An immunohistochemical characterization of the primitive and maturing neuroepithelial components in the OTT-6050 transplantable mouse teratoma.

The neuroepithelial component of the OTT-6050 mouse teratoma has previously been characterized as an experimental system for the study of differentiation and cytologic maturation in embryonal tumours of the human central nervous system. A number of transplantable tumours composed of primitive stem cells and of a neuroepithelial component displaying a spectrum of differentiation were previously produced by centrifugal elutriation of the dissociated OTT-6050 teratoma. These tumours have provided a reproducible cell population that has permitted the study of both the early and later stages of neoplastic neurocytogenesis. The purpose of the present study was to detect, by immunohistochemistry, the earliest stages of neurocytogenesis in these tumours as shown by the expression of neuron-associated microtubule proteins. This was correlated to the appearance and localization of other markers associated with neuronal and glial differentiation. The primitive neuroepithelial structures resembling neural tubes (medulloepithelial rosettes) contained single or small groups of cells which reacted with the monoclonal antibody TUJ1, specific for the neuron-associated class III beta-tubulin isotype. Immature neuroblasts and maturing polar neurons also showed immunoreactivity with TUJ1, whereas reactivity for microtubule-associated protein 2 (MAP2), tau, the 200 kilodalton isoform of neurofilament protein, neuron-specific enolase and synaptophysin was primarily seen in maturing neurons. By comparison, both medulloepithelial and ependymoblastic rosettes, neuroblasts and glial cells were immunopositive with monoclonal antibody TU27, which defines an antigenic site shared by most mammalian beta-tubulin isotypes. Astroglia were reactive with antisera to glial fibrillary acidic and S-100 proteins, but not with monoclonal antibody (MAb) TUJ1, or with MAbs to the other neuron-associated cytoskeletal proteins, MAP2, tau and the 200 kilodalton subunit of neurofilament protein. Our findings suggest that (1) expression of the class III beta-tubulin isotype is an early event during neoplastic neurocytogenesis, (2) this isotype is subsequently preserved in maturing neuronal populations, and (3) it is not present at detectable levels in stem cells or glial cells. The observation that morphologically undifferentiated neuroepithelial cells express a neuron-associated beta-tubulin isotype signifies the value of examining tubulin isotype expression in the characterization of normal and neoplastic neuroepithelial differentiation.