Plasma Galanin Research Articles

Galanin in human plasma

The neuropeptide galanin has important effects on hormone secretion from the hypothalamus and pituitary, and it may also be involved in central 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 26 healthy subjects, 14 women and 12 men, using two extraction methods and a specific radioimmunoassay for human galanin. Galanin-LI levels in unextracted plasma were higher (141-191 pmol/L) than after immunoextraction (3.4-30.7 pmol/L) and Sep Pak extraction (2.2-12.6 pmol/L). Galanin immunoreactivity after Sep Pak and immunoextraction correlated (r = 0.74, p<0.001). Galanin-LI levels were significantly higher in the men than in the women (p = 0.01) after Sep Pak extraction. A small increase in galanin-LI was seen with age in the women (r = 0.54, p < 0.05). The proportion of Sep Pak extracted galanin-LI increased with age in the women (r = 0.73, p < 0.05)) but not in the men.

Positive Correlation of Galanin With Glucose in Type 2 Diabetes

Compelling literature (1) has been accumulated regarding the action of galanin as an incretin on insulin secretion. Although galanin inhibits glucose-stimulated insulin release in animals, no such effect has been documented in humans (2). Actually, galanin administration in humans has been shown to suppress the initial postprandial rise in plasma concentration of glucose and insulin (3) with unaltered glucose-stimulated insulin release (2). Nevertheless, basal plasma galanin levels have been shown to diverge with obesity and hormonal status. We enrolled 21 patients with type 2 diabetes (HbA1c 7.8 ± 1.19%): 9 men (aged 54.6 ± 3.87 years, BMI 28.5 ± 1.6 …

Expression of galanin and galanin receptor-1 in normal bone and during fracture repair in the rat

The neuropeptide galanin (GAL) has recognized physiological actions in the nervous system and other tissues, but there is no documented evidence of GAL influencing normal or pathological bone metabolism. GAL expression, however, is upregulated in central and peripheral nerves following axotomy and is known to influence neural regeneration. Thus, severance of skeletal-associated nerves during fracture could similarly increase local GAL concentrations and thereby influence fracture healing. The initial aim of this study was therefore to identify the presence of GAL in normal bone and/or fracture callus by assessing the concentration and cellular localization of GAL in intact and/or fractured rat rib, using radioimmunoassay and immunohistochemistry, respectively. Groups of Sprague–Dawley rats (13 weeks old) had their left sixth ribs surgically fractured or underwent sham surgery and then calluses and nonfractured rib samples were analyzed at 1 and 2 weeks postsurgery ( n = 5–6 per group). Low (basal) concentrations of GAL were detected in control ribs, whereas at 1 and 2 weeks postfracture, callus samples contained markedly increased levels of peptide (∼32- and 18-fold increase, respectively, relative to controls; P < 0.01), revealing a strong upregulation during bone healing. Plasma GAL concentrations were also increased at 2 weeks postfracture ( P < 0.005). In normal (nonfractured) rib, minimal levels of GAL-like immunoreactivity (LI) were present in cortical bone, periosteum, endosteum, and surrounding skeletal muscle. In costal cartilage plates, intense GAL-LI was present in all chondrocytes of the hypertrophic zone and in a population of chondrocytes in the reserve zone. GAL-LI was not present, however, in chondrocytes in the proliferative zone of costal cartilage or skeletal muscle fibers. In fracture callus, levels of GAL-LI were moderate to intense in osteoprogenitor cells and osteoblasts, in some chondrocytes, and in cartilaginous, osseous, and periosteal matrices. Subsequent studies revealed the presence of galanin receptor-1-like immunoreactivity (GALR1-LI) in most cell types shown to contain GAL-LI, although the distribution of GALR1-LI was more extensive in reserve zone chondrocytes than that of GAL-LI; and GALR1-LI also appeared in late proliferative zone chondrocytes of costal cartilage. In summary, GAL concentrations were significantly increased in fracture callus and plasma of rats that underwent rib fracture. In addition, GAL- and GALR1-LI was also detected in specific cells and structures within costal cartilage, bone, and fracture callus. These results strongly implicate GAL in aspects of cartilage growth plate physiology and fracture repair, possibly acting in an autocrine/paracrine fashion via GALR1.

