Bertalan Dudás | Semmelweis University (original) (raw)

Papers by Bertalan Dudás

Injury-international Journal of The Care of The Injured, 2008

The sacrospinous (SS) and sacrotuberous (ST) ligaments of the pelvic ring are known as mechanical... more The sacrospinous (SS) and sacrotuberous (ST) ligaments of the pelvic ring are known as mechanical stabilisers of the pelvic girdle, primarily against rotational forces in the sagittal and horizontal planes. Earlier studies, however, raised the possibility that ST/SS ligaments possess significant proprioceptive function, while the mechanical role of these ligaments in maintaining the structural integrity of the pelvis is of less importance.The aim of this study is to determine whether the function of these ligaments is strictly to provide mechanical stability or if they have any additional functional properties, i.e., proprioception. In order to reveal the function of the SS/ST ligaments, biomechanical studies of cadaver pelvis were used along with the histological analysis of the ligaments. Following measurements to determine the accurate mechanical role of the pelvic ligaments, the strength of these ligaments was significantly less than we earlier expected. For this reason other functions of the SS/ST ligaments were considered, including the proprioceptive role. Indeed, histological studies revealed ramifying nerve terminals in the SS/ST ligaments. These terminals may represent the morphological substrate of the proprioceptive function associated with the ligaments.Our studies revealed that SS/ST ligaments might have a significant proprioceptive function providing information of the position of the pelvis. Consequently, the mechanical role of the ligaments in maintaining the structural integrity of the pelvis may be significantly less than previously assumed. Understanding the function of the SS/ST ligaments is crucial for providing more precise guidelines for patient management with injuries to the posterior pelvic region.

The present study determined whether the serotonin 2A (5-HT 2A ) receptors in the hypothalamic pa... more The present study determined whether the serotonin 2A (5-HT 2A ) receptors in the hypothalamic paraventricular nucleus mediate the neuroendocrine responses to a peripheral injection of the 5-HT 2A/2C receptor agonist (Ϫ)DOI [(Ϫ)1-(2,5-dimethoxy-4iodophenyl)-2-aminopropane]. The 5-HT 2A receptor antagonist MDL100,907 ((Ϯ)-␣-(2,3-dimethoxyphenyl)-1-[2-(4-fluorophenylethyl)]-4-piperidinemethanol), the 5-HT 2C receptor antagonist SB-242084 (6-chloro-5-methyl-1-[[2-[(2-methyl-3-pyridyl)oxy]-5pyridyl]carbamoyl]-indoline), or vehicle were microinjected bilaterally through a chronically implanted double-barreled cannula into the hypothalamic paraventricular nucleus 15 min before a peripheral injection of (Ϫ)DOI in conscious rats. (Ϫ)DOI significantly elevated plasma levels of oxytocin, prolactin, ACTH, corticosterone, and renin. Neither the 5-HT 2A receptor antagonist nor the 5-HT 2C receptor antagonist, injected alone, altered the basal levels of these hormones. MDL100,907 (0.748, 7.48, and 18.7 nmol) dose dependently inhibited the (Ϫ)DOI-induced increase in all of the hormones except corticosterone. In contrast, SB-242084 (10 nmol) did not inhibit (Ϫ)DOI-increased hormone levels. To confirm the presence of 5-HT 2A receptors in the hypothalamic paraventricular nucleus, 5-HT 2A receptors were mapped using immunohistochemistry. Densely labeled magnocellular neurons were observed throughout the anterior and posterior magnocellular subdivisions of the hypothalamic paraventricular nucleus. Moderately to densely labeled cells were also observed in parvicellular regions. Thus, it is likely that 5-HT 2A receptors are present on neuroendocrine cells in the hypothalamic paraventricular nucleus. These data provide the first direct evidence that neuroendocrine responses to a peripheral injection of (Ϫ)DOI are predominantly mediated by activation of 5-HT 2A receptors in the hypothalamic paraventricular nucleus.

Growth Hormone & Igf Research, 2010

Previous studies revealed that growth hormone-releasing hormone (GHRH)-immunoreactive (IR) neuron... more Previous studies revealed that growth hormone-releasing hormone (GHRH)-immunoreactive (IR) neurons form a circumscribed cell group in the basal infundibulum/median eminence of the human hypothalamus. GHRH from these neurons is released into the hypothalamo-hypophyseal portal circulatory system in a pulsatile manner. It is a common consensus that the pulsatile release of GHRH is the main driving force behind the pulsatile release of growth hormone (GH) and may contribute to the regulation of other hypothalamic functions. The pulsatile release of GHRH requires synchronized activity of GHRH-IR neurons. However, the morphological basis of this synchronization between the GHRH-IR neural elements has not been elucidated yet. Since the utilization of electron microscopy combined with immunohistochemistry is virtually impossible in the human brain due to the long post mortem period, immunohistochemistry, evaluated with oil immersion light microscopy, was used in order to reveal the associations between the GHRH elements. Numerous GHRH-GHRH juxtapositions have been detected in the infundibular area/median eminence, where GHRH-IR axonal varicosities often formed multiple contacts with GHRH-IR perikarya. Examination of these associations with high magnification oil immersion light microscopy revealed (1) axonal swellings at the site of the contacts and (2) no gaps between the contacting elements suggesting that these juxtapositions may be functional synapses. The large number of GHRH-GHRH juxtapositions in the infundibular area/ median eminence suggests that these synapse-like structures may represent the morphological substrate of the synchronized activity of GHRH neurons that in turn may result in the pulsatile release of GHRH in human.

Neuroscience, 2004

However, the morphological substrate of any similar modulation is not known in human. In the pres... more However, the morphological substrate of any similar modulation is not known in human. In the present series of experiments we first mapped the galanin-IR and LHRH-IR neural elements in human brain, utilizing single label immunohistochemistry. Then, following the superimposition of the maps of these systems, the overlapping sites were identified with double labeling immunocytochemistry and examined in order to verify the putative juxtapositions between galanin-IR and LHRH-IR structures. LHRH and galanin immunoreactivity were detected mainly in the medial basal hypothalamus, in the medial preoptic area and along the diagonal band of Broca. Careful examination of the IR elements in the overlapping areas revealed close, bi-directional contacts between galanin-IR and LHRH-IR structures, which have been verified in semithin plastic sections. These galanin-LHRH and LHRHgalanin juxtapositions were most numerous in the medial preoptic area and in the infundibulum/median eminence of the human diencephalon.

