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M A Matthews

Publications and source records attributed to M A Matthews.

At least 37 records · Page 2Linked to original sources
PubMed1989

Ulex europaeus agglutinin-I binding to dental primary afferent projections in the spinal trigeminal complex combined with double immunolabeling of substance P and GABA elements using peroxidase and colloidal gold.

Ulex europaeus agglutinin I (UEA-I) is a plant lectin with an affinity for L-fucosyl residues in the chains of lactoseries oligosaccharides associated with medium- and smaller-diameter dorsal root ganglion neurons and their axonal processes. These enter Lissauer's tract and terminate within the superficial laminae of the spinal cord overlapping projections known to have a nociceptive function. This implies that the surface coatings of neuronal membranes may have a relationship with functional modalities. The present investigation further examined this concept by studying a neuronal projection with a nociceptive function to determine whether fucosyl-lactoseries residues were incorporated in its primary afferent terminals. Transganglionic transport of horseradish peroxidase (HRP) following injection into tooth pulp chambers was employed to demonstrate dental pulp terminals in the trigeminal spinal complex, while peroxidase and fluorescent tags were used concomitantly to stain for UEA-I. Double immunolabeling for substance P (SP) and gamma-aminobutyric acid (GABA) using peroxidase and colloidal gold allowed a comparison of the distribution of a known excitatory nociceptive transmitter with that of UEA-I binding in specific subnuclei. Synaptic interrelationships between UEA-I positive dental pulp primary afferent inputs and specific inhibitory terminals were also examined. SP immunoreactivity occurred in laminae I and outer lamina II (IIo) of subnucleus caudalis (Vc) and in the ventrolateral and lateral marginal region of the caudal half of subnucleus interpolaris (Vi), including the periobex area in which Vi is slightly overlapped on its lateral aspect by cellular elements of Vc. The adjacent interstitial nucleus (IN) also showed an intense immunoreactivity for this peptide antibody. UEA-I binding displayed a similar distribution pattern in both Vc and Vi, but extended into lamina IIi and the superficial part of Lamina III in Vc. Dental pulp terminals were found to have a comparable distribution; however, many extended into the dorsal portion of the caudal half of Vi and the ventromedial quadrant of rostral Vi. Electron-microscopic analysis showed that transganglionically labeled dental pulp terminals contained ovoid, complex membrane-bound vacuoles laden with transported HRP. The preterminal axon and synaptic membranes of those dental pulp terminals located in zones of Vc and Vi displaying an affinity for UEA-I were usually characterized by a patchy, electron-dense coating of the peroxidase tag. SP was demonstrated ultrastructurally with Protein-A colloidal gold (3-nm particles), whereas GABA immunoreactivity was revealed by the avidin-biotin-peroxidase method.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

Resistance to Water Transport in Shoots of Vitis vinifera L. : Relation to Growth at Low Water Potential.

Apparent resistances to water transport in the liquid phase were determined from measurements of soil, root, basal shoot internode, shoot apex, and leaf water potentials and water flux in Vitis vinifera (cv White Riesling) during soil drying. Predawn water potential differences (DeltaPsi) in the shoots accounted for 20% of the total DeltaPsi between the soil and the shoot apex when plants were well-watered but increased to about 90% when shoot growth ceased. The DeltaPsi from soil to root was essentially constant during this period. At low water potential, the DeltaPsi in the shoot was persistent when transpiration was low (predawn) or completely prevented (plant bagging). The apparent hydraulic resistance between the basal shoot internode and most rapidly expanding leaf (or shoot apex) increased several-fold when water was withheld. Leaf and internode expansion both exhibited high sensitivity to increasing hydraulic resistance. Measurements of pneumatic resistance to air flow through frozen internode segments indicated progressive vapor-filling of vessels as soil drying progressed. From these observations and others in the literature, it was suggested that embolization may be a common occurrence and play an important role in the inhibition of shoot growth at moderate water deficits.

Journal Article↗
PubMed1988

GABA distribution in a pain-modulating zone of trigeminal subnucleus interpolaris.

