J.A. Narváez

Simultaneous depletion of neurokinin A, substance P and calcitonin gene- related peptide from the caudal trigeminal nucleus of the rat during electrical stimulation of the trigeminal ganglion

Abstract The central terminals of the primary sensory trigeminal ganglion (TG) neurons projecting into the caudal trigeminal nucleus (CTN) of the rat exhibit neurokinin A (NKA)‐, substance P (SP)‐, and calcitonin gene‐related peptide (CGRP)‐immunoreactivities (IRs). We stimulated the TG in the rat to induce some of the alterations which might occur during migraine (neurogenic inflammation). Under a stereotaxic apparatus and by means of a bipolar electrode, one‐side TG of the animals were electrically stimulated (7.5 Hz, 5 ms, 0.8–1.4 mA) with square pulses for 5 min. Then, using immunohistochemical methods, the lower medulla of each rat was studied for NKA‐, SP‐ and CGRP‐IRs. Light microscopic examination of brain‐stem sequencial sections revealed a simultaneous decrease in the immunoreactivities of all neuropeptides (NKA, SP and CGRP) in the CTN ipsilateral to TG stimulation in comparison with the other (not stimulated) side CTN. It is suggested that this decrease in immunoreactivity would be due to the co‐release of neuropeptides following noxious stimuli and that NKA, SP and CGRP might therefore act as co‐transmitters or co‐modulators at the first central synapses of the trigeminal sensory pathway.

Microinjections of subpicomolar amounts of NPY(13–36) into the nucleus tractus solitarius of the rat counteract the vasodepressor responses of NPY(1–36) and of a NPY Y1 receptor agonist

Microinjections of neuropeptide Y (NPY) (1-36) and of the NPY Y1 agonist [Leu31,Pro34]NPY into the caudal dorsomedial part of the nucleus tractus solitarius (Sol) in the anaesthetized rat led to the development of dose-related vasodepressor and bradycardic responses. The threshold dose of the NPY Y2 agonist NPY(13-36) (50 fmol) significantly counteracted the vasodepressor actions of a close to ED50 dose of NPY(1-36) (2.5 pmol) and of the NPY Y1 agonist (5 pmol). These results indicate that NPY Y1 receptor activation in the Sol leads to the development of a vasodepressor response, which can be counteracted by NPY Y2 receptor activation in the Sol. The results support the existence of a Y2/Y1 receptor-receptor interaction in the Sol, via which NPY Y2 receptors may reduce transduction over NPY Y1 receptors.

Distribution of neuropeptide Y-like immunoreactive cell bodies and fibers in the brain stem of the cat

By using intratissue injections of colchicine and an indirect immunoperoxidase technique, we studied the distribution of cell bodies and fibers containing neuropeptide Y-like immunoreactivity in the brain stem of the cat. The densest clusters of immunoreactive perikarya were observed in the following nuclei: anteroventral cochlear, lateral reticular (internal and external divisions), dorsal tegmental, inferior colliculus and dorsal nucleus of the lateral lemniscus. By contrast, the nuclei abducens, the nucleus of the trapezoid body, preolivary, interpeduncularis, infratrigeminal, gigantocellular tegmental field, coeruleus and dorsal motor nucleus of the vagus had the lowest density. Finally, a moderate density of neuropeptide Y-like immunoreactive cell bodies was found in the nuclei: lateral tegmental field, laminar spinal trigeminal, praepositus hypoglossi, superior colliculus, lateral vestibular and motor trigeminal. In addition, a mapping of the neuropeptide Y-like immunoreactive fibers was carried out. Thus, the densest network of immunoreactive fibers was observed in the laminar spinal trigeminal nucleus. The nuclei periaqueductal gray, inferior central, praepositus hypoglossi, postpyramidal raphe, dorsal raphe, incertus and medial vestibular contained a moderate density of immunoreactive fibers, whereas the nuclei interpeduncularis, inferior colliculus, superior central, gracile, retrorubral, Kölliker-Fuse, dorsal tegmental, ambiguus and alaminar spinal trigeminal had the lowest density of neuropeptide Y-like immunoreactive fibers. The anatomical location of neuropeptide Y-like immunoreactivity suggests that the peptide could play an important role in several physiological functions, e.g., those involved in cardiovascular, auditory, motor, visual, nociceptive and somatosensory mechanisms.