Neuroscience, 2010

Previous studies have demonstrated that catecholaminergic, tyrosine hydroxylase (TH)-immunoreacti... more Previous studies have demonstrated that catecholaminergic, tyrosine hydroxylase (TH)-immunoreactive (IR) perikarya and fibers are widely distributed in the human hypothalamus. Since TH is the key and rate-limiting enzyme for catecholaminergic synthesis, these IR neurons may represent dopaminergic, noradrenergic or adrenergic neural elements. However, the distribution and morphology of these neurotransmitter systems in the human hypothalamus is not entirely known. Since the different catecholaminergic systems can be detected by identifying the neurons containing the specific key enzymes of catecholaminergic synthesis, in the present study we mapped the catecholaminergic elements in the human hypothalamus using immunohistochemistry against the catecholaminergic enzymes, TH, dopamine beta-hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT). Only a few, PNMT-IR, adrenergic neuronal elements were found mainly in the infundibulum and the periventricular zone. DBH-IR structures were more widely distributed in the human hypothalamus occupying chiefly the infundibulum/infundibular nucleus, periventricular area, supraoptic and paraventricular nuclei. Dopaminergic elements were detected by utilizing double label immunohistochemistry. First, the DBH-IR elements were visualized; then the TH-IR structures, that lack DBH, were detected with a different chromogen. In our study, we conclude that all of the catecholaminergic perikarya and the majority of the catecholaminergic fibers represent dopaminergic neurons in the human hypothalamus. Due to the extremely small number of PNMT-IR, adrenergic structures in the human hypothalamus, the DBH-IR fibers represent almost exclusively noradrenergic neuronal processes. These findings suggest that the juxtapositions between the TH-IR and numerous peptidergic systems revealed by previous reports indicate mostly dopaminergic synapses.

Journal of Neuroendocrinology, 2006

It has been postulated that the stress response is associated with water balance via regulating v... more It has been postulated that the stress response is associated with water balance via regulating vasopressin release. Nausea, surgical stress and insulin-induced hypoglycaemia were shown to stimulate vasopressin secretion in humans. Increased vasopressin release in turn induces water resorption through the kidneys. Although the mechanism of the stress-mediated vasopressin release is not entirely understood, it is generally accepted that catecholamines play a crucial role in influencing water balance by modulating the secretion of vasopressin. However, the morphological substrate of this modulation has not yet been established. The present study utilised double-label immunohistochemistry to reveal putative juxtapositions between tyrosine hydroxylase (TH)-immunoreactive (IR) catecholaminergic system and the vasopressin systems in the human hypothalamus. In the paraventricular and supraoptic nuclei, numerous vasopressin-IR neurones received TH-IR axon varicosities. Analysis of these juxtapositions with high magnification combined with oil immersion did not reveal any gaps between the contacted elements. In conclusion, the intimate associations between the TH-IR and vasopressin-IR elements may be functional synapses and may represent the morphological basis of vasopressin release modulated by stressors. Because certain vasopressin-IR perikarya receive no detectable TH innervations, it is possible that additional mechanisms may participate in the stress-influenced vasopressin release.

Neuroscience, 2008

Previous studies revealed that stress is a pivotal factor in the regulation of growth. Psychologi... more Previous studies revealed that stress is a pivotal factor in the regulation of growth. Psychological harassment may result in psychosocial dwarfism with delayed puberty, short stature and depression. Growth hormone (GH) secretion is suppressed by stress, possibly via the attenuation of growth hormone-releasing hormone (GHRH) secretion. However, the morphological substrate of this phenomenon has not been elucidated yet.

Journal of Chemical Neuroanatomy, 2009

Journal of Neuroendocrinology, 2006

The gonadotrophin-releasing hormone (GnRH) represents the final common pathway of a neuronal netw... more The gonadotrophin-releasing hormone (GnRH) represents the final common pathway of a neuronal network that integrates multiple external and internal factors to control fertility. Among the many inputs GnRH neurones receive, oestrogens play the most important role. In females, oestrogen, in addition to the negative feedback, also exhibits a positive feedback influence upon the activity and output of GnRH neurones to generate the preovulatory luteinising hormone surge and ovulation. Until recently, the belief has been that the GnRH neurones do not contain oestrogen receptors and that the action of oestrogen upon GnRH neurones is indirect, involving several, oestrogen-sensitive neurotransmitter and neuromodulator systems that trans-synaptically regulate the activity of the GnRH neurones. Although this concept still holds for humans, recent studies indicate that oestrogen receptor-beta is expressed in GnRH neurones of the rat. This review provides three dimensional stereoscopic images of GnRH-immunoreactive (IR) and some peptidergic (neuropeptide Y-, substance P-, β-endorphin-, leu-enkaphalin-, corticotrophin hormone-releasing- and galanin-IR) and catecholaminergic neurones and the communication of these potential oestrogen-sensitive neuronal systems with GnRH neurones in the human hypothalamus. Because the post-mortem human tissue does not allow the electron microscopic identification of synapses on GnRH neurones, the data presented here are based on light microscopic immunocytochemical experiments using high magnification with oil immersion, semithin sections or confocal microscopy.

