Evidence for a critical period of neuronal trophism late in the development of the chick visual system

Chick embryos and young hatchling chicks have undergone unilateral retinal ablation and the brains were subsequently prepared for modified Fink-Heimer staining to detect degenerating axoplasm. Additional animals have been injected with tritiated proline or combinations of tritiated proline and tritiated fucose. In examining the patterns of degeneration and labelling produced, it appears that during the stage 40–44 period of embryonic life there is a critical period during which damage to retinal axons produces degeneration not only in the direct targets of the optic tract (primary centers), but also in a number of nuclei not projected to by the optic tract (secondary centers). By examining various survival times following retinal ablation and animals of varying embryonic and post-hatch age, we have concluded that retinal ablation during the critical period causes a rapid and fulminant degeneration in the secondary centers because they are all targets of the projection from the optic tectum, itself a prime target of the optic projection. Presumably, interruption of the retino-tectal projection during the critical period disrupts some as yet undefined trophic relationship between these two sets of neurons. Our studies show that the observed degenerative reaction in the secondary centers occurs most strongly in chick embryos between stage 40 and stage 44. This period coincides with the onset of synaptogenesis in the retino-tectal system (Rager, '76a,b). We therefore hypothesize that during this period some property or influence is passing from the retinal to the tectal neurons such that interruption of this process leads to death of the tectal neurons.     

Wulst efferents in the little owl Athene noctua: an investigation of projections to the optic tectum.

The efferent projections from the Wulst were studied in the little owl, Athene noctua, using anterograde migration of wheat-germ-agglutinin conjugated horseradish peroxidase (WGA-HRP). Wulst projections were distributed to telencephalic, diencephalic and mesencephalic targets in a general pattern similar to that previously described in other avian species. Our results on the organization of the Wulst-optic tectum pathway in the little owl reveal well defined and laminarly arranged terminal projections into the superficial tectal layers, with a distribution suggestive of topographical relationships between neurons of origin in the Wulst and termination fields in the optic tectum. In contrast to lateral-eyed birds, the little owl possesses conspicuous contralateral projections to the optic tectum. Ipsilateral and contralateral efferents are restricted to different tectal regions, i.e. ipsilateral projections to the caudo-dorsal and contralateral projections to the rostro-ventral optic tectum. In addition, the anterior and posterior Wulst differentially contribute to the ipsilateral and contralateral projections to the optic tectum. This differential organization of Wulst efferents, as well as the presence of substantial contralateral projections, might be related to the high degree of binocular overlap typical of frontal-eyed birds. At a functional level, electric potentials recorded in the optic tectum and evoked by visual stimulation showed that information from one eye can reach the ipsilateral optic tectum. After Wulst ablation, the amplitude of these potentials was significantly reduced, indicating that Wulst efferents may influence visually-evoked activity in the optic tectum.     

Development of the retinotectal system in normal quail embryos: cytoarchitectonic development and optic fiber innervation

The development of the optic tectum and the establishment of retinotectal projections were investigated in the quail embryo from day E2 to hatching day (E16) with Cresyl violet-thionine, silver staining and anterograde axonal tracing methods. Both tectal cytodifferentiation and retinotectal innervation occur according to a rostroventral-caudodorsal gradient. Radial migration of postmitotic neurons starts on day E4. At E14, the tectum is fully laminated. Optic fibers reach the tectum on day E5 and cover its surface on day E10. 'Golgi-like' staining of optic fibers with HRP injected in vitro on the surface of the tectum reveals that: growing fronts are formed exclusively by axons extending over the tectal surface; fibers penetrating the outer tectal layers are always observed behind the growing fronts; the penetrating fibers are either the tip of the optic axons or collateral branches; as they penetrate the tectum, optic fibers give off branches which may extend for long distances within their terminal domains; the optic fiber terminal arbors acquire their mature morphology by day E14. The temporal sequence of retinotectal development in the quail was compared to that already established for the chick, thus providing a basis for further investigation of the development of the retinotectal system in chimeric avian embryos obtained after xenoplastic transplantation of quail tectal primordia into the chick neural tube.     

