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Several chimeric peptides consisting of the N-terminal fragment of galanin (GAL) and C-terminal fragments of other bioactive peptides (e.g. substance P, bradykinin, neuropeptide Y, mastoparan) have been synthesized and reported as high-affinity galanin receptor antagonists. Recently we have synthesized a new chimeric peptide, GAL(1-13)-[Ala10,11]ET-1(6-21)-NH2, consisting of the N-terminal fragment of GAL and the C-terminal fragment of endothelin-1 (ET-1) analogue. This chimera was previously shown to be a moderate-affinity ligand to hypothalamic galanin receptors with a KD value of 205 nM. However, its biological action has been unknown so far. In our studies we characterized the biological properties of this new chimeric analogue, investigating its action on rat isolated gastric smooth muscles and influence on insulin secretion from rat isolated islets of Langerhans. Data acquired in the course of our studies suggest that analogue GAL(1-13)-[Ala10,11]ET-1(6-21)-NH2 does not seem to be a potent galanin receptor antagonist in the gastrointestinal tract.
Galanin is a peptide present in the nervous system and peripheral tissues which exerts a broad range of physiological functions. The influence of centrally administered galanin (Gal; 100 pM i.c.v.) on arginine vasopressin (AVP) and oxytocin (OT) content in the hypothalamus and neurohypophysis as well as on their blood plasma concentration was estimated in male Wistar rats drinking ad libitum 2% solution of natrium chloride per 48 hours. In euhydrated rats and subsequently applied i.c.v. with Gal a significant fall in the hypothalamic and neurohypophysial content of OT but not AVP was observed, however, without simultaneous changes in these neurohormones blood plasma concentration. On the contrary, i.c.v. injection of Gal to salt-loaded rats caused a marked raise in AVP and OT level in the hypothalamus and neurohypophysis with subsequent diminution of both neurohormones concentration in blood plasma. These results suggest that in euhydrated rats Gal has an inhibitory influence on the biosynthesis as well as axonal transport of OT, but not AVP. On the contrary, in salt-loaded rats galanin restricts secretion of both neurohormones into the systemic circulation.
The presence of choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), galanin (GAL), substance P (SP) and calcitonin gene-related peptide (CGRP) was studied in neurons and nerve fibers of the porcine otic ganglion. ChAT-positive neurons were very numerous while VAChT-positive nerve cells were moderate in number. The number of neurons containing NPY and VIP was lower and those containing SOM, GAL, SP or CGRP were observed as scarce, or single nerve cells. The above mentioned substances (except SOM) were present in nerve fibers of the ganglion. ChAT- and VAChT-positive nerve fibers were numerous, while the number of nerve terminals containing NPY, VIP and SP was lower. GAL- and CGRP-positive nerve fibers were scarce.
The effect of centrally administered galanin (Gal; 100 pM i.c.v.) on the hypothalamo-neurohypophysial storage as well as blood plasma level of vasopressin and oxytocin was estimated in haemorrhaged (1 ml per 100 g b.w.) male Wistar rats. Gal i.c.v. treatment did not alter vasopressin and oxytocin content both in the hypothalamus and neurohypophysis as well as their concentration in blood plasma of not haemorrhaged rats. Haemorrhage decreased the hypothalamic and neurohypophysial vasopressin and oxytocin storage but increased the neurohormones plasma level in animals injected with vehicle solution. During the haemorrhage, the increase in plasma vasopressin and oxytocin was inhibited in rats previously treated i.c.v. with galanin. The hypothalamic and neurohypophysial vasopressin as well as oxytocin content significantly increased in animals treated with galanin and subsequently haemorrhaged. These results suggest that galanin may have a regulatory role in the hypothalamo-neurohypophysial function especially under condition of hypovolemia.
Galanin (Gal) as a neuropeptide with widespread distribution in the central nervous system may be involved in the mechanisms of vasopressin (AVP) and oxytocin (OT) release from the hypothalamo-neurohypophysial system. Vasopressin and oxytocin content in the hypothalamus and neurohypophysis as well as plasma level of both neurohormones were studied after galanin treatment in euhydrated and dehydrated rats. In not dehydrated rats intracerebroventricular (i.c.v.) injections of Gal did not affect the hypothalamic and neurohypophysial OT content, however, distinctly increased plasma OT concentration. In the same animals Gal diminished the hypothalamic AVP content but was without the effect on neurohypophysial AVP storage; plasma AVP level then raised. Galanin, administered i.c.v. to rats deprived of water, distinctly inhibited AVP and OT release from the hypothalamo-neurohypophysial system. Simultaneously, plasma AVP and OT level was significantly diminished after Gal treatment in dehydrated rats. These results suggest that modulatory effect of galanin on vasopressin and oxytocin release depends on the actual state of water metabolism. Gal acts as an inhibitory neuromodulator of AVP and OT secretion under conditions of the dehydration but stimulates this process in the state of equilibrated water metabolism.
