Galanin-like Immunoreactivity Research Articles

Alterations in Galanin-Like Immunoreactivity in the Enteric Nervous System of the Porcine Stomach Following Acrylamide Supplementation.

In recent years, a significant increase in the consumption of products containing large amounts of acrylamide (e.g., chips, fries, coffee), especially among young people has been noted. The present study was created to establish the impact of acrylamide supplementation, in tolerable daily intake (TDI) dose and a dose ten times higher than TDI, on the population of galanin-like immunoreactive (GAL-LI) stomach neurons in pigs. Additionally, in the present study, the possible functional co-operation of GAL with other neuroactive substances and their role in acrylamide intoxication was investigated. Using double-labelling immunohistochemistry, alterations in the expression of GAL were examined in the porcine stomach enteric neurons after low and high doses of acrylamide supplementation. Generally, upregulation in GAL-LI immunoreactivity in both myenteric and submucous plexuses was noted in all stomach fragments studied. Additionally, the proportion of GAL-expressing cell bodies simultaneously immunoreactive to vasoactive intestinal peptide (VIP), neuronal nitric oxide synthase (nNOS) and cocaine- and amphetamine- regulated transcript peptide (CART) also increased. The results suggest neurotrophic or/and neuroprotective properties of GAL and possible co-operation of GAL with VIP, nNOS, CART in the recovery processes in the stomach enteric nervous system (ENS) neurons following acrylamide intoxication.

Plasmin: Its Role in the Extracellular Processing of Progalanin in Tumor Tissue

Galanin is a neuropeptide that is widely distributed in the central and peripheral nervous systems. In a previous study, we showed that a small cell lung carcinoma (SCLC) cell line, SBC-3A, released progalanin but not galanin, and that progalanin was then converted to galanin(1-20), the active form. Because the galanin(1-20) had undergone hydrolysis at Arg and Lys residues, the protease concerned was surmised to have a trypsin-like activity. The present study was performed to identify the trypsin-like protease which had previously been found to activate progalanin in this tumor tissue. The protease was isolated using chromatography and electrophoresis, and identified in tumor extracts from SBC-3A tumor-bearing mice; the major protease was found to be plasmin. We next confirmed that extracellular processing of progalanin occurs in SCLC tumor tissue (tumors produced by the implantation of SBC-3A cells into mice), and in two types of breast tumor tissue (obtained by implantation into mice of BT-549 and MDA-MB-436 cells). In cell culture, processed forms of progalanin were undetectable in SBC-3A, BT-549 or MDA-MB-436 cells. Conversely, gel filtration chromatography analysis of tumor extracts from SBC-3A, BT-549 and MDA-MB-436-bearing mice, revealed that galanin-like immunoreactivity (galanin-LI) in these tumor extracts was due to the presence of progalanin (14 kDa) and galanin(1-20) (2 kDa). Moreover, trypsin-like protease activity was elevated, and plasmin was expressed abundantly in SBC-3A, BT-549 and MDA-MB-436 tumors in mice. In addition, tranexamic acid, a plasmin inhibitor, inhibited progalanin conversion to galanin(1-20). The present study revealed that plasmin was present in tumor tissue, and that it was responsible for processing progalanin to galanin(1-20) in the extracellular environment.

Expression of galanin and galanin receptor mRNA in skin during the formation of granulation tissue

Galanin is a neuropeptide widely distributed in the central and peripheral nervous systems. Although its role in non-neural cells is poorly understood, it is known that during inflammation, the dermis layer of the skin produces and releases galanin. The aim of this report is to study the expression of galanin in granulation tissue. After inducing inflammation by cotton thread implantation, galanin-like immunoreactivity (galanin-LI) in plasma reached a maximum on the third day. Galanin-LI was observed in fibroblast-like cells occurring close to collagen fibers in developing granulation tissue. Furthermore, galanin receptor subtypes 1 and 2 (GALR1 and GALR2)-expressing cells were observed around microvessels and were found to produce desmin. Galanin was injected along the cotton threads immediately after implantation, resulting in rapid formation of granulation tissue, and an increase in the contents of microvessels, indicating a stimulatory effect of galanin on the process of angiogenesis in granulation tissue. The results demonstrate that some galanin was released from fibroblast-like cells during the formation of granulation tissue, and that it stimulated angiogenesis.

