Ultrastructural evidence for a separate, small synaptic vesicle (SSV) pathway in ligated bovine splenic nerves, incubated in vitro

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In sympathetic nerves the tubules of the axonal reticulum make up the immature elements of the neurosecretory apparatus. The formation of the mature large dense granules occurs via a less dense tubular intermediate, representing the maturing part. At a terminal small synaptophysin-positive vesicles are found intermingled with the dense granules. The biogenesis of these clear, small synaptic vesicles and their relationship with dense granules remains to be determined. In search for the small synaptic vesicles we undertook a careful ultrastructural examination of the axons proximal to a ligation in bovine splenic nerve incubated in vitro for 3 h. The distended axons were crowded with tubules, granulo-tubular elements and dense granules. Occasionally homogeneous clusters of small, uniform vesicles were detected. They were shown to be positive for synaptophysin and were negative for dopamine-β-hydroxylase, a marker for the granular pathway. The clusters of small vesicles could be found in close spatial relationship with the maturing and mature elements of granular secretion. Our findings argue for the presence of two separate neurosecretory pathways in sympathetic nerves and favour the idea that both small synaptic vesicles and dense granules are a differentiation product of the axonal reticulum. This configuration can explain the biogenesis of small synaptic vesicles and dense granules both in the cell body and at the nerve terminal.