Intracisternally injected galanin-(1-15) modulates the cardiovascular responses of galanin-(1-29) and the 5-HT1A receptor agonist 8-OH-DPAT

In view of the demonstration of specific binding sites for [125I]galanin-(1-15) in several brain areas including the nucleus of the solitary tract, possibly indicating the existence of multiple galanin receptor subtypes, the effects of intracisternal injections of galanin-(1-15) on cardiovascular parameters were studied. The effects of co-injections of galanin-(1-15) and galanin-(1-29) and co-injections of galanin-(1-15) and the 5-HT1A receptor agonist 8-OH-2-(di-n-propylamino)tetralin (8-OH-DPAT) were also evaluated. Galanin-(1-15) produced a significant increase in mean arterial blood pressure (maximum effect 10% at 3 nmol of galanin-(1-15)) and in heart rate (maximum effect 12% at 1 nmol). When threshold doses of galanin-(1-15) (0.1 nmol) and galanin-(1-29) (3 nmol) were injected simultaneously they elicited an increase in mean arterial blood pressure. The vasodepressor response induced by an ED50 dose of 8-OH-DPAT (6 nmol) was not modulated by a threshold dose of galanin-(1-15), but the increase in heart rate area induced by galanin-(1-15) alone was no longer observed. When threshold doses of both galanin-(1-15) and 8-OH-DPAT (0.3 nmol) were co-injected a vasodepressor response developed and on heart rate a tachycardic response was seen in the peak effects and the overall tachycardic response induced by galanin-(1-15) was sustained. The results show a different role for galanin-(1-15) as compared with galanin-(1-29) in central cardiovascular control.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of somatostatin-28 (1–12) in the cat brainstem: an immunocytochemical study

We studied the distribution of somatostatin-28 (1-12)-immunoreactive fibers and cell bodies in the cat brainstem. A moderate density of cell bodies containing the peptide was observed in the ventral nucleus of the lateral lemniscus, accessory dorsal tegmental nucleus, retrofacial nucleus and in the lateral reticular nucleus, whereas a low density of such perikarya was found in the interpeduncular nucleus, nucleus incertus, nucleus sagulum, gigantocellular tegmental field, nucleus of the trapezoid body, nucleus praepositus hypoglosii, lateral and magnocellular tegmental fields, nucleus of the solitary tract, nucleus ambiguous and in the nucleus intercalatus. Moreover, a moderate density of somatostatin-28 (1-12)-immunoreactive processes was found in the dorsal nucleus of the raphe, dorsal tegmental nucleus, accessory dorsal tegmental nucleus, periaqueductal gray and in the marginal nucleus of the brachium conjunctivum. Finally, few immunoreactive fibers were visualized in the interpeduncular nucleus, cuneiform nucleus, locus coeruleus, nucleus incertus, superior and inferior central nuclei, nucleus sagulum, ventral nucleus of the lateral lemniscus, nucleus praepositus hypoglosii, medial vestibular nucleus, Kölliker-Fuse area, nucleus ambiguous, retrofacial nucleus, postpyramidal nucleus of the raphe, nucleus of the solitary tract, dorsal motor nucleus of the vagus, lateral reticular nucleus and laminar and alaminar spinal trigeminal nuclei.

Evidence for an antagonitic angiotensin II/α2-adrenoceptor interaction in the nucleus tractus solitarii

Interactions between alpha 2-adrenoceptors and angiotensin II receptors were evaluated in the nucleus tractus solitarii of the rat by means of quantitative receptor autoradiography and cardiovascular analysis. In binding experiments using l-noradrenaline to compete for [3H]p-aminoclonidine binding sites, angiotensin II (1 nM) increased the IC50 value of l-noradrenaline by 50%. The angiotensin AT1 receptor antagonist, DUP753 (losartan), not only blocked this action but also decreased the IC50 value of l-noradrenaline. The modulatory effect of angiotensin II was also evaluated after addition of both DUP753 and PD123319, an angiotensin AT2 receptor antagonist, and counteraction of the reduction in the IC50 value of l-noradrenaline was observed. In saturation experiments angiotensin II increased the KD and Bmax values of [3H]p-aminoclonidine binding sites, compatible with possible uncoupling of the alpha 2-adrenoceptors. Cardiovascular analysis demonstrated that a threshold dose of angiotensin II (0.05 pmol) counteracted the vasodepressor effect produced by an ED50 dose of l-adrenaline, l-noradrenaline or clonidine coinjected in the nucleus tractus solitarii. DUP753 fully blocked this in vivo modulation of alpha 2-adrenoceptors by angiotensin II. These findings suggest the existence of an antagonistic angiotensin AT1/alpha 2-adrenoceptor interaction in the nucleus tractus solitarii. Therefore, it can be surmised that the activation of angiotensin II AT1 receptors may reduce the transduction of the alpha 2-adrenoceptors and thus the alpha 2-mediated vasodepressor responses.

Increased vasopressor actions of intraventricular neuropeptide Y-(13–36) in spontaneously hypertensive versus normotensive Wistar-Kyoto rats. Possible relationship to increases in Y2 receptor binding in the nucleus tractus solitarius

