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R M Gaze

Publications and source records attributed to R M Gaze.

At least 19 recordsLinked to original sources

Spatio-temporal patterns of retinal ganglion cell death during Xenopus development.

During development of the retina in mammals and birds, most retinal ganglion cells (RGC) that are produced are eliminated later in development by cell death. In lower vertebrates, however, such massive cell death has not been observed; total ganglion cell number increases linearly during most of development. Using 3H-thymidine or 5-bromodeoxyuridine labeling of retinal cell nuclei, we have been able to identify postmitotic RGC populations in Xenopus central retina at different developmental stages and follow their fate during development to postmetamorphic stages. RGC populations that become postmitotic between embryonic stages 32 and 49, during the initial stages of retinal growth, lose 40-77% of their cells during metamorphosis (approximately 4,000-5,000 cells). Twenty percent of the RGC present at stage 54, which later disappear, represent the same population of dying RGC that were present at stage 49. This suggests that the ganglion cells that became postmitotic between stage 49 and 53/54 show no apparent decline in numbers during metamorphosis. Since thyroxine is known to stimulate an increase in RGC number as well as the extent of fiber projection on the tectum, we suggest that this reduction in RGC numbers is not due to thyroxine-induced neuronal cell death. After stage 54, however, binocular vision develops in Xenopus (Keating, '74) and ipsilateral fibers begin to grow into thalamic visual neuropils (Hoskins and Grobstein, '85). We suggest, therefore, that as in mammals, in which RGC elimination correlates with binocular segregation of contralateral and ipsilateral retinal axons in visual centers, a similar process may occur in the frog among those RGC projecting to thalamic visual neuropils.

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Development of the tectum and diencephalon in relation to the time of arrival of the earliest optic fibres in Xenopus.

The development of the tectum and diencephalon in Xenopus has been investigated in relation to recent descriptions of the establishment of the retinotectal projection. Tritiated thymidine autoradiography and bromodeoxyuridine immunohistology were used to identify the stages at which cells became postmitotic. Cells in the diencephalon were found to become postmitotic before cells in the tectum. At the time of arrival of the first optic fibres (stage 37/38) no postmitotic cells appeared to be present in the tectal precursor region. The first postmitotic cells which could be definitely assigned to the tectum appeared between stages 41 and 45. The results suggest that the initial retinotopic ordering of optic fibres observed from stage 37/38 relates to the position of fibres in the optic tract and not the tectum.

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Regeneration in the Xenopus tadpole optic nerve is preceded by a massive macrophage/microglial response.

Changes in the optic nerve following a crush lesion and during axonal regeneration have been studied in Xenopus tadpoles, using ultrastructural and immunohistological methods. Degeneration of both unmyelinated and myelinated axons is very rapid and leads to the formation, within 5 days, of a nerve which consists largely of degeneration debris and cells. Immunohistological analysis with monoclonal antibody 5F4 shows that there is a rapid and extensive microglial/macrophage response to crush of the nerve. Regenerating axons have begun to enter the distal stump by 5 days and grow along the outer part of the nerve in close approximation to the astrocytic glia limitans. Between 5 and 10 days after nerve crush, regenerating axons reach and pass the chiasma. Macrophages are seen in the nerve at the site of the lesion within 1 h, and the response peaks between 3-5 days, just before axonal regeneration gets under way.

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Microglia in tadpoles of Xenopus laevis: normal distribution and the response to optic nerve injury.

We have studied the distribution of microglia in normal Xenopus tadpoles and after an optic nerve lesion, using a monoclonal antibody (5F4) raised against Xenopus retinas of which the optic nerves had been cut 10 days previously. The antibody 5F4 selectively recognizes macrophages and microglia in Xenopus. In normal animals microglia are sparsely but widely distributed throughout the retina, optic nerve, diencephalon and mesencephalon (other regions were not examined). After crush or cut of an optic nerve, or eye removal, there occurs an extensive microglial response along the affected optic pathway. Within 18 h an increase in the number of microglial cells in the optic tract and tectum can be detected. This response increases to peak at around 5 days after the lesion. At this time the nerve distal to the lesion contains many microglial cells; the entire optic tract is outlined by microglia, extended along the degenerating fibres; and the affected tectum shows a heavy concentration of microglia. This microglial response thereafter decreases and has mostly gone by 34 days. We conclude that the microglial response to optic nerve injury in Xenopus tadpoles starts early, peaks just before the regenerating optic nerve axons enter the brain, and is much diminished by the time the retinotectal projection is re-established. The timing is such that the microglial response could play a major role in facilitating regeneration.