Gonadal functions are modulated by corticotropin-releasing factor (CRF) in the rat via direct sup... more Gonadal functions are modulated by corticotropin-releasing factor (CRF) in the rat via direct suppression of LH-releasing hormone (LHRH) release. Although there is evidence of direct morphological contacts between the LHRH and CRF-immunoreactive (-IR) structures in the rat hypothalamus, little is known about the morphological base of CRF-influenced LHRH release in man. Thus, we studied the distribution of the CRF-IR and LHRH-IR systems in the human diencephalon and revealed putative CRF-LHRH juxtapositions using double label immunohistochemistry. LHRH-IR cells were present mainly in the infundibular region and the medial preoptic area. CRF-IR neuronal structures were observed in the periventricular area, paraventricular nucleus, infundibular region, and median eminence. CRF-LHRH juxtapositions were found mainly in the infundibulum and median eminence. Few juxtapositions were detected in the medial preoptic area. In these regions, black diaminobenzidine/silver-labeled CRF-IR fibers abutted fusiform brown diaminobenzidine-labeled LHRH neurons, usually forming multiple contacts. Examination of semithin sections of these close associations with the aid of oil immersion revealed no cleft between CRF-IR nerve terminals contacting LHRH-IR structures. These findings suggest that the juxtapositions between the LHRH-IR and CRF-IR neurons may be functional synapses forming the morphological substrate of the CRF-controlled LHRH secretion. Moreover, the wide distribution of CRF-IR elements suggests that CRF controls other diencephalic functions as well.

Neuropeptides, 2011

Neuropeptide Y (NPY) is a 36 amino acid peptide, which among others, plays a pivotal role in stre... more Neuropeptide Y (NPY) is a 36 amino acid peptide, which among others, plays a pivotal role in stress response. Although previous studies confirmed that NPY release is increased by stress in several species, the exact mechanism of the stress-induced NPY release has not been elucidated yet.

Neuroscience, 2010

Galanin and neuropeptide Y (NPY) are among the most abundant neuropeptides in the hypothalamus. T... more Galanin and neuropeptide Y (NPY) are among the most abundant neuropeptides in the hypothalamus. The role of NPY and galanin in the regulation of the secretory activity of the anterior pituitary has been well established. In addition, the two peptides interact with a number of neurons synthesizing the releasing and inhibiting hormones and a large number of other neuropeptides. The aim of the present studies was to explore if, as in rodents, NPY innervates galanin-immunoreactive (IR) neurons in the human diencephalon. Due to the long post mortem period and subsequent lack of optimal preservation of the cell membranes in the brain, electron microscopy could not be employed to show the presence of NPY-IR synapses on galanin-IR neurons. Therefore, we used light microscopic double label immunocytochemistry and high magnification microscopy with oil immersion to identify putative juxtapositions between NPY and galanin. Our studies show that similarly to rats, numerous NPY-IR nerve terminals surrounded galanin-IR neurons in the human hypothalamus. Among the hypothalamic regions, the infundibulum (infundibular or arcuate nucleus) contained the largest number of galanin-IR neurons heavily surrounded with NPY-IR nerve terminals. These en passant-type intimate associations between NPY-IR and galanin-IR neuronal elements may be functional synapses and may provide the morphological basis for the NPY-mediated galanin release. Consequently, NPY-galanin communication may mediate effects of NPY on neuronal systems innervated by galanin, and therefore may play a pivotal role in the regulation of reproduction, growth, energy and metabolism.

LHRH release is induced by substance P (SP) in the rat hypothalamus. Recent immunocytochemical st... more LHRH release is induced by substance P (SP) in the rat hypothalamus. Recent immunocytochemical studies indicate that SP-immunoreactive axons synapse on LHRH neurons in the diencephalon of the rat, but this phenomenon has not yet been demonstrated in human. Therefore, in the present study we visualized the SP- and LHRH-immunoreactive (IR) elements in the human diencephalon and evaluated the close juxtapositions between them. The distribution of LHRH- and SP-IR sites were investigated in diencephalic sections of six, postmortem human brains by means of double-labeling immunocytochemistry. The LHRH-containing perikarya were located in the diagonal band of Broca, lamina terminalis cinerea, preopticoseptal, medial preoptic, and infundibular areas of the brain. The SP-IR fibers formed a network in the periventricular zone in the infundibular region, median eminence, and corpus striatum. The SP-IR cell bodies were located mainly in the infundibular region, median eminence, basal part of the periventricular area, dorsomedial subdivision of the ventromedial nucleus, and basal perifornical area of the tuberal region. The juxtapositions between LHRH-IR cell bodies and SP-IR varicosities were detected in the infundibular and periventricular regions. In these sites black, silver-intensified, SP-IR fiber varicosities abutted on brown, DAB-labeled, LHRH-IR cell bodies. Similar structures were detected between the SP-IR fibers and SP-IR perikarya. These findings suggest that the juxtapositions between the SP and LHRH systems may be the morphological basis of SP-controlled LHRH release in the human diencephalon. Moreover, the intimate contacts between SP-IR fiber varicosities and SP-IR cell bodies or axons indicate direct control of SP on the diencephalic SP release.

Neuroscience, 2004

Endogenous opiates, such as ␤-endorphin, inhibit the release of luteinizing hormone (LH) release ... more Endogenous opiates, such as ␤-endorphin, inhibit the release of luteinizing hormone (LH) release in the pituitary gland of several species including rat, pig, sheep, and human. Although it is generally believed that ␤-endorphin influences gonadal functions via the regulation of hypothalamic LH-releasing hormone (LHRH) release, the morphological substrate underlying this regulation in humans remains elusive. In the present series of experiments the ␤-endorphinimmunoreactive (IR) and LHRH-IR neural elements, utilizing single label immunohistochemistry, were mapped. Following the superimposition of the maps of these systems, the overlapping sites were identified and examined in order to verify the putative juxtapositions between the ␤-endorphin-IR and LHRH-IR structures. LHRH-IR elements were detected mainly in the medial basal hypothalamus, in the medial preoptic area and along the diagonal band of Broca. ␤-Endorphin-IR perikarya were observed in the infundibular region/median eminence, whereas ␤-endorphin-IR axon varicosities were detected periventricularly in the preoptic and tuberal regions, in the medial basal hypothalamus and around the mamillary bodies. Careful examination of the immunoreactive elements in the overlapping areas revealed close contacts between ␤-endorphin-IR and LHRH-IR structures, which have been verified in semithin plastic sections. These putative ␤-endorphin-LHRH juxtapositions were most numerous in the medial preoptic area and in the infundibulum/median eminence of the human diencephalon. In conclusion, the present paper is the first study that revealed close juxtapositions between the ␤-endorphin-IR and LHRH-IR neural elements in the human diencephalon. These ␤-endorphin-LHRH contacts may be functional synapses, and they may be the morphological substrate of the ␤-endorphin control on gonadal functions in man.