Kainate-induced lesion in the optic tectum: dependency upon optic nerve afferents or glutamate

Kainic acid is known to induce characteristic lesions in neurons receiving an intact input with presumed glutamate-mediated neurotransmission. There are indications for glutamate as a transmitter of retinal afferent terminals in the pigeon optic tectum. After tectal injection of kainic acid (0.5–2.0 μg in 0.5 μl) the optic tectum was studied by light and electron microscopy and the following changes were observed: (a) within 1–48 h important neuropil vacuolization predominantly in lower part of layer 5. Such vacuoles were sometimes postsynaptic to identified retinal afferent terminals: (b) within 1 h to 21 days progressive neuronal cell loss throughout the tectal layers. These toxic effects were not observed 2–12 weeks after contralateral retinal ablation but could partially be restored by combined glutamate (0.2 mg) and kainate injection. Thus in the pigeon tectum, kainic acid neurotoxicity is dependent upon an intact retinal input, a finding consistent with a special role for glutamate — possibly as a transmitter — in retinal terminals.     

The development of the nucleus isthmi in Xenopus laevis. I. Cell genesis and the formation of connections with the tectum

The nucleus isthmi (NI) of the amphibian relays visual input from one tectum to the other tectum and thus brings a visual map from the eye to the ipsilateral tectum. This isthmotectal visual map develops slowly; it is first detected electrophysiologically at stages 60-62, the age at which the eyes begin their dorsalward migration and the region of binocular overlap beings to increase in extent. During this critical period of life, normal binocular visual input is required for establishment of normal topographic isthmotectal projections. In this study, we have used anatomical methods to trace cell birth, cell death, and formation of connections by the nucleus isthmi during the critical period. Tritiated thymidine labelling demonstrates that cells in the nucleus isthmi are generated throughout most of tadpole life (stages 29-62). Most cells conform to an orderly ventrodorsal gradient starting from stage 29 and extending to stages 56; later cells are inserted at apparently random locations in the nucleus. We have re-examined the hypothesis of Tay and Straznicky ('80) that the order of cell genesis in the NI and tectum could help establish proper isthmotectal connections, and we find that a timing mechanisms does not explain the two-dimensional topography of the isthmotectal map but that timing may aid in proper mediolateral positioning of isthmotectal axons at the points where they first enter the tectum. Horseradish peroxidase labelling was used to investigate whether anatomical projections from tectum to NI and from NI to tectum are present prior to the onset of eye migration. The results show that there are tectoisthmotectal projections by stage 52. Moreover, isthmotectal axons grow into as yet monocular tectal regions prior to the onset of eye migration. At stage 60, when binocular overlap begins, isthmotectal axons are visible throughout the tectum but are densely branched only at the rostral tectal margin, the location where they are predicted to occur on the basis of electrophysiological maps.     

Development of the mesencephalic nucleus of the trigeminal nerve in chick embryos: Target innervation,neurotrophin receptors,and cell death

