Bilateral serotonergic projections to the dorsal hippocampus of the rat: Simultaneous localization of ³H‐5HT and HRP after retrograde transport

Horseradish peroxidase (HRP, Sigma VI, 30–70 nl of a 10–15% solution in saline) or ^3H‐5HT (30 Ci/mmole, 2.5 × 10⁻³ M containing 3.3 × 10⁻³ M norepinephrine in saline, 50–100 nl) was injected unilaterally into the dorsal hippocampus in separate groups of rats. HRP‐labeled cells were seen in the hippocampus, medial septal nucleus, nucleus of the diagonal band, supramammilary nucleus, median raphe nucleus, interfascicular portion of the dorsal raphe nucleus, and the locus coeruleus. In contrast, ^3H‐5HT‐labeled cells were largely restricted to the raphe nuclei. In this nucleus an equal number of ipsilateral and bilateral cells were found. Occasionally, these labeled cells stretched across the midline (bridge pattern). In another series, the ³H‐5HT and HRP were injected into the same hippocampus either as a mixture or sequentially. This resulted in double labeling of the median and dorsal raphe neurons. A final group of rats received injections of ³H‐5HT and HRP into opposing hippocampi. Double‐labeled cells accounted for 10% of the neurons labeled. In addition, closely paired neurons composed of an HRP‐and ³H‐5HT‐containing cell were found. In summary, the serotonergic fibers may play a key role in harmonizing the electrical activity of the hippocampi by use of bilateral projections, paired neurons with differential projections, and bridging neurons stretching across the midline but with unilateral projections.