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Regeneration of optic fibres through the chiasma in Xenopus laevis tadpoles.

The path through the chiasma followed by regenerating optic nerve fibres in Xenopus tadpoles was studied at light- and electron-microscopic levels, and with horseradish peroxidase as a fibre label. Over the period (5-10 days) in which regenerating fibres reach and cross the chiasma, they did not follow residual deep fibres through the chiasma, nor were they associated with the trail of degeneration in the chiasma which represented the remains of the deeper (older) parts of the original projection. The regenerating optic fibres were always seen in close association with the inner surfaces of the ependymoglial endfeet, or in the extracellular spaces that lie close to the endfeet in the most superficial part of the chiasma, where newly-growing fibres from the retinal margin are normally to be found.

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The induction of an anomalous ipsilateral retinotectal projection in Xenopus laevis.

The conditions necessary for extensive regeneration of fibres from one optic nerve to both tecta in Xenopus have been investigated. The effects of various types and positions of nerve lesion on the distribution of regenerated projections were examined by labelling the regenerated projections with either horseradish peroxidase or tritiated proline. The only types of nerve lesion which consistently gave rise to extensive regeneration to both the contralateral and the ipsilateral tectum were those close to the chiasma, liable to have caused damage to the nerve entry point. However, all other types of lesion studied (near the eye; near the skull; crush or cut) frequently led to regeneration of a very few fibres to the ipsilateral tectum. These fibres gained access to the ipsilateral tectum by various routes: in some cases via the optic tract; but more commonly either across the posterior commissure or by a complex pathway following the oculomotor nerve root. Over time periods of between one and seven months, the distribution of the regenerated fibres following each type of lesion showed little change. We conclude that the regeneration of bilateral retinotectal projections in Xenopus is caused by tissue damage to the region of the chiasma resulting in misrouting of fibres.

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The course of regenerating retinal axons in the frog chiasma: the influence of axons from the other eye.

There is evidence, from a variety of species, that axons arising from the two eyes interact as they pass through the optic chiasma. In this paper we examine the effect of altering the composition of the normal chiasmatic environment on the course taken by regenerating axons in Xenopus. We show that the usual contralateral route of such axons in the optic chiasma is not affected by the absence of axons from the other eye. We have not observed any influence of embryonic enucleation, or eye removal before the onset of metamorphosis, on either the normal ipsilaterally projecting axons or the contralateral route taken by regenerating axons. Further, we have found no effect of the degeneration of the other projection, either for long or short periods of time, suggesting that axonal debris does not influence the course of regenerating axons. The simultaneous regeneration of the two projections also has no effect on the trajectory of regenerating axons. We conclude that in spite of the close association of axons from the two eyes in the chiasma, there is no interaction involved in Xenopus in the generation of the decussation pattern.

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A developmental and ultrastructural study of the optic chiasma in Xenopus.

The structure of the optic chiasma in Xenopus tadpoles has been investigated by light and electron microscopy. Where the optic nerve approaches the chiasma, a tongue of cells protrudes from the periventricular cell mass into the dorsal part of the nerve. Glial processes from this tongue of cells ensheath fascicles of optic axons as they enter the brain. Coincident with this partitioning, the annular arrangement of axons in the optic nerve changes to the laminar organization of the optic tract. Beyond the site of this rearrangement, all newly growing axons accumulate in the ventral-most part of the nerve and pass into the region between the periventricular cells and pia which we have called the 'bridge'. This region is characterized by a loose meshwork of glial cell processes, intercellular spaces and the presence of both optic and nonoptic axons. In the bridge, putative growth cones of retinal ganglion cell axons are found in the intercellular spaces in contact with both the glia and with other axons. The newly growing axons from each eye cross in the bridge at the midline and pass into the superficial layers of the contralateral optic tracts. As the system continues to grow, previous generations of axon, which initially crossed in the existing bridge, are displaced dorsally and caudally, forming the deeper layers of the chiasma. At their point of crossing in the deeper layers, these fascicles of axons from each eye interweave in an intimate fashion. There is no glial segregation of the older axons as they interweave within the chiasma.

