Vasopressinergic innervation of the mouse suprachiasmatic nucleus: an immuno-electron microscopic analysis

Attempts are being made to unravel the local circuitry of the suprachiasmatic nucleus, with a view toward eventually correlating specific neuronal systems with circadian events. Hence, the vasopressinergic innervation of this nucleus in the laboratory mouse has been analyzed immunocytochemically at the light and electron microscopical levels. Monoclonal antineurophysin and polyclonal antivasopressin were used on aldehyde-fixed brains. Serial vibratome sections of the appropriate forebrain region were prepared for pre-embedding immunoperoxidase staining and/or postembedding immunogold labeling. Immunoreactive somata, processes, varicosities, and synaptic terminals were found throughout the suprachiasmatic nucleus, their ratio and density varying at different locations. The predominant type of vasopressinergic soma was ovoid to rounded (7-10 microns), containing secretory granules (85-120 nm), a large proportion of which were immunoreactive. Axon terminals, both nonimmunoreactive and immunoreactive, impinged upon vasopressinergic somata and processes, often displaying synaptic specializations. Vasopressinergic terminals, containing secretory granules and microvesicles, were found throughout the nucleus, particularly within the dorsomedial neuropil. These labeled terminals varied in size (0.4-3.4 microns 2) and shape, ranging from compact boutons to pleomorphic profiles, some deeply indented by postsynaptic spines, either dendritic or somatic. Approximately 65% of the vasopressin-containing terminals were axodendritic and 30% axosomatic; about 5% appeared to be axoaxonic. At least a quarter of all vasopressinergic synaptic terminals were axospinous. Other forms of interneuronal contact involving vasopressinergic elements (somata, dendrites) included extensive membrane to membrane appositional sites, and multiple puncta adhaerentia. The versatility of interconnections between vasopressin-containing neurons in the mouse suprachiasmatic nucleus suggests a highly active and coordinated network, which contributes substantially to local intranuclear circuitry. In addition, a dense efferent vasopressinergic output is directed dorsally towards the periventricular hypothalamus, where direct associations may be established with diverse hypothalamic neuroendocrine systems.