Are birds pseudoteeth and denticulations related to touch papilla? An investigation in parrot, goose, and chicken.

We first studied the morphology and the development of goose denticulations, which develop mainly by a ripple process, and the touch papillae of the bill tip organ, which appears through an evagination process at the end of the beak. During their development, we observed the specific expression of PAX9, PITX2, and BMP4, while SHH was expressed mainly in the basal layer of the epithelium in a non-specific manner. Adult goose denticulations are associated with numerous columns. The goose denticulations and columns were filled with numerous Herbst and Grandry corpuscles, as well the touch papillae of the bill tip organ. Histological analysis of adult parrot pseudoteeth revealed that the osseous pseudoteeth were extended by similar columns filled with Herbst and Grandry corpuscles. We also examined adult and embryonic chicken beaks. During ontogeny, we observed a process of rostral evagination with folding associated with discrete ripples in the anterior part of the beak rudiment, in which PAX9, PITX2, and BMP4 are expressed. In the corresponding adult areas, there were numerous sensory corpuscles with rostral columns, which were similar to the features observed in goose. These observations support the hypothesis that pseudoteeth and denticulations constitute sensory organs, and that the touch papillae exhibit some similarities with pseudoteeth.

Effects of HSP90 inhibition on primordial germ cells migration: A study in the gonad of the chick embryo.

INTRODUCTION: Primordial Germ Cells (PGCs) differentiate into spermatozoa or oocytes. They appear early during embryonic development before migrating to the gonadal ridges. Because of their long migration, PGCs have been proposed as a valuable model to study long distance cell migration. Some species also present a vascular phase in the migration of the germline and could therefore be compared to metastatic migration. HSP90 is a heat shock protein involved in the stabilization of several client-proteins, including oncoproteins. HSP90 inhibition has been proved to decrease PGCs migration in mouse and zebrafish. MATERIAL AND METHODS: We investigated the effect of geldanamycin on PGCs migration in a species with a vascular phase, the chicken. Geldanamycin was injected in the egg at 48h of incubation, PGC's were detected in blood using of blood smears, and in the embryo by immunohistochemistry using anti-HSP90 antibody. RESULTS: The effects of the treatment were similar to those observed in mouse and zebrafish. We show the presence of ectopic germs cells in the vasculature and in the dorsal mesentery, and some deformities of the gonads. CONCLUSION: Inhibition of HSP90 decreases the migration of PGCs and proposed the migration of PGCs in the chick embryo as an interesting model to study metastatic invasion.

Is Vasa such a highly specific marker for primordial germ cells? A comparison of VASA and HSP90 proteins expression in young chicken embryos.

INTRODUCTION: Primordial germ cells (PGCs) have been studied since the 19th century with several different methods. The earliest works were based on the morphological criteria of these cells associated or not with a particular staining. Different markers have been proposed in immunohistochemistry among which we can quote the Stage-specific embryonic antigene-1 (SSEA-1), the embryonic mouse antigen-1 (EMA-1) or the heat shock protein 90. Unfortunately, none of them are germline specific. The VASA protein is considered as one of the most reliable marker for PGCs by some authors with its expression being considered to limited to the germ cells. However, other studies have reported its expression in somatic cells. Here, we described the expression of the heat shock protein, HSP90, and the VASA protein in the early chick embryo. MATERIAL AND METHODS: Embryos from stages Hamburger-Hamilton (HH) 19, 21 and 28 were collected. Embryos were dissected and fixed in Serra's medium. Sections were placed on slides for PAS staining and for double immunohistochemistry with HSP90 and VASA. RESULTS: VASA and HSP90 expression have been observed in germ cells but as well in other cell lineages with a spatio-temporal gradient in respect to the characteristics of development of each organ. The conclusion is that VASA expression is not limited to the germ line in chick embryo.

A reviewed chronology of primordial germ cells migration in the chick embryo.

INTRODUCTION: Primordial Germ Cells (PGCs) are present in all sexually reproducing animals. They differentiate into spermatozoa or oocytes and are therefore responsible for the transmission of genetic and epigenetic information across generations. In birds, PGCs are first observed in the center of the blastodisc at stage Eyal-Giladi X. With the formation of the primitive streak, germ cells are translocated anteriorly to the germinal crescent. At stage Hamburger- Hamilton 10-12, they enter the vasculature before migrating through the dorsal mesentery towards the genital ridges. MATERIAL AND METHODS: Embryos from stages Hamburger-Hamilton (HH) 16 to 22 were collected. Blood samples were taken from the dorsal aorta and from the heart in order to perform blood smears and PAS staining. Embryos were dissected and fixed in Serra's medium. Sections were placed on slides for PAS staining. A sample of each embryo was collected for DNA extraction and PCR in order to determine the sex of the embryos. RESULTS: PGCs were observed in blood circulation until stage HH 20 on blood smears and until stage HH 19 on histological sections. The first PGCs arrived in the genital ridges were observed from stage HH 17. A few germ cells were still migrating in the dorsal mesentery at stage HH 22. The aim of this study was to review the chronology of the migration of PGCs in chick embryos.

