Annick Mauger

Early regionalization of the somitic mesoderm as studied by the development of the axial skeleton of the chick embryo

Abstract The excision of a portion of unsegmented somitic mesoderm from the thoracic region has no or little effect on the development of vertebrae and ribs. Its orthotopic replacement by thoracic somitic mesoderm obtained from another embryo does not cause important deficiencies of the rib basket. But its replacement by nonsomitic tissue (neural tube, midgut, somatic mesoderm) not only causes the formation of a costal gap within the thoracic region, but also leads to the production of a defective spine on the operated side. Segmented or unsegmented somitic mesoderm from the cervical region, implanted in place of unsegmented thoracic somitic mesoderm, differentiates according to its origin: no ribs develop in the host embryo at the operation level. Conversely, segmented or unsegmented somitic mesoderm from the thoracic region, implanted in place of unsegmented cervical somitic mesoderm, also differentiates according to its origin, giving rise to supernumerary ribs in front of the hosts rib basket. These extra ribs, however, always lack a sternal component. It is concluded that the level-specific morphogenetic capacity of the cervical and thoracic somitic mesoderm is determined before metamerization occurs. Despite this early determination, the unsegmented somitic mesoderm is endowed with regulative properties.

Immunofluorescent localization of collagen types I and III, and of fibronectin during feather morphogenesis in the chick embryo.

Abstract Collagen types I and III were purified from the skin of 3- or 7-week chickens and fibronectin from human serum. Antibodies were raised in rabbits and used in indirect immunofluorescence on frozen sections of 5- to 16-day chick embryo feather-forming skin. Prior to the formation of dense feather-forming dermis, anticollagen fluorescence was confined to a thin underlining of the dermal-epidermal junction (DEJ), while antifibronectin label was retained on loosely dispersed material in the predermal mesenchyme. Dense feather-forming dermis was characterized by loosening of the anti-collagen label along the DEJ, by its spreading throughout the thickness of dermis, and by an overall densification of antifibronectin label. Feather formation coincided with a decrease of anti-collagen and an increase of antifibronectin label density in the dermal feather condensations and in the core of outgrowing feather buds. By contrast, density of anti-collagen-labeled material was highest and anti-fibronectin-labeled material was lowest in interplumar and glabrous skin. In slanting feather buds and feather filaments, distribution of anti-collagen-labeled material exhibited a type-specific cranial-caudal asymmetry. The microheterogeneous distribution of extracellular matrix components might constitute part of the morphogenetic message that the dermis is known to transmit to the epidermis during the formation of cutaneous appendages.

Migratory and organogenetic capacities of muscle cells in bird embryos

SummaryThe migratory and organogenetic capacities of muscle cells at different stages of differentiation were tested in heterospecific chick/quail recombinants. Grafts containing muscle cells were taken from the premuscular masses from 4- to 5-day quail embryos, from the limb or trunk muscles of 12-day embryonic and 4-day post-natal quails, and from experimentally produced bispecific premuscular masses in which the myoblasts are of quail origin and the connective tissue cells of chick origin. Grafts were implanted into 2-day chick embryos in place of the somitic mesoderm at the limb level. Hosts were examined 4 to 7 days after operation.After implantation of a piece of premuscular mass, quail cells were found at and around the site of the graft in the truncal region and within the limb as far as the autopod. Quail cells participated predominantly in the trunk and limb musculature, which contained a number of quail myotubes and of bispecific quail/chick myotubes. Apart from skeletal muscles, quail cells contributed sporadically to nerve envelopes and blood vessel walls in the limb.When the graft was of bispecific constitution, quail nuclei in the limb and the trunk were found exclusively in monospecific and bispecific myotubes.After implantation of differentiated embryonic or post-natal muscle tissue, quail cells in the limb contributed only sporadically to nerve envelopes and blood vessel walls, while in the trunk they also participated in the formation of muscles and tendons.It is concluded that the myogenic cells in 4 to 5-day quail premuscular masses are still able to undergo an extensive migration into the limb buds and there participate in the formation of myotubes and anatomically normal muscles. They display developmental potentialities equivalent to those of the somitic myogenic stem cells. These capacities are lost in 12-day embryonic muscles.