Inflammation- and axotomy-induced changes in galanin-like immunoreactive (GAL-LI) nerve structures in the porcine descending colon

This study reports on changes caused by chemically driven inflammation and axotomy in galanin-like immunoreactive (GAL-LI) nerve structures in the porcine descending colon. The distribution pattern of GAL-LI structures was studied using the immunofluorescence technique in the circular muscle layer, the myenteric (MP), outer submucous (OSP) and inner submucous plexuses (ISP), and also in the mucosal layer. Under physiological conditions GAL-LI perikarya were shown to constitute 3.68 +/- 0.32%, 7.02 +/- 0.93% and 10.99 +/- 0.71% in MP, OSP and ISP, respectively. Both colitis and axotomy caused an increase in GAL-like immunoreactivity, which was different in particular parts of the bowel segment studied. The numbers of GAL-LI perikarya increased to 14.16 +/- 0.49%, 16.78 +/- 1.09% and 37.46 +/- 1.18% during colitis and 7.92 +/- 0.72%, 10.44 +/- 0.71% and 16.20 +/- 0.96% after axotomy in MP, OSP and ISP, respectively. Both these processes caused an increase in the number of GAL-LI nerve fibres in the circular muscle and mucosal layers as well as the appearance of a population of GAL-LI cells in the mucosa.

Presence of Galanin‐Like Immunoreactivity in Mesenchymal and Neural Crest Origin Tissues During Embryonic Development in the Mouse

Galanin is a highly conserved neuropeptide with a wide range of biological effects. Recently, through transcriptome analysis, galanin was identified in undifferentiated mouse embryonic stem cells as one of the most abundant transcripts. We have examined the developmental expression of galanin-like immunoreactivity in mice from embryonic day 10 (E10) to embryonic day 15 (E15). At E10, galanin was readily detected in the undifferentiated head and trunk mesenchyme of both mesodermal and neural crest origin. There was also strong immunoreactivity in the mesenchymal spiral ridges of the outflow tract of the heart and the endocardial cushions. The highest level of galanin detected was at E13 in the craniofacial mesenchyme and proliferating chondrocytes in bones of both neural crest and mesoderm origin. Dorsal root ganglia and trigeminal ganglia contained galanin immunoreactive cells as well. These data indicate the presence of galanin peptide during periods of morphogenesis and thus a developmental role for the peptide in mesenchymal and neural crest origin tissues in the mouse embryo. Whether galanin has a growth and/or differentiating role, still remains to be demonstrated.

Proliferative enteropathy (PE): Induced changes in galanin-like immunoreactivity in the enteric nervous system of the porcine distal colon

The aim of this study was to investigate the changes of galanin (GAL) like immonoreactivity in the porcine descending colon during proliferative entheropathy (PE). Accordingly, the distribution pattern of GAL - like immunoreactive (GAL-LI) nerve structures was studied by the immunofluorescence technique in the circular muscle layer, myenteric (MP), outer submucous (OSP) and inner submucous plexuses (ISP), as well as in the mucosal layer of the porcine descending colon under physiological conditions and during PE. In control animals GAL-LI perikarya have been shown to constitute 4.03 ± 0.1%, 6.67 ± 0.3% and 11.20 ± 0.5% in MP, OSP and ISP, respectively. PE caused changes in the GAL - like immunoreactivity, which differed in particular parts of the studied bowel segment. During PE the number of GAL-LI perikarya amounted to 2.90±0.5%, 8.42±1.0% and 21.72±1,4% within the MP, OSP and ISP, respectively. Moreover PE caused an increase in the number of GAL-LI nerve fibers in the colonic circular muscle and mucosal layers, as well as in all intramural plexuses, especially in ISP, where nearly every ganglion contained a very dense meshwork of the GAL-positive nerve fibers under the studied pathological factor. This study for the first time reports on changes in GAL-LI nerve structures of the porcine descending colon during Lawsonia intracellularis infection.