Catecholamines have been shown to modulate gonadal functions via interactions with hypothalamic L... more Catecholamines have been shown to modulate gonadal functions via interactions with hypothalamic LH-releasing hormone (LHRH)-synthesizing neurons. To reveal the morphological background of this phenomenon, the distribution of LHRH neurons and tyrosine hydroxylase (TH)-immunoreactive (IR), catecholaminergic structures were mapped in the human diencephalon. First, the location of LHRH and TH-IR neuronal elements was analyzed, and then the relationship between the two different systems was examined. The LHRH-IR cell bodies were mainly present in the medial preoptic and infundibular areas. The TH-IR perikarya were located in the periventricular, paraventricular, and supraoptic hypothalamic nuclei and also in the median eminence. The TH-IR fibers were numerous in septal, infundibular, periventricular, and lateral hypothalamic regions. The brown, diaminobenzidine-labeled LHRH-containing perikarya were found to receive black, silver-intensified, TH-positive axon terminals in the infundibular and medial preoptic areas. However, in the preoptic and caudal parts of the diencephalon, only a few juxtapositions were noted. The present results indicate that hormone released from diencephalic LHRH-IR neurons in humans may be influenced by the central catecholaminergic system via direct synaptic mechanisms.

Brain Research, 2003

Previous studies revealed that cholinergic neurons possessing long axons are extremely sensitive ... more Previous studies revealed that cholinergic neurons possessing long axons are extremely sensitive to ethylcholine aziridinium ion (AF64A) administration [Neuropharmacology 31 (1992) 397]. In the present paper we examined the effect of AF64A on the cholinergic elements of the cingulum bundle. Seven days after AF64A administration choline acetyltransferase (ChAT)-immunoreactive fibers were extensively damaged on the dorsal part of cingulum bundle. These findings are the first reporting damage by AF64A to this brain region. 

Thrombosis Research, 2002

Although heparin-derived oligosaccharide(s) (HDO) have been clinically used for the management of... more Although heparin-derived oligosaccharide(s) (HDO) have been clinically used for the management of neurological disorders, such as stroke and Alzheimer's disease (AD), very little information on the mechanism of their therapeutic action is known. To test the hypothesis that HDO may pass through the blood -brain barrier (BBB) to mediate their effects, a pharmacodynamic (PD) model was developed and the presence of HDO in the cerebrospinal fluid (CSF) was used as a BBB accessibility index. Rats were treated with an ultralow molecular weight (MW) heparin fragment C3 via the intravenous or subcutaneous routes at 5 -10 mg/kg. At varying periods, the plasma, CSF, and brain samples were collected, and functional anti-factor Xa activities were measured to quantitate the CSF/plasma ratios (CPR) and the brain uptake. C3 showed CPR of 1.7% and 0.8% after intravenous and subcutaneous injections, respectively. These findings were verified by intravenous administration of tritium-labeled C3 followed by detection of the radioactivity in the CSF and brain homogenates. These data suggest that ultralow MW HDO may pass through the BBB. D

Brain Research, 2005

ABSTRACT Ethylcholine aziridinium (AF64A) induces cholinergic lesion in animal models of AD. Alth... more ABSTRACT Ethylcholine aziridinium (AF64A) induces cholinergic lesion in animal models of AD. Although higher concentrations of AF64A are known to induce nonspecific, cholinergic, and non-cholinergic lesions, low concentrations are believed to be selectively cholinotoxic. However, morphological evidence of this phenomenon has not been demonstrated yet. The present study demonstrates that while AF64A damaged septal cholinergic fibers, periventricular GnRH-immunoreactive fibers remained intact, confirming the highly selective cholinotoxicity of AF64A at appropriate concentrations.

Thrombosis Research, 2002

This study was designed to characterize a heparin-derived oligosaccharide (HDO), C3, using chemic... more This study was designed to characterize a heparin-derived oligosaccharide (HDO), C3, using chemical and biochemical methods. Although previous studies have suggested C3 as a promising compound in the treatment of Alzheimer's disease (AD), its molecular and biochemical properties are still unknown. In this study, the molecular profiles and anticoagulant effects of C3 were investigated. To characterize the molecular and biochemical properties of C3, gel permeation chromatography (GPC), polyacrylmide gel electrophoresis (PAGE), radiolabeling and anticoagulant assays, such as activated partial thromboplastin time (APTT), Heptest, and anti-factor Xa assay, were used. The GPC profile revealed that C3 was an ultra-low-molecular-weight (MW) heparin mixture. The multiple components in C3 were studied with PAGE analysis. Tritium-labeled C3 exhibited similar biological properties as nonlabeled materials. The biological assays showed that C3 and its components exhibited weak anticoagulant effect. These results demonstrated the applicability of the combination of GPC, PAGE, and coagulation assays to characterize the molecular and biochemical profile of HDO. In addition, the low anticoagulant effect of C3 suggests that this compound could be a relatively low-risk adjunct in the treatment of AD. D

Journal of Neuroscience Methods, 2000

The guinea pig (Ca6ia porcellus) is a species frequently used in neuromorphological and neurophys... more The guinea pig (Ca6ia porcellus) is a species frequently used in neuromorphological and neurophysiological studies. Some experimental data suggest that the guinea pig might also be used to develop an animal model of Alzheimer's disease. These studies would require microsurgical manipulations of the nervous system. The present paper describes a method for ventral stereotaxic intrusions in the guinea pig brain through the oval foramen at the skull base. The topographic relationships of the bony landmarks to major parts of the central nervous system and the cranial nerves are analysed, and the results are tested by intrahippocampal injection of horseradish peroxidase.

