Extrasynaptic neurotransmission in the modulation of brain function. Focus on the striatal neuronal glial networks

Extrasynaptic neurotransmission is an important short distance form of volume transmission (VT) and describes the extracellular diffusion of transmitters and modulators after synaptic spillover or extrasynaptic release in the local circuit regions binding to and activating mainly extrasynaptic neuronal and glial receptors in the neuroglial networks of the brain. Receptor-receptor interactions in G protein-coupled receptor (GPCR) heteromers play a major role, on dendritic spines and nerve terminals including glutamate synapses, in the integrative processes of the extrasynaptic signaling. Heteromeric complexes between GPCR and ion-channel receptors play a special role in the integration of the synaptic and extrasynaptic signals. Changes in extracellular concentrations of the classical synaptic neurotransmitters glutamate and GABA found with microdialysis is likely an expression of the activity of the neuron-astrocyte unit of the brain and can be used as an index of VT-mediated actions of these two neurotransmitters in the brain. Thus, the activity of neurons may be functionally linked to the activity of astrocytes, which may release glutamate and GABA to the extracellular space where extrasynaptic glutamate and GABA receptors do exist. Wiring transmission (WT) and VT are fundamental properties of all neurons of the CNS but the balance between WT and VT varies from one nerve cell population to the other. The focus is on the striatal cellular networks, and the WT and VT and their integration via receptor heteromers are described in the GABA projection neurons, the glutamate, dopamine, 5-hydroxytryptamine (5-HT) and histamine striatal afferents, the cholinergic interneurons, and different types of GABA interneurons. In addition, the role in these networks of VT signaling of the energy-dependent modulator adenosine and of endocannabinoids mainly formed in the striatal projection neurons will be underlined to understand the communication in the striatal cellular networks

Enhanced vascular contractility in alpha1-adrenergic receptor-deficient mice.

AIM: Alpha1-adrenergic receptors (alpha1-ARs) are classified into three subtypes: alpha1A-AR, alpha1B-AR, and alpha1D-AR. Triple disruption of alpha1A-AR, alpha1B-AR, and alpha1D-AR genes results in hypotension and produces no contractile response of the thoracic aorta to noradrenalin. Presently, we characterized vascular contractility against other vasoconstrictors, such as potassium, prostaglandin F2alpha (PGF(2alpha)) and 5-hydroxytryptamine (5-HT), in alpha1A-AR, alpha1B-AR, and alpha1D-AR triple knockout (alpha1-AR triple KO) mice. MAIN METHODS: The contractile responses to the stimulation with vasoconstrictors were studied using isolated thoracic aorta. KEY FINDINGS: As a result, the phasic and tonic contraction induced by a high concentration of potassium (20 mM) was enhanced in the isolated thoracic aorta of alpha1-AR triple KO mice compared with that of wild-type (WT) mice. In addition, vascular responses to PGF(2alpha) and 5-HT were also enhanced in the isolated thoracic aorta of alpha1-AR triple KO mice compared with WT mice. Similar to in vitro findings with isolated thoracic aorta, in vivo pressor responses to PGF(2alpha) were enhanced in alpha1-AR triple KO mice. Real-time reverse transcription-polymerase chain reaction analysis and western blot analysis indicate that gene expression of the 5-hydroxytryptamine 2A (5-HT(2A)) receptor was up-regulated in the thoracic aorta of alpha1-AR triple KO mice while the prostaglandin F2alpha receptor (FP) was unchanged. SIGNIFICANCE: These results suggest that loss of alpha1-ARs can lead to enhancement of vascular responsiveness to the vasoconstrictors and may imply that alpha1-ARs and the subsequent signaling regulate the vascular responsiveness to other stimulations such as depolarization, 5-HT and PGF(2alpha).