Differential galanin upregulation in dorsal root ganglia and spinal cord after graded single ligature nerve constriction of the rat sciatic nerve

Single ligature nerve constriction (SLNC) is a newly developed animal model for the study of neuropathic pain. SLNC of the rat sciatic nerve induces pain-related behaviors, as well as changes in the expression of neuropeptide tyrosine and the Y 1 receptor in lumbar dorsal root ganglia (DRGs) and spinal cord. In the present study, we have analyzed the expression of another neuropeptide, galanin, in lumbar DRGs and spinal cord after different degrees of constriction of the rat sciatic nerve. The nerve was ligated and reduced to 10–30, 40–80 or 90% of its original diameter (light, medium or strong SLNCs). At different times after injury (7, 14, 30, 60 days), lumbar 4 and 5 DRGs and the corresponding levels of the spinal cord were dissected out and processed for galanin-immunohistochemistry. In DRGs, SLNC induced a gradual increase in the number of galanin-immunoreactive (IR) neurons, in direct correlation with the degree of constriction. Thus, after light SLNC, a modest upregulation of galanin was observed, mainly in small-sized neurons. However, following medium or strong SLNCs, there was a more drastic increase in the number of galanin-IR neurons, involving also medium and large-sized cells. The highest numbers of galanin-IR neurons were detected 14 days after injury. In the dorsal horn of the spinal cord, medium and strong SLNCs induced a marked ipsilateral increase in galanin-like immunoreactivity in laminae I–II. These results show that galanin expression in DRGs and spinal cord is differentially regulated by different degrees of nerve constriction and further support its modulatory role on neuropathic pain.

The galaninergic enteric nervous system of pleuronectiformes (Pisces, Osteichthyes): An immunohistochemical and confocal laser scanning immunofluorescence study

The galaninergic enteric nervous system of three flatfishes was studied using immunohistochemical and immunofluorescence methods. Galanin-like immunoreactivity was mainly detected within the enteric intramural neurons of the stomachs and the proximal intestines. The sole, Solea solea L. and the flounder, Platichthys flesus L. showed a similar occurrence and relative distribution of galaninergic intramural neurons. Rare nervous fibre immunoreactive to the anti-galanin serum were observed in the muscular layers of the oesophagus of the turbot, Psetta maxima L. The presence and relative abundance of galanin-like immunoreactive neurons in the remaining organs of the alimentary canal of the turbot showed a different pattern in comparison to those observed in the sole and the flounder. A galanin-like peptide was detected in nerve fibres running through the exocrine parenchyma of the pancreas of all three species. It is conceivable that the galaninergic system in these species plays a role in regulating gut muscle activity, and in controlling pancreatic secretion. Galanin and choline acetyltransferase were co-localized within the same neurons of the stomach and intestine. The result confirms the hypothesis that galanin in the gut of fish functions as a cholinergic modulator. Differently from other fish species, immunoreactive endocrine cells were not detected in the studied pleuronectiformes.

Galanin-like immunoreactivity in the brain of the snake Bothrops jararaca

The distribution of galanin-like immunoreactive perikarya and nerve fibers in the brain of the snake Bothrops jararaca was studied by means of immunohistochemistry using an antiserum against porcine galanin. Immunoreactive neurons were only detected in the infundibular recess nucleus. Immunoreactive fibers were found in the telencephalic, diencephalic and mesencephalic areas such as the dorsal cortex, nucleus accumbens, lamina terminalis, preoptic area, mediodorsal region of the supraoptic nucleus, subfornical organ, nucleus of the paraventricular organ, subcommisural organ and periventricular grey region. The habenula, paraventricular nucleus, infundibular recess nucleus and hypothalamo-hypophyseal tract presented denser innervations. The outer layer of the median eminence displayed numerous fibers located close to the portal system, while scarce fibers were seen in the inner median eminence and neural lobe of the hypophysis. The distribution of labelled neurons in the brain of this snake was more restricted than that described in a turtle. The wide hypothalamic and extrahypothalamic distribution of labelled fibers suggests that galanin peptides may have hypophysiotropic, neuromodulator and neurotransmitter roles in the snake B. jararaca.