The C-terminal NPY fragment (13-36)[NPY-(13-36)], a Y2 receptor agonist, elicits vasopressor responses upon central administration. The cardiovascular responses of NPY-(13-36) together with the distribution of NPY receptor subtypes within the nucleus tractus solitarius (nTS) have therefore been studied in spontaneously hypertensive rats (SHR). NPY-(13-36) was injected intracerebro-ventricularly in different doses (7.5 to 3000 pmol) in awake, unrestrained rats to evaluate the cardiovascular effects. NPY receptor subtypes were studied by autoradiography using [125I]peptide YY ([125I]PYY) as a radioligand and by masking the NPY Y1 and Y2 receptor subtypes with unlabelled [Leu31,Pro43]NPY and NPY-(13-36) respectively. In both male SHR and age-matched male normotensive Wistar-Kyoto rats (WKY) NPY-(13-36) injections elicited vasopressor effects. In WKY this effect was dose-dependent and became significant at doses from 75 pmol, whereas in the SHR the vasopressor effect had a longer duration than in the WKY and became significant at lower doses (25 pmol) but associated with the development of an early ceiling effect. The heart rate was unaffected in both groups of rats. Total specific [125I]PYY binding in the nTS was 25% higher in SHR than in WKY rats. By masking the Y1 and Y2 receptor subtypes respectively it could be shown that this difference was due to an increase in Y2 receptor binding within the nTS. The present results give evidence for an increased potency but not an increased efficacy of NPY-(13-36) in inducing a pressor response in the SHR associated with a longer duration as compared with the WKY rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Alterations in neurokinin A-, substance P- and calcitonin gene- related peptide immunoreactivities in the caudal trigeminal nucleus of the rat following electrical stimulation of the trigeminal ganglion

We have carried out an immunohistochemical study on the presence of neurokinin A (NKA) and substance P (SP) in the rat caudal trigeminal nucleus (CTN) after electrical stimulation of the trigeminal ganglion (TG), used as an experimental model to induce alterations, some of which may occur during migraine attacks (release of vasoactive peptides from perivascular trigeminal axons and neurogenic inflammation). Both unilateral, 30 min electrical stimulation (5 Hz, 5 ms, 0.1-1 mA) of the TG and 5 min stimulation with a slight increase in the stimulating parameters (7.5 Hz, 5 ms, 1.4 mA) caused a significant depletion of the NKA and SP immunoreactivities (-IR) of the TG nerve central terminals in the ipsilateral CTN. Calcitonin gene-related peptide (CGRP)-IR of the ipsilateral CTN was also studied in the CTN using the increased stimulating parameters and a marked depletion of CGRP-IR was also observed following TG stimulation. Such depletion may be due to the release of neuropeptides from the trigeminal central terminals. These findings suggest that NKA, SP and CGRP could act as neurotransmitters at the first central synapses of the trigeminal nociceptive pathway to transmit the sensory stimuli to the higher brain centers.

Coinjections of NPY(1–36) or [Leu31,Pro34]NPY with adrenaline in the nucleus tractus solitarius of the rat counteract the vasodepressor responses to adrenaline

The cardiovascular effects of adrenaline microinjected alone or together with neuropeptide Y (NPY) receptor agonists into the nucleus tractus solitarius (Sol) of the anaesthetized rat have been investigated in order to evaluate NPY/adrenergic receptor interactions. In the dose range 0.05-20 nmol, adrenaline microinjected unilaterally into the Sol produced significant dose-related reductions in mean arterial blood pressure and heart rate. The vasodepressor action of a close to ED50 dose of adrenaline (0.5 nmol) was significantly counteracted by a threshold dose of NPY (1-36) (1 pmol) and of the NPY Y1 receptor agonist [Leu31,Pro34]NPY (2.5 pmol) microinjected into the Sol, but not by a threshold dose of NPY(13-36)(50 fmol), a selective Y2 receptor agonist. The present study provides evidence for an antagonistic NPY Y1/adrenergic receptor interaction in the Sol of the rat, involved in cardiovascular regulation.

Somatostatin-28 (1–12)-like immunoreactivity in the cat diencephalon

Using an indirect immunoperoxidase technique, the location of somatostatin-28 (1-12)-like immunoreactive fibres and cell bodies in the cat diencephalon was studied. The hypothalamus was richer in somatostatin-28 (1-12)-like immunoreactive structures than the thalamus. A high density of immunoreactive fibres was observed in the nuclei habenularis lateralis, paraventricularis anterior (its caudal part), filiformis, hypothalami ventromedialis, and regio praeoptica, whereas a moderate density was found in the nuclei paracentralis, supraopticus, supra chiasmaticus, hypothalamus posterior and area hypothalamica dorsalis. The nuclei lateralis dorsalis, lateralis posterior, medialis dorsalis, rhomboidens, centralis medialis, ventralis medialis, reuniens, anterior dorsalis, parataenialis, interanteromedialis, hypothalamus lateralis, hypothalamus dorsomedialis and arcuatus had the lowest density of immunoreactive fibres. In addition, a high or moderate density of somatostatin-28 (1-12)-like immunoreactive cell bodies was observed in the nuclei paraventricularis hypothalami, supraopticus, supra chiasmaticus, area hypothalamics dorsalis, subparafascicularis, hypothalamus posterior and hypothalamus anterior, whereas scarce immunoreactive perikarya were visualized in the nuclei lateralis dorsalis and parafascicularis. The distribution of somatostatin-28 (1-12)-like immunoreactive structures is compared with the location of other neuropeptides in the cat diencephalon.