Growth factor-induced neurite growth in primary neuronal cultures of dogs with neuronal ceroid lipofuscinosis.

Neuronal ceroid lipofuscinosis (NCL) is a type of lysosomal storage disease resulting in the progressive deterioration of neuronal function. Little is known about the genetics, pathophysiology and biochemical basis of this disease. This is, in part, due to the complexity of the central nervous system and the lack of an in vitro model. In this report, we describe the conditions to establish neuronal cells in primary culture from the brains of newborn English setters with NCL, a canine model for this disease. Over 80% of the neuronal cells from normal dog brain establish well-developed interconnecting networks of long neurites. On the contrary, approximately 50% of the neurons cultured from NCL dog brains do not assemble neurites. Of those NCL neurons with processes, the neurites are routinely shorter and fewer in number than those seen in normal cultures. In addition, the characteristic inclusion bodies, pathological markers for this disease in vivo, are prevalent in the soma of cultured neuronal cells isolated from NCL dog brain. A time-dependent maturation of the inclusion bodies suggests a progression of the disease state in culture. The reduced ability of the NCL neurons to establish neurites prompted us to examine the effects of growth factors on neurite assembly. Our data show that insulin-like growth factor I, epidermal growth factor and platelet-derived growth factor are capable of stimulating neurite outgrowth of NCL neurons. We report the establishment and morphological characterization of neuronal cultures from normal and NCL dog brains. The abnormal morphology of cultured NCL neurons can, in part, be alleviated by supplementing the medium with growth factors. The results suggest that this cellular model of NCL will be useful to study the molecular and physiological mechanisms of NCL disease, as well as to test potential therapeutic agents and candidate genes.