Transplantation of M213-2O cells with enhanced GAD67 expression into the inferior colliculus alters audiogenic seizures.

The purpose of the present study was to examine the effects of GABA-producing cell transplants on audiogenic seizures (AGS). The M213-2O cell line was derived from fetal rat striatum and has GABAergic properties. This cell line was further modified to express human GAD(67) and produce elevated levels of GABA. The present study compares the effects of parent M213-2O cell transplants with those of GAD(67)-modified M213-2O cells in AGS-prone Long-Evans rats. Two weeks following implantation of engineered cells, latency to AGS-typical wild running was increased compared to nonimplanted subjects. Survival of the transplanted cells was confirmed by immunochemical labeling of GAD(67) and Epstein-Barr virus nuclear antigen. These findings support the use of GABA-producing cell lines to modify seizure activity.

Effects of tectal grafts on sound localization deficits induced by inferior colliculus lesions in hooded rats.

The goals of this research were to examine the role of the inferior colliculus (IC) in mediating sound localization behavior and the ability of tectal grafts to restore function after IC ablation in the Long-Evans rat. Previous work has suggested that the IC is a major center for processing of information used in localizing sound sources in space. Adult rats were trained on a lick suppression paradigm to discriminate the location of the second pulse in a noise burst pair presented in the horizontal interaural plane. Following baseline testing, rats received bilateral IC lesions, bilateral lesions followed in 1 week by bilateral tectal grafts, or were sham operated. Sound localization ability was then tested 15 to 30 days and 40 to 50 days following surgical procedures. Performance across experimental groups was statistically the same during baseline testing. During the first operative test period lesion-only and grafted animals showed deficits in sound localization ability relative to controls. By the second postoperative test period control and grafted animals did not differ statistically in sound localization ability and performance of both groups was superior to that of lesion-only animals. Histology revealed a similar extent of IC damage in lesion-only and lesion-graft animals and revealed the presence of implanted tectal tissue in all grafted animals. There was significant neuron loss in the dorsal nucleus of lateral lemniscus (DNLL) in lesion-only animals relative to grafted rats and sham controls. Behavioral results suggest that the IC of pigmented rat is important for sound localization ability. The sparing of DNLL neurons in grafted animals suggests that the tectal grafts may directly integrate into or aid intrinsic recovery of the host auditory pathway.

The inferior colliculus: calbindin and parvalbumin immunoreactivity in neural grafts.

The inferior colliculus was selected as a brain stem site for study of neural grafting and identification of calcium binding proteins. Unilateral ablation sites of eight midbrain inferior colliculus in adult Long-Evans rats were implanted with E17-18 caudal tectum. After 2 to 9 months animals were sacrificed and sections reacted using antibodies for calbindin and parvalbumin. The central nucleus of normal inferior colliculus shows high density of neuronal and fiber staining for parvalbumin. Typical graft cores had similar staining distributions including discoid and stellate neuron populations. Graft cores showed low densities of reactivity for calbindin comparable to central nucleus. In surrounding graft regions there was substantive-neuronal and fiber labeling for calbindin and parvalbumin including stellate neuron populations normally found in the dorsal and lateral nuclei of inferior colliculus. These results demonstrate that the expression of calcium binding proteins in tectal grafts resembles that of inferior colliculus.

Structural features of neurons in whole grafts of the rat inferior colliculus.

The inferior colliculus (IC) is a midbrain structure that receives ascending auditory input from brainstem nuclei via the lateral lemniscus, sends efferent fibers to the medial geniculate body of thalamus and receives descending projections from auditory cortex. In the rat, the IC consists of dorsal and external cortices surrounding the central nucleus of IC (CNIC) which is populated by discoid and stellate neurons; the CNIC has a laminar appearance arising from organization of lemniscal fibers and processes of discoid cells. The IC of adult rats was chosen for implantation of whole grafts of E16-17 caudal tectum into unilateral lesion sites. Dendritic and somal architecture of graft neurons was examined 1 to 4.5 months following implantation using rapid Golgi, HRP and Nissl methods. The CNIC of rat is dominated by principal neurons with relatively flattened dendritic fields. In grafts of caudal tectum the most common neuron class observed possesses flattened dendritic arbors which often parallel one another. These neurons also resemble CNIC neurons of host tissue adjacent to the graft border. Spine formations appear on both proximal and distal dendrites of this neural type in both normal and implanted tissues. In addition, comparable somal features of graft neurons include ovoid or fusiform shapes with regular nuclear membranes as found in the normal colliculus. In Golgi stained material fewer stellate class neurons appear as in the normal CNIC, although stellate cell classes are more abundant in the pericentral areas of normal tissue. Both neuron populations are retrogradely labelled in graft and normal IC after HRP injection into the medial geniculate body. These features suggest that the graft core typically consists of prototypic CNIC cells. Other features of neuron and glial cell density vary in graft material which also shows a complex network of vasculature. These results demonstrate that whole grafts of caudal tectum placed into the inferior colliculus can form organized neural architecture similar to the normal CNIC. The somal, dendritic and spine features of these neurons form a potential substrate for connectional and functional properties which establish this preparation as suitable for further investigation as a model for development and recovery of function in the central auditory system.