Plasma galanin response to head-up tilt in normal subjects and patients with recurrent vasovagal syncope

Neurohumoral factors may contribute to cardiovascular changes associated with vasovagal syncope (VVS). Galanin (GAL) is a neuropeptide, widely distributed in the central and peripheral nervous systems, that interacts with both sympathetic and vagal systems as well as with neurotransmitters, such as serotonin. We investigated the changes in plasma GAL and catecholamine levels during head-up tilt (HUT) test in patients with recurrent VVS. Twenty-two patients (11 women, aged 33.1 ± 4.2 years) with a history of VVS and 10 healthy subjects (5 women, aged 38.0 ± 5.8 years) underwent HUT test (60°, 45 minutes). GAL and catecholamine plasma levels were measured in the supine position, during HUT and, in patients with positive response, at presyncope, syncope, and after recovery of consciousness. Thirteen patients developed syncope during HUT, whereas no healthy subjects had a positive response. In healthy subjects, GAL did not change during HUT. By contrast, in patients with a history of VVS and a negative response to tilting (no syncope), GAL significantly (P <.001) increased in response to tilting (supine, 10.2 ± 0.6 pmol/L; tilting, 18.1 ± 1.1 pmol/L at 45 minutes) and correlated positively with the increases in blood pressure (BP) and heart rate (HR). In patients with a positive response, GAL did not change either before the loss of consciousness or during syncope. In patients with a positive response, norepinephrine (NE) significantly (P <.001) increased during tilting and then remained practically unchanged during syncope, whereas epinephrine (E) significantly (P <.001) increased during tilting and then showed further significant increases at presyncope and syncope. In conclusion, this study shows that circulating GAL levels progressively increase in correlation with the cardiovascular parameters during a negative HUT in patients with a history of VVS, whereas they remain unchanged in healthy subjects. Moreover, in the patients with tilting-induced syncope GAL does not change either before or during loss of consciousness. These data suggest a role for endogenous GAL in the adaptive responses to acute orthostatic stress preventing syncope in susceptible individuals. Copyright 2003, Elsevier Science (USA). All rights reserved.

Evaluation of the circadian profile of peripheral plasma galanin concentrations in normal subjects

Galanin administration can influence pituitary function principally resulting in an increase in GH secretion. However, the role of circulating GAL levels in human endocrine function is still unknown. In the present study we simultaneously measured the circadian profiles of GAL, ACTH and GH in peripheral blood of ten adult subjects. Plasma samples were collected through an intravenous catheter at 0800, 1200, 1600, 2000, 2200, 2400, 0200, 0400 hours. The results were statistically evaluated by the cosinor analysis technique. A significant circadian rhythm of both plasma ACTH (p < 0.001) and GH levels (p < 0.03) was found with acrophases occurring at 0753 hrs and 0131 hrs for ACTH and GH, respectively. On the contrary, no significant rhythm was found in plasma GAL levels, indicating that no correlations exist between GAL and either GH or ACTH circadian profiles. Furthermore, the simultaneous assay of both GAL and GH plasma levels during a nocturnal frequent sampling performed in four volunteers showed the presence of peaks in GAL levels which, however, were not concomitant to the peaks in GH levels. These data demonstrate the lack of rhythmicity in the circadian profile of plasma GAL levels in healthy human subjects. The role of GAL in human endocrine function remains unknown and these results suggest that, in spite of the well documented increase in plasma GH concentrations following the intravenous administration of GAL, physiologically circulating levels of GAL are likely not involved in the regulation of GH secretion.

Galanin is released by adrenocorticotropin-secreting pituitary adenomas in vivo and in vitro.

Galanin, a brain-gut peptide, is also synthesized and released by the pituitary. In man, galanin-like immunoreactivity and galanin messenger RNA have been detected specifically in normal and tumoral corticotropes, but little is known about the production and release of galanin by the human pituitary. We evaluated galanin release by 5 ACTH-secreting pituitary adenomas in culture and plasma galanin concentrations in the inferior petrosal sinuses (IPSs) of 15 patients with Cushing's disease before and after CRH administration. For comparison, the galanin response to CRH was evaluated in 8 normal controls. Galanin secretion by pituitary tumor cultures ranged from 30-230 pmol/4 h. Incubation with CRH induced an increase in galanin concentrations (100 pM CRH: 151 +/- 32%; 1 nM CRH: 232 +/- 43%; 10 nM CRH: 246 +/- 35%; and 100 nM CRH: 270 +/- 44% unstimulated levels at 24 h, P < 0.05). The stimulatory effect of CRH seemed to be dose-dependent. Basal and CRH-stimulated ACTH and galanin concentrations also exhibited a strong positive correlation in single tumor cultures. At IPS sampling, mean basal plasma galanin concentrations in the dominant IPS were somewhat higher than those registered at the periphery (18.6 +/- 1.94 vs. 15.8 +/- 1.60 pmol/L, P = 0.05). Administration of CRH induced a modest but significant increase in galanin concentrations at all three sampling sites. No correlations were found between ACTH and galanin levels in the IPSs and at the periphery. Different from what was observed in patients with Cushing's disease, CRH did not modify plasma galanin concentrations in normal subjects. In conclusion, this study demonstrates that galanin is released by human tumoral corticotropes and responds to CRH. The role of locally produced galanin is, as yet, unknown but may possibly be that of a autocrine/paracrine modulator.