The goal of this study was to determine whether processes of neurons in the mesencephalic nucleus of the trigeminal nerve (Mes V) of chick embryos arrive in their peripheral target prior to the period of developmental cell death, and to determine whether neurons with early target contact survive to a greater extent than neurons with processes that reach their peripheral target later. The arrival of Mes V nerve fibers in the masticatory muscles was determined by injecting the fluorescent tracer DiI, and the position of labeled and unlabeled neurons was mapped in subdivisions of the Mes V nucleus. Developmental changes in the numerical configuration of Mes V subdivisions were studied in DiI-labeled as well as Nissl-stained material. The expression of law-affinity (p75) neurotrophin receptors was investigated throughout development of the Mes V nucleus with in situ hybridization to assess whether and how levels of expression of this trophic receptor may relate to target innervation and cell death. The extent of cell death was evaluated by counting pyknotic nuclei. Processes of Mes V neurons invade their peripheral target between 5 and 7 days of incubation (E5–7). At E7–12, between 800 and 1,400 labeled Mes V neurons were distributed throughout the two main subdivisions of the Mes V nucleus, the tectal commissure and the optic tectum. Only few Mes V neurons were labeled in the posterior commissure or outside the brain. Cell counts in Nissl-stained material from E7–13 revealed that the numbers of Mes V neurons in the optic tectum decreased to about 40–60%, and in the tectal commissure to 20–25%, whereas Mes V neurons in the posterior commissure disappeared almost entirely. Few Mes V neurons remained in the leptomeninges at E8–10, but a considerable number was found outside the midbrain at E11, indicating ongoing migration of some Mes V neurons. Neurotrophin receptors were differentially expressed in the Mes V nucleus: Before and after the period of cell death, 90–100% of Mes V neurons expressed neurotrophin receptors, whereas during, and immediately preceding the period of developmental cell death (E9–E13), merely 70% of Mes V neurons expressed this receptor. These findings are consistent with the hypothesis that early target contact may provide an advantage for the survival of Mes V neurons and that competition for trophic factors may occur in the peripheral target of this nucleus prior to the period of cell death. © 1993 Wiley-Liss, Inc.     

An atlas of the primary visual projections in the brain of the chick Gallus gallus

The localisation of the primary visual centres in the chick mesencephalon and diencephalon was determined by autoradiographic anterograde transport and degeneration techniques. Strong visual projections were found in the tectum, lateral anterior thalamic nucleus, lateroventral geniculate nucleus, superficial synencephalic nucleus, external nucleus, ectomammillary nucleus, tectal grey, dorsolateral anterior thalamus, rostrolateral part, and the pretectal optic area. Weaker retinal projections were found in the ventrolateral thalamus, two subregions of the dorsolateral anterior thalamus, lateral part, diffuse pretectal nucleus, dorsolateral anterior thalamus, magnocellular part, and the hypothalamus. An atlas of the retinal projections was constructed from sections.     

Tectal mosaic: organization of the descending tectal projections in comparison to the ascending tectofugal pathway in the pigeon

The optic tectum of vertebrates is an essential relay station for visuomotor behavior and is characterized by a set of connections that comprises topographically ordered input from the eyes and an output that reaches premotor hindbrain regions. In the avian tectofugal system, different ascending cell classes have recently been identified based on their dendritic and axonal projection patterns, although comparable information about the descending cells is missing. By means of retrograde tracing, the present study describes the detailed morphology of tectal output neurons that constitute the descending tectobulbar and tectopontine pathways in pigeons. Descending cells were more numerous in the dorsal tectum and differed in terms of 1) their relative amount of ipsi- vs. contralateral projections, 2) the location of the efferent cell bodies within different tectal layers, and 3) their differential access to visual input via dendritic ramifications within the outer retinorecipient laminae. Thus, the descending tectal system is constituted by different cell classes presumably processing diverse aspects of the visual environment in a visual field-dependent manner. We demonstrate, based on a careful morphological analysis and on double-labeling experiments, that the descending pathways are largely separated from the ascending projections even when they arise from the same layers. These data support the concept that the tectum is arranged as a mosaic of multiple cell types with diverse input functions at the same location of the tectal map. Such an arrangement would enable the tectal projections onto diverse areas to be both retinotopically organized and functionally specific.     

Topographic and morphometric effects of bilateral embryonic eye removal on the optic tectum and nucleus isthmus of the leopard frog

Rana pipiens were raised through metamorphosis after extirpation of both eye primordia at Shumway embryonic stage 17 (Shumway '40). The visual connections between the isthmic nuclei and the optic tectum were examined in these animals using horseradish peroxidase (HRP) histochemistry. Isthmo-tectal projections are normally aligned with the primary retinotectal map. We asked whether these connections would develop normal topographic organization in the absence of normal retinal input. HRP was formed into a solid pellet (? 200–500 μm diameter) and inserted into one tectal lobe on the tip of a fine metal probe. The procedure produced relatively restricted retrograde label in somas and dendrites in both isthmi nuclei. In the nucleus isthmus ipsilateral to the tectal lobe receiving the HRP pellet, processes of tecto-isthmi neurons were labeled by anterograde transport. The topography of the isthmo-tectal and tecto-isthmic projections were identical in the developmentally enucleated animals and in normal frogs, even though eye removal severely reduced the volume of the optic tecta and the isthmi nuclei. Thus our analyses indicate that retinal contacts do not play an active role in the development of the positional or polarity cues that are involved in “mapping” projections between central visual nuclei. These results are discussed in the context of peripheral specification of central connections and in terms of models that have recently been proposed to explain the development of the retinotectal system.     