Alternation of galanin in nociceptive modulation in the central nervous system of rats during morphine tolerance: A behavioral and immunohistochemical study

The present study was performed to investigate the alternation of galanin in nociceptive modulation and galanin-like immunoreactivity in the central nervous system of rats after morphine tolerance. The hindpaw withdrawal latencies to both thermal and mechanical stimulation increased significantly after intracerebroventricular injection of 3 nmol of galanin in opioid-naive rats. The antinociceptive effect induced by galanin was attenuated remarkably at the same dose in morphine-tolerant rats. Furthermore, an up-regulation of galanin-like immunoreactivity in the arcuate nucleus of hypothalamus of morphine-tolerant rat was observed by immunohistochemical methods, whereas no significant changes were detected in periaqueductal gray. The present study demonstrated that thereare alternations in both galanin-induced antinociception and galanin-like immunoreactivity in the brain of rat after morphine tolerance. The results suggest an involvement of galanin in the central nervous system in morphine tolerance.

Phenotyping of sensory and sympathetic ganglion neurons of a galanin-overexpressing mouse—Possible implications for pain processing

The distribution of galanin was studied in the lumbar 5 dorsal root ganglia (DRGs) and spinal cord, superior cervical ganglia (SCGs), and skin of transgenic mice overexpressing galanin under the dopamine β-hydroxylase (DBH) promoter (GalOE-DBH mice) and in wild type (WT) mice. The DRGs and spinal cord were analysed before and after a unilateral, complete transection (axotomy) of the sciatic nerve and after dorsal rhizotomy. Both galanin protein and transcript were studied by, respectively, immunohistochemistry and in situ hybridization. Increased galanin expression was observed in several small, medium-sized and large DRG neuron profiles (NPs) in the naïve transgenic mouse, frequently in neurons lacking calcitonin gene-related peptide (CGRP) and isolectin B4-binding. This lack of coexistence was particularly evident in the medium-sized/large NPs. In the dorsal horn of the spinal cord, no differences were detected between GalOE-DBH and WT mice, both displaying a strong galanin-positive neuropil in the superficial laminae of the dorsal horn, but the transgenic mice showed a more abundant galanin-positive innervation of the ventral horn. A 12-day dorsal rhizotomy, surprisingly, failed to alter the galanin staining patterns in the dorsal (and ventral) dorsal horn. Unilateral axotomy induced upregulation of galanin in DRG NPs of all sizes in both types of mouse. In the hindpaw skin, a profuse galanin-positive fiber plexus was observed in sweat glands and around blood vessels of the transgenic mice, being much more restricted in WT mice. Finally, GalOE mice exhibited a strong galanin-like immunoreactivity in most SCG NPs. The overexpression of the peptide in DRGs and SCGs was paralleled by increased mRNA levels. The present results show that overexpression of galanin under the control of the DBH promoter does not only occur, as expected in these mice, in noradrenline/adrenaline neurons but also in DRG neurons, particularly in large and medium-sized NPs. To what extent and how this overexpression pattern is related to the previously shown elevated pain threshold under normal and lesion conditions is discussed [Grass, S., Crawley, J.N., Xu, X.J., Wiesenfeld-Hallin, Z., 2003a. Reduced spinal cord sensitization to C-fibre stimulation in mice over-expressing galanin. Eur. J. Neurosci. 17, 1829–1832; Hygge-Blakeman, K., Brumovsky, P., Hao, J.X., Xu, X.J., Hökfelt, T., Crawley, J.N., Wiesenfeld-Hallin, Z., 2004. Galanin over-expression decreases the development of neuropathic pain-like behaviour in mice after partial sciatic nerve injury. Brain Res. 1025, 152–158].