A recent model for control of spinal and medullary nociceptive neurons (Basbaum and Fields, 1984) incorporates a gamma-aminobutyric acid-ergic (GABA-ergic) cell into this circuitry and indicates that such elements could act as one substrate for presynaptic inhibition of primary afferents. This concept is supported by a variety of pharmacological and electrophysiological studies. We therefore examined the distribution of GABA-ergic activity in trigeminal subnucleus interpolaris (Vi) by focusing on the types of cells, together with dendritic and synaptic profiles, that are immunocytochemically labeled with an antiserum against glutamic acid decarboxylase (GAD). GAD occurred throughout Vi but was most concentrated in the ventrolateral quadrant and interstitial nucleus. It was localized to groups of small neurons with two to three primary dendrites, and within numerous punctate profiles suggestive of synaptic elements. Electron microscopy revealed labeled dendrites, some of which were postsynaptic to scalloped terminals of presumptive primary afferents. Other labeled dendritic elements, which were quite variable in size, engaged both GAD-labeled and unlabeled synapses. Most GAD synapses displayed clear round vesicles and formed contacts with unlabeled perikarya and a variety of dendritic processes. Numerous GAD-positive synapses were also incorporated into axoaxonic clusters, in which the GAD element was presynaptic to scalloped terminals. Others engaged in serial arrays with other unlabeled terminals, which, in turn, were presynaptic to dendrites. Occasionally, GAD synapses formed contacts with GAD-positive dendrites. These data show that GABA is localized to a variety of neuronal elements in ventrolateral Vi and the interstitial nucleus. These occur in spatial arrangements providing an anatomical substrate for postsynaptic modulation of activity in this area. GABA terminals also appear to be involved in a presynaptic inhibitory mechanism, which may, in some instances, affect transmission in primary afferents.

Animals↗
PubMed1987

Effects of intraocular tetrodotoxin on the postnatal development of the dorsal lateral geniculate nucleus of the rat: a Golgi analysis.

The postnatal development of the rat dorsal lateral geniculate nucleus (dLGn) during tetrodotoxin (TTX)-induced monocular impulse blockade was investigated by quantitative Golgi techniques. Beyond 14 days postnatal (dpn), the effectiveness of TTX was monitored by loss of the pupillary light reflex. By 21 dpn, Golgi analysis indicated that TTX had no effect on the pattern of dendritic branching of class A or class B neurons, although the number of dendritic spinous protrusions was reduced. No evidence of any TTX-induced loss of optic axons or neuronal degeneration in the dLGn was found, despite a 16% decrease in the size of the nucleus, suggesting a reduction in the growth of neuropil. These data indicate that optic impulses are important in mediating the proper growth of postsynaptic specializations in the dLGn during ontogenesis, but that the postnatal development of the dendritic arbor of neurons in the dLGn appears to be independent of retinal impulse activity.

Animals↗
PubMed1987

Immunocytochemistry of enkephalin and serotonin distribution in restricted zones of the rostral trigeminal spinal subnuclei: comparisons with subnucleus caudalis.

The spinal trigeminal subnucleus caudalis processes nociceptive input from the head. However, physiological and behavioral studies in monkeys and humans indicate that painful stimuli from the central face and oral cavity also project through trigeminal nuclei rostral to the spinal subnucleus caudalis. Both enkephalin (ENK) and serotonin (5-HT) are present in rostral trigeminal nuclei and these regions receive inputs from the raphe complex. Thus, it appears that elements of pain-modulating circuitry proposed by Basbaum and Fields (Annu. Rev. Neurosci., 7:309-338, 1984) for the spinal and medullary dorsal horn may also exist in this region. In order to begin an exploration of this circuitry, the present study combines the techniques of retrograde transport of HRP from the ventral posteromedial thalamic nucleus (VPM) of the cat's thalamus to label trigeminothalamic relay cells. Secondarily, immunocytochemical techniques are employed to define the distribution patterns of ENK and 5-HT cells and terminals in relationship to both labeled and nonlabeled neurons in each of the subnuclei of the spinal trigeminal nucleus. Trigeminothalamic relay cells were observed in laminae I and II, the magnocellular region, and the interstitial nucleus (IN) of subnucleus caudalis (Vc). ENK was found in axodendritic and axosomatic terminals, together with a population of small fusiform neurons in all these same areas except the magnocellular region. ENK axosomatic contacts innervated approximately 30% of labeled relay cells, chiefly in lamina I and the IN, or small unlabeled neurons in the same area. Serotonin activity occurred principally in lamina I and the IN and was confined almost exclusively to axodendritic terminals. Examination of subnucleus interpolaris (Vi) revealed relay cells distributed throughout the length of the nucleus and increasing in numbers at rostral levels. A rostral extension of the IN was found just ventrolateral to the main body of Vi and contained numerous labeled cells. The distribution of ENK activity was restricted to the ventral part of Vi and the IN and occurred in axodendritic and axosomatic terminals. These latter elements innervated 30-40% of labeled relay cells in Vi, particularly those located in the IN. Cells containing ENK generally resembled the fusiform cells found in Vc and were distributed in ventral Vi and the IN. Some ENK cells were larger, displayed several dendrites, and occurred only in the ventral Vi. Serotonin within Vi and Vc was confined principally to axodendritic terminals.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1987

