Hippocampal regulation of the survival and morphological development of locus coeruleus neurons in dissociated cell culture

Abstract

The influence of target structures on neural development, originally described for the peripheral nervous system, has more recently been investigated in the central nervous system. We sought to determine whether targets regulate the development of the locus coeruleus, with its diffuse and complex projections in marked contrast to the simpler neural circuits of the peripheral nervous systems. Dissociated locus coeruleus cells were grown alone or with the hippocampus in serum-free, chemically defined medium that minimized non-neuronal growth. Coculture with the hippocampus resulted in a significant increase in locus coeruleus tyrosine hydroxylase activity. Elevated tyrosine hydroxylase activity was accompanied by a commensurate increase in the number of tyrosine hydroxylase-immunoreactive cells, suggesting hippocampal enhancement of locus coeruleus survival. Furthermore, when hippocampal cells were added to locus coeruleus dissociates at zero time, or after two days, tyrosine hydroxylase-positive cell number was increased only by hippocampal cells added initially, suggesting that the target does indeed foster survival. The apparent target enhancement of locus coeruleus survival seems to be selective since total protein and total neuron number, estimated with neuron-specific enolase immunocytochemistry, were not affected by the hippocampus. Coculture with the hippocampus also increased the length and complexity of locus coeruleus cell processes. Neither the increase in tyrosine hydroxylase cell number nor the changes in morphology could be attributed to nonspecific effects of the increased cell density in cocultures. Our observations thus suggest that the target hippocampus influences the survival and neurite elaboration of afferent locus coeruleus neurons.