Photoreceptor morphogenesis and retinal degeneration: lessons from Drosophila

Cells exhibit an amazingly wide range of different forms, and in most cases the shape of a cell is crucial for performing its specific function(s). But how does a cell obtain its particular shape during development, how can the shape be adapted to different environmental conditions, and what are the consequences if morphogenesis is impaired? An ideal cell type to study these questions is the photoreceptor cell, a photosensitive cell present in most metazoa, highly specialised to transform the energy from the light into a visual response. In the last few years, studies in the Drosophila eye have led to a considerable increase in understanding of the genetic control of photoreceptor morphogenesis; lessons, which may apply to other cell types as well. Most of the genes involved have been conserved during evolution, and mutations in several of them result in retinal degeneration, both in flies and humans. This makes the fly eye an attractive model to unravel the genetic, molecular and cell biological basis of the mechanisms that prevent retinal dystrophies.     

Invagination and evagination: a hydromechanical model

It has been proposed that processes of invagination and evagination be considered in the context of a single, "hydromechanical," model. The model is based on the assumption that the structures demonstrating invaginations and evaginations are closed systems capable of changing their intracavitary pressure in an autonomous regime. The mass, which occupies the cavity, should overcome the resistance of the surrounding membrane during its growth. In places of the weakest resistance, the inner mass expands especially quickly: it bulges out. If the pressure under the membrane is less than the exterior one, the process goes in the opposite direction: the outer mass intrudes (invaginates) into the cavity, causing its wall to sag in the weakest regions.     

The mechanism of evagination of imaginal discs of Drosophila melanogaster. V. Evagination of disc fragments.

Experiments designed to test the hypothesis that imaginal disc evagination may be accomplished by a continuous circumferential constriction of the basally located microfilaments in disc cells [Fristrom, D., and Fristrom, J. W. (1975). Develop. Biol.43, 1–23] are presented. Discs were cut in various planes so as to interrupt the hypothesized lines of constriction and the resulting fragments tested for the ability to evaginate in vitro. Fragments from each type of cut were able to undergo evagination without resealing of the would surfaces. Furthermore, sections of evaginated cut fragments showed no evidence for a general contraction of the basal cell surface. We conclude that the ability to evaginate is an intrinsic property of each fragment independent of its attachment to the rest of the disc.     

Murine lambda gene rearrangements: the stochastic model prevails over the ordered model.

The ontogeny of the immunoglobulin (Ig) gene rearrangement in mammalian B cells seems to be ordered. Heavy chain gene segments rearrange first, followed by light chain gene segments, kappa before lambda. The genomic organization of murine lambda locus does not preclude the simultaneous expression of two subtypes from the same chromosome. In order to distinguish between an ordered and a stochastic model of rearrangement, a panel of 67 B cell hybridomas secreting either lambda 1, lambda 2, lambda 3 or lambda x (recently described) were analysed for V lambda J lambda rearrangements. The results show that in 97% of cases, a single rearrangement occurred, favouring the stochastic model over the ordered one. Strikingly, the possibility of having a productive rearrangement if the first try results in an aberrant one is rare. We propose therefore, that the lambda Ig is not necessarily required to ensure allelic and subtypic exclusion mechanisms. Moreover, in 97% of the cases, at least one kappa allele is rearranged. Furthermore, the RS recombination has been detected in 77% of the cases. This suggests that, although the stimulation of kappa precedes that of lambda locus, the RS recombination acts as a transacting albeit dispensable lambda activator.     

Microtubule assembly and phage morphogenesis: new results and classical paradigms

The classical analyses of phage morphogenesis provide experimental paradigms for dissecting other assembly pathways. A new set of results, derived from examination of the quantitative controls of tubulin levels and microtubule assembly in Saccharomyces cerevisiae, evokes the 'balance of components' hypothesis. These results show that balanced levels of the tubulin proteins are crucial for microtubule assembly. Imbalances leading to excess beta tubulin have far more deleterious consequences than those leading to excess alpha tubulin, including dramatic cellular toxicity, quantitative depolymerization of cellular microtubules, and self-aggregation of the excess beta tubulin. These and other results suggest that beta tubulin may possess a unique ability to interact with a component of microtubule nucleating sites, and provide a rationale for the universal polarity of nucleated microtubules.     