Injury-international Journal of The Care of The Injured, 2008

The sacrospinous (SS) and sacrotuberous (ST) ligaments of the pelvic ring are known as mechanical... more The sacrospinous (SS) and sacrotuberous (ST) ligaments of the pelvic ring are known as mechanical stabilisers of the pelvic girdle, primarily against rotational forces in the sagittal and horizontal planes. Earlier studies, however, raised the possibility that ST/SS ligaments possess significant proprioceptive function, while the mechanical role of these ligaments in maintaining the structural integrity of the pelvis is of less importance.The aim of this study is to determine whether the function of these ligaments is strictly to provide mechanical stability or if they have any additional functional properties, i.e., proprioception. In order to reveal the function of the SS/ST ligaments, biomechanical studies of cadaver pelvis were used along with the histological analysis of the ligaments. Following measurements to determine the accurate mechanical role of the pelvic ligaments, the strength of these ligaments was significantly less than we earlier expected. For this reason other functions of the SS/ST ligaments were considered, including the proprioceptive role. Indeed, histological studies revealed ramifying nerve terminals in the SS/ST ligaments. These terminals may represent the morphological substrate of the proprioceptive function associated with the ligaments.Our studies revealed that SS/ST ligaments might have a significant proprioceptive function providing information of the position of the pelvis. Consequently, the mechanical role of the ligaments in maintaining the structural integrity of the pelvis may be significantly less than previously assumed. Understanding the function of the SS/ST ligaments is crucial for providing more precise guidelines for patient management with injuries to the posterior pelvic region.

The present study determined whether the serotonin 2A (5-HT 2A ) receptors in the hypothalamic pa... more The present study determined whether the serotonin 2A (5-HT 2A ) receptors in the hypothalamic paraventricular nucleus mediate the neuroendocrine responses to a peripheral injection of the 5-HT 2A/2C receptor agonist (Ϫ)DOI [(Ϫ)1-(2,5-dimethoxy-4iodophenyl)-2-aminopropane]. The 5-HT 2A receptor antagonist MDL100,907 ((Ϯ)-␣-(2,3-dimethoxyphenyl)-1-[2-(4-fluorophenylethyl)]-4-piperidinemethanol), the 5-HT 2C receptor antagonist SB-242084 (6-chloro-5-methyl-1-[[2-[(2-methyl-3-pyridyl)oxy]-5pyridyl]carbamoyl]-indoline), or vehicle were microinjected bilaterally through a chronically implanted double-barreled cannula into the hypothalamic paraventricular nucleus 15 min before a peripheral injection of (Ϫ)DOI in conscious rats. (Ϫ)DOI significantly elevated plasma levels of oxytocin, prolactin, ACTH, corticosterone, and renin. Neither the 5-HT 2A receptor antagonist nor the 5-HT 2C receptor antagonist, injected alone, altered the basal levels of these hormones. MDL100,907 (0.748, 7.48, and 18.7 nmol) dose dependently inhibited the (Ϫ)DOI-induced increase in all of the hormones except corticosterone. In contrast, SB-242084 (10 nmol) did not inhibit (Ϫ)DOI-increased hormone levels. To confirm the presence of 5-HT 2A receptors in the hypothalamic paraventricular nucleus, 5-HT 2A receptors were mapped using immunohistochemistry. Densely labeled magnocellular neurons were observed throughout the anterior and posterior magnocellular subdivisions of the hypothalamic paraventricular nucleus. Moderately to densely labeled cells were also observed in parvicellular regions. Thus, it is likely that 5-HT 2A receptors are present on neuroendocrine cells in the hypothalamic paraventricular nucleus. These data provide the first direct evidence that neuroendocrine responses to a peripheral injection of (Ϫ)DOI are predominantly mediated by activation of 5-HT 2A receptors in the hypothalamic paraventricular nucleus.

Growth Hormone & Igf Research, 2010

Previous studies revealed that growth hormone-releasing hormone (GHRH)-immunoreactive (IR) neuron... more Previous studies revealed that growth hormone-releasing hormone (GHRH)-immunoreactive (IR) neurons form a circumscribed cell group in the basal infundibulum/median eminence of the human hypothalamus. GHRH from these neurons is released into the hypothalamo-hypophyseal portal circulatory system in a pulsatile manner. It is a common consensus that the pulsatile release of GHRH is the main driving force behind the pulsatile release of growth hormone (GH) and may contribute to the regulation of other hypothalamic functions. The pulsatile release of GHRH requires synchronized activity of GHRH-IR neurons. However, the morphological basis of this synchronization between the GHRH-IR neural elements has not been elucidated yet. Since the utilization of electron microscopy combined with immunohistochemistry is virtually impossible in the human brain due to the long post mortem period, immunohistochemistry, evaluated with oil immersion light microscopy, was used in order to reveal the associations between the GHRH elements. Numerous GHRH-GHRH juxtapositions have been detected in the infundibular area/median eminence, where GHRH-IR axonal varicosities often formed multiple contacts with GHRH-IR perikarya. Examination of these associations with high magnification oil immersion light microscopy revealed (1) axonal swellings at the site of the contacts and (2) no gaps between the contacting elements suggesting that these juxtapositions may be functional synapses. The large number of GHRH-GHRH juxtapositions in the infundibular area/ median eminence suggests that these synapse-like structures may represent the morphological substrate of the synchronized activity of GHRH neurons that in turn may result in the pulsatile release of GHRH in human.

Neuroscience, 2004

However, the morphological substrate of any similar modulation is not known in human. In the pres... more However, the morphological substrate of any similar modulation is not known in human. In the present series of experiments we first mapped the galanin-IR and LHRH-IR neural elements in human brain, utilizing single label immunohistochemistry. Then, following the superimposition of the maps of these systems, the overlapping sites were identified with double labeling immunocytochemistry and examined in order to verify the putative juxtapositions between galanin-IR and LHRH-IR structures. LHRH and galanin immunoreactivity were detected mainly in the medial basal hypothalamus, in the medial preoptic area and along the diagonal band of Broca. Careful examination of the IR elements in the overlapping areas revealed close, bi-directional contacts between galanin-IR and LHRH-IR structures, which have been verified in semithin plastic sections. These galanin-LHRH and LHRHgalanin juxtapositions were most numerous in the medial preoptic area and in the infundibulum/median eminence of the human diencephalon.

