Expression of Hex during feather bud development

We studied proline-rich divergent homeobox gene Hex/Prh expression in the dorsal skin of chick embryo during feather bud development. Hex mRNA expression was first observed in the dorsolateral ectoderm and mesenchyme at 5 days, then in the epithelium and the dermis of the dorsal skin before placode (primordium of feather bud) formation and then was restricted to the placode and the dermis under the placode. Afterward, Hex expression was seen in the epidermis and the dermis of the posterior region of short bud. In accordance with Hex mRNA expression in the placode, Hex protein was observed in the epidermis as well as in the dermis of the placode. Immunoelectron microscopic study indicated that the protein located both in the nuclei and cytoplasm of the epidermis and the dermis at the short bud stage. The Wnt signaling pathway plays an essential role in the early inductive events in hair (Wnt3a and 7a) and feather (Wnt7a) follicles. The pattern of Hex expression in the epidermis was similar to that of Wnt7a, while little, if any, expression of Wnt7a was detected in the dermis under the placode or the dermis of the short bud compared with that of Hex, suggesting that Hex plays an important role in the initiation of feather morphogenesis.     

Fine structure ofDrosophila wing imaginal discs during early stages of metamorphosis

Summary Fine structural changes are described which occur in wing imaginal discs taken fromDrosophila prepupae at 120–128 hours of age. The first signs of cuticle formation appear at 124 hours in the tips of microvilli which extend into the peripodial cavity of the disc. A continuous epicuticular layer is evident at 126 hours, and the beginning of laminated endocuticle at 128 hours. Electron-lucid spaces appear in the apical regions of the cells at 122 and 124 hours. These are often clearly intercellular and contain membranous material, but sometime appear to be intracellular, or vacuoles. Their frequency decreases sharply after 122 hours, and begins to rise again at 126 hours. Junctional complexes between cells are present in all ages studied, consisting predominantly of long zonulae adhaerentes in the 120 h disc, of various attachment specializations, including septate desmosomes in the 128 h disc. Inclusions interpreted as lipid are found randomly scattered in the cytoplasm at 120 hours. They change in number, size, arrangement and location in later stages.Microtubules are widely distributed in discs of all ages reported; their function is thought to be cytostructural. A method of precise staging of prepupae based on ultrastructural criteria is proposed.This research was supported by NIH Training Grant HD-139 and NSF Grant GB 7803.     

Localization of HB9 homeobox gene mRNA and protein during the early stages of chick feather development

We performed in situ hybridization and immunohistochemical analysis of HB9 homeobox gene mRNA and protein, respectively, during chick feather development. HB9 mRNA was highly expressed in epidermal basal cells and dermal cells of the placodes and feather buds, but not in those of the interplacodes and interbud regions. HB9 protein was predominantly expressed in dermal cells of the symmetric short buds and decreased after the asymmetric bud stage when the feather bud had become elongated along the anterior-posterior (A-P) and proximal-distal (P-D) axis. These results suggest that HB9 gene is regulated in a spatiotemporal manner during feather development, and may be involved in early feather bud morphogenesis.     

Corticosterone inhibits feather growth: potential mechanism explaining seasonal down regulation of corticosterone during molt

Corticosterone (CORT) is seasonally modulated in many passerines, with plasma CORT concentrations lowest during the prebasic molt when all feathers are replaced. To explain why, we proposed that the birds downregulate natural CORT release during molt in order to avoid CORT's degradative effects on proteins and its inhibition of protein synthesis. If CORT exerted these effects during molt, it could slow protein deposition during feather production and potentially result in a longer period of degraded flight performance. To test this hypothesis, either empty or CORT-filled silastic implants were inserted into captive European starlings (Sturnus vulgaris) and white-crowned sparrows (Zonotrichia leucophrys) undergoing induced (feather replacement after plucking) and natural molts. We then measured the rate of feather re-growth by regularly measuring the length of primary, secondary, and tail feathers. CORT implanted birds showed a significantly decreased rate of feather growth compared to control animals. Basal CORT concentrations of induced molt and non-molting birds were also compared but no difference was noted. The results suggest a tradeoff; a complete set of new feathers may be more important to the survival of a bird than the ability of CORT to respond maximally to a stressor.     

