Comparative studies on mammalian Müller (retinal glial) cells.

Müller cells from 22 mammalian species were subjected to morphological and electrophysiological studies. In the 'midperiphery' of retinae immunocytochemically labeled for vimentin, estimates of Müller cell densities per unit retinal surface area, and of neuron-to-(Müller) glia indices were performed. Müller cell densities were strikingly similar among the species studied (around 8000-11,000 mm-2) with the extremes of the horse (< or = 5000 mm-2) and the tree shrew (> or = 20,000 mm-2). By contrast, the number of neurons per Müller cell varied widely, being clustered at 6-8 (in retinae with many cones), at about 16, and at up to more than 30 (in strongly rod-dominated retinae). Isolated Müller cell volumes were estimated morphometrically, and cell surface areas were calculated from membrane capacities. Müller cells isolated from thick vascularized retinae (carnivores, rats, mice, ungulates) were longer and thinner, and had smaller volumes but higher surface-to-volume ratios than cells from thin paurangiotic (i.e. with blood vessels only near the optic disc) or avascular retinae (rabbits, guinea pigs, horses, zebras). In whole-cell voltage-clamp studies, Müller cells from all mammals studied displayed two dominant K+ conductances, inwardly rectifying currents and delayed rectifier currents. TTX-sensitive Na+ currents were recorded only in some species. Based on these data, the following hypotheses are presented, (a) neuron-to-(Müller) glia indices are determined by precursor cell proliferation rather than by metabolic demands; (b) Müller cell volumes depend on available space rather than on the number of supported neurons; and (c) it follows that, the specific metabolic activities of Müller cells must differ greatly between species, a difference that may contribute to distinct patterns of retinal vascularization.

LiCl-induced malformations of the eyes and the rostral CNS in Xenopus laevis.

Ocular malformations such as synophthalmia and cyclopia occur in all vertebrates including humans. We induced fused eye anlagen by exposure of Xenopus laevis embryos to dissolved LiCl. Three basic forms of malformations were observed in the rostral CNS; these represent major steps in a continuous sequence of ocular abnormalities: (i) "hour-glass eyes", (ii) synophthalmia, and (iii) cyclopia vera. The type of abnormity induced seemed to depend on the time and/or the dosage of LiCl exposure. In all cases studied, the histology of the retinae was normal. We reconstructed olfactory organs, eyes, and rostral portions of the CNS from serial sections of 16 Xenopus larvae. Our results confirm that defective bilateralization affects all parts of the rostral CNS and suggest that the telencephalon displays the weakest, and the eyes have the highest, resistance against midline fusion. Defective bilateralization also involves the pineal anlagen, as duplication or enlargement of the pineal organs occurred in most cases with ocular fusion. We conclude that LiCl-induced interferences with ontogenetic bilateralization of prosencephalic structures are the consequence of an elimination of an, as yet, undefined bilateralization signal from (ventral) midline structures of the neural plate and tube.

Development of the rabbit retina. V. The question of 'columnar units'.

A qualitative and quantitative description of the columnar units in the mammalian retina, and a discussion of their ontogeny and putative functions is given. Columnar arrangements of cells exist in the developing retina which can be observed by means of scanning electron microscopy. In the adult retina, each Müller cell ensheaths a columnar group of neuronal cells. Counting the number of cells in radial H/E stained sections at various developmental stages reveals a constant ratio of neuronal cells per Müller cell, independent of the developmental stage (after postnatal day 9), and independent of the retinal topography. Such groups of cells always consist of one Müller cell, 11 rod photoreceptor cells, about 2 bipolar cells, and 1 to 2 amacrine cells. Retinal ganglion cells, cone photoreceptor cells, and horizontal cells are more sparsely distributed in the retina than these units; since they are known to arise earlier in the ontogenesis than other cell types they are considered to exist independently of the columnar units. It is suggested that the units arise by migration of groups of preneurons along a common Müller (precursor) cell; these preneurons and the corresponding Müller cell may be clonally related. In the adult retina, such columns might constitute metabolic and functional units.