Immunofluorescent localization of collagen types I, III, IV, fibronectin and laminin during morphogenesis of scales and scaleless skin in the chick embryo

SummaryCollagen types I and III were purified from the skin of 3-or 7-week-old chickens, collagen type IV from bovine skin or EHS mouse tumour, fibronectin from human serum, and laminin from EHS mouse tumour. Antibodies were produced in rabbits or sheep, and used in indirect immunofluorescence on frozen sections of 9-to 16-day-old normal or mutant (scaleless) chick-embryo foot skin. In normal scale-forming skin and inscaleless skin, the distribution of anti-laminin and anti-type IV collagen label was uniform along the dermal-epidermal junction and showed no stage-related variations, except for fluorescent granules located in the dermis of early scale rudiments. By contrast, in normal scale-forming skin, the density of anti-types I and III label decreased in the dermis within scale rudiments, whereas it gradually increased in interscale skin. Conversely, anti-fibronectin label accumulated at a higher density within scale rudiments than in interscale skin. In the dermis of thescaleless mutant, anti-types I and III label and antifibronectin label were distributed evenly: the density of anti-collagen label increased with age, while that of antifibronectin decreased and almost completely vanished in 16-day-old skin, except around blood vessels. The microheterogeneous distribution of some extracellular matrix components, namely interstitial collagen types I and III and fibronectin, is interpreted as part of the morphogenetic message that the dermis is known to transmit to the epidermis during the formation of scales. The even distribution of these components in mutantscaleless skin is in agreement with this view. Basement membrane constituents laminin and type-IV collagen do not appear to be part of the dermal morphogenetic message.

Peridermal cell patterning in the feather-forming skin of the chick embryo

Abstract The shape, distribution, and orientation of peridermal cells were examined in the dorsolumbar skin of 7 1 2 - day chick embryos. Most feather rudiments of middorsal and lateral rows showed a marked cephalocaudal polarity. A similar polarity was found in the prospective rudiments of skin areas lateral to the last-formed row. On the cranial slope and apex of rudiments, cells are convex and predominantly elongated at right angles with respect to the cephalocaudal axis, whereas on the caudal slope, most cells are flat, polygonal, surrounded by a border-line ridge, and oriented predominantly with their long axis parallel to the cephalocaudal axis. The significance of this pattern is discussed in view of the fact that the epidermis is the determinant tissue in feather orientation.

Effect of hydrocortisone on skin development in the chick embryo: ultrastructural, immunohistological, and biochemical analysis.

The effect of hydrocortisone on the development of dorsal skin was analyzed in the chick embryo by (1) transmission electron microscopy, (2) indirect immunofluorescence histology of extracellular matrix components (collagen types I, III, and IV; fibronectin; and laminin), and (3) quantitative determination of collagen content and proline incorporation, between administration of the drug at 6 or 6.5 days and final retrieval of skin pieces at 11 days of incubation. Treatment caused the formation of featherless skin areas which exhibited an early maturation of the epidermis, a uniform distribution of interstitial collagen and rarefaction of fibronectin in the dermal extracellular matrix, and a significant increase of collagen content and proline incorporation in collagen noncollagen proteins, characterized by an increased hydroxyproline-to-proline ratio. The distribution of type IV collagen and of laminin was unchanged. The absence of feather formation in hydrocortisone-induced apteria is interpreted as resulting primarily from an early extinction of epidermal morphogenetic competence, and secondarily from modifications in the amount and distribution of extracellular matrix components in the dermis.

Rôle du tube neural dans le développement du plumage dorsal de l'embryon de Poulet

Experiments were performed on 2- to 2 1/2-day chick embryos in order to study the role of the neural tube in the development of the dorsal plumage. Pieces of neural tube were excised or replaced by various living or inanimate implants. In other experiments, a screen was interposed between neural tube and somitic mesoderm.The excision resulted in the non-differentiation of a portion of the spinal pteryla: absence of several feather chevrons without disruption of the hexagonal feather pattern, or formation of an apterium or an irregularly feathered area.The replacement by a piece of agar or tantalum or by another tissue (gut, somatopleural mesoderm, somites) led to the same type of dorsal plumage deficiencies as those which were produced by the excisions. Similarly, the replacement by a fragment of neural tube treated at 100° C severely interfered with the development of dorsal plumage. On the contrary, when the neural tube had been exposed to a temperature of 60° C only, the spinal feather pattern was normal or nearly so.The interposition of a solid screen 0.8 to 2 mm in length (tantalum, egg shell membrane, Millipore filter) between neural tube and somitic mesoderm resulted in the formation of a featherless triangular notch in one side of the spinal pteryla. The screen prevented the development of the feathers in the feather field lateral to the screen.These experiments show that the neural tube is indispensable for the differentiation of the dorsal feather pattern. The neural tube cannot be replaced by inanimate objects or by any of the tested tissues, not even by feather-forming ones like somites. Its presence is likely to be necessary for the transformation of dermatomes into predermal cells. When it is absent, dense feather-forming dermis does not form at that level and, consequently, the corresponding portion of the spinal pteryla cannot develop.