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T R Tokarski

Publications and source records attributed to T R Tokarski.

8 recordsLinked to original sources

Photopigment gene expression and rhabdom formation in the crayfish (Procambarus clarkii).

This study examines the expression of the photopigment gene in the developing retina of the freshwater crayfish Procambarus clarkii(Crustacea, Malacostraca, Decapoda). Both sense and anti-sense RNA probes were used for in situ hybridization (ISH) of whole embryos collected at various stages during development. A characteristic of retinal development is the formation of screening pigment in the retinular cells of the retinal ommatidia. This pigmentation is seen as a band that begins at the lateral side of the retinal field and progresses medially. At hatching the retina is approximately 50% pigmented. ISH of whole embryos shows that expression of the photopigment gene by the retinular cells correlates with the extent of the screening pigment band in the retina and with the presence of rhabdoms within the ommatidia. Sections taken through embryos after being hybridized indicate that staining is localized in the cytoplasm of the retinular cells and in the axonal region below the basement membrane. No staining reaction was seen in the rhabdoms of older ommatidia. ISH staining was also seen at the anterior midline of the protocerebrum where extraretinal photoreceptors have been reported. The data presented here show a close correlation of opsin expression within the retinular cells of the ommatidia and the formation of the very early rhabdoms, similar to Drosophila. The results will be discussed in relation to recent studies in Drosophila that suggest rhodopsin plays a role in effecting the organization of the terminal web-like cytoskeleton at the base of the developing rhabdom microvilli.

Animals↗

Retinal development in the lobster Homarus americanus. Comparison with compound eyes of insects and other crustaceans.

Pattern formation and ommatidial differentiation were examined in the developing retina of the lobster Homarus americanus using light and electron microscopy. In the lobster the retina differentiates from the surface ectoderm that covers the optic primordia. Initially a single band of proliferation moves across this surface ectoderm. Immediately following this wave of proliferation, rows of ommatidial cell clusters appear. The earliest cell clusters are often seen adjacent to dividing cells of the proliferation band. The changing organization of the first seven rows of ommatidial clusters, visible at the surface of the retina, reveals events in early ommatidial differentiation. A rosette-like cluster of 18 cells forms the first row. Each stage following the rosette clusters occurs in a separate staggered row. Developing ommatidia have a central cluster of retinula cells, whose organization changes at each stage. Four cone cells enclose the retinula cells in each cluster and extend to the surface. In the seventh row, rhabdome formation begins and the retinula cells recede, leaving only cone cells visible at the retinal surface. This change initiates the two-tiered organization of the adult ommatidium. In 70% embryos, asymmetries in the position of the R8 axon around R7 create an equatorial line separating the dorsal and ventral halves of the retina. Possible mechanisms for the formation of these asymmetries are discussed. Postembryonic growth of the retina continues in stage VI juvenile animals along the ventral edge of the retina.

Age Factors↗

Morphogenesis and pattern formation in the retina of the crayfish Procambarus clarkii.

Pattern formation and ommatidial differentiation in the crayfish retina were analyzed using confocal, light and electron microscopy. Optic primordia first appear in the embryo as round elevations covered by a surface epithelial layer. Retinal differentiation begins with a wave of mitotic activity that moves across this epithelium from lateral to medial. Ommatidial cell clusters are visible at the surface along a transition zone, which lies at the interface of the medial undifferentiated retina and the lateral patterned retina. This zone is 8-10 cells wide and composed of small uniform cell profiles. Lateral to the transition zone the initial ommatidial cell clusters form staggered rows across the surface. Each first row cluster contains eight retinula cells surrounded by four cone, two corneagenous and two distal pigment cells. Ommatidial clusters in the first nine rows show significant changes in their organization, which are visible at the surface of the retina. In row 10 the retinula cells recede from the surface and the cone cells close in above them creating a constant cell pattern at the surface. Rhabdome development begins distally and extends downward as the retinula cluster recedes from the surface. Movement of the retinula cells inward and enlargement of the cone and corneagenous cells at the surface creates a two-tiered organization characteristic of each ommatidium. Comparison of retinal pattern formation and differentiation in the crayfish with retinal morphogenesis in Drosophila and other insects show several similarities between the two arthropod groups.

Actins↗

Localization of actin in the retina of the crayfish Procambarus clarkii.