Plasticity in the chick ventral lateral geniculate nucleus

The chick ventral lateral geniculate nucleus (GLv) receives topographically corresponding projections from the retina and optic tectum. Tectal lesions produced on the day of hatching removed the tectogeniculate input to the GLv region corresponding to the tectal lesion and also severed some retinotectal axons. Following a survival period of 3 to 10 weeks, a patch of augmented retinogeniculate projection was noted in the GLv segment that corresponds topographically to the damaged area of the tectum. Changing the site of the tectal lesion led to changes in the locus of heavy retinal projection to the GLv predictable from topographic maps. Nuclei which received retinal but not tectal projections did not appear to have regions of augmented retinal termination nor did nuclei which received tectal but not retinal innervation. It is unlikely that the increased retinogeniculate termination is due to rerouting of growing retinotectal axons since the chick retinofugal pathway is well established by the time of hatching. Furthermore, there was no evidence of a projection from the ipsilateral eye to the affected GLv. On the basis of these light microscopic studies, it would appear that retinogeniculate terminals have sprouted in the GLv and that competition for terminal space, conservation of terminal space, proximity, and perhaps other factors are necessary for the augmented projection to occur.     

Frog tectal efferent axons fail to regenerate within the CNS but grow within peripheral nerve implants

Tectal efferent axons, located adjacent to the optic tract, fail to regenerate past diencephalic lesions in Rana pipiens even though optic axons regenerate after the same injury (M. J. Lyon and D. J. Stelzner, J. Comp. Neurol. 255: 511-525). We tested the possibility that tectal efferent axons can regenerate within peripheral nerve implants. A 6- to 8-mm segment of autologous sciatic nerve was implanted into the anterolateral (N = 23) or centrolateral (N = 22) portion of the dorsal surface of the tectum. Frogs survived for 6 (N = 16) or 12 weeks (N = 29) before the free end of the nerve was recut and HRP applied. A control group had the nerve crushed prior to the HRP application. Neurons within the tectum, near and medial to the implant site, were retrogradely labeled from the nerve graft in most experimental operates but no neurons were labeled in controls. In addition, neurons were also labeled in nuclei which projected to the tectum in a number of cases. Three times as many neurons were labeled in 12-week operates (42 +/- 46) as in 6-week operates (15 +/- 12). The morphology and location of labeled neurons in the tectum was similar to tectal efferent neurons except that the somal area of neurons labeled from the graft was significantly larger (41%) than normal tectal efferent neurons. The basic finding is similar to experiments using the same paradigm in the mammalian central nervous system (CNS). One difference is the minimal glial reaction at the graft insertion site.(ABSTRACT TRUNCATED AT 250 WORDS)     

Localization of α-bungarotoxin binding sites to the goldfish retinotectal projection

The optic tectum of the goldfishCarassius auratus is a rich source of α-bungarotoxin (α-Btx) binding protein. In order to determine whether some fraction of these receptors is present at retinotectal synapses, we have compared the histological distribution of receptors revealed by the use of [¹²⁵Iα-Btx radioautography to the distribution of optic nerve terminals revealed by the use of cobalt and horseradish peroxidase (HRP) techniques. The majority of α-Btx binding is concentrated in those tectal layers containing primary retinotectal synapses. The same layers contain high concentrations of acetylcholinesterase (AChE), revealed histochemically. Following enucleation of one eye, there is a loss of α-Btx binding in the contralateral tectum, observed both by radioautography and by a quantitative binding assay of α-Btx binding. Approximately 40% of the α-Btx binding sites are lost within two weeks following enucleation. By contrast, no significant change in AChE activity could be demonstrated up to 6 months enucleation. These results are discussed in light of recent studies which show that the α-Btx binding protein and the nicotinic acetylcholine receptor are probably identical in goldfish tectum. We conclude that the 3 main classes of retinal ganglion cells projecting to the goldfish tectum are nicotinic cholinergic and that little or no postdenervation hypersensitivity due to receptor proliferation occurs in tectal neurons following denervation of the retinal input.     