External and computed tomography analysis of a strophocephalic lamb.

CONTEXT: The Museum of Anatomy and Embryology Louis Deroubaix attached to the Laboratory of Anatomy, Biomecanics and Organogenesis, ULB, Brussels, possesses in its liquid collections a cephalic extremity of a lamb suffering from strophocephaly. The origins have not been determined. The trunk and the limbs are resected. MATERIAL AND METHODS: The piece has been studied and photographed. A volumic computed tomography acquisition has been performed with a Siemens Volume Zoom. For pedagogic and museological purposes, surface reconstructions and 3D printing have been obtained. RESULTS: An otocephaly is observed. Both ears are located in place of the oral cavity. The mandible is welded to the braincase. The eyeballs are close together (synophtalmia) which confirms the presence of a cyclotocephaly. They are surmounted by a rudimentary snout rather than a proboscis. The presence of this muzzle allows the anomaly to be classified as a strophocephaly, a malformation already described in sheeps. CT slices of the brain show a semi-lobar holoprosencephaly with incomplete division of the cerebral hemispheres and ventricules. DISCUSSION AND CONCLUSION: The CT examination allows the facial anomalies to be allocated to a holoprosencephaly. The singularity of this case, compared to the human cyclotocephalies, is the presence of a differentiated muzzle rather than a simple proboscis. The holoprosencephaly is uncomplete. Such anomalies have been associated with an entire absence of cerebral differentiation but with a complete absence of the muzzle. The tridimensional printing represents an interesting educational tool easily transportable in contrast to the original specimen.

Expression of Wnt and beta-catenin in mouse tooth germs and chick facial primordia: a comparison / Expresión de Wnt y beta-catenina en gérmenes de dientes de ratón y primordios faciales de polluelos: una comparación

In order to compare Wnt/ beta-catenin expression in mouse and chick facial primordia during development, we performed an immunohistochemical analysis of both protein expressions in E12 to E17 mouse embryos and in E3 and E10 chick embryos. During odontogenesis, from bud to bell stage, both proteins exhibit similar fixation patterns, with epithelial and mesenchymal immunoreactivity, consistant with literature data. Double labelling demonstrates that the same cells express both antigens, even in undifferentiated mesenchyme. The enamel knot, and the ameloblastic and odontoblastic layers are stained at the same manner. In the chick, Wnt and beta-catenin are diffusely present on craniofacial mesenchyme. In both species, premuscular blastemata express Wnt and b-catenin, but Wnt is specifically expressed on the perichondrium and ossification centers, suggesting a role independent from beta-catenin pathway.

Geldanamycin administration reduces the amount of primordial germ cells in the mouse embryo.

INTRODUCTION: Heat shock proteins (HSPs) are expressed or overexpressed in response to exposure to stress. They act as molecular chaperones, ensuring the correct folding of numerous client proteins. HSP90 is one of the most conserved HSPs. Its role extends beyond stress tolerance. HSP90 also contributes to development, differenciation, apoptosis and oncogenesis. Numerous tumors are associated with an overexpression of HSP90 and this expression can be used to evaluate its metastatic capacity. Primordial germ cells (PGCs) exhibit HSP90 expression under normal conditions. PGCs arise early in development and migrate by a combination of passive and active movements towards the gonads. The aim of this work was to study the impact of an inhibition of HSP90 on the migration of the PGCs. Geldanamycin, a well established HSP90 inhibitor with potent antitumor properties was used to achieve this inhibition. MATERIEL AND METHODS: 5mg of Geldanamycin were administered to E8 pregnant mice. E17 embryos were removed and fixed for staining and Immunohistochemistry with anti-HSP90 and anti-VASA antibodies. RESULTS: Geldanamycin-treated mouse embryos exhibited less VASA-immunopositive cells compared to the non-treated ones. These results suggest that geldanamycin administration at the time of PGCs migration reduces the number of PGCs in the gonads. HSP90 and VASA stainings were identical. We therefore expressed the idea that HSP90 could be used as a reliable marker for PGCs.