Neuropeptide Y, leptin, galanin and insulin in women with polycystic ovary syndrome

It has been reported that polycystic ovary syndrome (PCOS) is very frequently associated with obesity, insulin resistance and hyperinsulinemia. However, metabolic disorders may lead to suppression of reproductive hormone secretion during undernutrition and in obesity. Some neuropeptides, such as neuropeptide Y (NPY) and galanin, modulate the control of appetite and play an important role in the mechanism of luteinizing hormone-releasing hormone (LHRH) secretion. NPY and galanin regulate appetite via both central and peripheral mechanisms. The interaction between central and peripheral signals for the control of food intake is due to leptin. Leptin can modulate the activity of NPY and other peptides in the hypothalamus that are known to affect eating behavior.In order to evaluate the relationship between NPY, galanin and leptin, 28 women with PCOS, 32 obese women (non-PCOS) and 19 lean healthy women (control group) were investigated. Obese women with PCOS were divided into two groups: PCOS (A) overweight (body mass index, BMI 26-30 kg/m2), and PCOS (B) obese (BMI 31-40 kg/m2). Plasma NPY, galanin and leptin concentrations were measured by radioimmunoassay.Plasma leptin levels in obese women with PCOS (groups A and B) were significantly higher than those in the control group (p < 0.05, p < 0.05, respectively). A significant positive correlation between plasma leptin and BMI in women with PCOS was found (r = 0.427, p < 0.01). A positive correlation was demonstrated between leptin and testosterone in PCOS (r = 0.461, p < 0.01). Plasma galanin concentrations in PCOS were higher than in the control group but the differences were not significant. Plasma NPY levels were significantly elevated in both non-obese (normal) and obese women with PCOS (group A) (p < 0.01, p < 0.005, respectively). However, in obese non-PCOS women plasma NPY levels gradually increased with increase in BMI. No significant correlations were found between galanin, NPY and percentage change in response of LH to LHRH, as well as between NPY and insulin, and galanin and testosterone. Plasma insulin concentrations in women with PCOS (group B) were significantly higher than in the control group (p < 0.001).Increased plasma NPY levels are found in both obese and non-obese women with PCOS. The increase in NPY is independent of the increase in BMI. In obese women with PCOS, plasma leptin is increased compared with control lean women. Serum insulin concentration is increased in obese women with PCOS. A positive correlation exists between leptin and BMI as well as between leptin and testosterone in women with PCOS.

Neuropeptide Y, galanin, and leptin release in obese women and in women with anorexia nervosa

The study objective was to determine circulating levels of the appetite-controlling neuropeptides, neuropeptide Y (NPY), galanin, and leptin, in subjects with eating disorders. The study group consisted of 48 obese women aged 19 to 45 years, 15 women with anorexia nervosa aged 18 to 23 years, and 19 lean healthy women aged 18 to 42 years (control group). The obese women were divided into four groups: (A) body mass index (BMI) = 25 to 30 kg/m2, n = 9 (overweight); (B) BMI = 31 to 40 kg/m2, n = 23 (moderate obesity); (C) BMI greater than 40 kg/m2, n = 9 (severe obesity); and (D) BMI = 31 to 40 kg/m2, n = 7 (moderate obesity + non—insulin-dependent diabetes mellitus [NIDDM]). Plasma NPY, galanin, and leptin concentrations were measured in peripheral blood samples with radioimmunoassay methods. Plasma NPY levels in obese women (groups A, B, C, and D) were significantly higher as compared with the control group (P < .01, P < .001, P < .001, and P < .001, respectively). The highest plasma NPY concentrations were observed in obese women with NIDDM. Plasma galanin levels were significantly higher in groups B, C, and D (P < .001, P < .001, and P < .001, respectively). Plasma leptin concentrations were significantly higher in groups C and D as compared with the control group (P < .001 and P < .001, respectively). Plasma NPY and galanin concentrations in women with anorexia nervosa did not differ from the levels in the control group. However, plasma leptin concentrations were significantly lower in anoractic women than in the control group (P < .01). Our results indicate that inappropriate plasma concentrations of NPY, galanin, and leptin in obese women may be a consequence of their weight status, or could be one of many factors involved in the pathogenesis of obesity.