Tests of the regenerative capacity of tectal efferent axons in the frog, Rana pipiens

Experiments were designed to determine if neurons of the ranid optic tectum, a major target of the optic nerve, possess the same regenerative potential as optic axons. Normal tectal efferent (TE) projections were reexamined by using the anterograde transport of 3H-proline and autoradiography (n = 18), bulk-filling damaged TE axons with horseradish peroxidase (HRP; n = 18) and anterogradely transporting wheat germ agglutinin-HRP (n = 8) to label TE axons. Results were similar to reports that used degeneration methods (Rubinson: Brain Behav. Evol. 1:529-561, '68; Lazar: Acta. Biol. Hung. 20:171-183, '69). Following a brainstem hemisection just caudal to the nucleus isthmi (1-20 weeks), the ipsilateral descending TE pathway was autoradiographically examined (n = 20). While all other TE projections appeared normal, there was no detectable ipsilateral descending projection beyond the lesion site. Ascending TE axons were cut at the anterior tectal border by hemisecting the left diencephalon (LDH)--a lesion that also cuts optic axons projecting to the left tectum. There was no indication of TE axonal regeneration with the aid of autoradiography or HRP histochemistry 1-30 weeks postlesion (n = 48) even when the medial diencephalon was intentionally left intact (n = 4). However, in all four cases examined, optic axons regenerated following the same LDH where TE axonal regeneration failed (also see Stelzner, Lyon, and Strauss: Anat. Rec. 205:191A-192A, '83). Local effects of LDH should be similar for both the cut optic and cut TE axons. Other factors were tested that may contribute to the lack of TE axonal regeneration. Our results indicate that optic regeneration itself (n = 8), postaxotomy retrograde cell death of TE neurons (n = 6), deafferentation of the tectum of optic axons, and potential sprouting within tectal targets by intact contralateral TE axons (n = 10) are not critical factors aborting TE axonal regeneration. TE axons filled with HRP at chronic periods after LDH (n = 4) terminate anomalously near the LDH border. Many of these endings are similar to reactive endings or terminal clubs seen after axonal injury in the mammalian CNS. Our results suggest that this disparity in regenerative ability of optic and TE axons may be related to a difference in the responsive ability of these cell types to initiate or maintain axonal elongation after axotomy within the amphibian CNS environment.     

Regulation of retinal ganglion cell axon arbor size by target availability: Mechanisms of compression and expansion of the retinotectal projection

The ability of pre- and postsynaptic populations to achieve the proper convergence ratios during development is especially critical in topographically mapped systems such as the retinotectal system. The ratio of retinal ganglion cells to their target cells in the optic tectum can be altered experimentally either by early partial tectal ablation, which results in an orderly compression of near-normal numbers of retinal projections into a smaller tectal area, or by early monocular enucleation, which results in the expansion of a reduced number of axons in a near-normal tectal volume. Our previous studies showed that changes in cell death and synaptic density consequent to these manipulations can account for only a minor component of this compensation for the population mismatch. In this study, we examine other mechanisms of population matching in the hamster retinotectal system. We used an in vitro horseradish peroxidase labeling method to trace individual retinal ganglion cell axons in superior colliculi partially ablated on the day of birth, as well as in colliculi contralateral to a monocular enucleation. We found that individual axon arbors within the partially lesioned tectum occupy a smaller area, with fewer branches and fewer terminal boutons, but preserve a normal bouton denstiy. In contrst, ipsilaterally projecting axon arbors in monoculary enucleated animals occupy a greater area than in the normal condition, with a much larger arbor length and greater number of boutons and branches compared with normal ipsilaterally projecting cells. Alteration of axonal arborization of retinalganglion cells is the main factor responsible for matching the retinal and tectal cell populations within the tectum. This process conserves normal electrophysiological function over a wide range of convergence ratios and may occur through strict selectivity of tectal cells for their normal number of inputs. © 1994 Wiley-Liss, Inc.     