The distribution of actin in the retina of the crayfish was investigated at the LM level using FITC-phalloidin. Fluorescent staining was associated with the main rhabdom and eighth cell rhabdom, the zonula adherens junctions between retinula cells, and the basement membrane of the retina. EM and S1 decoration were used to confirm the presence of actin and identify its structural relationships. Phalloidin staining of the rhabdom and S1 decoration of actin filaments in the rhabdom microvilli confirmed earlier findings that actin is a component of the microvillus cytoskeleton in the crayfish. At the zonula adherens junctions, actin filaments, identified by S1 decoration, run longitudinally within the plaque of the junction. At the extreme proximal end of the rhabdom, actin filaments associated with the junctions fill each small area of retinula cell cytoplasm. In the basement membrane, EM and S1 decoration show that basilar cells contain large bundles of actin filaments which are associated with cell-matrix adherens junctions. Foot cells which lie immediately below the rhabdom also contain similar junctions and actin is tentatively identified in these cells. The functional role of actin at these various locations is discussed in relation to retinal organization in the crayfish and other invertebrates.

Actin Cytoskeleton↗

Changes in the microvillus cytoskeleton during rhabdom formation in the retina of the crayfish Procambarus clarkii.

Changes in the microvillus cytoskeleton during the formation of the light-receptive rhabdom in the crayfish retina were examined at four structurally distinct stages. The cytoskeleton of microvilli in early rhabdoms is composed of a regularly packed bundle of 12-25 actin filaments. The polarity of S1 decorated filaments indicates that the plus end of the actin filaments is located at the microvillus tip. The hexagonal packing of filaments within the bundle, their spacing, and the presence of cross-striations along the bundle in longitudinal sections indicate the filaments are held together by cross-linking proteins. Electron microscopic observations and data from three-dimensional reconstructions of individual microvilli indicate that the filaments arise from a concentration of dense material at the tip of the microvillus and extend into the cytoplasm as a rootlet. Over the four developmental stages examined there is an increase in the number of microvilli forming the rhabdomeres and a 50% decrease in the mean cross-sectional area of individual microvilli. During this same period the number of actin filaments forming the microvillus cytoskeleton also decreases. Following this decline, microvilli of late stage rhabdoms, which are structurally similar to adults, contain only two to four filaments. These changes are discussed in relation to the three phases of growth described for stereocilia and brush border microvilli.

Actin Cytoskeleton↗

The diurnal pattern of protein and photopigment synthesis in the retina of the crayfish, Procambarus clarkii.

The interrelationship between the diurnal cycle of membrane loss and synthesis of new rhabdom components remains a key element in forming a complete picture of the turnover of photopigment-containing membrane in the crayfish photoreceptor cell. In order to examine this aspect of the turnover process, the diurnal pattern of photopigment synthesis was examined using an in vitro incubation system for incorporation of 3H-leucine into photoreceptor protein. The incorporation of 3H-leucine into total protein and photopigment specifically was measured in photoreceptors isolated from incubated retinas. The results indicate that for both total protein and photopigment there is no significant variation in the rate of synthesis during the 12-12 light-dark cycle. These data combined with earlier data on diurnal membrane loss from the rhabdom suggest that light-stimulated rhabdom membrane loss is superimposed on a diurnally constant level of synthesis and assembly of new rhabdom constituents.

Animals↗

Regional morphological variations within the crayfish eye.

The existence of structural asymmetries has been quantitatively demonstrated in the crayfish compound eye. Variations in the size of the rhabdomes and corneal facets, as well as the size and extent of the accessory reflecting pigment cells, have been found. It was determined that the mean rhabdome diameter within a 70 degrees arc in the dorsal quadrant of the retina is 11-19% smaller than the mean rhabdome diameter in the remaining areas of the eye. Also, the extent of the accessory reflecting pigment cells is diminished over an area corresponding generally to the dorsal region of smaller rhabdomes. Corneal facet size and shape vary over the surface of the cornea, with smaller facets occurring in the dorsal region. Both the mean rhabdome diameter and the mean corneal facet area for whole eyes increases linearly in animals ranging in size from 3.9-12 cm. The estimated number of corneal facets, and therefore the number of rhabdomes, increases from an average of 4700 in the 3-6.9 cm size range to about 6000 in 7-12 cm animals. These data indicate that structural asymmetries and various size-related parameters exist in the crayfish eye and should be considered in any quantitative analysis of this structure.

Animals↗