Sugarbeet leaf disc culture: an improved procedure for inducing morphogenesis

In preparation for gene transfer experiments we investigated factors that might affect the production of shoots and somatic embryos from the wound callus of cultured sugarbeet leaf discs. A complex interaction was found between the leaf disc plating density, the disc culture medium, the source-shoot culture medium and the frequency of disc transfer to fresh medium. The most productive protocol utilized: source shoots maintained on MS medium containing 0.25 mg 1^-1 BA; multiple leaf discs (ten 4-mm discs/plate) plated onto an enriched modification of MS medium (RV) containing 1.0 mg 1^-1 BA and solidified with 0.3% Gelrite (not permitted to dry during hardening); and transfer of the discs to fresh medium every two weeks during the first month. This standard protocol produced more callus per plate and higher rates of morphogenesis per unit dry weight of callus than did the one-step method of Saunders and Doley. Water availability considerations were found to be critical to obtaining high morphogenic rates. Root induction frequency and quality was superior on shoots transplanted to MS medium containing 1 mg 1^-1 NAA as the sole growth regulator compared to IAA at the same concentration.Abbreviations BA N⁶-benzyladenine - IAA indole-3-acetic acid - NAA -naphthaleneacetic acid     

器官成形素调控果蝇翅芽细胞形貌的研究进展

果蝇翅芽是研究细胞形貌发生的模式系统。在果蝇翅芽的发育过程中,器官成形素由浓度高的区域(成形素表达细胞)向浓度低的区域(接收细胞)移动,形成动态的浓度梯度。器官成形素信号通路的激活调控翅芽细胞的形貌发生、存活、生长和分化。目前已鉴定的在翅芽细胞表达的器官成形素包括Hedgehog(Hh),Decapentaplegic(Dpp)和Wingless(Wg)。结合国际最新研究进展,本文综述了3种器官成形素在翅芽细胞形貌发生过程中的重要作用,讨论了细胞形貌发生的分子机制。     

Euglena: The Photoreceptor System for Phototaxis*

SYNOPSIS. The photoreceptor structures (eyespot-paraflagellar body-flagellum) for Euglena phototaxis were investigated by electron microscopy. The paraflagellar body—the photoreceptor—is a highly ordered crystalline lamellar structure. Optical diffraction of the electron micrographs and resulting filtered images of the paraflagellar body suggest that it is formed of rods in a helical arrangement. The action spectra for phototaxis, the in situ spectrum by microspectrophotometry of the paraflagellar body, and flavin analysis of the organism indicate that the photoreceptor molecule is a flavoprotein. The phototaxis action spectrum is similar to the spectrum for O₂ evolution and implies that similar molecules participate in the photo-processes. As a result, a photochemical scheme is suggested in which a photo-excited flavin and a cytochrome participate in the photoprocess. The photochemistry and photoreceptor structures for Euglena phototaxis are likened to a photoneuro sensory cell.     

The evagination of the genital imaginal discs ofDrosophila melanogaster

Summary The morphology of the evaginating female genital disc ofDrosophila melanogaster was examined at different stages of metamorphosis. The observations show that the internal genital organs are derived from the anterior half of the disc and that their morphogenesis is mainly a protrusion of the different primordial areas of the disc epithelium. The external genital and anal derivatives originate from the posterior half of the disc, which undergoes complex rearrangements during metamorphosis. The disc opens along the posterior margin and the dorsal and ventral epithelia evert and thereby completely reverse their anteroposterior orientation. Dramatic elongation has been observed during the formation of the seminal receptacle. The cells of the repressed male genital primordium do not form any recognizable structures and are assumed to be eliminated during metamorphosis.     

Pattern formation and morphogenesis: A reaction-diffusion model

The problem of cellular differentiation and consequent pattern generation during embryonic development has been mathematically investigated with the help of a reaction-diffusion model. It is by now a well-recognized fact that diffusion of micromolecules (through intercellular gap junctions), which is dependent on the spatial parameter (r), serve the purpose of ‘positional information’ for differentiation. Based on this principle the present model has been constructed by coupling the Goodwin-type equations for RNA and protein synthesis with the diffusion process. The homogeneous Goodwin system can exhibit stable periodic solution if the value of the cooperativity as measured by the Hill coefficient (ρ) is greater than 8, which is not biologically realistic. In the present work it has been observed that inclusion of a negative cross-diffusion can drive the system into local instability for any value of ρ and thus a time-periodic spatial solution is possible around the unstable local equilibrium, eventually leading to a definite pattern formation. Inclusion of a negative cross-diffusion thus makes the system biologically realistic. The cross-diffusion can also give rise to a stationary wave-like dissipative structure.     