On the existence and function of galanin receptor heteromers in the central nervous system

Galanin receptor (GalR) subtypes 1-3 linked to central galanin neurons may form heteromers with each other and other types of G protein-coupled receptors in the central nervous system (CNS). These heteromers may be one molecular mechanism for galanin peptides and their N-terminal fragments (gal 1-15) to modulate the function of different types of glia-neuronal networks in the CNS, especially the emotional and the cardiovascular networks. GalR-5-HT1A heteromers likely exist with antagonistic GalR-5-HT1A receptor-receptor interactions in the ascending midbrain raphe 5-HT neuron systems and their target regions. They represent a novel target for antidepressant drugs. Evidence is given for the existence of GalR1-5-HT1A heteromers in cellular models with trans-inhibition of the protomer signaling. A GalR1-GalR2 heteromer is proposed to be a galanin N-terminal fragment preferring receptor (1-15) in the CNS. Furthermore, a GalR1-GalR2-5-HT1A heterotrimer is postulated to explain why only galanin (1-15) but not galanin (1-29) can antagonistically modulate the 5-HT1A receptors in the dorsal hippocampus rich in gal fragment binding sites. The results underline a putative role of different types of GalR-5-HT1A heteroreceptor complexes in depression. GalR antagonists may also have therapeutic actions in depression by blocking the antagonistic GalR-NPYY1 receptor interactions in putative GalR-NPYY1 receptor heteromers in the CNS resulting in increases in NPYY1 transmission and antidepressant effects. In contrast the galanin fragment receptor (a postulated GalR1-GalR2 heteromer) appears to be linked to the NPYY2 receptor enhancing the affinity of the NPYY2 binding sites in a putative GalR1-GalR2-NPYY2 heterotrimer. Finally, putative GalR-α2-adrenoreceptor heteromers with antagonistic receptor-receptor interactions may be a widespread mechanism in the CNS for integration of galanin and noradrenaline signals also of likely relevance for depression

Secondary metabolismin in Annonaceae: potencial source of drugs

Several species of Annona (Annonaceae) are used in traditional Mexican medicine by their anti-anxiety, anticonvulsant and tranquilizing properties. It has been reported that the alkaloids isolated from some species of the Annona have affinity to serotonergic 5-HT1A receptors and modulate dopaminergic transmission, which is involved in depressive disorders. In this review it is showed the results of the antidepressant-like effect of an alkaloid extract from the aerial parts of Annona cherimola (TA) in mice. The antidepressant-like effect was evaluated in the forced swimming test. To elucidate a possible mechanism of action, experiments of synergism with antidepressant drugs, such as imipramine (IMI), clomipramine (CLIMI), and fluoxetine (FLX), were carried out. The neurotransmitter content (DA: dopamine, 5HT: serotonin and its metabolites, HVA: homovanillic acid and 5HIAA:5-hydroxyindoleacetic) in the whole brain of mice were also determined by HPLC method. The results showed that repeated treatment with TA produced antidepressant-like effects in mice. This effect was not related to an increase in locomotor activity. Administration of TA facilitated the antidepressant effect of IMI and CLIMI as well as increased the turnover of DA and 5-HT. The alkaloids: 1,2-dimethoxy-5, 6.6 to 7-tetrahydro-4H-dibenzoquinoline-3,8,9,10-tetraol, anonaine, liriodenine, and nornuciferine were the main constituents of TA.

The study of genetic polymorphisms related to serotonin in Alzheimer's disease: a new perspective in a heterogenic disorder