Heterogeneous Forms of Immunoreactive Galanin in Human Plasma; Higher Levels in Men than in Women

Galanin, a neuropeptide, has important effects on hormone secretion from the hypothalamus and pituitary, and may also be involved in important biological processes such as pain, memory, and food intake. Yet, there is limited knowledge about how these processes are reflected by circulating galanin. To study the levels and molecular forms of galanin in the human circulation, plasma was analysed from 27 healthy subjects, 14 women and 13 men, using two extraction methods and a specific radioimmunoassay for human galanin. After extraction on Sep Pak C-18 columns, plasma galanin-like immunoreactivity (galanin-LI) in the healthy men was 6.3 +/- 2.5 pmol/l (mean +/- SD, n = 12), which was higher than in the women, 4.1 +/- 1.5 pmol/l (n = 14, p = 0.010). A small increase in galanin-LI was seen with age in the women (r = 0.54, p < 0.05) but there was no significant difference between pre- and postmenopausal women. Galanin immunoreactivity after Sep Pak and immunoextraction correlated (r = 0.74, p < 0.001) the levels being higher after immunoextraction (p < 0.001). Gel chromatography disclosed heterogeneity of circulating galanin-LI with the majority eluting as homologs with a molecular weight higher than synthetic human galanin. Homologs smaller than galanin were also found. Sep Pak C-18 extraction eliminated the majority of the higher molecular forms. In conclusion, circulating galanin-LI was found to be higher in men and to be present mainly as molecular forms larger than synthetic galanin.

Distribution of galanin‐like immunoreactivity in the brain of the Siberian sturgeon (Acipenser baeri)

Galanin is a 29-amino acid peptide widely distributed in the central nervous system of vertebrates. The organization of galaninergic systems is well known in teleosts, the most advanced actinopterygians, but no data are available on primitive bony fish. To extend the evolutionary analysis of galaninergic systems we studied the distribution of galanin-like immunoreactive (GAL-ir) cells and fibers in the sturgeon brain, since chondrosteans are among the most primitive extant actinopterygians. Double-immunolabeling experiments were performed to compare the distribution of galanin with that of neurophysin, tyrosine hydroxylase, and serotonin. Numerous GAL-ir cells of cerebrospinal fluid-contacting (CSF-C) type were found in the ventral telencephalon, preoptic area, and in the tuberal and caudal hypothalamus. The distribution of GAL-ir elements in the sturgeon brain shows many similarities to that observed in other vertebrates, but also important differences, such as the abundance of GAL-ir CSF-C cells, which appear to be a primitive characteristic. GAL-ir neurons observed in the sturgeon telencephalic hemispheres perhaps represent the basic organization of common ancestors of bony fishes and tetrapods. In the preoptic-hypophyseal system, GAL-ir cells appeared to be related not only with neurophysin-expressing neurons (in the tuberal hypothalamus) but also with serotoninergic and catecholamines-synthesizing neurons (in preoptic and tuberal nuclei). Numerous GAL-ir fibers were observed in the median eminence and neural lobe of the hypophysis, indicating that galanin may play a role in the modulation of hypophyseal secretion. GAL-ir neurons were absent from the sturgeon brainstem, suggesting that their presence in other vertebrates could represent an evolutionary recent acquisition.

Estrogen induces a rapid increase in galanin levels in female rat hippocampal formation − possibly a nongenomic/indirect effect

Administration of 17beta-estradiol to ovariectomized rats increased the concentrations of galanin-like immunoreactivity (LI) in the hippocampal formation by 215% (P < 0.001) within 1 h. An increase of 125% (P < 0.05) was observed in the same brain region in the proestrous phase of a normal estrous cycle. Tamoxifen did not block the 17beta-estradiol-induced increase in the concentration of galanin-LI but resulted in a 62% decrease in the hypothalamus within 1 h. In vivo microdialysis in the dorsal hippocampal formation showed a decrease of extracellular galanin-LI (P < 0.001) 1-2 h after treatment with 17beta-estradiol, indicating a decreased release of galanin. For comparison, we studied the concentrations of neuropeptide Y, which were not influenced significantly in any of the regions studied. Taken together our results suggest that 17beta-estradiol inhibits galanin release, presumably from noradrenergic nerve terminals, and primarily via a nongenomic/indirect action, not necessarily involving the classical nuclear receptors ER-alpha or ER-beta. These rapid estrogen-induced changes in galanin release could influence transmitter signalling and plasticity in the hippocampal formation.