Neuroscience, 2010

Previous studies have demonstrated that catecholaminergic, tyrosine hydroxylase (TH)-immunoreacti... more Previous studies have demonstrated that catecholaminergic, tyrosine hydroxylase (TH)-immunoreactive (IR) perikarya and fibers are widely distributed in the human hypothalamus. Since TH is the key and rate-limiting enzyme for catecholaminergic synthesis, these IR neurons may represent dopaminergic, noradrenergic or adrenergic neural elements. However, the distribution and morphology of these neurotransmitter systems in the human hypothalamus is not entirely known. Since the different catecholaminergic systems can be detected by identifying the neurons containing the specific key enzymes of catecholaminergic synthesis, in the present study we mapped the catecholaminergic elements in the human hypothalamus using immunohistochemistry against the catecholaminergic enzymes, TH, dopamine beta-hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT). Only a few, PNMT-IR, adrenergic neuronal elements were found mainly in the infundibulum and the periventricular zone. DBH-IR structures were more widely distributed in the human hypothalamus occupying chiefly the infundibulum/infundibular nucleus, periventricular area, supraoptic and paraventricular nuclei. Dopaminergic elements were detected by utilizing double label immunohistochemistry. First, the DBH-IR elements were visualized; then the TH-IR structures, that lack DBH, were detected with a different chromogen. In our study, we conclude that all of the catecholaminergic perikarya and the majority of the catecholaminergic fibers represent dopaminergic neurons in the human hypothalamus. Due to the extremely small number of PNMT-IR, adrenergic structures in the human hypothalamus, the DBH-IR fibers represent almost exclusively noradrenergic neuronal processes. These findings suggest that the juxtapositions between the TH-IR and numerous peptidergic systems revealed by previous reports indicate mostly dopaminergic synapses.

Journal of Neuroendocrinology, 2006

It has been postulated that the stress response is associated with water balance via regulating v... more It has been postulated that the stress response is associated with water balance via regulating vasopressin release. Nausea, surgical stress and insulin-induced hypoglycaemia were shown to stimulate vasopressin secretion in humans. Increased vasopressin release in turn induces water resorption through the kidneys. Although the mechanism of the stress-mediated vasopressin release is not entirely understood, it is generally accepted that catecholamines play a crucial role in influencing water balance by modulating the secretion of vasopressin. However, the morphological substrate of this modulation has not yet been established. The present study utilised double-label immunohistochemistry to reveal putative juxtapositions between tyrosine hydroxylase (TH)-immunoreactive (IR) catecholaminergic system and the vasopressin systems in the human hypothalamus. In the paraventricular and supraoptic nuclei, numerous vasopressin-IR neurones received TH-IR axon varicosities. Analysis of these juxtapositions with high magnification combined with oil immersion did not reveal any gaps between the contacted elements. In conclusion, the intimate associations between the TH-IR and vasopressin-IR elements may be functional synapses and may represent the morphological basis of vasopressin release modulated by stressors. Because certain vasopressin-IR perikarya receive no detectable TH innervations, it is possible that additional mechanisms may participate in the stress-influenced vasopressin release.

Neuroscience, 2008

Previous studies revealed that stress is a pivotal factor in the regulation of growth. Psychologi... more Previous studies revealed that stress is a pivotal factor in the regulation of growth. Psychological harassment may result in psychosocial dwarfism with delayed puberty, short stature and depression. Growth hormone (GH) secretion is suppressed by stress, possibly via the attenuation of growth hormone-releasing hormone (GHRH) secretion. However, the morphological substrate of this phenomenon has not been elucidated yet.

Journal of Chemical Neuroanatomy, 2009

Journal of Neuroendocrinology, 2006

The gonadotrophin-releasing hormone (GnRH) represents the final common pathway of a neuronal netw... more The gonadotrophin-releasing hormone (GnRH) represents the final common pathway of a neuronal network that integrates multiple external and internal factors to control fertility. Among the many inputs GnRH neurones receive, oestrogens play the most important role. In females, oestrogen, in addition to the negative feedback, also exhibits a positive feedback influence upon the activity and output of GnRH neurones to generate the preovulatory luteinising hormone surge and ovulation. Until recently, the belief has been that the GnRH neurones do not contain oestrogen receptors and that the action of oestrogen upon GnRH neurones is indirect, involving several, oestrogen-sensitive neurotransmitter and neuromodulator systems that trans-synaptically regulate the activity of the GnRH neurones. Although this concept still holds for humans, recent studies indicate that oestrogen receptor-beta is expressed in GnRH neurones of the rat. This review provides three dimensional stereoscopic images of GnRH-immunoreactive (IR) and some peptidergic (neuropeptide Y-, substance P-, β-endorphin-, leu-enkaphalin-, corticotrophin hormone-releasing- and galanin-IR) and catecholaminergic neurones and the communication of these potential oestrogen-sensitive neuronal systems with GnRH neurones in the human hypothalamus. Because the post-mortem human tissue does not allow the electron microscopic identification of synapses on GnRH neurones, the data presented here are based on light microscopic immunocytochemical experiments using high magnification with oil immersion, semithin sections or confocal microscopy.