Genetic and environmental factors have been implicated in the development of Alzheimer's disease (AD), the most common form of dementia in the elderly. Mutations in 3 genes mapped on chromosomes 21, 14 and 1 are related to the rare early onset forms of AD while the e4 allele of the apolipoprotein E (APOE) gene (on chromosome 19) is the major susceptibility locus for the most common late onset AD (LOAD). Serotonin (5-hydroxytryptamine or 5-HT) is a key neurotransmitter implicated in the control of mood, sleep, appetite and a variety of traits and behaviors. Recently, a polymorphism in the transcriptional control region upstream of the 5-HT transporter (5-HTT) gene has been studied in several psychiatric diseases and personality traits. It has been demonstrated that the short variant(s) of this 5-HTT gene-linked polymorphic region (5-HTTLPR) is associated with a different transcriptional efficiency of the 5-HTT gene promoter resulting in decreased 5-HTT expression and 5-HT uptake in lymphocytes. An increased frequency of this 5-HTTLPR short variant polymorphism in LOAD was recently reported. In addition, another common polymorphic variation in the 5-HT2A and 5-HT2C serotonin receptor genes previously analyzed in schizophrenic patients was associated with auditory and visual hallucinations in AD. These observations suggest that the involvement of the serotonin pathway might provide an explanation for some aspects of the affective symptoms commonly observed in AD patients. In summary, research on genetic polymorphisms related to AD and involved in receptors, transporter proteins and the enzymatic machinery of serotonin might enhance our understanding of this devastating neurodegenerative disorder.

Effect of the intracerebroventricular administration of GR 113808, a selective 5-HT4 antagonist, on water intake during hyperosmolarity and hypovolemia

We demonstrate here that acute third ventricle injections of GR 113808, a highly selective 5-HT4 antagonist, decrease water intake induced by a previous salt load while potentiating drinking elicited by hypovolemia induced by previous subcutaneous administration of polyethylene glycol in male Wistar rats (200 ± 20 g). At the dose of 160 nmol/rat, third ventricle injections of GR 113808 induced a significant reduction of water intake in salt-loaded animals after 120 min as compared to salt-loaded animals receiving third ventricle injections of saline (salt load + GR = 3.44 ± 0.41 ml, N = 12; salt load + saline = 5.74 ± 0.40 ml, N = 9). At the dose of 80 nmol/rat, GR 113808 significantly enhanced water intake in hypovolemic animals after 120 min as compared to hypovolemic animals receiving third ventricle injections of saline (hypovol + GR = 4.01 ± 0.27 ml, N = 8; hypovol + saline = 2.41 ± 0.23 ml, N = 12). We suggest that central 5-HT4 receptors may exert a positive drive on water intake due to hyperosmolarity and a negative input on drinking provoked by hypovolemia.

Effect of the 5-HT4 receptor and serotonin transporter on visceral hypersensitivity in rats

Visceral hypersensitivity plays an important role in motor and sensory abnormalities associated with irritable bowel syndrome, but the underlying mechanisms are not fully understood. The present study was designed to evaluate the expression of the 5-HT4 receptor and the serotonin transporter (SERT) as well as their roles in chronic visceral hypersensitivity using a rat model. Neonatal male Sprague-Dawley rats received intracolonic injections of 0.5% acetic acid (0.3-0.5 mL at different times) between postnatal days 8 and 21 to establish an animal model of visceral hypersensitivity. On day 43, the threshold intensity for a visually identifiable contraction of the abdominal wall and body arching were recorded during rectal distention. Histological evaluation and the myeloperoxidase activity assay were performed to determine the severity of inflammation. The 5-HT4 receptor and SERT expression of the ascending colon were monitored using immunohistochemistry and Western blot analyses; the plasma 5-HT levels were measured using an ELISA method. As expected, transient colonic irritation at the neonatal stage led to visceral hypersensitivity, but no mucosal inflammation was later detected during adulthood. Using this model, we found reduced SERT expression (0.298 ± 0.038 vs 0.634 ± 0.200, P < 0.05) and increased 5-HT4 receptor expression (0.308 ± 0.017 vs 0.298 ± 0.021, P < 0.05). Treatment with fluoxetine (10 mg·kg-1·day-1, days 36-42), tegaserod (1 mg·kg-1·day-1, day 43), or the combination of both, reduced visceral hypersensitivity and plasma 5-HT levels. Fluoxetine treatment increased 5-HT4 receptor expression (0.322 ± 0.020 vs 0.308 ± 0.017, P < 0.01) but not SERT expression (0.219 ± 0.039 vs 0.298 ± 0.038, P = 0.654). These results indicate that both the 5-HT4 receptor and SERT play a role in the pathogenesis of visceral hypersensitivity, and its mechanism may be involved in the local 5-HT level.

