Ultrastructural and histochemical evidence for differentiation of intraocular locus coeruleus grafts and invasion of the host iris by central neurites and glia

Publisher Website
Google Scholar
Abstract
Intraocular grafts of dorso-lateral pons, including the noradrenaline-containing cell group locus coeruleus, have been studied with ultrastructural and histochemical techniques. Also, the invasion of neuronal and glial constituents from the grafts into the iris of the host animal is described. In mature brain grafts, aggregates of locus coeruleus neurons were easily discernible with monoamine histofluorescence. These cells had an ultrastructural appearance very similar to thatin situ. Numerous somatic spines were frequently associated with synaptic specializations, and monoamine-containing vesicles could be found scattered in the cytoplasm of the locus coeruleus cells. Large neurons of the nucleus tractus mesencephalici nervi trigemini were also found. These cells were neurofilament-immunoreactive just asin situ, and were ultrastructurally characterized by size, distribution of the granular endoplasmic reticulum and abundant large terminals in synaptic contact with their somata and processes. All grafts showed a vigorous astroglial proliferation, evidenced both with immunohistochemistry of glial fibrillary acidic protein and electron microscopy. The astroglial cells were more numerous, larger and with more processes than in adultin situ counterparts. At the attachment site of the brain stem grafts, the iris dilator plate was entirely changed ultrastructurally by a vigorous invasion of neuronal and astrocytic processes. The normal, loose connective tissue stroma of the iris was replaced by layers of almost exclusively central nerve fibres and astrocytes respectively. Monoamine histofluorescence demonstrated an extreme adrenergic hyperinnervation of the iris at the attachment site of the graft, compared to the normal sympathetic ground plexus, whereas neurofilament immunohistochemistry did not visualize any substantial ingrowth of such positive central nerve fibres. Immunohistochemistry of glial fibrillary acidic protein strongly supported the ultrastructural evaluation, showing profound astroglial invasion deep into the iris stroma. Electron microscopic identification of central nerve fibres in the iris showed numerous adrenergic locus coeruleus fibres with small dense-core vesicles. Also, bundles of thin, central, unmyelinated axons were found deep in the iris as well as occasional dendrites. Both large dense-cored and small clear vesicles were encountered in the iris fibres of brain graft origin. Axo-dendritic synaptic specializations formed by locus coeruleus-derived adrenergic fibres were found in the iris. The present results show that there is a profound and intimate contact established between the iris and an immature brain stem area upon intraocular transplantation. The detailed interactions between peripheral iris nerves and supportive glia on one hand, and corresponding central nerves and glia of the grafts on the other hand, can hereby be scrutinized with further histochemical and ultrastructural investigations.