Dynamic Relation between Expansion and Cellular Turgor in Growing Grape (Vitis vinifera L.) Leaves.

Measurements of the growth and water relations of expanding grape (Vitis vinifera L.) leaves have been used to determine the relationship between leaf expansion rate and leaf cell turgor. Direct measurement of turgor on the small (approximately 15 micrometer diameter) epidermal cells over the midvein of expanding grape leaves was made possible by improvements in the pressure probe technique. Leaf expansion rate and leaf water status were perturbed by environmentally induced changes in plant transpiration. After establishing a steady state growth rate, a step decrease in plant transpiration resulted in a rapid and large increase in leaf cell turgor (0.25 megapascal in 5 minutes), and leaf expansion rate. Subsequently, leaf expansion rate returned to the original steady state rate with no change in cell turgor. These results indicate that the expansion rate of leaves may not be strongly related to the turgor of the leaf cells, and that substantial control of leaf expansion rate, despite changes in turgor, may be part of normal plant function. It is suggested that a strictly physical interpretation of the parameters most commonly used to describe the relationship between turgor and growth in plant cells (cell wall extensibility and yield threshold) may be inappropriate when considering the process of plant cell expansion.

Journal Article↗
PubMed1985

Intraocular kainic acid injection suppresses fast axonal transport in the developing rat optic nerve.

A single intraocular injection of 1 or 3 nmol kainic acid (KA) into the right eye of rats aged 5 days postnatal (5 dpn) significantly reduced the incorporation of 3H-proline into retinal proteins and suppressed the amount of 3H-proline-labeled materials fast axonally transported in the optic nerve for at least 2 weeks thereafter. Intraocular KA injection within this dose range had no adverse effect on the optic axon population compared to normal nerves determined at 21 dpn; however, doses above 3 nmol (i.e., 6 nmol) caused significant axonal degeneration. Although partially recovered by 21 dpn this effect of KA on protein synthesis and axonal transport suggests that, as in the adult, retinal ganglion cells are also KA-sensitive during postnatal development.

Animals↗
PubMed1985

Transplantation of fetal lateral geniculate nucleus to the occipital cortex: connectivity with host's area 17.

The developing lateral geniculate complex was excised from fetal albino rats at 18 days of gestation and implanted into the occipital cortex of host animals at 5 days of postnatal age. Groups of host animals were sacrificed at 10, 20 and 30 days following this procedure. The transplant tissue of selected animals was stereotaxically lesioned 2 days prior to scheduled sacrifice and their brains subjected to either Fink-Heimer or electron microscopic analysis of the distribution and density of degenerating efferents from the transplant. The remaining animals were analysed by means of Bodian, Golgi-Cox or electron microscopic techniques. Transplanted neurons displayed typical dendritic branching patterns of geniculate relay neurons by 20 days following implantation. Intrinsic neurons, characterized by a small ovoid soma and two main stem dendrites, only became evident in transplant tissue by 30 days and were much reduced in number. Synapses developed by 10 days and rapidly increased in number by 20 and 30 days. Most complexes were simple axo-dendritic, asymmetric junctions. Multiple serial and reciprocal complexes, as well as the characteristic glomerular complex, failed to appear. Analysis of Bodian stained material revealed a dense network of fibers coursing about the transplant. Distinct bundles of these fibers were observed extending from the medial edge of the transplant into area 17 by 20 days following implantation. A Fink-Heimer analysis of animals whose transplants were stereotaxically lesioned revealed degeneration in Layers II-VI of the primary visual cortex but the majority of these fibers terminated within the lateral two-thirds of Layer IV. Few degenerated fibers could be found in the underlying white matter indicating that efferents from the transplant found their way to their "correct" target zone by growing through a complex neuropil which provided minimal physical substrates to guide such growth. Most of the contacts formed by these fibers were simple junctions along the shafts of dendrites with a wide range in diameter. It is concluded that the nearby host visual neurons, which are the correct target cells for the afferents arising in the transplant, induced a directed growth of these fibers.

