Biomedical subjects

T Chan-Ling

Publications and source records attributed to T Chan-Ling.

31 records · Page 2Linked to original sources

PubMed1995

Development of retinal vasculature is mediated by hypoxia-induced vascular endothelial growth factor (VEGF) expression by neuroglia.

We have studied the role of the hypoxia-inducible angiogenic growth factor vascular endothelial growth factor (VEGF) in the induction and control of vessel growth in the developing retina of rats and cats, using in situ hybridization techniques. VEGF is expressed successively in two layers of neural retina, the innermost (axon) layer and the inner nuclear layer (INL). In the axon layer, VEGF is expressed transiently by astrocytes as they spread across the layer, closely preceding the formation of superficial vessels. In the INL, VEGF is expressed transiently by somas at the middle of the layer (presumably Müller cells), closely preceding the formation of the deep layer of retinal vessels. We propose that hypoxia caused by the onset of neuronal activity is detected by strategically located populations of neuroglia, first astrocytes, then Müller cells. In response they secrete VEGF, inducing formation of the superficial and deep layers of retinal vessels, respectively. As the vessels become patent, they relieve the hypoxic stimulus, so vessel formation is matched to oxygen demand. This hypothesis was tested experimentally in three ways. Expression of the high affinity flk-1 receptor for VEGF was demonstrated in newly formed retinal vessels, confirming that the secreted VEGF acts on the vessels, in a paracrine fashion. Direct hypoxic regulation of VEGF expression by macroglia was demonstrated in primary cultures of astrocytes and in cells of a glioma line. Hypoxic regulation of VEGF expression in the intact developing retina was demonstrated by showing that oxygen-enriched atmospheres that inhibit vessel formation also suppress endogenous VEGF production.

Animals↗

PubMed1995

The effect of oxygen on vasoformative cell division. Evidence that 'physiological hypoxia' is the stimulus for normal retinal vasculogenesis.

PURPOSE: To assess the role of oxygen in normal retinal vasculogenesis. METHODS: A new preparation for studying cytogenesis in retinal wholemounts was developed. Nuclei of dividing cells were labeled with a monoclonal antibody to bromodeoxyuridine (BrdU), and vascular cells were covisualized with Griffonia simplicifolia lectin. The topography and time course of vasculogenic cell division and vessel formation were determined in the kitten retina during normal development and under experimental hyperoxia and hypoxia. RESULTS: During normal development, vasculogenic cell division was maximal at the leading edge of the forming vessels. Normal vessel formation was initially proliferative, and cell division was high. However, after vessel formation occurred, which presumably relieved tissue hypoxia, the mitogenic process was markedly reduced, and many excess capillary segments underwent retraction. The rate of vasculogenic cell division and vessel formation increased when the inner layers of the retina were made avascular after exposure to hyperoxia, and it decreased when there was an increase in inspired oxygen. CONCLUSIONS: The authors have shown that between 17% and 45% oxygen, the extent of vasculogenic cell division is inversely proportional to the level of oxygen in the inspired gas mixture. They have further shown that dividing vascular cells have a peak density in a region proximal to the edge of the forming vasculature. The density is maximal between P7 and P8, a time when formation of photoreceptor outer segment begins, only a few days before the onset of retinal function. These results led the authors to conclude that the stimulus for normal vasculogenesis is a transient but physiological level of hypoxia induced by the increasing activity of retinal neurons.

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PubMed1993

Contact-spacing among astrocytes is independent of neighbouring structures: in vivo and in vitro evidence.

We have examined the morphology of astrocytes and the arrays they form in two situations, in retinas from which ganglion cells and blood vessels have been caused to degenerate, and in vitro. These observations were made to test whether the regularity of the spacing of astrocytes within normal central nervous tissue results from interaction among astrocytes, or from interaction between astrocytes and other elements of that tissue. Both in the partially degenerated cat retina, and in cultures of astrocytes from neonatal rat cortex, astrocytes make and maintain contact with neighbouring astrocytes, yet space their somas apart, giving regularity to the arrays. These results support the hypothesis that the regularity observed in arrays of astrocytes in intact tissue results from an interaction among astrocytes, independent of neighbouring structures, and lead us to suggest that the cell-cell interactions involved in contact spacing serve to distribute astrocytes through the central nervous system, and may, in other tissues, underlie the formation of epithelia.