Plasma Galanin Immunoreactivity in the Rat After Swimming

The purpose of our experiment was to study plasma immunoreactive galanin in the rat after swimming. Four groups of rats were used. At rest, one group was studied after an intravenous injection of D-glucose while another group received a corresponding saline injection. The two remaining groups, treated respectively with glucose and saline, were investigated after a 30-min swimming session. After 30 min of rest in their respective cages, or after swimming, the animals were anesthetized for immediate blood sampling. The main observation was that plasma galanin was higher after swimming than after rest only in glucose-treated rats, in the vena cava (11.82 ± 2.90 vs. 5.05 ± 1.65 pM) and the portal vein (15.75 ± 3.74 vs. 6.58 ± 1.75 pM). Both saline- and glucose-treated groups had a significant increase of plasma norepinephrine from rest to swimming in the vena cava. A decrease of plasma insulin was observed in the portal vein of exercised glucose-treated rats, while plasma glucose was higher in the portal vein of this group. In conclusion, after swimming, plasma galanin level was increased only in glucose-treated rats. A significant correlation was also observed between plasma galanin and glucose ( r = 0.69, p < 0.01), suggesting that the presence of glucose is necessary to induce galanin liberation during exercise.

Evaluation of circulating galanin levels after exercise-induced pituitary hormone secretion in man

The neuropeptide galanin (GAL) is widely distributed in the central and peripheral nervous systems, anterior pituitary, and adrenal medulla. GAL is colocalized with corticotropin (ACTH) in the human pituitary and with epinephrine (E) and norepinephrine (NE) in chromaffin cells of the adrenal medulla. The function of GAL in peripheral tissues is not known, although the presence of the peptide in corticotrophs and the adrenal gland suggest that it participates in stress responses. In the present study, we investigated whether GAL is cosecreted with ACTH during activation of corticotrophs by an acute physical exercise test. Circulating levels of GAL and pituitary hormones were measured in healthy exercise-tested and control male subjects. Blood samples were collected during basal conditions, maximal power output (MPO), and the recovery period. Control subjects were sampled during the resting condition. The pituitary response to exercise was characterized by a significant increase in ACTH plasma levels (peak value 13.28 ± 2.19 v 6.68 ± 1.01 pmol/L, P < .05) and growth hormone (GH) serum levels (peak value, 14.53 ± 5.59 v 0.29 ± 0.1 μg/L, P < .02), with the peak in hormone levels detected 15 minutes after the end of exercise. No change in circulating prolactin (PRL) levels was detected. An expected significant increase in plasma levels of both E (peak value, 1,574.41 ± 403.31 v 267.44 ± 66.03 pmol/L, P < .01) and NE (peak value, 7,275.25 ± 955.80 v 961.51 ± 168.40 pmol/L, P < .01) was also observed. Plasma GAL levels were not affected by the acute exercise test, with the levels being comparable to baseline during the exercise test and the recovery phase. At any sample time, GAL values were comparable between exercise-tested and control subjects. These data show that despite the colocalization of GAL and ACTH within the same pituitary cells, the two peptides are not coreleased in response to stress resulting from acute physical exercise. Furthermore, pituitary GAL seems not to be involved in the, stimulation of GH secretion in exercise-tested subjects. The results also indicate that GAL is not coreleased with E or NE in response to the exercise-induced stress condition.

Regulation of galanin gene expression in the rat anterior pituitary gland by the somatostatin analog SMS 201-995.