Gonadal functions are modulated by corticotropin-releasing factor (CRF) in the rat via direct sup... more Gonadal functions are modulated by corticotropin-releasing factor (CRF) in the rat via direct suppression of LH-releasing hormone (LHRH) release. Although there is evidence of direct morphological contacts between the LHRH and CRF-immunoreactive (-IR) structures in the rat hypothalamus, little is known about the morphological base of CRF-influenced LHRH release in man. Thus, we studied the distribution of the CRF-IR and LHRH-IR systems in the human diencephalon and revealed putative CRF-LHRH juxtapositions using double label immunohistochemistry. LHRH-IR cells were present mainly in the infundibular region and the medial preoptic area. CRF-IR neuronal structures were observed in the periventricular area, paraventricular nucleus, infundibular region, and median eminence. CRF-LHRH juxtapositions were found mainly in the infundibulum and median eminence. Few juxtapositions were detected in the medial preoptic area. In these regions, black diaminobenzidine/silver-labeled CRF-IR fibers abutted fusiform brown diaminobenzidine-labeled LHRH neurons, usually forming multiple contacts. Examination of semithin sections of these close associations with the aid of oil immersion revealed no cleft between CRF-IR nerve terminals contacting LHRH-IR structures. These findings suggest that the juxtapositions between the LHRH-IR and CRF-IR neurons may be functional synapses forming the morphological substrate of the CRF-controlled LHRH secretion. Moreover, the wide distribution of CRF-IR elements suggests that CRF controls other diencephalic functions as well.

Neuropeptides, 2011

Neuropeptide Y (NPY) is a 36 amino acid peptide, which among others, plays a pivotal role in stre... more Neuropeptide Y (NPY) is a 36 amino acid peptide, which among others, plays a pivotal role in stress response. Although previous studies confirmed that NPY release is increased by stress in several species, the exact mechanism of the stress-induced NPY release has not been elucidated yet.

Neuroscience, 2010

Galanin and neuropeptide Y (NPY) are among the most abundant neuropeptides in the hypothalamus. T... more Galanin and neuropeptide Y (NPY) are among the most abundant neuropeptides in the hypothalamus. The role of NPY and galanin in the regulation of the secretory activity of the anterior pituitary has been well established. In addition, the two peptides interact with a number of neurons synthesizing the releasing and inhibiting hormones and a large number of other neuropeptides. The aim of the present studies was to explore if, as in rodents, NPY innervates galanin-immunoreactive (IR) neurons in the human diencephalon. Due to the long post mortem period and subsequent lack of optimal preservation of the cell membranes in the brain, electron microscopy could not be employed to show the presence of NPY-IR synapses on galanin-IR neurons. Therefore, we used light microscopic double label immunocytochemistry and high magnification microscopy with oil immersion to identify putative juxtapositions between NPY and galanin. Our studies show that similarly to rats, numerous NPY-IR nerve terminals surrounded galanin-IR neurons in the human hypothalamus. Among the hypothalamic regions, the infundibulum (infundibular or arcuate nucleus) contained the largest number of galanin-IR neurons heavily surrounded with NPY-IR nerve terminals. These en passant-type intimate associations between NPY-IR and galanin-IR neuronal elements may be functional synapses and may provide the morphological basis for the NPY-mediated galanin release. Consequently, NPY-galanin communication may mediate effects of NPY on neuronal systems innervated by galanin, and therefore may play a pivotal role in the regulation of reproduction, growth, energy and metabolism.

LHRH release is induced by substance P (SP) in the rat hypothalamus. Recent immunocytochemical st... more LHRH release is induced by substance P (SP) in the rat hypothalamus. Recent immunocytochemical studies indicate that SP-immunoreactive axons synapse on LHRH neurons in the diencephalon of the rat, but this phenomenon has not yet been demonstrated in human. Therefore, in the present study we visualized the SP- and LHRH-immunoreactive (IR) elements in the human diencephalon and evaluated the close juxtapositions between them. The distribution of LHRH- and SP-IR sites were investigated in diencephalic sections of six, postmortem human brains by means of double-labeling immunocytochemistry. The LHRH-containing perikarya were located in the diagonal band of Broca, lamina terminalis cinerea, preopticoseptal, medial preoptic, and infundibular areas of the brain. The SP-IR fibers formed a network in the periventricular zone in the infundibular region, median eminence, and corpus striatum. The SP-IR cell bodies were located mainly in the infundibular region, median eminence, basal part of the periventricular area, dorsomedial subdivision of the ventromedial nucleus, and basal perifornical area of the tuberal region. The juxtapositions between LHRH-IR cell bodies and SP-IR varicosities were detected in the infundibular and periventricular regions. In these sites black, silver-intensified, SP-IR fiber varicosities abutted on brown, DAB-labeled, LHRH-IR cell bodies. Similar structures were detected between the SP-IR fibers and SP-IR perikarya. These findings suggest that the juxtapositions between the SP and LHRH systems may be the morphological basis of SP-controlled LHRH release in the human diencephalon. Moreover, the intimate contacts between SP-IR fiber varicosities and SP-IR cell bodies or axons indicate direct control of SP on the diencephalic SP release.

Neuroscience, 2004

Endogenous opiates, such as ␤-endorphin, inhibit the release of luteinizing hormone (LH) release ... more Endogenous opiates, such as ␤-endorphin, inhibit the release of luteinizing hormone (LH) release in the pituitary gland of several species including rat, pig, sheep, and human. Although it is generally believed that ␤-endorphin influences gonadal functions via the regulation of hypothalamic LH-releasing hormone (LHRH) release, the morphological substrate underlying this regulation in humans remains elusive. In the present series of experiments the ␤-endorphinimmunoreactive (IR) and LHRH-IR neural elements, utilizing single label immunohistochemistry, were mapped. Following the superimposition of the maps of these systems, the overlapping sites were identified and examined in order to verify the putative juxtapositions between the ␤-endorphin-IR and LHRH-IR structures. LHRH-IR elements were detected mainly in the medial basal hypothalamus, in the medial preoptic area and along the diagonal band of Broca. ␤-Endorphin-IR perikarya were observed in the infundibular region/median eminence, whereas ␤-endorphin-IR axon varicosities were detected periventricularly in the preoptic and tuberal regions, in the medial basal hypothalamus and around the mamillary bodies. Careful examination of the immunoreactive elements in the overlapping areas revealed close contacts between ␤-endorphin-IR and LHRH-IR structures, which have been verified in semithin plastic sections. These putative ␤-endorphin-LHRH juxtapositions were most numerous in the medial preoptic area and in the infundibulum/median eminence of the human diencephalon. In conclusion, the present paper is the first study that revealed close juxtapositions between the ␤-endorphin-IR and LHRH-IR neural elements in the human diencephalon. These ␤-endorphin-LHRH contacts may be functional synapses, and they may be the morphological substrate of the ␤-endorphin control on gonadal functions in man.

