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Structure and development of neuronal connections in isogenic organisms: transient gap junctions between growing optic axons and lamina neuroblasts.

We previously showed that the growth of each bundle of eight optic fibers from one ommatidium into the optic lamina of Daphnia occurs in such a way that one of the eight fibers precedes the others into the lamina. The growth cone of this lead fiber makes surface contact with undifferentiated neuroblasts near the midplane. This is followed by a glial-like wrapping of each neuroblast around the fiber. In this report, gap junctions are shown to form for a short period of time between the growing lead fiber and the neuroblast that is wrapping around it. It is proposed that these junctions may represent a morphological correlate of informational exchange between axon and neuroblast. This signaling would then reflect the fact that the sequence of axon proliferation by the lamina neuroblasts within an optic cartrdige, ultimately composed of five lamina neurons and eight optic fibers, parallels the order in which the neuroblasts undergo the wrapping reaction with the lead fiber.

Animals↗

Thyroxine influences neuronal connectivity in the adult frog brain.

In contrast to results of earlier investigations the influence of thyroxine on CNS connectivity is not restricted to circum-metamorphic stages in frogs. Neuroanatomical findings in adult Xenopus treated with thyroxine reveal a spread of the ipsilateral retino-tectal projection. Sprouting fibers establish a tectal innervation pattern similar to the one found in primitive fish. The question arises, whether thyroxine also has morphogenetic effects in the mature CNS of other species.

Animals↗

Bidirectional neuronal connections between the cerebellar interpositus nucleus and the brainstem (an autoradiographic study).

Ortho- and retrograde axonal labelings were demonstrated in several brainstem nuclei of cat after [3H]-leucine and [3H]-lysine microinjections into the cerebellar interpositus nucleus. Retrogradely labeled perikarya in the lateral reticular and solitary tract nuclei, locus coeruleus, substantia nigra and the raphe nuclei (midbrain raphe, dorsal, pontine and magnus) indicate the existence of possible aminergic afferents to the interpositus nucleus. The anterograde labeling of the neuropil in the above brainstem regions suggests the termination of axons or axon-collaterals originating from the interpositus nucleus.

Animals↗

Tracing neuronal connections with radioisotopes applied extracellularly.

Methodological and technical problems of the autoradiographic-neuroanatomical tracing (ARNT) technique are discussed. The size of the labeled area after a tritiated amino acid injection varies directly with the volume of isotope, the rate of injection, and the length of exposure of the secretion to the emulsion. Frozen sections can be used for autoradiography if they are mounted on subbed slides, dehydrated in ethanol, defatted for 1 hour in xylene, rehydrated through ethanol and water, and dried before coating with emulsion. Brushes used for mounting frozen sections should be used only for this purpose and dipped in boiling distilled water before use to avoid chemoreduced streaks in the emulsion due to contamination from the brushes. Excessive dilution of Kodak emulsion can leave less than a monolayer of grains over certain types of sections; the emulsion thickness should be checked by exposing coated test sections to a brief flash of light and developing immediately. A high intensity safelight recommended for the ARNT darkroom is the Thomas Duplex Super monochromatic sodium vapor bulb safelight; for Kodak NTB-2, -3 and Ilford L4 emulsions the red-banded and yellow-banded filters are used. A useful stain combination for ARNT is Luxol fast blue stained before coating and cresyl violet stained after developing which demonstrates both neuronal cell bodies and myelinated tracts in the same section.

Amino Acids↗

Digoxigenylated wheat germ agglutinin visualized with alkaline phosphatase-labeled anti-digoxigenin antibodies--a new, sensitive technique with the potential for single and double tracing of neuronal connections.

For double tracing experiments, wheat germ agglutinin (WGA) molecules labeled with two different haptens are desirable. In the present report the suitability of digoxigenylated WGA (DIG-WGA) for retrograde tracing was investigated. For this purpose the new tracer was pressure injected into rat brains and the transported DIG-WGA visualized via its digoxigenyl group with an alkaline phosphatase linked anti DIG antibody in permanently stained sections of high quality. With fixatives containing 2.5% glutaraldehyde only few positive cells were found. However, at milder fixation conditions (4% paraformaldehyde, 0.05% glutaraldehyde 0.2% picric acid, 30 min) retrogradely labeled cells were detected with a sensitivity comparable to tetramethylbenzidine protocols for conventional WGA-HRP (horseradish peroxidase) tracing. Preliminary experiments suggest excellent suitability for double labeling.