Substance P, récepteurs NK1 et neurones à sérotonine : relations anatomiques et fonctionnelles dans le noyau raphe dorsalis

Nous avons étudié les relations anatomiques entre les systèmes de neurotransmission à substance P (SP) et à sérotonine (5-hydroxytryptamine, 5-HT) dans le noyau du raphé dorsal (NRD) du rongeur, afin de mieux comprendre les interactions entre ces systèmes durant la régulation de l’humeur. Le NRD reçoit une innervation SP provenant de l’habenula, et le blocage pharmacologique des récepteurs neurokinine-1 (rNK1) de la SP aurait des effets antidépresseurs. Chez le rongeur, le traitement par les antagonistes des rNK1 s’accompagne d’une désensibilisation des autorécepteurs 5-HT1A de la 5-HT et d’une hausse de l’activité des neurones 5-HT dans le NRD, suggérant des interactions locales entre ces deux systèmes. Dans un premier temps, nous avons démontré par doubles marquages immunocytochimiques en microscopies optique, confocale et électronique, la présence du rNK1 dans une sous-population de neurones 5-HT du NRD caudal. Lors de l’analyse en microscopie électronique, nous avons pu constater que les rNK1 étaient principalement cytoplasmiques dans les neurones 5-HT et membranaires sur les neurones non 5-HT du noyau. Grâce à d’autres doubles marquages, nous avons aussi pu identifier les neurones non-5-HT porteurs de rNK1 comme étant GABAergiques. Nous avons ensuite combiné l’immunomarquage de la SP avec celui du rNK1, dans le but d’examiner les relations entre les terminaisons (varicosités *) axonales SP et les neurones 5-HT (pourvus de rNK1 cytoplasmiques du NRD caudal. En simple marquage de la SP, nous avons pu estimer à 41% la fréquence avec laquelle les terminaisons SP font synapse. Dans le matériel doublement marqué pour la SP et son récepteur, les terminaisons SP ont été fréquemment retrouvées en contact direct ou à proximité des dendrites munies de rNK1 cytoplasmiques, mais toujours éloignées des dendrites à rNK1 membranaires. Pour tester l’hypothèse d’une internalisation soutenue des rNK1 par la SP dans les neurones 5-HT, nous avons ensuite examiné la localisation subcellulaire du récepteur chez le rat traité avec un antagoniste du rNK1, le RP67580. La densité du marquage des rNK1 a été mesurée dans le cytoplasme et sur la membrane des deux types de dendrites (5-HT: rNK1 cytoplasmiques; non 5-HT: rNK1 membranaires). Une heure après une injection unique de l’antagoniste, la distribution du rNK1 est apparue inchangée dans les deux types de neurones (5-HT et non 5-HT). Par contre, après un traitement quotidien de 7 ou 21 jours avec l’antagoniste, nous avons mesuré une augmentation significative des densités cytoplasmique et membranaire du rNK1 dans les neurones 5-HT, sans aucun changement dans les neurones non 5-HT. Ces traitements ont aussi augmenté l’expression du gène rNK1 dans le NRD. Enfin, nous avons mesuré une hausse de la densité membranaire du rNK1 dans les neurones 5-HT, sans hausse de densité cytoplasmique, par suite d’une lésion bilatérale de l’habenula. Ces résultats confortent l’hypothèse d’une activation et d’une internalisation soutenues des rNK1 par la SP dans les neurones 5-HT du NRD caudal. Ils suggèrent aussi que le trafic des rNK1 dans les neurones 5-HT du NRD représente un mécanisme cellulaire en contrôle de l’activation du système 5-HT par les afférences SP en provenance de l’habenula.

