The pupal cuticle of Drosophila: differential ultrastructural immunolocalization of cuticle proteins

Precise ultrastructural localization of Drosophila melanogaster pupal cuticle proteins (PCPs) was achieved by the immunogold labeling of frozen thin sections. PCPs were found in lamellate cuticle and intracellular vesicles but, curiously, were absent from the assembly zone of the cuticle. Antibodies that distinguish between the two classes of PCPs--low molecular weight (L-PCPs) and high molecular weight (H-PCPs)--revealed that the morphologically distinct outer lamellae contained L-PCPs and the inner lamellae contained H-PCPs. The sharp boundary between these two antigenic domains coincides with the transition from the outer to the inner lamellae, which in turn is correlated with the cessation of L-PCP synthesis and the initiation of H-PCP synthesis in response to 20-hydroxyecdysone (Doctor, J., D. Fristrom, and J.W. Fristrom, 1985, J. Cell Biol. 101:189-200). Hence, differences in protein composition are associated with differences in lamellar morphology.     

Studies on plant cuticle

The cuticles of plants of the Saxifragaceae, Rosaceae and Leguminosae are compared by chemical methods. Wide differences occur in the deposits of surface wax and cuticular membrane even within species of one genus. The relative proportions of four hydroxy-fatty acids in the cutin acids of plants of the families are assessed and the value of cutin analysis as a taxonomic criterion is discussed.     

Freeze-fracture studies on tardigrade cuticle

The cuticles of the heterotardigrade Echiniscus testudo and the eutardigrades Macrobiotus hufelandi and Milnesium tardigradum have been studied using freeze-fracture technique. Most of the layers seen in conventional TEM micrographs can be visualized. There is no clear evidence that the trilaminar components of the cuticle such as the outer epicuticle and the tripartite layer separating epi- and intracuticle or procuticle (whose membranous origin has been suggested by previous authors) fracture like a lipid bilayer. Microfibres not resolved or only poorly resolved by TEM can be recognized in the procuticle of all three species. Obviously their visualization depends upon the fracture angle. In Echiniscus testudo and Milnesium tardigradum the intracuticle or at least parts of it show a wavy arrangement of microfibres. Parts of the ventral intracuticle of E. testudo fracture in an obviously non-random pattern revealing distinct sublayers.     

The stabilization of locust cuticle

Cuticle from the metathoracic femur of adult locusts (Locusta migratoria) is characterized with respect to changes in water content and in protein extractability during maturation. The swelling behaviour and extractability of fully-sclerotized cuticle are compared to those of chemically-modified, unsclerotized cuticle.It is concluded that although dehydration may contribute to the stabilization of cuticle, it cannot account for the observed differences. The properties of mature cuticle can best be explained by the assumption that covalent cross-links are present between protein molecules.     

Structure and expression of a Manduca sexta larval cuticle gene homologous to Drosophila cuticle genes

A genomic clone was isolated from the tobacco hornworm, Manduca sexta, by virtue of its similarity to a Drosophila larval cuticle gene. RNA analysis shows that this clone, B311, is expressed at times appropriate for a larval cuticle gene. Hybrid-selection experiments using B311 DNA show that it encodes a 14 x 10(3) Mr protein, LCP-14, which is precipitated by an antiserum to Manduca larval cuticle. We have sequenced both genomic and cDNA clones for the LCP-14 gene. A conceptual translation of the cDNA sequence shows that the LCP-14 protein is similar not only to another Manduca cuticle protein, but also to Drosophila, Sarcophaga and Hyalophora cecropia cuticle proteins. Since these proteins are found in flexible cuticle and have similar sequences, we conclude they are encoded by homologous genes.     

An improved water content model for Periplaneta cuticle: Effects of epidermis removal and cuticle damage

All previously reported integumental permeabilities to water for Periplaneta at 20°C have been shown to be overestimates by at least an order of magnitude. Cuticle damage, apparently due to mechanical contact, is a common occurrence in cockroach cultures. Damage is increased by handling during experimental manipulation. Repair of the damage is readily observed in isolated individuals as cuticle permeability declines over a few days. Repaired cuticle permeability is not humidity dependent, but damage affecting a few per cent of the cuticle area, renders it so. Cuticle-water contents are slightly influenced by sex related size differences. Cuticle-water contents are unaffected by variations in ambient activity. Epidermal removal initiates a drop in cuticle-water content, apparently not involving active processes. Cuticle-water contents can be accurately predicted in any ambient vapour pressure using a two-layered model combining equilibrium water contents and component permeabilities. Incorporating a parallel unprotected endocuticle pathway into the model successfully simulates the observed humidity dependence of damaged permeabilities. Epidermal permeability cannot be less than 2× 10⁻⁵ cm.s⁻¹; 2000 times more permeable than the overlying cuticle.     

An overview on plant cuticle biomechanics

Plant biomechanics combines the principles of physics, chemistry and engineering to answer questions about plant growth, development and interaction with the environment. The epidermal-growth-control theory, postulated in 1867 and verified in 2007, states that epidermal cells determine the rate of organ elongation since they are under tension, while inner tissues are under compression. The lipid cuticle layer is deposited on the surface of outer epidermal cell walls and modifies the chemical and mechanical nature of these cell walls. Thus, the plant cuticle plays a key role in plant interaction with the environment and in controlling organ expansion. Rheological analyses indicate that the cuticle is a mostly viscoelastic and strain-hardening material that stiffens the comparatively more elastic epidermal cell walls. Cuticle stiffness can be attributed to polysaccharides and flavonoids present in the cuticle whereas a cutin matrix is mainly responsible for its extensibility. Environmental conditions such as temperature and relative humidity have a plasticizing effect on the mechanical properties of cuticle since they lower cuticle stiffness and strength.The external appearance of agricultural commodities, especially fruits, is of great economic value. Mechanical properties of the cuticle can have a positive or negative effect on disorders like fruit cracking, fungal pathogen penetration and pest infestation. Cuticle rheology has significant variability within a species and thus can be subjected to selection in order to breed cultivars resistant to pests, infestation and disorders.