Contributions of feather microstructure to eider down insulation properties

Insulation is an essential component of nest structure that helps provide incubation requirements for birds. Many species of waterfowl breed in high latitudes where rapid heat loss can necessitate a high energetic input from parents and use down feathers to line their nests. Common eider Somateria mollissima nest down has exceptional insulating properties but the microstructural mechanisms behind the feather properties have not been thoroughly examined. Here, we hypothesized that insulating properties of nest down are correlated to down feather (plumule) microstructure. We tested the thermal efficiency (fill power) and cohesion of plumules from nests of two Icelandic colonies of wild common eiders and compared them to properties of plumules of wild greylag goose Anser anser. We then used electron microscopy to examine the morphological basis of feather insulating properties. We found that greylag goose down has higher fill power (i.e. traps more air) but much lower cohesion (i.e. less prone to stick together) compared to common eider down. These differences were related to interspecific variation in feather microstructure. Down cohesion increased with the number of barbule microstructures (prongs) that create strong points of contact among feathers. Eider down feathers also had longer barbules than greylag goose down feathers, likely increasing their air‐trapping capacity. Feather properties of these two species might reflect the demands of their contrasting evolutionary history. In greylag goose, a temperate, terrestrial species, plumule microstructure may optimize heat trapping. In common eiders, a diving duck that nests in arctic and subarctic waters, plumule structure may have evolved to maximize cohesion over thermal insulation, which would both reduce buoyancy during their foraging dives and enable nest down to withstand strong arctic winds.     

Histochemical localization of skin glycosaminoglycans during feather development in the chick embryo

Summary The distribution of glycosaminoglycans (GAGs) was studied in embryonic chick skin, using alcian blue staining with critical electrolyte concentration and glycanase treatment, immunofluorescence and transmission electron microscopy. Light microscopy revealed an uneven distribution of sulphated and non-sulphated GAGs at all stages of feather development. Along the dermal-epidermal junction and throughout the depth of the dermis, staining was stronger inside the feathers than in the interplumar skin. With increasing MgCl₂ concentration, the decrease in stain intensity along the dermal-epidermal junction was stronger in interplumar skin than inside feather structures, indicating that sulphated GAGs are more abundant within feathers than in interplumar skin. The same differential sensitivity to electrolyte concentration was noted in the dermis, except at the feather placode stage, when labelling inside the dermal condensation was virtually wiped out at 0.6 M MgCl₂ and higher concentrations, whereas it persisted in the surrounding dermis up to 0.8 M MgCl₂, indicating that the dermal condensation contains a larger amount of hyaluronate than non-feather-forming dermis. Enzyme treatment of sections with Streptomyces hyaluronidase as compared with those treated with chondroitinase ABC corroborated these findings. Immunofluorescent detection of heparan sulphate proteoglycan revealed the presence of the antigen along the dermal-epidermal junction at all stages of feather development, with peaks of brightness in discrete spots of feather structures. Electron microscopy revealed the presence of ruthenium red and tannic acid positive material in the dermal-epidermal junctional zone and inside the dermis. The density of marked granules was somewhat higher in intraplumar than in interplumar regions. These observations demonstrate that certain sulphated and non-sulphated GAGs are distributed in a microheterogeneous manner, which appears to be related to the morphogenetic events of feather development. They are discussed in view of the possible role these components might play in dermal-epidermal interactions. They strengthen the notion, already gained from previous studies on the localization of interstitial collagens and fibronectin, that extracellular matrix components play an important structural and informative role in organogenesis.     

Fine structure of the sporocysts of Ascosphaera apis during development as revealed by the scanning electron microscope

The fine morphology of the sporocysts of Ascosphaera apis (Maassen ex Claussen) Olive & Spiltoir, an entomopathogenic fungus of the immature honey bee has been studied using the scanning electron microscope. During the third day of mycelial growth in the culture medium, numerous short, branched hyphae were formed. The tip of the branch-hypha gradually expanded to form immature sporocysts. The immature sporocysts contained a large number of globules of varying sizes. The walls of the immature sporocysts were finely wrinkled, becoming smooth as the sporocyst matured. Both exterior and interior surfaces of the sporocyst wall possessed numerous papillae. Some globules in the developing sporocyst began to form immature spores which aggregated to form spore balls. The fully formed spore balls were not enveloped by a membrane.     

Deduced primary structure of a Xenopus proteasome subunit XC3 and expression of its mRNA during early development

Proteasome is a non-lysosomal proteinase complex ubiquitously distributed in eukaryotic cells. We isolated here the cDNA clone for one of the proteasome subunits (XC3) from Xenopus ovary cDNA libraries using rat RC3 cDNA as a prove. The cDNA is 885 bp long and encodes 234 amino acids. The deduced amino acid sequence is highly homologous (95.3%) to those of rat RC3 and human HC3 subunits. The mRNA for XC3 is one of the maternal mRNAs and detected at all the embryonic stages investigated, but its level changes in a characteristic way especially at the gastrula stage. We suggest that the highly conserved XC3 subunit plays an essential role in proteasome function and also that during Xenopus embryogenesis mRNA for XC3 subunit is replaced from maternal to newly-synthesized one probably around the gastrula stage.