Generation and phenotypic characterization of a galanin overexpressing mouse

In most parts of the peripheral nervous system galanin is expressed at very low levels. To further understand the functional role of galanin, a mouse overexpressing galanin under the platelet-derived growth factor-B was generated, and high levels of galanin expression were observed in several peripheral tissues and spinal cord. Thus, a large proportion of neurons in autonomic and sensory ganglia were galanin-positive, as were most spinal motor neurons. Strong galanin-like immunoreactivity was also seen in nerve terminals in the corresponding target tissues, including skin, blood vessels, sweat and salivary glands, motor end-plates and the gray matter of the spinal cord. In transgenic superior cervical ganglia around half of all neuron profiles expressed galanin mRNA but axotomy did not cause a further increase, even if mRNA levels were increased in individual neurons. In transgenic dorsal root ganglia galanin mRNA was detected in around two thirds of all neuron profiles, including large ones, and after axotomy the percentage of galanin neuron profiles was similar in overexpressing and wild type mice. Axotomy reduced the total number of DRG neurons less in overexpressing than in wild type mice, indicating a modest rescue effect. Aging by itself increased galanin expression in the superior cervical ganglion in wild type and transgenic mice, and in the latter also in preganglionic cholinergic neurons projecting to the superior cervical ganglion. Galanin overexpressing mice showed an attenuated plasma extravasation, an increased pain response in the formalin test, and changes in muscle physiology, but did not differ from wild type mice in sudomotor function. These findings suggest that overexpressed galanin in some tissues of these mice can be released and via a receptor-mediated action influence pathophysiological processes.

Increase of galanin-like immunoreactivity in rat hypothalamic arcuate neurons after peripheral nerve injury

Galanin and galanin receptors are widely distributed within the central nervous system, and may play important roles in pain signaling and modulation. In the present study, we examined the galanin immunoreactivity (IR) in the hypothalamus and the amygdala following peripheral nerve injury. Four weeks after the operation, the ipsilateral mechanical threshold in the spared nerve injury (SNI) group (0.87 ± 0.33 g) was significantly lower than that in the sham group (12.53 ± 3.41 g; P < 0.05). In the SNI group, the number of galanin-IR neurons per section in the arcuate nucleus (Arc) of the hypothalamus was 10.2 ± 1.7, significantly higher than that in the sham group (5.6 ± 1.0; P < 0.05). These data suggest that the galanin-ergic neurons in the Arc may be involved in the functional modulation of descending pain modulation system following peripheral nerve injury.

Selective reduction in ventral hippocampal acetylcholine release in awake galanin-treated rats and galanin-overexpressing transgenic mice

The neuropeptide galanin is an inhibitory modulator of hippocampal acetylcholine (ACh) release and cognitive functions. Anatomical evidence demonstrated some differences between the dorsal and ventral hippocampi notably in the expression of galanin receptor subtypes, and the neuronal population on which galanin-like immunoreactivity is expressed. This is suggestive of a differential role for this peptide in these two areas of the hippocampal formation. Using in vivo microdialysis, we investigated the role of galanin on ACh release in the dorsal and ventral hippocampi. Two models were studied: galanin-administered rats and transgenic mice over-expressing galanin (GAL-tg). In rats, galanin (2.0 and 10.0 μM) infused locally through the dialysis probe induced a significant decrease in ACh release in the ventral hippocampus, confirming previous findings, while no effect was seen in the dorsal hippocampus. Using the no net flux method, a significant reduction in ACh levels was noted only in the ventral hippocampus of GAL-tg compared to wild-type littermates. These results suggest that excess endogenous galanin can suppress basal ACh release, with anatomical specificity, to the ventral hippocampus. These results are of interest in the context of galanin receptor subtypes in the dorsal and ventral hippocampus, and the differential alterations of hippocampal subregions in neurological diseases such as Alzheimer's dementia.