Catecholamines have been shown to modulate gonadal functions via interactions with hypothalamic L... more Catecholamines have been shown to modulate gonadal functions via interactions with hypothalamic LH-releasing hormone (LHRH)-synthesizing neurons. To reveal the morphological background of this phenomenon, the distribution of LHRH neurons and tyrosine hydroxylase (TH)-immunoreactive (IR), catecholaminergic structures were mapped in the human diencephalon. First, the location of LHRH and TH-IR neuronal elements was analyzed, and then the relationship between the two different systems was examined. The LHRH-IR cell bodies were mainly present in the medial preoptic and infundibular areas. The TH-IR perikarya were located in the periventricular, paraventricular, and supraoptic hypothalamic nuclei and also in the median eminence. The TH-IR fibers were numerous in septal, infundibular, periventricular, and lateral hypothalamic regions. The brown, diaminobenzidine-labeled LHRH-containing perikarya were found to receive black, silver-intensified, TH-positive axon terminals in the infundibular and medial preoptic areas. However, in the preoptic and caudal parts of the diencephalon, only a few juxtapositions were noted. The present results indicate that hormone released from diencephalic LHRH-IR neurons in humans may be influenced by the central catecholaminergic system via direct synaptic mechanisms.

Brain Research, 2003

Previous studies revealed that cholinergic neurons possessing long axons are extremely sensitive ... more Previous studies revealed that cholinergic neurons possessing long axons are extremely sensitive to ethylcholine aziridinium ion (AF64A) administration [Neuropharmacology 31 (1992) 397]. In the present paper we examined the effect of AF64A on the cholinergic elements of the cingulum bundle. Seven days after AF64A administration choline acetyltransferase (ChAT)-immunoreactive fibers were extensively damaged on the dorsal part of cingulum bundle. These findings are the first reporting damage by AF64A to this brain region. 

Thrombosis Research, 2002

Although heparin-derived oligosaccharide(s) (HDO) have been clinically used for the management of... more Although heparin-derived oligosaccharide(s) (HDO) have been clinically used for the management of neurological disorders, such as stroke and Alzheimer's disease (AD), very little information on the mechanism of their therapeutic action is known. To test the hypothesis that HDO may pass through the blood -brain barrier (BBB) to mediate their effects, a pharmacodynamic (PD) model was developed and the presence of HDO in the cerebrospinal fluid (CSF) was used as a BBB accessibility index. Rats were treated with an ultralow molecular weight (MW) heparin fragment C3 via the intravenous or subcutaneous routes at 5 -10 mg/kg. At varying periods, the plasma, CSF, and brain samples were collected, and functional anti-factor Xa activities were measured to quantitate the CSF/plasma ratios (CPR) and the brain uptake. C3 showed CPR of 1.7% and 0.8% after intravenous and subcutaneous injections, respectively. These findings were verified by intravenous administration of tritium-labeled C3 followed by detection of the radioactivity in the CSF and brain homogenates. These data suggest that ultralow MW HDO may pass through the BBB. D

Brain Research, 2005

ABSTRACT Ethylcholine aziridinium (AF64A) induces cholinergic lesion in animal models of AD. Alth... more ABSTRACT Ethylcholine aziridinium (AF64A) induces cholinergic lesion in animal models of AD. Although higher concentrations of AF64A are known to induce nonspecific, cholinergic, and non-cholinergic lesions, low concentrations are believed to be selectively cholinotoxic. However, morphological evidence of this phenomenon has not been demonstrated yet. The present study demonstrates that while AF64A damaged septal cholinergic fibers, periventricular GnRH-immunoreactive fibers remained intact, confirming the highly selective cholinotoxicity of AF64A at appropriate concentrations.

Thrombosis Research, 2002

This study was designed to characterize a heparin-derived oligosaccharide (HDO), C3, using chemic... more This study was designed to characterize a heparin-derived oligosaccharide (HDO), C3, using chemical and biochemical methods. Although previous studies have suggested C3 as a promising compound in the treatment of Alzheimer's disease (AD), its molecular and biochemical properties are still unknown. In this study, the molecular profiles and anticoagulant effects of C3 were investigated. To characterize the molecular and biochemical properties of C3, gel permeation chromatography (GPC), polyacrylmide gel electrophoresis (PAGE), radiolabeling and anticoagulant assays, such as activated partial thromboplastin time (APTT), Heptest, and anti-factor Xa assay, were used. The GPC profile revealed that C3 was an ultra-low-molecular-weight (MW) heparin mixture. The multiple components in C3 were studied with PAGE analysis. Tritium-labeled C3 exhibited similar biological properties as nonlabeled materials. The biological assays showed that C3 and its components exhibited weak anticoagulant effect. These results demonstrated the applicability of the combination of GPC, PAGE, and coagulation assays to characterize the molecular and biochemical profile of HDO. In addition, the low anticoagulant effect of C3 suggests that this compound could be a relatively low-risk adjunct in the treatment of AD. D

Journal of Neuroscience Methods, 2000

The guinea pig (Ca6ia porcellus) is a species frequently used in neuromorphological and neurophys... more The guinea pig (Ca6ia porcellus) is a species frequently used in neuromorphological and neurophysiological studies. Some experimental data suggest that the guinea pig might also be used to develop an animal model of Alzheimer's disease. These studies would require microsurgical manipulations of the nervous system. The present paper describes a method for ventral stereotaxic intrusions in the guinea pig brain through the oval foramen at the skull base. The topographic relationships of the bony landmarks to major parts of the central nervous system and the cranial nerves are analysed, and the results are tested by intrahippocampal injection of horseradish peroxidase.