Alkaline Phosphatase↗

Neuronal connections between the cerebellar nuclei and hypothalamus in Macaca fascicularis: cerebello-visceral circuits.

The purpose of this study was to identify the basic pattern of interconnections between the cerebellar nuclei and hypothalamus in Macaca fascicularis. The distribution of retrogradely labeled cells and anterogradely filled cerebellofugal axons in the hypothalamus of M. fascicularis was investigated after pressure injections of a horseradish peroxidase mixture (HRP + WGA-HRP) in the cerebellar nuclei. Following injections in the lateral, anterior, and posterior interposed cerebellar nuclei retrogradely labeled cells were present in the following areas (greatest to least concentration): lateral and dorsal hypothalamic areas, dorsomedial nucleus, griseum periventriculare hypothalami, supramammillary and tuberomammillary nuclei, posterior hypothalamic area, ventromedial nucleus and periventricular hypothalamus, around the medial mammillary nucleus, lateral mammillary nucleus, and infundibular nucleus. Cell labeling was bilateral with an ipsilateral preponderance. In these same experiments anterogradely labeled cerebellar efferent fibers terminated in the contralateral posterior, dorsal and lateral hypothalamic areas, and the dorsomedial nucleus. In these regions retrogradely labeled hypothalamic cells were occasionally found in areas that also contained anterogradely filled cerebellar axons. This suggests a partial reciprocity in this system. In addition, sparse numbers of labeled cerebellar fibers recross in the hypothalamus to distribute to homologous areas ipsilateral to the injection site. Subsequent to an injection in the medial cerebellar nucleus (NM), cell labeling was present in more rostral hypothalamic levels including the lateral and dorsal hypothalamic areas, the dorsomedial nucleus, around or in fascicles of the column of the fornix, and in the periventricular hypothalamic area. Although no fastigiohypothalamic fibers were seen in this study, on the basis of information available from the literature it is likely that such a connection exists in primates. In summary, hypothalamic projections to NM originated mainly from rostral to midhypothalamic levels, whereas those projections to the lateral three cerebellar nuclei came from mid and more caudal levels. The existence of direct hypothalamic projections to cerebellar nuclei in M. fascicularis and of cerebellofugal projection to some hypothalamic centers indicates that circuitry is present through which the cerebellum may influence visceral functions. Furthermore, the fact that projections to NM versus the other cerebellar nuclei originate from somewhat different regions of the hypothalamus would suggest that the visceral functions modulated by each pathway is not the same.

Animals↗

[Neuronal connectivity and chemical mediators involved in olfactory message transmission].

In this review, we discuss some of the neural processes involved in the perception of odors which, together with audition and vision, provide essential information for analyzing our surroundings. We shall see how odor detection and learning induce substantial structural and functional changes at the first relay of the olfactory system, i.e., the main olfactory bulb. Among the mechanisms which participate in these modifications are changes in the cell's responses to a transmitter and the persistence of a high level of interneuron neurogenesis within the adult olfactory bulb. Our goal is to present some observations related to these two phenomena that may aid in understanding the neural mechanisms of sensory perception and shed light on the cellular basis of olfactory learning. To this purpose, we summarize the current ideas concerning the molecular mechanisms and organizational strategies used by the olfactory system to transduce, encode, and process information at various levels in the olfactory sensory pathway. Due to space constraints, this review focuses exclusively on the olfactory systems of vertebrates and primarily those of mammals.

Electrophysiology↗

The development of neuronal connection patterns in the visual systems of insects.