Animals↗
PubMed1985

Effects of neonatal intraocular colchicine on synaptogenesis and on the retention of the ipsilateral retinofugal projection within the superior colliculus.

A normally transient ipsilateral retinofugal projection exists in the rat but is retained following eye removal because of the loss of competitive interaction between crossed and uncrossed fibers. To further explore this phenomenon, colchicine (10(-3)M) was injected into the right eye of newborn albino rats to partially suppress axonal transport in optic fibers, alter the developmental time course of retinofugal synaptic terminals and determine if this would in turn extend the period of survival of the ipsilateral projection. Measurements of the number of fibers in the nerve were also made to insure that colchicine was not lethal to the retinofugal projection. Projections into the superior colliculus were demonstrated by anterograde movement of HRP from the left eye. TMB histochemistry revealed dense labeling of the contralateral retino-recipient layers at 5 dpn in untreated or saline-injected controls. The ipsilateral projection was seen as a lighter band of activity across the colliculus which was most concentrated in the antero-medial quadrant. This pathway was transient and degenerated by 10 dpn, except for a few antero-medial fibers. Animals treated with colchicine demonstrated a retention of this pathway through 20 dpn. A concomitant quantitative analysis of synaptic development within the superior colliculus revealed populations of boutons with round (R) and flattened (F) vesicles, as well as multiple junctional (MJ) and serial (S) complexes, most of which were specialized R boutons. The various synaptic categories displayed specific ratios unique to the different stages of maturation. Intraocular colchicine reduced the ratio of R, MJ and S boutons to F terminals between 5-15 dpn (P less than 0.01). By 20 dpn, the proportions of MJ and S boutons remained depressed but the normal ratio of R to F boutons was restored. Areal determinations of each synaptic profile included in the counts revealed a significant reduction in the size of MJ synaptic profiles examined in colchicine-treated animals and this may have been reflected in the slight loss of tectal volume (6-9%). Removal of the left eye and assessment of degenerating boutons showed that the expanded ipsilateral projection was not sufficiently dense to produce such a restoration. It thus appears that colchicine delayed the growth of the R population, but the effect was reversible and this category of boutons continued to develop, albeit on a later time course.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1985

The postnatal development of the rat primary visual cortex during optic nerve impulse blockade by intraocular tetrodotoxin: a quantitative electron microscopic analysis.

The effect of tetrodotoxin (TTX)-induced monocular impulse blockade on various parameters of synaptogenesis during the first 3 postnatal weeks of the developing rat visual cortex was investigated by quantitative electron microscopy. During the injection period, beyond 14 days postnatal (dpn), the effectiveness of TTX in blocking optic nerve impulses was monitored by loss of the pupillary light reflex. Between 5 and 21 dpn, TTX treatment reduced the number of type I axodendritic synapses by approximately 23%, when compared to sham-injected controls. These reductions were found in layers III, IV, and the superficial region of layer V. Layer IV exhibited the greatest decrease (24%) while layers III and V showed reductions of 20% and 18%, respectively. At 21 dpn, the number of type II axodendritic synapses decreased by 19% in the same layers, but no reductions were found at earlier ages. TTX also reduced the mean number of synaptic vesicles within type I and type II terminals by 27% and 15%, respectively. At 9 dpn, reductions were first found in layers IV and V, but by 21 dpn significant decreases were found in layers II/III, IV and V. TTX had no effect on the length of the postsynaptic density of both synaptic types or on cortical thickness at any age. These data indicate that optic impluses are important mediators of synaptogenesis in the developing visual cortex, the loss of which induces localized and specific synaptic alterations, possibly due to a change in cortical circuitry.

Animals↗
PubMed1985

Effects of intraocular tetrodotoxin on dendritic spines in the developing rat visual cortex: a Golgi analysis.

