The Corticotectal Projection of the Rat In Vitro: Development, Anatomy and Physiological Characteristics

Publisher Website
Google Scholar
Abstract
In this study, the formation of the corticotectal projection of the rat in organotypic slice culture was investigated, using both anatomical and physiological approaches. The establishment of fibre connections from visual cortex to superior colliculus explants was monitored after 3, 6, 14, 20 and 30 days in vitro by cortical injections of Dil. As in cortical cultures without cocultured colliculus, fibres anterogradely labelled by this procedure spread radially from the injection site into the surroundings of the explant, without displaying any directional preference. Especially, layer V pyramidal cells could be seen to extend processes not only to the collicular target, but also in the opposite direction, suggesting that no axonal guidance was exerted by the projection target. The total number of fibres projecting in the direction of the colliculus was not higher than of those projecting in the opposite direction. However, fibres projecting into the colliculus were significantly longer. This was also the case when the colliculus was placed next to the pial side of the cortical explant, indicating that outgrowth direction was not related to this observation. We therefore assume a chemotrophic rather than a chemotactic influence of the projection target on cortical axons, which is based on direct contact of axons to the target tissue. It cannot be excluded, however, that the failure to detect chemotactic guidance was caused by the lack of diffusion gradients in our culture system. Innervation of the collicular slice exclusively originated from layer V pyramidal cells, irrespective of the position of the collicular target. Fibre courses suggested that discrimination of the projection target was achieved upon encounter with the collicular surface by direct membrane contact. Inside the collicular tissue, fibre arborizations occurred preferredly in up to three layers perpendicular to the surface. Even after the smallest tracer injections, termination fields were diffusely distributed over the collicular slice. Also, the spatial distribution of retrogradely stained projection neurons did not differ statistically from an equal distribution. Thus, a high degree of convergence and divergence was observed anatomically in the corticotectal projection formed in vitro, corresponding to the immature state in vivo. The functionality of the corticotectal projection was assessed by intracellular recordings from collicular neurons. Electrophysiological properties, such as membrane potential (‐68 ± 11 mV), membrane resistance (35.4 ± 27.7 Mω) and the time constant (3.0 ± 2.1 ms) were comparable to reference values, confirming the viability of our culture preparation. The functionality of corticotectal transmission was revealed by intracellularly recorded responses of collicular cells to extracellular cortical stimulation. Most responses were excitatory (90%), although inhibitory responses were also encountered (10%). High‐frequency stimulation suggested polysynaptic transmission in all cases tested. Responses from single collicular cells could always be elicited from various cortical stimulation sites, which were usually distributed over the whole cortical explant, confirming the high degree of convergence suggested by the anatomical results. Conduction velocities of corticotectal fibres were estimated to be ∼0.3 m/s, indicating that the fibres of the corticotectal connection in vitro were probably unmyelinated.