Effet de la N-acétylcystéine sur la dysfonction endothéliale des artères coronaires épicardiques associée à une hypertrophie ventriculaire gauche dans un modèle porcin

Effet positif de la N-acétylcystéine sur la dysfonction endothéliale des artères coronaires épicardiques associée à une hypertrophie ventriculaire gauche dans un modèle porcin A. A. HORN, M-C AUBIN, YF SHI, J-C TARDIF, M. CARRIER , L. P. PERRAULT INSTITUT DE CARDIOLOGIE DE MONTRÉAL, MONTRÉAL, CANADA, Objectif : Il a été démontré dans le laboratoire que dans notre modèle d’hypertrophie ventriculaire gauche, la dysfonction endothéliale est secondaire à une diminution de la biodisponibilité du NO, celle-ci étant causée par une augmentation du stress oxydant tel que démontré par Malo et al. (2003) et Aubin et al. (2006). Le but de la présente étude est d’étudier l’effet d’un traitement chronique de la N-acétylcystéine (NAC) (un antioxydant) sur la dysfonction endothéliale associée à une hypertrophie ventriculaire gauche (HVG). Méthodologie: L’HVG a été induite par cerclage aortique (CA) chez vingt-et-un porcelets âgés de deux mois qui furent divisés aléatoirement en quatre groupes expérimentaux. Le groupe témoin (groupe 1) a été soumis à une thoracotomie antérolatérale gauche sans cerclage aortique (n=3). Le groupe 2 a été soumis à un cerclage aortique pour une période de 60 jours (n=6). Le groupe 3 a subi un cerclage aortique et a reçu un traitement oral de N-acétylcystéine de 1000 mg/jour per os pendant 60 jours commençant le jour de la chirurgie (n=6). Le groupe 4 a été soumis à un cerclage aortique et a reçu un traitement oral de N-acétylcystéine : 1000 mg/par jour pendant 30 jours commençant le jour 30 (post-chirurgie) (n=6). L’hypertrophie fut évaluée par échocardiographie. La réactivité vasculaire fut étudiée à l’aide de chambres d’organes par la construction des courbes concentration-réponse à la sérotonine (5-HT: relaxations induites par les récepteurs 5-HT1D, couplés aux protéines Gi) et à la bradykinine (BK: relaxations induites par les récepteurs B2, couplés aux protéines Gq). Les quantités de nitrites/nitrates et la production basale de GMPc ont été mesurées pour évaluer la fonction endothéliale. Le stress oxydant a été étudié en quantifiant les concentrations plasmatiques d’hydroperoxydes lipidiques et de glutathion réduit, ainsi que l’activité plasmatique des enzymes antioxydantes peroxydase du glutathion et dismutase du superoxyde. Résultats: Le rapport masse ventricule gauche/masse corporelle était significativement plus élevé pour le groupe 2 comparativement au groupe 1 (p<0,05) confirmant la présence d’une HVG. Le développement de l’HVG dans le groupe 3 a pu être prévenu par la NAC et sa progression fut atténuée dans le groupe 4 (p<0,05 versus groupe 2). La présence de la dysfonction endothéliale a été confirmée chez le groupe 2, tel qu’illustré par une diminution significative des relaxations maximales à la 5-HT et à la BK comparativement au groupe témoin. Le traitement à la NAC a significativement potentialisé les relaxations maximales (p<0,05) induites par la sérotonine et par la bradykinine, chez les deux groupes traités. Cette amélioration des relaxations dépendantes de l’endothélium peut être la conséquence d’une augmentation significative (p<0,05) de la biodisponibilité du monoxyde d’azote pour les cellules musculaires lisses, tel que suggéré par l’augmentation du ratio nitrites/nitrates et de la production basale de GMPc chez les groupes 3 et 4 comparativement au groupe 2. Cette augmentation du facteur relaxant peut résulter d’une augmentation de sa production par les cellules endothéliales ou d’une diminution de sa neutralisation par les espèces réactives oxygénées. De fait, les concentrations d’hydroperoxydes lipidiques étaient significativement inférieures (p<0,05) et associées à une augmentation des concentrations de l’antioxydant glutathion réduit et de l’activité de la peroxydase du glutathion chez les deux groupes traités par rapport au groupe 2. Conclusion: Le traitement à la NAC prévient le développement de la dysfonction endothéliale coronaire ainsi que l’HVG qui lui est associée.