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The discontinuous visual projections on the Xenopus optic tectum following regeneration after unilateral nerve section.

The establishment of retinotectal projections following transection of one optic nerve in developing Xenopus has been investigated. Between 3 weeks and 11 months after the operation, the nerve fibre tracer horseradish peroxidase (HRP) was applied to either the operated or the unoperated nerve, and the brains were prepared for examination as whole mounts. In most cases fibres from the operated nerve innervated both tecta, with the result that one tectum was doubly innervated and one tectum singly innervated. Two months after transection of the optic nerve in tadpole life, between stages 50 and 54, this nerve usually made a uniform projection on the contralateral tectum and a striped projection on the ipsilateral, doubly innervated, tectum. The projection made by the unoperated nerve on this tectum was a similar pattern of stripes, which ran generally rostrocaudally. Two months after transection of the optic nerve of newly metamorphosed animals, the projection formed by the operated nerve on the doubly innervated tectum was usually a pattern of spots or spots mixed together with stripes in no particular orientation superimposed on a roughly uniform background. In a small number of cases the projections made by the same nerve on the two tecta were approximately complementary; that is, the presence of label on one tectum corresponded with its absence on the other tectum. The results are examined in the context of the development of the retina and of the tectum. It is suggested that the consistently oriented stripes which result from nerve transection at a stage at which only a small proportion of the retinal fibres had reached the tectum are formed by the interaction of two equally matched sets of developing fibres, stripe orientation being determined by the mode of growth of the optic tectum. The formation of patterns of spots or spots mixed together with stripes following nerve transection after the end of the main phase of tectal histogenesis, and when 50% of the optic fibres had already reached the tectum, is attributed to an unequal competition between the two sets of fibres.

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The distribution of fibres in the optic tract after contralateral translocation of an eye in Xenopus.

In Xenopus embryos of stage 30 the right eye was translocated, without rotation, to a left host orbit. Shortly after metamorphosis the visuotectal projection through the operated eye was mapped electrophysiologically and shown to be normal dorsoventrally but reversed nasotemporally. Labelling of small groups of retinal axons with HRP showed that the fibre trajectories from dorsal and ventral retina were normal, whereas fibres from nasally placed retina had diencephalic pathways and tectal terminations typical and temporal fibres, and fibres from temporally placed retina had diencephalic pathways and tectal terminations typical of nasal fibres. Thus from just beyond the chiasma the fibres had already achieved the major uniaxial rearrangement necessary to establish a normal tract distribution despite the eye translocation. The fibre rearrangement required to permit the formation of a nasotemporally inverted visuotectal projection appears, therefore, to occur not on the tectum or in the optic tract, but either within the nerve or at the chiasma.

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The effects of the fibre environment on the paths taken by regenerating optic nerve fibres in Xenopus.

The paths taken by fibres regenerating to the tectum from various parts of the Xenopus retina were investigated in whole-mount preparations, after localized retinal labelling with HRP. The effects of different environments on the fibres were studied by comparing contralateral with ipsilateral regeneration, in the presence of the other eye or after it had been removed in embryonic life. Under all conditions fibres from the various parts of the retina regenerated to the corresponding appropriate parts of the tectum, but they took a variety of pathways, some grossly abnormal, to get there. Contralaterally regenerating fibres tended to behave less abnormally than ipsilateral fibres; and regeneration in the absence of the other eye tended to be more abnormal than in its presence. In any one category of regeneration the most nearly normal pathways were those of fibres from temporal retina, followed by ventral, nasal and dorsal fibres. Fibres regenerating from all parts of the retina, in the presence of the other eye, tended to become gathered into the medial brachium as they approached the tectum. All regenerating fibres approached their tectal terminations by one or more of three main pathways: round one or both brachia, thus encircling the tectum to get to their terminal zone; directly across the tectum; or by passing on to the tectum before changing course. The changes of direction required to enable fibres wrongly positioned in the tract to reach their correct terminal zones were frequently sudden and considerable, and took place on the tectum or at the tectodiencephalic junction. The results are discussed in relation to the differing substrates over which the fibres regenerate.

