTUC] and renal cancer) in patients referred to secondary care with haematuria, adjusted for established patient risk markers and geographical variation. Patients and Methods This was an international multicentre prospective observational study. We included patients aged ≥16 years, referred to secondary care with suspected urinary tract cancer. Patients with a known or previous urological malignancy were excluded. We estimated the prevalence of bladder cancer, UTUC, renal cancer and prostate cancer; stratified by age, type of haematuria, sex, and smoking. We used a multivariable mixed-effects logistic regression to adjust cancer prevalence for age, type of haematuria, sex, smoking, hospitals, and countries. Results Of the 11 059 patients assessed for eligibility, 10 896 were included from 110 hospitals across 26 countries. The overall adjusted cancer prevalence (n = 2257) was 28.2% (95% confidence interval [CI] 22.3–34.1), bladder cancer (n = 1951) 24.7% (95% CI 19.1–30.2), UTUC (n = 128) 1.14% (95% CI 0.77–1.52), renal cancer (n = 107) 1.05% (95% CI 0.80–1.29), and prostate cancer (n = 124) 1.75% (95% CI 1.32–2.18). The odds ratios for patient risk markers in the model for all cancers were: age 1.04 (95% CI 1.03–1.05; P < 0.001), visible haematuria 3.47 (95% CI 2.90–4.15; P < 0.001), male sex 1.30 (95% CI 1.14–1.50; P < 0.001), and smoking 2.70 (95% CI 2.30–3.18; P < 0.001). Conclusions A better understanding of cancer prevalence across an international population is required to inform clinical guidelines. We are the first to report urinary tract cancer prevalence across an international population in patients referred to secondary care, adjusted for patient risk markers and geographical variation. Bladder cancer was the most prevalent disease. Visible haematuria was the strongest predictor for urinary tract cancer

“Ondes, photons et cancer : vers de nouvelles thérapies ?”

International audienceLa problématique sanitaire de la lutte contre le cancer conduit les scientifiques à concevoir des stratégies de recherche interdisciplinaire afin de développer de nouveaux traitements plus puissants, sans effets aversifs, pour contrer la résistance de certains cancers aux chimiothérapies et radiothérapies. La piste bioélectrique est ainsi explorée à l'Institut de recherche XLIM, grâce, notamment, aux avancées technologiques ayant permis l’arrivée de générateurs d’impulsions de l’ordre de la nanoseconde et de forte intensité (kV/cm), on parle alors de nsPEF (nanosecond pulsed electric field). Des outils d'imagerie multiphotonique intravitale sont utilisés en renfort pour comprendre comment ces nsPEF affectent la physiologie des cellules tumorales dans leur microenvironnement et ainsi décrypter les acteurs moléculaires intracellulaires sous-jacents

Organización morfológica y citogenética del hipotálamo del pollo sobre base de mapas moleculares / Sylvia Marianne Bardet Coste; director Luis V. Puelles López.

Tesis-Universidad de Murcia.Consulte la tesis en: BCA. GENERAL. ARCHIVO UNIVERSITARIO. T.M. 3519

PB-94 [14:30] Visualisation of an nsPEF induced calcium wave using the genetically encoded calcium indicator GCaMP in U87 human glioblastoma cells

International audienceThis study looks at the application of genetically encoded calcium indicators (GECIs), to investigate the spatiotemporal nature of nsPEF-induced calcium signals, using fluorescent live cell imaging. GECIs can be targeted to different cellular locations and, by using a plasma membrane expressed GECI, we demonstrate calcium entry into the cell as a wave, initiated at the cathode side, following nsPEF treatment. This wave was masked by commercially available chemical, cytosolic, calcium indicators

Sexually differentiated cell proliferation in the medial preoptic nucleus of Japanese quail