Observations on the projections and intrinsic organization of the pigeon optic tectum: an autoradiographic study based on anterograde and retrograde, axonal and dendritic flow.

Three aspects of the labelling pattern seen after the injection of 13 different radioactive amino acids into the pigeon optic tectum have been described: The efferent projections of the optic tectum; the specific labelling of two pathways; and the dendritic organisation of tectal layer III neurons based on the retrograde and anterograde movement of label within these dendrites. Discrete injections of tritiated amino acid that involved all or only the superficial tectal layers suggested that layer III gave rise to the massive non-topographically organised and bilateral projections (fibers crossing within the decussato supraoptica ventrlis) upon the nuclei rotundus, subpraetectalis and interstitio-praetecto-subpraetectalis and to the ipsilaterally directed pathways terminating within the nuclei praetectalis, triangularis, subrotundus, dorsolateralis anterior thalami, posteroventralis and ventrolateralis thalami. Layer III neurons may also be the source of efferents to the posterior dorsolateral thalamus (the layer III pathway), the pontine grey and, bilaterally to the reticular formation and of the layer IV or tectal commisural pathway terminating within the contralateral tectal cortex. In contrast projections originating from layer II were generally topographically organised and terminated either within certain of the isthmic nuclei (n. isthmi pars parvocellularis, n. isthmo-opticus and n. semilunaris) or ran within layer I (layer I pathways) to end in the pretectum (griseum tectale) and ventral thalamus (n. ventrolateralis thalami, n. geniculatus, pars ventralis). A small projection from layer II upon the ipsilateral nucleus rotundus may also be present. Triated serine and tyrosine were found to be particularly effective in labeling perikarya as well as axons and terminals. The layer I pathway could be selectively labelled after tectal injections of 3H-GABA while the cell bodies of Ipc neurons were labelled in a retrograde fashion after tectal injections of 3H-glycine, serine or alanine. Intrinsic tectal labelling was found by correlation with Golgi material to reflect both anterograde and retrograde transport of label within dendrites of layer III cells. Anterograde movement of label indicated that the terminal portions of layer III cell dendrites ended in an orderly radial arrangement within sublayers IIb and IId, while the retrograde movement of label resulted in the labelin of layer III perikarya outside the injection field.     

The response of optic tract glia during regeneration of the goldfish visual system. II. Tectal factors stimulate optic tract glia

After transection, retinal ganglion cell axons of the goldfish will regenerate by growing into a primary target tissue, the optic tectum. To determine what role the target tissue may play in regulating glial cell growth, we measured biosynthetic activity of optic tract glia following excision of the optic tectum and compared it to activity of glia found in the regenerating visual system. Ablation of the tectum reduced glial incorporation of both [³H]thymidine and [³⁵S]methionine. Tectal ablation also led to nearly 80% reduction of amino acids incorporated by oligodendroglia as well as a decrease in the amount of newly synthetized protein found within multipotential glia and within cytoplasmic projections of astroglia. Since the tectal influence upon optic tract glia was detected at a time when tract and tectum are physically separated, we sought to determine if the optic tectum contained soluble glia-promoting factors. A soluble fraction recovered from tecta of the regenerating visual system increased amino acid incorporation within optic tract glia at 2–3-fold above preparations incubated with fractions from control, intact tecta. Comparisons of radiolabeled proteins separated by sodium dodecyl polyacrylamide gel electrophoresis from regenerating and factor-stimulated optic tract were similar and indicated that a soluble tectal fraction promoted biosynthesis of specific glial proteins. Our findings suggest that during regeneration of the goldfish visual system glia are influenced by humoral factor(s) released from the synaptic target site.     