Impact des polymorphismes 5-HTTLPR et rs4675690 sur les corrélats neuronaux de la tristesse chez les enfants

Les avancées technologiques rendent maintenant possible d’étudier l’impact de gènes spécifiques sur les corrélats cérébraux des psychopathologies. En rapport avec ce nouveau champ de recherche, la présente thèse décrit l’impact du 5-HTTLPR et du rs4675690 – deux polymorphismes semblant jouer un rôle dans le trouble dépressif majeur - sur les corrélats neuronaux de l’expérience subjective de la tristesse chez les enfants. Après une mise en contexte (Chapitre 1), les deux études incluses dans cette thèse seront décrites (Chapitre 2 et 3). Ces études d’imagerie par résonance magnétique fonctionnelle visaient à comparer les patrons d’activation – durant un état temporaire de tristesse - chez des enfants de 8 ans porteurs d’allèles différents. La tristesse était induite grâce au visionnement passif d’extraits d’un film triste. Les résultats suggèrent que les deux polymorphismes ont un impact sur les activations cérébrales associées à la tristesse. Spécifiquement, les enfants porteurs des allèles associés à un plus grand risque de développer un trouble dépressif majeur, soit les porteurs de l’allèle court du 5-HTTLPR et les individus homozygotes pour l’allèle T du rs4675690, ont présenté des activations cérébrales altérées dans des régions qui sont impliquées dans la tristesse normale et pathologique. Le Chapitre 4 présente la mise en perspective des résultats dans le cadre de la littérature actuelle.

Decreased 5-HT2C receptor binding in the cerebral cortex and brain stem during pancreatic regeneration in rats

The purpose of this study was to investigate the role of central 5-HT2C receptor binding in rat model of pancreatic regeneration using 60-70% pancreatectomy. The 5-HT and 5-HT2c receptor kinetics were studied in cerebral cortex and brain stem of sham operated, 72 h pancreatectomised and 7 days pancreatectomised rats. Scatchard analysis with [3H] mesulergine in cerebral cortex showed a significant decrease (p < 0.05) in maximal binding (B^,ax) without any change in Kd in 72 h pancreatectomised rats compared with sham. The decreased Bmax reversed to sham level by 7 days after pancreatectomy. In brain stem , Scatchard analysis showed a significant decrease (p < 0.01) in Bax with a significant increase (p < 0.01) in Kd. Competition analysis in brain stem showed a shift in affinity towards a low affinity. These parameters were reversed to sham level by 7 days after pancreatectomy. Thus the results suggest that 5-HT through the 5-HT2C receptor in the brain has a functional regulatory role in the pancreatic regeneration. (Mol Cell Biochem 272: 165-170, 2005)

Decreased 5-F{T1fti Receor Gene Expression and 5-F4T11 Receptor Protein in the Cerebra'' Cortex and Brain Stem during Pancreatic Regenera tion in Rats

The present study was to investigate the rote of central 5-11T and 5-HT,:v receptor Lindin4o and acne expression in it 'at mo(lel of pancreatic regeneration using 60" -, pancreatcutumy. The pancreatic regeneration was evaluated by 5-HT content and 5-HT,,receptor gene expression in the cerebral cortex (CC) and brain stem MS) of Alain opcrate,t, 7 It utd 7 (.lays panereatectomised rats. 5-11T content significantly increased in the CC' (I' 1.1)11 and 13S (P 0.05) of 72 Ii p.ntcreateetomiscd rats. Sympathetic activity was decreased as indicated by the significantly decreased norcpiuephrine (NIi) and epinephrine (FTI) Icvcl (1' < 0.001 and P < 0.05) in the plasma of 72 h panereateetomised rats. 5-111 ,^, receptor density and affinity was decreased in the CC (P < 0.01) and BS (P < 0.01). These rh:)nge; correlated with a diminished 5-IITIA receptor mRNA expression in the brain region. studied. Our resuils suggest that the brain 5-11T through 5-HTin receptor has it funcuon:0 rule iii 11w pi+ncreatic regcner:ttion through the sympathetic regulation.

