EVIDENCE FOR DIFFERING ORIGINS OF THE SEROTONERGIC INNERVATION OF MAJOR CEREBRAL-ARTERIES AND SMALL PIAL VESSELS IN THE RAT

We have studied the nature and origin of the serotonergic innervation of two distinct anatomical cerebrovascular compartments, namely, small pial vessels and major cerebral arteries, in the rat. To this end, the levels of serotonin [5-hydroxytryptamine (5-HT)] and 5-hydroxyindoleacetic acid (5-HIAA) were measured by HPLC in both cerebrovascular compartments after either bilateral sympathectomy or destruction of the ascending serotonergic pathways, which originate from the raphe nuclei. We first showed that the small pial vessel samples were not contaminated by underlying cortical tissues through the use of an immunohistochemical approach that revealed the glia limitans, the most superficial cortical layer. Superior cervical ganglionectomy caused a marked decrease in noradrenaline concentrations in major cerebral arteries (-77%), although the reduction was less pronounced (-34%) in small pial vessels. Sympathectomy decreased by 33% 5-HT concentrations in the major cerebral arteries but was without effect on 5-HT levels in the small pial vessels. Destruction of the ascending serotonergic pathways (via local administration of 5,7-dihydroxytryptamine into the ventral tegmental area) produced a dramatic fall in 5-HT and 5-HIAA concentrations in both vascular compartments. To establish the authenticity of the serotonergic innervation, the synthesis of 5-HT [as assessed by measuring the accumulation of 5-hydroxytryptophan (5-HTP) after decarboxylase inhibition] was measured in the two vascular beds under control conditions and after destruction of the ascending serotonergic pathways. The rate of accumulation of 5-HTP was higher in the small pial vessels than in major cerebral arteries, an observation that indicates an important de novo synthesis of 5-HT in small pial vessels. The neurotoxic lesion of the ascending serotonergic fibers caused a marked reduction of 5-HTP accumulation in small pial vessels (-67%) and a slight but significant decrease (-24%) in major cerebral arteries. These results collectively point to the existence of a true serotonergic innervation of small pial vessels that originates in the rostral raphe nuclei; in contrast, most of the 5-HT present in major cerebral arteries seems to originate from sources other than these nuclei.