The effect of tetrodotoxin (TTX)-induced monocular impulse blockade on the growth of dendritic spines in the developing rat primary visual cortex was analysed by quantitative Golgi techniques. Between 5 and 21 days postnatal (dpn), rats were injected with TTX every 2 days into the right eye to chronically eliminate optic impulses. Effectiveness of TTX was monitored by loss of the pupillary light reflex. At 21 dpn, the number of spines located on the portion of the apical dendrite within layers III, IV and the superficial region of layer V was reduced by approximately 26%. These decreases were found on the apical dendrites of both large and medium sized pyramidal cells. TTX also reduced the number of spines on the proximal portion of oblique dendrites in layer IV by 16%, yet did not change the number of spines on basilar dendrites. No evidence of transneuronal degeneration was seen following long-term TTX treatment. These data indicate that dendritic spine development in the visual cortex is sensitive to the loss of optic impulses and that the decrease in spine population is principally due to a reduction in spine growth.

Animals↗
PubMed1985

The effect of intraocular injection of tetrodotoxin on fast axonal transport of [3H]proline- and [3H]fucose-labeled materials in the developing rat optic nerve.

The fast axonal transport of [3H]proline-labeled proteins and [3H]fucose-labeled glycoproteins delivered to the dorsal lateral geniculate nucleus in the developing rat optic nerve was investigated during tetrodotoxin-induced monocular impulse blockade. Repeated intraocular injections of various dosages of tetrodotoxin or citrate buffer vehicle were made every two days in rats aged 5-21 days postnatal, and the accumulation of rapidly transported radioactivity in the lateral geniculate nucleus measured between three and twelve hours post-injection at each age. The effectiveness of prolonged tetrodotoxin treatment was monitored by loss of the pupillary light reflex and the level of cytochrome oxidase activity in the contralateral superior colliculus and dorsal lateral geniculate nucleus. Numbers of optic axons proximal to the chiasm and the frequency of retinal ganglion cells per unit distance from the optic disc were examined for signs of tetrodotoxin-induced degeneration of the retinofugal pathway. Tetrodotoxin-treatment reduced the amount of fucosyl glycoproteins, but not proline-labeled proteins, axonally transported to the lateral geniculate nucleus during the first three weeks of postnatal development. Other studies indicated that tetrodotoxin significantly reduced the incorporation of [3H]fucose into retinal proteins indicating that the reduction in transport was probably due to a decrease in precursor incorporation into retinal ganglion cells. Electron microscopy of ganglion cells at 21 days revealed dilated and vacuolated Golgi cisternae associated with tetrodotoxin treatment, suggesting that tetrodotoxin may alter fucose metabolism by secondarily disrupting Golgi organization. Other protein synthetic machinery in these cells, including ribosomes and rough endoplasmic reticulum, appeared normal throughout tetrodotoxin treatment. These data indicate that Na+-dependent optic impulse activity may be indirectly related to the axonal transport of glycoproteins during early postnatal development by mediating the incorporation of precursor into glycoproteins at the Golgi apparatus and their subsequent entrance into the fast transport system.

Animals↗
PubMed1984

Inhibition of axoplasmic transport in the developing visual system of the rat: IV. Quantitative Golgi, electron microscopic, and histochemical analyses of the maturation of the visual cortex.