Immunocytochemical demonstration of astrocytes and microglia in the whale brain.

Whale brains have attracted the attention of neuroscientists but there are only sparse studies on whale glial cells. Here we report on immunolabeling of astrocytes by antibodies to glial fibrillary acidic protein (GFAP) or protein S-100 beta (both by the streptavidin/biotin technique), and labeling of microglial cells by Griffonia simplicifolia agglutinin (GSA I-B4, coupled to horseradish peroxidase), in the neocortex of a harbour porpoise Phocoena phocoena L. Many subpial and perivascular astrocytes were stained; they differed greatly in thickness and length of their processes. Subpial astrocytes were coarse with a few stout stem processes, whereas perivascular astrocytes deeper in the brain had many long and slender processes. Additionally, some long radial astrocytes were observed. Microglia were labeled throughout the brain, and showed similar features as 'resting' (ramified) microglia in the brain of other mammals.

A set of early-born neurons is distinctly labeled by several defined antibodies in the adult rabbit retina.

Retinal ganglion cells, cone photoreceptor cells, and horizontal cells arise earlier in ontogenesis than the other retinal cell types. Although during the first postnatal week of life much cell proliferation occurs in the rabbit retina, 3H-thymidine labeling shows that these particular neurons are already postmitotic in neonatal animals. We show here that, in the adult retina, these early-born neurons express antigens as neuron-specific enolase, HNK-1 epitope of N-CAM, and PGP 9.5 antigen, which are not expressed by the neurons later born. It is concluded that the mammalian retina contains two genotypically different sets of neuronal cells which can be distinguished by immunocytochemistry.

Effect of in vivo application of the ginkgo biloba extract EGb 761 (Rökan) on the susceptibility of mammalian retinal cells to proteolytic enzymes.

Lesions, inflammations, or degenerative insults of the human retina are accompanied by the release of proteolytic enzymes. Their deleterious effect may be enhanced by the release of free radicals. Ginkgo biloba extracts are known to exert protective influences against the action of free radicals, and this prompted us to ask whether the application of such extracts might protect retinal tissue against proteolytic damage. Eighteen adult rabbits were fed for 3 weeks (+/- 3 days) with 40 mg/kg of G. biloba extract (EGb 761) or a terpene-free fraction of this extract, dissolved in their drinking water. Twelve control rabbits received no G. biloba extract. The animals were then euthanatized and their retinae isolated. After appropriate enzymatic treatment, the tissue was dissociated and the number of isolated Müller cells counted as an indication of the strength of the proteolytic effects. There was a significant protective action of EGb 761: in an average control rabbit 5,200 cells per milligram retinal tissue were isolated; application of EGb 761 markedly reduced this number to 2,500 (terpene-free fraction; CP 205) or 3,050 (terpene-containing fraction). It is concluded that G. biloba extracts may have a significant therapeutic value in cases of retinal damage.

Optic tectum in congenitally monophthalmic fishes and chicks.

Morphometry of optic tectum layers was performed in fishes and birds that were congenitally monophthalmic (or had one very small or malformed eye). The optic chiasm was studied with regard to possible anomalies of the fiber distribution. In two very young fish larvae with only one ocular rudiment, no differences were found between the two tecta. In fishes and chicks with one normal and one eye of reduced size, the volume of superficial tectal layers contralateral to the small eye was significantly more affected (reduction by 40 to 70%) than that of the deep layers (reduction by 10 to 30%). Particularly pronounced differences between the superficial tectal layers of both sides were found in a fish where a thin optic nerve, originating in an unusually small eye, projected to the ipsilateral side of the brain. It is concluded that (1) the development of superficial tectal layers depends more strongly on optic innervation than that of deep layers; (2) obvious differences between tecta with and without optic innervation develop rather late in ontogeny; (3) there seems to be no measurable difference between effects of a poor (less than 25%) innervation of the tectum and a complete lack of this projection.