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The central pathways of optic fibres in Xenopus tadpoles.

A cobalt chloride impregnation technique was applied to the optic nerve in Xenopus tadpoles and the central optic pathways were examined in cleared, whole-mounted preparations, and in thick sections. The overall plan of the optic input was visualized in relation to the outlines of the parts of the brain and details of the structure of the tectal optic neuropil, the neuropil of Bellonci and the basal optic neuropil were seen. The fibres in the main retinotectal tract maintained an orderly disposition with respect to each other, in contrast to the fibres of the basal optic tract, in which no order was apparent. Optic fibres were seen passing caudally from the region of the basal optic neuropil.

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Selection of appropriate medial branch of the optic tract by fibres of ventral retinal origin during development and in regeneration: an autoradiographic study in Xenopus.

The formation of the branches of the optic tract has been studied with the use of [3H]proline autoradiography, during development and during regeneration of the optic nerve in Xenopus with one compound ventral (VV) eye made by the embryonic fusion of two ventral eye fragments. The formation of the optic pathway was abnormal in that the lateral branch failed to develop, suggesting that fibres from a VV retina selectively entered the tectum via the medial branch during development. Three months after section of the optic nerve of a VV eye, regenerated fibres were present both in the contralateral and ipsilateral tecta. On the ipsilateral side regenerated fibres entered the tectum via the medial branch only. Retinal fibres entered the contralateral tectum through both branches in some animals and through the medial branch only in others. It is concluded that mechanical factors alone are insufficient to explain the phenomenon of selection of the appropriate medial branch by fibres of ventral retinal origin either during development or in regeneration. Some form of fibre-substrate interaction seems to be necessary; and this ability of fibres from a VV eye to take the path appropriate for ventral retina argues strongly that the VV eye is not a regulated system in terms of cell specificities.

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The relationship between retinal and tectal growth in larval Xenopus: implications for the development of the retino-tectal projection.

The modes of retinal and tectal histogenesis, as well as the patterns of terminal distribution of optic axons in larval Xenopus were studied, using anatomical techniques. We show that the retina grows by adding strips of cells at its ciliary margin. The pattern of retinal growth is asymmetrical along the dorso-ventral axis of the retina. On the other hand, the tectum grows by adding newly formed cells caudo-medially. The most rapid tectal growth takes place between stages 50 and 53, and thereafter only a small proportion of cells are added near the midline of the caudal tectum. Despite such incongruent modes of retinal and tectal growth, retinotopic order is maintained throughout larval life. We present here further evidence supporting the idea that connexions between the arrays of retinal and tectal cells shift progressively caudo-medially on the tectum during the period of growth. When the temporal pole of the retina is destroyed at various developmental stages, the density of degenerating synapses is always highest in the rostral pole of the tectum. Moreover, optic terminals stemming from the central retina spread caudally, invading newly maturing regions of the tectum. Quantitative analysis of the terminal distributions of optic axons suggest that orderly shifts in synaptic contacts between optic axons and tectal dendrites take place in the course of development. Ultrastructural observations on the maturing tectal neuropil are consistent with this view.

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The orientation of the visuotectal map in Xenopus: developmental aspects.

Rotations and translocations of the eye anlage were performed in Xenopus embryos of stages ranging from 21/22 to 30. Some of the operations involved grafting wild-type eye anlagen into albino host orbits. Operations were performed under a variety of operating media and conditions. In later larval life, or after metamorphosis, the visuotectal maps from the operated eyes were recorded electrophysiologically. Results fell into two classes. In the majority, the orientation of the visuotopic map corresponded to the orientation of the eye at the time of recording. In the minority the visuotopic maps were 'compound', consisting of two parts each with its own independent orientation. The organization of the compound maps was such that one component was oriented in correspondence with the orientation of the eye, while the other component was normally oriented. Histological analysis and observations on genetically marked grafts indicated that the component parts of the compound eye were of dual cellular origin. The component giving the rotated (or translocated) map belonged to the originally operated eye tissue; whereas the component giving the normally oriented map was derived from newly grown eye tissue coming from the optic stalk. In no case was a normally oriented map obtained from a rotated or translocated eye. The results are discussed in relation to mechanisms proposed to account for the determination of map-related retinal specificity.

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