5-HT1R OR 5-HT1D SITES - EVIDENCE FOR 5-HT1D BINDING-SITES IN RABBIT BRAIN

Radioligand binding studies were performed in membranes of rabbit whole brain and striatum using the novel iodinated radioligand for 5-hydroxytryptamine 5-HT1B and 5-HT1D sites, Serotonin-5-O-Carboxy-methyl-glycyl[I-125]Tyrosinamide ([I-125]GTI). [I-125]GTI labelled a finite number of high affinity sites in rabbit brain membranes, B(max) = 191 +/- 47 fmol/mg protein, pK(D) (-log mol/l) = 8.50 +/- 0.13, n = 5. The pharmacological profile of [I-125]GTI binding was fully comparable to that reported previously in human and other brain preparations known to possess 5-HT1D sites (using either [H-3]5-HT or [I-125]GTI) and displayed a characteristic rank order of affinity: 5-carboxamidotryptamine > 5-HT = dihydroergotamine greater-than-or-equal-to ergotamine greater-than-or-equal-to sumatriptan greater-than-or-equal-to CGS 12066 greater-than-or-equal-to metergoline > yohimbine greater-than-or-equal-to methysergide > ICYP > 8-OH-DPAT greater-than-or-equal-to CP93129 > (-)pindolol > ketanserin > isamoltane > mesulergine > corynanthine > buspirone > MDL 72222. Autoradiographic studies were performed on rabbit brain slices using [H-3]5-HT in the presence of 100 nmol/l 8-OH-DPAT and mesulergine (in order to mask 5-HT1A and 5-HT1C binding sites) and [I-125]CYP (iodocyanopindolol) in the presence of 3-mu-mol/l isoprenaline and 100 nmol/l 8-OH-DPAT (in order to mask beta-adrenoceptor and 5-HT1A binding sites). There was no detectable specific binding of [I-125]CYP through the brain, thus excluding the presence of 5-HT1B sites in rabbit brain. By contrast, [H-3]5-HT labelled a high density of sites in globus pallidus, substantia nigra and superior colliculus. Other regions displaying labelling include the striatum, the dentate gyrus of the hippocampus and the periaqueductal grey matter. This pattern of distribution is compatible with that reported for 5-HT1D sites in other species. The present data strongly suggest that rabbit brain has recognition sites with the pharmacological profile and distribution characteristic of the 5-HT1D recognition site. These findings are in agreement with results obtained by Limberger et al. (1991), suggesting the terminal 5-HT autoreceptor of the rabbit brain to belong to the 5-HT1D subtype. Except for limited species variations (see Bruinvels et al. 1992), the pharmacology of these sites does not justify an addition to the existing nomenclature ("5-HT1R"), in contrast to what has been proposed by Xiong and Nelson (1989). The rabbit represents another laboratory species in which 5-HT1D receptor mediated effects could be studied.