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PubMed1993

The role of Müller cells in the formation of the blood-retinal barrier.

We have compared the ability of Müller cells and astrocytes to induce the formation of barrier properties in blood vessels. Müller cells cultured from the rabbit retina, and astrocytes and meningeal cells cultured from the rat cerebral cortex, were injected into the anterior chamber of the rat eye, where they formed aggregates on the iris. We have examined the barrier properties of the vessels in those aggregates and, for comparison, the barrier properties of vessels in the retina, ciliary processes and iris. Two tracers were perfused intravascularly to test barrier properties. The movement of Evans Blue was assessed by light microscopy, and the movement of horseradish peroxidase by light and electron microscopy. Our results indicate that Müller cells share the ability of astrocytes to induce the formation of barrier properties by vascular endothelial cells, and we suggest that Müller cells play a major role in the formation of barrier properties in retinal vessels.

Animals↗

PubMed1993

Comparisons between microvascular changes in cerebral and non-cerebral malaria in mice, using the retinal whole-mount technique.

CBA/T6 mice inoculated with Plasmodium berghei ANKA strain (PbA) exhibited cerebral symptoms and died from cerebral malaria 6-8 days p.i. whereas DBA/2J mice developed (around days 6-9) a non-fatal cerebral malaria, with milder cerebral symptoms, and died between days 15 and 22 from other malaria-related complications. When inoculated with P. berghei K173 (Pb) these mouse strains did not develop cerebral malaria. These mouse/parasite strain combinations were used, in conjunction with the retinal whole-mount technique, to elucidate factors critical in the pathology of murine cerebral malaria. CBA/T6 mice infected with PbA (PbA-CBA mice) demonstrated mild changes in vascular permeability as early as days 2-3, prior to the appearance on day 5 of cerebral symptoms, whereas mice with non-cerebral malaria did not show any vascular permeability changes until the very late stage of the disease (days 14-22). In the PbA infections, progressive deterioration of endothelial barrier properties, demonstrated by Evans' Blue leakage both generally and from specific focal areas, as well as a developing monocytosis and adherence of mononuclear cells to the endothelium of the retinal vessels continued until death (in CBA/T6 mice) or resolution (in DBA/2J mice). Adherent monocytes, particularly in PbA-CBA mice, were associated with reduced Hoechst staining of individual endothelial cells and a banking up proximally of both parasitized and non-parasitized blood cells in the small blood vessels, often with accompanying focal leakage of Evans' Blue from the retinal vessels. The occurrence and severity of these early changes in the microcirculation correlated with the subsequent development of cerebral symptoms. Monocyte margination appeared to be the most significant factor associated with the development of cerebral symptoms.

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PubMed1992

Vascular changes and their mechanisms in the feline model of retinopathy of prematurity.

This study documents changes to retinal vasculature during the feline form of retinopathy of prematurity (ROP). The authors describe the closure and obliteration of retinal vessels during exposure to high oxygen, the pattern and tempo of growth of proliferative vasculature, which, after the return of the animal to room air, extends from the optic disc in a spectacular "rosette" pattern, the formation of preretinal vascular growths, and an initial lack of barrier properties in the new vessels. Finally, the response of the vasculature to the relief of hypoxia is reported, including the gradual establishment of barrier properties in the intraretinal vessels, the partial normalization of the proliferative vessels, and the abnormalities that persist. It is suggested that the vascular changes occur in successive stages: closure and obliteration during hyperoxia, vasoproliferation induced by hypoxia, and normalization after the relief of hypoxia with distinct cellular mechanisms and stimuli. It is argued that the same stages can be seen in the human form of ROP; two possible stimuli for the fibroplasia that damages the retina in human ROP are discussed.