The retina of the compound eye contains a patterned spatial array of receptor cell groups whose symmetry and component numerical constancy allow the unique identification of individual cells. The axon connection patterns arising from the retina similarly are uniquely identifiable and relate by simple consistnet transpositons to the retinal position of their somata. The coincidence of these features and the epidermal origin or retinula cells provides a model system for the developmental analysis of neural patterns. It is known that the retina develops by a wave of determinative mitoses and subsequent cellular differentiation, transmitted inductively from a posteriorly situated differentiation centre, which passes over the prospective eye field independently of the growing axon connections with the optic lobe. The ganglion cells of the optic lobe are similarly produced in a temporal sequence but their subsequent differentiation depends on centripetal innervation and proceeds as a wave initiated in the retina. Axon growth between the retina and optic neuropiles occurs in this temporal sequence, appears largely non-exploratory and dependent on mechanical guidance. By contrast, individual axon connections within the neuropile form probabilistically by extensive filopod explorations between cones. Both growth between and growth within the neuropile depend upon features in the pattern of retinal development which are discussed.

Animals↗

Mode of growth of retinal axons within the tectum of Xenopus tadpoles, and implications in the ordered neuronal connection between the retina and the tectum.

Retinal axons of Xenopus tadpoles at various stages of larval development were filled with horseradish peroxidase (HRP), and their trajectories and the patterns of branching within the tectum were analyzed in wholemount preparations. To clarify temporal and spatial modes of growth of retinal axons during larval development, special attention was directed to labeling a restricted regional population of retinal axons with HRP, following reported procedures (H. Fujisawa, K. Watanabe, N. Tani, and Y. Ibata, Brain Res. 206:9-20, 1981; 206:21-26, 1981; H. Fujisawa, Dev. Growth Differ 26:545-553, 1984). In developing tadpoles, individual retinal axons arrived at the tectum, without clear sprouting. Axonal sprouting first began when growing tips of each retinal axon had arrived at the vicinity of its site of normal innervation within the tectum. Thus, the terminals of the newly added retinal axons were retinotopically aligned within the tectum. The retinotopic alignment of the terminals may be due to an active choice of topographically appropriate tectal regions by growth cones of individual retinal axons. The stereotyped alignment of the newly added retinal axons was followed by widespread axonal branching and preferential selection of those branches. Each retinal axon was sequentially bifurcated within the tectum, and old branches that had inevitably been left at ectopic parts of the tectum (owing to tectal growth) were retracted or degenerated in the following larval development. The above mode of axonal growth provides an adequate explanation of cellular mechanisms of terminal shifting of retinal axons within the tectum during development of retinotectal projection. Selection of appropriate branches may also lead to a reduction in the size of terminal arborization of retinal axons, resulting in a refinement in targeting.

Animals↗

Development of intertectal neuronal connections in xenopus: the effects of contralateral transposition of the eye and of eye removal.

The development of intertectal neruonal connections has been investigated in Xenopus laevis. Contralateral eye grafts and enucleations were performed in embryos and the resultant visual projections to the optic tecta were mapped electrophysiologically after metamorphosis. In enucleated animals the ipsilateral projections were found to be normally organised retinotopically but consisted of visual units with abnormally large multi-unit receptive fields. In 10 animals with contralaterally grafted eyes a normal ipsilateral projection had developed from the abnormal eye and an abnormal projection from the normal eye, to produce congruent maps via the two eyes to one tectum. All the maps in these animals were retinotopically organised. In another 11 animals the ipsilateral projection from the operated eye was fragmentary or absent, while that from the unoperated eye resembled the pattern found after enucleation. Retinotopically abnormal contralateral projections had developed in 5 animals of this group. These results suggest that prefunctional specification determines the initial development of diffuse intertectal visual connections but these may be modified by a process of binocular interaction in the presence of a normal primary contralateral input.

Animals↗

A simple computer model of excitable synaptically connected neurons.

The space-lumped two-variable neuron model is studied. Extension of the neural model by adding a simple synaptic current allows the demonstration of neural interactions. The production of synchronous burst activity in this simple two-neuron excitatory loop is modeled, including the influence of random background excitatory input. The ability of the neuron model to integrate inputs spatially and temporally is shown. Two refractory periods after stimuli were identified and their role in burst cessation is demonstrated. Our findings show that simple neural units without long-lasting membrane processes are capable of generating long lasting patterns of activity. The results of simulation of simple background activity suggest that an increase in background activity tends to cause decreased activity of the network. This phenomenon, as well as the existence of two refractory periods, allows for burst cessation without inhibition in this simple model.

Neural Networks, Computer↗