The androgen-depend sequence of male copulatory behaviors is sexually differentiated in Japanese quail like in many other vertebrates. This behavior disappears within a week after castration of males and is rapidly restored following treatment with testosterone. In contrast, ovariectomized females treated with testosterone never show these behaviors. This sex-specific responsiveness to testosterone results from organizational effects of embryonic ovarian estrogens. The behavioral phenotype of male and female quail is completely reversed by treatment, before embryonic day 12 (E12), of male embryos with estrogens or of female embryos with an inhibitor of aromatase, the enzyme converting testosterone into estradiol. In the quail brain, the medial preoptic nucleus (POM) is a necessary and sufficient site for the activation of sexual behavior by testosterone. It can be identified by a dense population of aromatase-immunoreactive neurons and, importantly, aromatase activity in this region is sexually differentiated (males>females) and this difference is maintained even when birds are treated with a same dose of testosterone. Aromatase is thus, like sexual behavior, a sex-specific feature regulated by testosterone in adult quail and presumably organized in early life by steroid action. The cellular basis of these organizational processes have not been identified. We analyzed the ontogeny of POM cells by injecting 5-bromo-2-deoxyuridine (BrdU) in eggs at different embryonic (E) stages (E8, E10, E12, E14 and E16) and quantifying BrdU-labeled cells at postnatal (PN) day 56. Large numbers of BrdU-positive cells were observed throughout the POM of males and females injected on E8-E10 but most cells were post-mitotic in both sexes on E14-E16. E12 injections resulted in a larger number of BrdU cells in females than in males at PN56. However, male and female embryos injected on E12 and killed on E13, PN1 or PN15 had similar numbers of BrdU positive cells. Furthermore, BrdU injections on E14 labeled very few cells at PN 56 suggesting that the POM is essentially post-mitotic at that age. Together these data suggest that a sex-specific apoptosis must occur between PN15 and PN56. Double-label immunohistochemistry for BrdU and for the neuron-specific marker Hu indicated that BrdU-positive cells born between E8 and E16 are not neurons and are thus presumably glial cells. This sex-specific cell proliferation occurring around the end of the critical period of sexual differentiation may have an important impact on brain and behavior differentiation. The phenotype of these cells and the mechanisms mediating their differential development are currently under investigation

Patrón de expresión de Sonic Hedgehog en la placa alar del prosencéfalo secundario de Pollo.

International audienc

Ontogeny of sex differences in steroid-sensitive regions in the quail brain (Coturnix Japonica)

Sex differences affecting the expression of sexual behavior are observed in many species. In quail, expression of the male-typical copulatory pattern is androgen-dependent. This behavior disappears within a week after castration and is restored after a few days of treatment with exogenous testosterone. In contrast, ovariectomized females treated with testosterone never show the sequence of male-typical copulatory behavior. This sex difference in responsiveness to testosterone results from organizational effects of embryonic estrogens secreted by the female ovary. The behavioral phenotype can be completely reversed by treatment, before embryonic day 12, of male embryos with estrogens or of female embryos with an aromatase inhibitor. In the quail brain, the medial preoptic nucleus (POM) is a necessary and sufficient site for the activation by testosterone of sexual behavior. Aromatase, the enzyme converting testosterone into estradiol, is densely expressed in POM and its activity is sexually differentiated (males>females) even when birds are treated with a same dose of testosterone. Aromatase and other neuroendocrine systems are thus, like sexual behavior, differentially activated by testosterone in adult quail but the cellular basis of these sexually differentiated features presumably organized in early life by steroid action have not been identified. To analyze the ontogeny of steroid sensitive regions that control behavioral sex differences in the quail brain, we injected 5-bromo-2-deoxyuridine (BrdU) in eggs at different stages of the embryonic (E) development (E8, E10, E12, E14 and E16) and sacrificed the animals at postnatal (PN) day 56. Large numbers of BrdU-positive cells were observed throughout the POM of males and females injected on E8-E10 but most cells were post-mitotic in both sexes on E14-E16. E12 injections resulted in a larger number of BrdU cells in females than in males. This differential number of BrdU-positive cells seen at PN56 in birds injected on E12 could result from a) a difference in the age at which cells become post-mitotic (males before females or alternatively females before males, so that male cells labeled by BrdU on E12 dilute their label in subsequent divisions) or b) a differential apoptosis between E13 and PN56. However, no sex differences in the number of BrdU positive cells was observed in embryos injected with BrdU on E12 and killed on E13. Furthermore, BrdU injections on E14 labeled very few cells at PN 56 suggesting that the POM is essentially post-mitotic at that age. The sex difference observed in birds injected at E12 should result from a differential apoptosis after E13. Double-label immunohistochemistry for BrdU and the neuronal marker Hu (C/D) indicated that all BrdU-positive cells born between E8 and E16 are not neurons (no double label) suggesting that these are glial cells. This sex difference in (glial?) proliferation around the end of the critical period of sexual differentiation may play a key role in the differentiation of brain and behavior. The specific phenotype of these cells and the mechanisms mediating their differential development are currently under investigation