Intraocularly injected colchicine suppresses axonal transport within the developing rat's optic nerve throughout the critical period of visual system development. This results in a stunting of retinofugal terminals and relay neurons in the lateral geniculate nucleus. The present study focuses upon the effects of this unique form of developmental deprivation on the maturation of the visual cortex. Colchicine, in concentrations of from 10(-5) to 10(-2) M, was injected into the eyes of albino rats at birth or at 5, 10, or 15 days of age. Litters were killed at 5 to 50 days after this single injection, and the brains were processed for Nissl, rapid Golgi, histochemical, or electron microscopic analysis. The following results were obtained: Planimetry of coronal sections of the striate cortex revealed a reduction in the thickness of the cortex and in the ratio of neuropil area to neuronal soma area contralateral to the injected eye which was confined principally to layer IV, lower layer III, and upper layer V. This effect was inversely related to postnatal age at injection and directly proportional to colchicine concentration. A rapid Golgi analysis of 51 pairs of layer V pyramidal neurons in control and experimental cortex demonstrated a reduction in the number and size of spines along the portion of the apical dendrite passing through lower layer III and IV following colchicine administration at birth or 5 or 10 days of age but no significant change in the branching pattern of the entire dendritic arbor. Electron microscopy revealed a reduction in the number of small, asymmetric synaptic complexes with the result that the average size of remaining profiles was increased in layers III and IV. Histochemical analysis of cortical succinic dehydrogenase and cytochrome oxidase revealed a distinct band of intense enzyme activity in lower layers III and IV in normal cortex at 20-30 days of age. This band was significantly reduced in intensity after neonatal injection of colchicine as shown by densitometric measurements and comparison of experimental and control cortex. It is concluded that the geniculocortical projection, while not affected directly by colchicine administration, is altered by the secondary effects of axonal transport suppression, leading to an alteration in the establishment of cortical synaptic patterns and arborizations of their postsynaptic neurons whose dendrites are located in those layers recipient to this projection.

Animals↗
PubMed1984

Acclimation of photosynthesis to low leaf water potentials.

Photosynthesis is reduced at low leaf water potentials (Psi(l)) but repeated water deficits can decrease this reduction, resulting in photosynthetic acclimation. The contribution of the stomata and the chloroplasts to this acclimation is unknown. We evaluated stomatal and chloroplast contributions when soil-grown sunflower (Helianthus annuus L.) plants were subjected to water deficit pretreatments for 2 weeks. The relationship between photosynthesis and Psi(l), determined from gas-exchange and isopiestic thermocouple psychometry, was shifted 3 to 4 bars towards lower Psi(l), in pretreated plants. Leaf diffusive resistance was similarly affected. Chloroplast activity, demonstrated in situ with measurements of quantum yield and the capacity to fix CO(2) at all partial pressures of CO(2), and in vitro by photosystem II activity of isolated organelles, was inhibited at low Psi(l) but less in pretreated plants than in control plants. The magnitude of this inhibition indicated that decreases in chloroplast activity contributed more than closure of stomata both to losses in photosynthesis and to the acclimation of photosynthesis to low Psi(l).

Journal Article↗
PubMed1984

Kok effect and the quantum yield of photosynthesis : light partially inhibits dark respiration.

The linear response of photosynthesis to light at low photon flux densities is known to change abruptly in the vicinity of the light compensation point so that the quantum yield seems to decrease as radiation increases. We studied this ;Kok effect' in attached sunflower (Helianthus annuus L. cv IS894) leaves using gas exchange techniques. The effect was present even though respiration was constant in the dark. It was observed at a similar photon flux density (7 to 11 micromole photons per square meter per second absorbed photosynthetically active radiation) despite a wide range of light compensation points as well as rates of photosynthesis. The effect was not apparent when photorespiration was inhibited at low pO(2) (1 kilopascal), but this result was complicated because dark respiration was quite O(2)-sensitive and was partially suppressed under these conditions. The Kok effect was observed at saturating pCO(2) and, therefore, could not be explained by a change in photorespiration. Instead, the magnitude of the effect varied as dark respiration varied in a single leaf, and was minimized when dark respiration was minimized, indicating that a partial suppression of dark respiration by light is responsible. Quantum yields measured at photon flux densities between 0 and 7 to 11 micromole photons per square meter per second, therefore, represent the combined yields of photosynthesis and of the suppression of a component of dark respiration by light. This leads to an overestimate of the quantum yield of photosynthesis. In view of these results, quantum yields of photosynthesis must be measured (a) when respiration is constant in the dark, and (b) when dark respiration has been inhibited either at low pO(2) to eliminate most of the light-induced suppression of dark respiration or at photon flux densities above that required to saturate the light-induced suppression of dark respiration. Significant errors in quantum yields of photosynthesis can result in leaves exhibiting this respiratory behavior if these principles are not followed.

Journal Article↗
PubMed1982

An ultrastructural analysis of the development of foetal rat retina transplanted to the occipital cortex, a site lacking appropriate target neurons for optic fibres.

