ABSTRACT
The growth patterns of axons and dendrites differ with respect to their number, length, branching, and spatial orientation; therefore, it is likely that these processes differ in their growth requirements. To examine this hypothesis, we have been analyzing the responses of cultured rat sympathetic neurons to three types of stimuli: large structural proteins of the extracellular matrix, matrix-associated growth factors, and neurotrophins. Purified structural proteins such as laminin and collagen IV have been found to promote only axonal growth; whereas the matrix associated growth factor, osteogenic protein-1, selectively stimulates dendritic growth. In contrast, nerve growth factor modulates the growth of both types of processes. These data suggest that process-specific interactions with the extracellular environment may be critical determinants of cell shape in neurons. Perinatal rat sympathetic neurons grown in culture in the absence of serum or glial cells extend a single process which is axonal in nature. Exposure to osteogenic protein-1 causes the formation of additional processes which express the morphological, cytoskeletal, and ultrastructural characteristics of dendrites. Consistent with observations on the regulation of dendritic growth in sympathetic neurons in situ, the dendrite-promoting activity of osteogenic protein-1 is independent of synaptic or electrical activity, but is modulated by nerve growth factor. In the presence of optimal concentrations of osteogenic protein-1 and nerve growth factor, the size of the dendritic arbor extended by cultured sympathetic neurons approximates that seen in situ at comparable developmental stages. Osteogenic protein-1 does not promote dendritic growth in cultured neurons obtained from embryonic ciliary, dorsal root, trigeminal or nodose ganglia, suggesting that its morphogenetic effects are cell selective. Since mRNA for osteogenic protein-1 is expressed in mature as well as embryonic target tissues of the sympathetic nervous system, we also examined the effects of osteogenic protein-1 on cultures of sympathetic neurons derived from adult rats. Consistent with results obtained with perinatal neurons, osteogenic protein-1 selectively promoted dendritic growth in adult neurons. These data suggest that this matrix-associated growth factor could play a role not only in the morphogenesis of the developing nervous system, but also in the maintenance and remodeling of dendritic structures in the mature animal.
Subject(s)
Bone Morphogenetic Proteins/physiology , Extracellular Matrix/physiology , Neurons/physiology , Sympathetic Nervous System/physiology , Transforming Growth Factor beta/physiology , Animals , Bone Morphogenetic Protein 7 , CHO Cells , Cell Differentiation/physiology , Cricetinae , Culture Techniques , Dendrites/physiology , Dendrites/ultrastructure , Female , Humans , Immunohistochemistry , Neurons/ultrastructure , Peripheral Nervous System/cytology , Peripheral Nervous System/embryology , RNA/biosynthesis , RNA/metabolism , Rats , Superior Cervical Ganglion/cytology , Superior Cervical Ganglion/embryology , Superior Cervical Ganglion/physiology , Sympathetic Nervous System/cytology , Sympathetic Nervous System/embryologyABSTRACT
The increase in the invertase activity of extracts from freshly cut potato (Solanum tuberosum L.) by "foaming," caused by selective denaturation of an endogenous invertase inhibitor, did not occur in extracts made from thin disks 2 days after slicing. Rather, foaming such extracts decreased invertase activity. Apparently, the inhibitor disappeared after slicing, and the enzyme became more labile to foaming. Such disappearance of inhibitor could account for up to 15% of the dramatic increase in total invertase activity that had occurred within 2 days after slicing. The difference between extracts from 0-day and 2-day slices was mainly in the first of two peaks of invertase activity eluted from diethylaminoethyl-cellulose columns. This peak was increased by foaming 0-day extracts, but even when foamed was much smaller than in 2-day extracts. The apparent loss in inhibitor was not caused by a decreasing susceptibility of the enzyme to the inhibitor. Both the increase in total invertase activity and the apparent loss of inhibitor after slicing were partially blocked by actinomycin D and completely blocked by cycloheximide.The presence of the inhibitor can lead to serious errors in the usual whole disk method of assay for invertase in slices. Ethyl acetate treatment reduces the solubility of the enzyme but does not inactivate the inhibitor.