In addition to inducing pituitary tumors in rats, estrogen (E2) markedly increases galanin and PRL gene expression. We previously showed that galanin secretion from pituitary cells in vitro is inhibited by dopamine and somatostatin and stimulated by TRH. The objectives of these in vivo studies were to assess whether the long-acting somatostatin analog SMS 201-995 alters 1) immunoreactive galanin or PRL levels in the anterior pituitary, neurointermediate lobe, hypothalamus, or plasma, 2) pituitary galanin and PRL mRNA levels, and 3) the development of E2-induced pituitary tumors. Ovariectomized Fischer 344 rats were implanted with E2-filled or empty Silastic capsules and treated with or without SMS 201-995 (1.5 mg) via Alzet miniosmotic pumps. Two or 6 weeks later, immunoreactive galanin and PRL levels were determined by RIA. In ovariectomized rats, the somatostatin analog lowered the anterior pituitary content of galanin by 50%, but had no effect on PRL concentrations. E2 increased galanin and PRL levels in the anterior pituitary by 220- and 4-fold, respectively. Concomitant E2 and SMS 201-995 treatment further increased galanin and PRL in the anterior pituitary by 60-80%, but decreased plasma galanin and PRL levels. Likewise, the administration of SMS 201-995 for 2 and 6 weeks inhibited the E2-induced growth of the anterior pituitary. Galanin and PRL mRNA levels were quantified by solution hybridization. Galanin mRNA levels were reduced to undetectable levels in ovariectomized rats treated with SMS 201-995. Furthermore, a 10-fold increase in galanin mRNA levels seen in the presence of E2 was inhibited 80% by SMS 201-995. PRL mRNA levels in E2-treated rats were unchanged by SMS 201-995. We conclude that SMS 201-995 1) lowers plasma galanin and PRL levels in E2-treated rats, 2) elevates the anterior pituitary contents of galanin and PRL in E2-exposed rats, probably through decreased secretion of the hormones, and 3) reduces galanin mRNA levels in E2-treated and untreated ovariectomized rats. Overall, these results establish the differential regulation of galanin and PRL gene expression in vivo by SMS 201-995. Moreover, the data demonstrate that somatostatin receptor agonists may have therapeutic potential for some prolactinomas.

Dopaminergic Regulation of Galanin Gene Expression in the Rat Anterior Pituitary Gland

Estrogen dramatically increases galanin mRNA and peptide levels in the rat anterior pituitary gland. We recently reported that galanin secretion in vitro from estrogen-exposed anterior pituitary cells is regulated by hypothalamic factors; dopamine and somatostatin inhibit galanin secretion, and thyrotrophin-releasing hormone stimulates galanin release. To determine whether galanin is regulated by a dopaminergic mechanism in vivo, we used ovariectomized Fischer 344 rats treated with 17ß-estradiol-containing or empty Silastic capsules. Rats were also administered bromocriptine, a dopamine receptor agonist, haloperidol, a dopamine receptor antagonist, or placebo for 2 weeks. Galanin peptide levels were measured in the anterior pituitary, neurointermediate lobe, medial basal hypothalamus, and plasma by radioimmunoassay. Plasma and pituitary prolactin levels were also determined. Bromocriptine decreased gaianin peptide levels in the anterior pituitary gland of ovariectomized rats by 30%, but had no effect on galanin in the neurointermediate lobe or medial basal hypothalamus. In contrast, haloperidol had no effect on galanin in the anterior pituitary or medial basal hypothalamus of ovariectomized rats, but decreased galanin peptide levels in the neurointermediate lobe. In the anterior pituitary gland of estrogen-treated rats, bromocriptine increased and haloperidol decreased both galanin and prolactin levels. Galanin mRNA levels were quantified in the anterior pituitary gland by solution hybridization. Bromocriptine increased galanin mRNA levels 3-fold in the anterior pituitary, whereas haloperidol had no effect. Galanin mRNA levels in the anterior pituitary were elevated 10-fold by estrogen. Bromocriptine reduced galanin mRNA levels in the pituitary by 50% in estrogen-treated rats, where again haloperidol had no effect. Estrogen increased plasma galanin levels 4-fold compared to ovariectomized rats and this effect was reduced 60% by bromocriptine and increased 20% by haloperidol. We conclude 1) galanin synthesis and release from the estrogen-exposed anterior pituitary gland is inhibited by a dopaminergic mechanism in vivo, 2) dopamine regulates galanin gene expression in the ovariectomized rat, 3) the changes in galanin peptide levels in the anterior pituitary of rats treated with estrogen and dopamine receptor ligands are primarily due to alterations in peptide secretion, and 4) galanin release from the neurointermediate lobe may also be regulated by a dopaminergic mechanism in vivo. These data, in conjunction with previous studies, provide evidence for the co-regulation of galanin and prolactin in estrogen-treated rats, and further discriminate between the regulation of galanin in the hypothalamus and pituitary gland.