Foetal retina was removed from donor rats at 15 days of gestation and transplanted to the occipital cortex of neonatal host rats. The purpose of this procedure was to examine the development of retinal neurons and photoreceptors, and document synaptic patterns during maturation of the transplanted retina in an environment lacking a normal target for optic axons. Host animals were sacrificed at 5, 10, 15, 20 and 30 days and samples of cortex containing the transplant were subjected to a light and electron microscopic analysis. During early stages of development, (5 days) the retina assumes a radial orientation with the scleral (outer) surface located centrally and the vitreal (inner) surface occupying the periphery. Numerous mitotic figures are found at the centre of the transplant and columns of primitive neuroblasts appear to radiate out from this zone. By 10 to 15 days after transplantation the retinal tissue contains numerous small rosettes each of which displays a histotypic organization with recognizable layers of sensory cells and their centrally-projecting processes, an outer limiting membrane, made up of a network of zonulae adherentes, and a rudimentary outer and inner plexiform layer which delineate the cells of the inner nuclear layer. Ultrastructural analysis of such rosettes confirmed the presence of typical bipolar, amacrine, horizontal and ganglion cells, but revealed that while the plexiform layers were occupied by numerous processes from these neurons, few if any, of these exhibited synaptic vesicles. By 20 to 30 days following transplantation sensory cells have completely differentiated, giving rise to prominent inner and outer segments which display typical cilia, centrioles and basal bodies, together with numerous stacked lamellae of photoreceptors which were contorted, presumably due to growth in an abnormal site. It should be further emphasized that these structures developed in the absence of pigment cells. Synaptic development ensues during this period to form characteristic dyads within the outer and inner plexiform layers. Additionally, clusters of amacrine to amacrine contacts occurred in the inner plexiform layer and were found to be increased relative to other types of junctions. In general, synaptogenesis takes place in the outer and inner plexiform layers and all categories of retinal synapses are established, but the process was found to be significantly delayed in comparison to normal retina at the same stage of development. Quantitative analysis revealed a reduced number of presumptive ganglion cells in proportion to the other categories of neurons. Optic fibres remained small and failed to myelinate. It is suggested that lack of an appropriate target for optic axons induced this alteration and may be indirectly related to the delay in the onset of synaptic development.

Animals↗
PubMed1982

Inhibition of axoplasmic transport in the developing visual system of the rat-L Structural changes in the retina and optic nerve with graded doses of intraocular colchicine.

In order to study the role of axonal transport in the mediation of transneuronal metabolic stimulations upon a population of differentiating neurons, colchicine, a potent inhibitor of rapid and slow phases of axonal transport, was injected into the eye of albino rats at 1, 3, 5, 10, 15 and 20 days postnatal in concentrations ranging from 10-5 M to 2 X 10-2 M and in quantities of 0.3 to 0.5 microliter. Quantitative light and electron microscopy were subsequently employed to assess reactive alterations in the developing retina and optic nerve. Application of colchicine severely retarded the development of the sensory elements, with disappearance of synaptic ribbons of sensory cell axons, a significant reduction in the thickness of the inner plexiform layer, due to the presence of numerous shruken synaptic elements and the appearance of rosettes of sensory cells displaced to the inner nuclear layer. These alterations were found to be dose-dependent. Counts of ganglion cell populations at various times after application of colchicine demonstrated optimal concentrations which could be injected at each postnatal age without causing ganglion cell degeneration. Ultrastructural examination of such cells revealed varying degrees of disorganization and dissolution of the endoplasmic reticulum with the formation of occasional small cytoplasmic vacuoles. Higher concentrations of colchicine caused extensive vacuole formation in all classes of retinal neurons, scattered hyperchromic cells and widespread degeneration and autolysis. The diameter of the optic nerve was reduced to 60-95% of normal following intraocular colchicine, depending on the concentration employed, but electron microscopy revealed normal patterns of distribution of axoplasmic microtubules and filaments in control and experimental animals and quantitative analysis revealed no significant loss of axons. While no reactive changes took place in individual elements, the periphery of the nerve was often indented by a highly-folded glia limitans. Maximal doses of intraocular colchicine for each age level were established by this study. These were: 1 day. 10-3 M: 5 days, 5 x 10-3 M; 10 days, 5 X 10-3 M; 15 and 20 days, 10-2 M. The information derived from this morphological analysis provides the foundation for subsequent measurements of axonal transport inhibition in the developing visual system to be reported in the second article of this series.

Animals↗