Animals↗

PubMed1992

Degeneration of astrocytes in feline retinopathy of prematurity causes failure of the blood-retinal barrier.

This study addresses the role of astrocytes in the genesis of retinopathy of prematurity, examined in the feline model of this condition. Evidence is presented that the hypoxia of retinopathy of prematurity, in addition to inducing vasoproliferation, damages the retina directly. Retinal neurons survive the hypoxia, but the astrocytes, which are involved in the formation of the glia limitans of the retinal vessels, degenerate. Astrocytes subsequently recolonize the retina after a delay that matches the period of leakiness of the proliferative vasculature (described in the companion article). Given the evidence from other studies that the barrier properties of vessels are induced by their glia limitans, the authors suggest that the initial lack of barrier properties in the new vasculature is caused by the degeneration of astrocytes and that the subsequent formation of those properties is induced by the astrocytes that recolonize the retina some days later. The observation that astrocytes are more sensitive to hypoxia than neurons, at least in developing tissue, was unexpected. The literature reporting on the damage caused to central nervous tissue by hypoxia is consistent in assessing neurons as more sensitive and glial changes as a reaction to neuronal damage. The sensitivity of astrocytes found in this study and earlier in vitro research suggests that degenerated astrocytes can be replaced and their structural and functional relationships reestablished.

Animals↗

PubMed1991

Factors determining the migration of astrocytes into the developing retina: migration does not depend on intact axons or patent vessels.

Astrocytes migrate into the cat retina from the optic nerve, beginning from embryonic day (E) 52. Once they have entered the retina they concentrate along major axon bundles and fail to enter regions of the retina with high densities of neurones, in particular the area centralis region of the ganglion cell layer. These nonuniformities appear as the astrocytes spread over the retina during development, and in this study we have examined factors that might control their spread. First we examined astrocytes in a retina in which the axon bundles had degenerated following an optic nerve lesion at birth. The area over which astrocytes had spread was normal, suggesting that their spread does not depend on the presence of intact axons. Second, we noted that, despite the degeneration of all ganglion cells following the nerve lesion, astrocytes still did not spread over the area centralis. Their spread is apparently not inhibited by concentrations of neurones. Third, we examined astrocytes in retinas of animals raised in an atmosphere containing 70-80% oxygen, which prevents the formation of retinal vessels. Again, the area over which the astrocytes had spread was normal, suggesting that their spread does not depend on the presence of patent blood vessels. These negative findings led us to compare the distribution of spindle cells (precursors of retinal vasculature) and astrocytes in the cat during development. The close correspondence in their topographical distribution and the earlier spread of the spindle cells lead us to suggest that spindle cells provide a basal lamina component that may guide the migration of astrocytes.

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PubMed1989

Sensitivity and neural organization of the cat cornea.

The innervation of the adult cat cornea was investigated both psychophysically and histologically. Mean corneal touch threshold (CTT) for 25 adult domestic cats was 43 +/- 9 mg in the center of the cornea and 100 +/- 32 and 94 +/- 33 mg in the superior and inferior cornea, respectively. Gold chloride impregnation showed that the cat cornea is innervated by 16-20 radial nerve trunks that enter the mid-posterior stroma at various sites around the corneal circumference. As these trunks travel anteriorly toward the center of the cornea they give off collaterals that form the anterior stromal and subepithelial plexus. Fibers from the subepithelial plexus penetrate the epithelial basement membrane and give off numerous long fibers that ramify in the basal epithelial layer. Intraepithelial terminals arise from these, penetrating between the epithelial cells, ending with a terminal enlargement at the wing cell level. A distinct pattern of neural organization was found in the periphery of the cat cornea. This consisted of finer nerve fibers that entered the cornea at the subepithelial and basal epithelial levels at numerous sites around the corneal circumference. These fibers branched after a short distance in the cornea and appeared to innervate the anterior stroma and epithelium in the periphery of the cornea. This study thus provides direct evidence of two types of neural organization in the cornea of the domestic cat. Stromal nerves appear to be the main source of innervation to the epithelium in the center of the cornea while conjunctival nerves supply the peripheral epithelium.