Substance P (SP), vasoactive intestinal polypeptide (VIP) and galanin (GAL), present in primary sensory neurons, are involved in transmission of nociceptive signaling from the peripheral to central nervous system. In this study we investigated the effect of GAL on SP-induced or VIP-induced evoked tongue jerks (ETJ) in response to noxious tooth pulp stimulation during perfusion of the cerebral ventricles with SP or VIP solutions. The experiments were carried out on rats under chloralose anesthesia. It was shown that both, SP and VIP, perfused through the cerebral ventricles enhanced the ETJ amplitude as compared with control, but the effect produced by SP was stronger. The intracerebroventricular perfusion of GAL 5 minutes before SP caused a dose-dependent inhibition of SP-induced ETJ, whereas GAL perfused through the cerebral ventricles 5 minutes before VIP did not reduce the excitatory effect of VIP on ETJ. These results indicate that the antinociceptive effect of GAL perfused through the cerebral ventricles, tested on the trigemino-hypoglossal reflex in rats, is specifically mediated by the SP-ergic system.
The uterine cervix-projecting neurons located in the lumbar paravertebral ganglia were identified by retrograde tracing. These contained immunoreactivity to TH and DBH. No immunoreactivity to GAL, VIP and SP was found in the neurons. Extirpation of the uterus reduced the expression of TH and induced the expression of GAL in the neurons. Expression of other substances studied was unchanged.
The aim of the study was to investigate the expression of biologically active substances in intramural neurons supplying the ileum and large intestines (caecum, spiral colon and descending colon) in normal (control) pigs and in pigs suffering from dysentery. Higher numbers of galanin (GAL)-, vasoactive intestinal polypeptide (VIP)- and calcitonin gene-related peptide (CGRP)-immunoreactive (-IR) neuronal somata were found in the myenteric (MP), and outer (OSP) and inner submucus (ISP) intestinal nerve plexuses in dysenteric pigs as compared to control animals. Additionally, the density of substance P (SP)- and VIP-IR nerve fibres in the studied tissues was higher in dysenteric than in controls animals, whereas the number of CGRP-IR nerve fibres remained unchanged, or even was lower in the experimental pigs. The number of SP-IR nerve cell bodies in the MP of all intestinal segments studied was comparable in dysenteric and control pigs. An increased number of SP-IR perikarya were observed in OSP and ISP of the ileum, cecum and centripetal turns; whereas the number of SP-IR somata was lower in the plexuses of centrifugal turns and the descending colon. The number of nerve fibres found in all layers of the intestinal wall was lower in dysenteric pigs. Each of the intramural plexuses in all the intestinal segments studied contained less than 1% of neuropeptide Y (NPY)-IR neurones and this characteristic was similar both in dysenteric and control pigs. The number of NPY-IR nerve fibres increased slightly in the plexuses as well as in both muscular layers and mucosa.
Molecular probes were generated to study the expression of tyrosine hydroxylase (TH), choline acetyltrans- ferase (ChAT), galanin (GAL), substance P (SP) and calcitonin gene-related peptide (CGRP) in the peripheral nervous system of the pig with in situ hybridization. Application of the probes to the sections from the inferior mesenteric ganglion (IMG) in control animals revealed numerous neurons displaying a hybridization signal for TH, while less numerous nerve cells exhibited the hybridization signal for GAL and ChAT. In the spinal ganglia L1-L3 the hybridization signal for SP and CGRP was detected in the vast majority of neurons, while the hybridization signal for ChAT was found in much smaller population of ganglionic nerve cells. After injection of retrograde tracer Fast Blue (FB) into the porcine ovary and subsequent ovariectomy, FB-positive neurons were detected in the ipsilateral IMG and spinal ganglia L1-L3. Application of a molecular probe for GAL to sections from IMG in ovariectomized animals revealed that the majority of FB-positive neurons displayed the hybridization signal what may suggest increased de-novo synthesis of the peptide in the injured neurons.
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