The presence of a galanin-like peptide in the gut neuroendocrine system of Lampetra fluviatilis and Acipenser transmontanus: an immunohistochemical study

Galanin is a brain-gut neuropeptide present in the central and peripheral nervous systems of vertebrates. In the present survey, the galaninergic and the diffuse endocrine systems of the alimentary canal of the river lamprey, Lampetra fluviatilis, and the white sturgeon, Acipenser transmontanus, were studied by immunohistochemistry. The results show the presence of galanin-like immunoreactive endocrine cells in the gut of L. fluviatilis. In addition, a galanin-like immunoreactivity was detected in enteric intramural neurons of both species. It is conceivable that the galaninergic system plays in both species a role in the regulation of the gut muscle contractility and in the modulation of mucosal secretive/absorptive processes. In A. transmontanus, the presence of galanin-like immunoreactive nerve fibres associated with components of the gut associated-lymphoid tissue is possibly correlated with a control of the defensive events at this site. The presence of a galanin-like immunoreactivity in the neuroendocrine system of these two ancient fishes confirms the hypothesis on the early occurrence of this regulative molecule in the gastro-enteric system of vertebrates.

Innervation of the Uvea by Galanin and Somatostatin Immunoreactive Axons in Macaques and Baboons

The neuropeptide galanin has not been localized previously in the primate uvea, and the neuropeptide somatostatin has not been localized in the uvea of any mammal. Here, the distribution of galanin-like and somatostatin-like immunoreactive axons in the iris, ciliary body and choroid of macaques and baboons using double and triple immunofluorescence labeling techniques and confocal microscopy was reported. In the ciliary body, galanin-like immunoreactive axons innervated blood vessels and the ciliary processes, particularly at their bases. In the iris, the majority of these axons was associated with the loose connective tissue in the stroma. Somatostatin-like immunoreactive axons were found in many of the same areas of the uvea supplied by cholinergic nerves. In the ciliary body, there were labelled axons within the ciliary processes and ciliary muscle. They were also found alongside blood vessels in the ciliary stroma. In the iris, somatostatin-like immunoreactive axons were abundant in the sphincter muscle and less so in the dilator muscle. A unilateral sympathectomy had no effect on the distribution of somatostatin-like or galanin-like immunoreactive axons, and these axons did not contain the sympathetic marker tyrosine hydroxylase. They did not contain the parasympathetic marker choline acetyltransferase, either. The galanin-like immunoreactive axons contained other neuropeptides found in sensory nerves, including calcitonin gene-related peptide, substance P and cholecystokinin. Somatostatin-like immunoreactive axons did not contain any of these sensory neuropeptides or galanin-like immunoreactivity, and they were neither labelled with an antibody to 200kDa neurofilament protein, nor did they bind isolectin-IB4. Nevertheless, they are likely to be of sensory origin because somatostatin-like immunoreactive perikarya have previously been localized in the trigeminal ganglion of primates. Taken together, these findings indicate galanin and somatostatin are present in two different subsets of sensory axons in primate uvea.

Increased density of galanin binding sites in the dorsal raphe in a genetic rat model of depression

The Flinders Sensitive Line rats showed an increased immobility response by 104% (P<0.01) in the forced swim test with respect to the Flinders Resistant Line rats (control). The Flinders Sensitive Line rats also had an increase of [125I]galanin (porcine) binding in the dorsal raphe (55±3.3 fmol/mg protein) with respect to the Flinders Resistant Line rats (38±3.6 fmol/mg protein) (42%, P<0.05) without changes in galanin receptor affinity and with a significant reduction (32.3%, P<0.05) of galanin-like immunoreactivity in the dorsal raphe. The results indicate that enhancement of galanin receptor function in the dorsal raphe rich in 5-HT neurons could be a mechanism involved in the production of depressive-like activity in this animal model of depression.