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PubMed1988

Symptoms in VDU operators.

The investigation of the role of visual and ocular characteristics of visual display unit (VDU) operators in the causation of subjective visual fatigue has not been reported previously. A retrospective study in which the visual and ocular characteristics of symptomatic and asymptomatic VDU operators was carried out in order to determine whether such a relation could be demonstrated. Three groups totaling 1013 VDU operators from two semigovernment organizations were assessed using either a full eye examination or a modified clinical screening technique. Before examination, the operators completed a short questionnaire on the symptoms they experienced when using VDU's. Significant differences between the symptomatic and asymptomatic operators in some groups were found for over-refraction (spherical equivalent and cylindrical) and near horizontal heterophoria. Despite the existence of this relation, no basis for valid visual standards could be established. In particular, no basis was found for the automatic prescribing of low plus corrections and, in the absence of clearly definable visual standards, the involvement of clinical judgment in the assessment of operators was found to be necessary.

Adolescent↗

PubMed1988

Changes in corneal endothelial morphology in cats as a function of age.

A cross-sectional study of changes in cat corneal endothelial cell morphology with age was conducted. The central corneal endothelium of 12 kittens and 70 adult cats was photographed using specular microscopy. Endothelial cell density (ECD), coefficient of variation of cell size (used as an index of polymegethism), and cellular shape factor (perimeter 2/area) were determined for each animal and analysed as a function of age. We found a rapid non-linear decrease in ECD and polymegethism in the first nine months of post-natal life. Subsequently there was a slight central cell loss of 11 cells/mm2 or 0.37% per year during adult life which was not statistically significant. However, polymegethism increased significantly with age during adult life. The shape factor for endothelial cells was 13.61 throughout adult life, indicating that the cat corneal endothelium consists predominantly of six-sided cells. Possible explanations for the finding of no significant decrease in cell density with age could include the higher peripheral ECD in the cat which may compensate for central loss, the short life expectancy of the cat and the large individual variation in corneal diameter.

Aging↗

PubMed1987

Long-term neural regeneration in the rabbit following 180 degrees limbal incision.

Penetrating 180 degrees superior limbal incisions were made on the right eye of four adult New Zealand albino rabbits. The contralateral eye served as control. Corneal touch thresholds (CTT) for the central, superior and inferior cornea (2-3 mm from limbus) were determined 3, 9, 15, 24 and 30 months after surgery. In all animals, the CTT was significantly elevated in the superior region of the cornea throughout the measurement period. CTT was elevated in the central and inferior cornea 3 months following surgery and was not affected in the inferior cornea on all other occasions. The animals were then sacrificed and the corneas subjected to histochemical demonstration of acetylcholinesterase corneal nerves. All rabbits showed a reduction in the number of histochemically detectable stromal nerve trunks in the operated region. These stromal nerve trunks showed regenerative changes including abnormally curved course and a subnormal number of axons within a nerve trunk. Epithelial nerve fiber defects included absence or distorted architecture of the basal epithelial plexus and intra-epithelial terminals. These results indicate that although extensive stromal reinnervation had occurred, the extent and quality of stromal nerves was inadequate to restore a normal epithelial plexus and corneal sensitivity.

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PubMed1985

Diurnal variation of corneal thickness in the cat.

Central corneal thickness of both eyes of seven cats was measured hourly for 72 hr using ultrasonic pachometry. The mean corneal thickness was 569 +/- 36 micron (mean +/- SD), and the diurnal variation was 49 +/- 14 micron (8.6% of corneal thickness). In a separate experiment, the corneal thickness of one eye of each of five cats was measured following 2 hr of natural sleep; 2 1/4 hr after eye opening, the corneas had thinned an average of 43 +/- 22 micron. The authors conclude that corneal swelling induced by eye closure during periods of sleep is the prime determinant of the diurnal variation in cat corneal thickness. In studies where corneal thickness is to be monitored over a period of time, it is possible to control against this large diurnal variation by ensuring that the cat is active for a period of two hours prior to pachometry measurements.

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