The Dorsocross T-box transcription factors promote tissue morphogenesis in the Drosophila wing imaginal disc

The Drosophila wing imaginal disc is subdivided into notum, hinge and blade territories during the third larval instar by formation of several deep apical folds. The molecular mechanisms of these subdivisions and the subsequent initiation of morphogenic processes during metamorphosis are poorly understood. Here, we demonstrate that the Dorsocross (Doc) T-box genes promote the progression of epithelial folds that not only separate the hinge and blade regions of the wing disc but also contribute to metamorphic development by changing cell shapes and bending the wing disc. We found that Doc expression was restricted by two inhibitors, Vestigial and Homothorax, leading to two narrow Doc stripes where the folds separating hinge and blade are forming. Doc mutant clones prevented the lateral extension and deepening of these folds at the larval stage and delayed wing disc bending in the early pupal stage. Ectopic Doc expression was sufficient to generate deep apical folds by causing a basolateral redistribution of the apical microtubule web and a shortening of cells. Cells of both the endogenous blade/hinge folds and of folds elicited by ectopic Doc expression expressed Matrix metalloproteinase 2 (Mmp2). In these folds, integrins and extracellular matrix proteins were depleted. Overexpression of Doc along the blade/hinge folds caused precocious wing disc bending, which could be suppressed by co-expressing MMP2RNAi.     

Plant morphogenesis: A geometrical model for the ramification

A geometrical model is proposed that describes the emergence of a primordium at the shoot apex in Dicotyledons. It is based on recent fundamental results on plant morphogenesis, viz.:

the emergence is preceded by the reorganization of the microtubules of the cortical cytoskeleton, leading to a new orientation of the synthesis of the cell wall microfibrils;

the resulting global stress is related to the general orientation of the cell growth;

The model sums up the continuous interactions that link the microtubules, the microfibrils and the cell growth axis. The paper tries to answer three essential questions:

Why does the principal stem shifts its growth direction after each lateral emergence?

Why do the three axes involved in any ramification (namely the old and the new principal stems and the lateral emergence) exhibit a plane configuration whereas this is an essentially three dimensional phenomenon?

Does phyllotaxis exclusively depend upon the local emergence of a primordium?

An interactive procedure between empirical botanical knowledge and the mathematical model leads to an insight of the compatibility mechanisms that link the various microtubules and microfibrils networks, and the apical dome restoration. A geometrical formalism allows a redefinition of both the “generating centre” and the “organizing centre”, and their field effect.     

The role of the peripodial membrane in the morphogenesis of the eye-antennal disc ofDrosophila melanogaster

Summary The early morphogenesis of the eye-antennal disc ofDrosophila in response to 20-hydroxy ecdysone involves the curling of the eye anlagen dorsally over the antenna. During this process, the area of the peripodial membrane is substantially reduced. The peripodial membrane is taut at this stage, and if it is cut the curling of the disc cannot continue, and the eye anlagen returns to its original position within one minute of the operation. In contrast, cutting the columnar epithelium between the eye and antennal anlagen does not disrupt curling, but actually facilitates it. During curling, the cells of the peripodial membrane appear healthy, and exhibit basal extensions. We suggest that the curling of the eye is mediated by the conversion of cuboidal peripodial membrane cells into pseudostratified columnar epithelium at the edges of the peripodial membrane. Subsequently, cells of the peripodial membrane secrete first a pupal cuticle, and then an imaginal cuticle.     

The Ventral Photoreceptor Cells of Limulus : I. The microanatomy

The ventral photoreceptor cells of Limulus polyphemus resemble the retinular cells of the lateral eyes both in electrical behavior and in morphology. Because of the great size of the ventral photoreceptor cells they are easy to impale with glass capillary micropipettes. Their location along the length of the ventral eye nerve makes them easy to dissect out and fix for electron microscopy. Each cell has a large, ellipsoidal soma that tapers into an axon whose length depends upon the distance of the cell from the brain. The cell body contains a rich variety of cytoplasmic organelles with an especially abundant endoplasmic reticulum. The most prominent structural feature is the microvillous rhabdomere, a highly modified infolding of the plasmalemma. The microvilli are tightly packed together within the rhabdomere, and quintuple-layered junctions are encountered wherever microvillar membranes touch each other. Glial cells cover the surface of the photoreceptor cell and send long, sheet-like projections of their cytoplasm into the cell body of the photoreceptor cell. Some of these projections penetrate the rhabdomere deep within the cell and form quintuple-layered junctions with the microvilli. Junctions between glial cells and the photoreceptor cell and between adjacent glial cells are rarely encountered elsewhere, indicating that there is an open pathway between the intermicrovillous space and the extracellular medium. The axon has a normal morphology but it is electrically inexcitable.