Late developmental expression of DARPP-32 in the chick optic tectum

Immunohistochemical techniques reveal that the dopamine- and cAMP-regulated phosphoprotein DARPP-32 is detectable in neurons of the chick optic tectum starting on embryonic day 13. The expression levels then increase steadily from embryonic day 15 through the first posthatching day. After 15 days posthatching, expression of DARPP-32 reaches the adult pattern, with many labeled cells in tectal layers 11 and 12. These cells exhibit a bipolar shape, with long processes directed both to the deep and superficial layers. These results suggest that DARPP-32 is present in specific neuronal populations of the chick tectum and that this protein may not have a function in early ontogenetic processes.     

Differences in the collateralization of neuronal projections from the dorsal column nuclei and lateral cervical nucleus to the thalamus and tectum in the cat: An anatomical study using two different double-labeling techniques

The dorsal column nuclei (DCN) and the lateral cervical nucleus (LCN) project to the diencephalon and tectum. In order to determine which neurons project to these targets and whether any of them have collateral projections, the double-retrograde labeling techniques developed by Hayes and Rustioni¹⁷ and Kuypers et al.²² were used.Both strategies produced similar results. Within DCN, neurons which projected to the diencephalon were located differently and had a different morphology than those which projected to the tectum. The diencephalic-projecting neurons, which were preferentially located within the middle part of the DCN complex were mainly large (> 15 μm diameter) and round. The tectal-projecting neurons, often located along the edges of the gracile and cuneate nuclei or between them, were most often found outside of the middle parts of the DCN complex (i.e. predominantly rostrally and frequently also caudally). These tectal-projecting neurons varied in size and were usually oval or fusiform in shape.In contrast to DCN, neurons in LCN which projected either to the tectum or diencephalon did not differ morphologically and in both groups were of various sizes and shapes. They were intermixed haphazardly, predominantly within the lateral two-thirds of the nucleus. In addition, another class of neurons, constituting more than 40% of the labeled population in some experiments, had collateral projections to both terminal targets. These double-labeled neurons were haphazardly mixed with other labeled neurons.These results demonstrate differences in the divergence patterns of DCN and LCN neurons. Considering these anatomical results together with the electrophysiological results of other investigators, it appears likely that DCN neurons which have different projections may also have different functional properties. The evidence for LCN neurons, however, is not consistent with such a suggestion.     

Tangential neuronal migration in the avian tectum: cell type identification and mapping of regional differences with quail/chick homotopic transplants.

This paper is a sequel to a previous report, using quail/chick chimeras with partial tectal transplants, in which a tangential invasion of host (chick) tectal territories by cells originating in the quail graft was demonstrated. The cells displaying this secondary tangential migration appeared restricted to two strata (stratum griseum centrale (SGC) and stratum griseum et fibrosum superficiale (SGFS)). Here we describe the morphology of the tangentially displaced neurons, as well as their overall distribution in the host tectal lobe, by means of an antibody that specifically recognizes quail cells, staining them in a Golgi-like manner. Neurons that migrated into the SGC are identified as multipolar projection neurons, typical of this stratum. The majority of cells that migrated into the SGFS correspond to horizontal neurons, as was also corroborated by observations in Golgi-impregnated material. These horizontal cells are concentrated in laminae b, d and f, where their processes form well delimited axonal plexuses. In confirmation of previous results, SGC neurons have a limited range of migration, whereas SGFS cells translocate across much longer distances. In reconstructions of appropriate cases, a remarkable polarity was noted. Significant invasion of chick tectum by quail cells mostly occurred in the rostral half of the host tectum. The long-range migration of superficial horizontal cells frequently reached, but did not cross, the rostral tectal boundary. Conversely, tangential migration in the caudal half of the host tectum was scarce and coincided with a typical arrangement of quail-derived radial columns interdigited with chick-derived columns. These findings are discussed in relation to existing data on immature neuronal populations, molecular marker distribution and polarity of the avian optic tectum.