Plasma neuropeptide Y (NPY) and galanin before and during exercise in type 1 diabetic patients with autonomic dysfunction

Plasma neuropeptide Y (NPY), plasma galanin and plasma catecholamines were determined before and during an ergometer exercise test in 11 type 1 diabetic patients (age 19–36 years, mean 30; duration of diabetes 2–18 years, mean 9) with autonomic dysfunction and in 13 age-matched healthy controls (age 24–36 years, mean 29). Before exercise, plasma NPY (100 ± 6 pmol/l vs 144 ± 7 pmol/l; P < 0.001) and plasma galanin (54 ± 3 pmol/l vs 77 ± 5 pmol/l; P < 0.005) were significantly lower in patients than in controls. During exercise, plasma NPY, plasma adrenaline, and plasma noradrenaline increased in patients and controls while galanin only increased in patients. Since there was a direct correlation between plasma NPY before exercise and the increment (Δ80%) in noradrenaline during exercise ( r = 0.54; P < 0.01), it is suggested that plasma NPY determined in the basal situation may be a useful marker of sympathetic nerve failure in diabetic patients.

Circulating galanin: origin, metabolism, and pharmacokinetics in anesthetized pigs.

The concentration of galanin-like immunoreactivity (GAL-LI) was 12.9 +/- 0.9 pmol/l in porcine arterial plasma (n = 9) and ranged from 1 to 14 pmol/g in extracts of porcine gastrointestinal tract (n = 5), the colon being the richest gut segment. A significant (P less than 0.05) arteriovenous concentration difference of circulating endogenous GAL-LI occurred across the kidney (15.1 +/- 2.3 vs. 6.2 +/- 0.5 pmol/l) and a hind leg (15.7 +/- 2.5 vs. 10.2 +/- 1.0 pmol/l), whereas a negative gradient was observed across the intestine (12.5 +/- 2.0 vs. 17.7 +/- 3.3 pmol/l) of anesthetized pigs. Passage through the brain, liver, or lungs did not change the concentration of endogenous GAL-LI significantly. During basal circumstances, the major source of circulating GAL-LI is therefore the gut. During infusion of 20 pmol.kg-1.min-1 of synthetic porcine galanin, a significant extraction occurred across the kidney (64.8 +/- 4.3%), hind leg (20.3 +/- 3.8%), and liver (19.7 +/- 4.3%). The overall metabolic clearance rate was 37.8 +/- 3.7 ml.min-1.kg-1. The half-life of galanin in plasma was 4.6 +/- 0.3 min, and the apparent distribution space was 255.6 +/- 31.4 ml/kg. Incubation studies in vitro showed that the concentration of galanin, added to blood and plasma at 37 degrees C, was halved in 1 h, unless stabilized with EDTA and aprotinin.

Localization and Molecular Forms of Galanin in Human Adrenals: Elevated Levels in Pheochromocytomas

Galanin immunoreactivity was measured by RIA, using antibodies directed against both the non-C- and C-terminal positions of porcine galanin, in tissue extracts of normal adrenals and pheochromocytomas and also in the plasma of normal subjects and patients with pheochromocytomas. No C-terminal galanin-like immunoreactivity was detected in plasma or tissue, suggesting differences in the amino acid sequence of human compared with porcine galanin. A non-C-terminally directed antibody was, therefore, used to characterize human galanin immunoreactivity by gel permeation chromatography and reverse phase high pressure liquid chromatography and to localize it by immunocytochemistry. The galanin content of whole adrenal gland was 2.6 +/- 0.9 (+/- SEM) pmol/g (n = 5). In contrast, however, pheochromocytomas had much greater concentrations (21 +/- 2.3 pmol/g; n = 16). Gel chromatography and reverse phase high pressure liquid chromatography revealed 2 molecular forms of galanin immunoreactivity with identical elution positions in both normal adrenals and tumors. The concentration of galanin in plasma from both normal subjects and pheochromocytoma patients was below the detection limit of the assay (less than 10 pmol/liter). Using immunocytochemistry, galanin was localized to scattered cells or clusters of tumor cells in 5 of 11 pheochromocytomas and only a few chromaffin cells and cortical nerve fibers in normal adrenals.