Increased 5-HT2C Receptor Binding in the Brain Stem And Cerebral Cortex During Liver Regeneration And Hepatic Neoplasia In Rats

In the present study, serotonin 2C (5-HT2c) receptor binding parameters in the brainstem and cerebral cortex were investigated during liver generation after partial hepatectomy (PH) and N-nitrosodiethylamine (NDEA) induced hepatic neoplasia in male Wistar rats. The serotonin content increased significantly (p<0.01) in the cerebral cortex after PH and in NDEA induced hepatic neoplasia. Brain stem serotonin content increased significantly (p<0.05) after PH and (p<0.001) in NDEA induced hepatic neoplasia. The number and affinity of the 5-HT2c receptors in the crude synaptic membrane preparations of the brain stem showed a significant (p<0.001) increase after PH and in NDEA induced hepatic neoplasia. The number and affinity of 5-HT2c receptors increased significantly (p<0.001) in NDEA induced hepatic neoplasia in the crude synaptic membrane preparations of the cerebral cortex. There was a significant (p<0.01) increase in plasma norepinephrine in PH and (p<0.001) in NDEA induced hepatic neoplasia, indicating sympathetic stimulation. Thus, our results suggest that during active hepatocyte proliferation 5-HT2c receptor in the brain stem and cerebral cortex are up-regulated which in turn induce hepatocyte proliferation mediated through sympathetic stimulation.

ALPHA2 Adrenergic and High Affinity Serotonergic Receptor Changes in the Brain Stem of Streptozotocin induced Diabetic Rats

The brain stems (13S) of streptozotocin (STZ)-diabetic rats were studied lo see the changes in neurotransmitter content and their receptor regulation. The norepinephrine (NE) content determined in the diabetic brain stems did ^ control. an E showed la while PI turnover hri content increased significantly compared N^r eNveFa o the recep significant increase. The alpha2 adrenergic receptor IneP utisoulinntreat d ratsetheNE contentt dec^ sled was significantly reduced during diabetes. in versedcto reanorm sed ulcrea e tK reatment the state. while EPI content remained increased as in die diabetic B,, for a]pha2 adrenergic receptors slw^nificantly while Unlabelled clonidine inhibited [31-I]NE binding in BS of control, diabetic and insulin treated ulations bindi diabetic rats showed that alpha2 adrenergicre^ punks cojnidiabetic animal the ligand bound sites with Hill slopes significantly away from unity. weaker to the low affinity site than in controls. Insulin treatment reversed[ this allumbmn to control levels. The displacement analysis using (-)-epinephrine age in control and diabetic animals revealed two populations of receptor affinidtyo=tat ss. In control animals, when GTP analogue added with epinephrine, the curve nagnlde caofnfitnroit yS model; but in the diabetic BS this effect `not aobserved. In bintact oth the diabetic data thus showlthat the effects of monovalent cations on affinity alphaz adrenergic receptors have a reduced affinity v due in stem ialtered Itscppeomson(5- regulation. The serotonin (5-HT) coat hydroxy) tryptophan (5-HTP) showed an increase and its breakdown metabolite (5-hydroxy) indoleacetic acid (5-I{IAA) showed a significant decrease. This showed that in serotonergic which l nerves there is a disturbance in both synthetic and breankduomwnbers pretma'med ana increased 5-HT. The high affinity serotonin receptor um ese serotonerg decrease in the receptor affinity. The insulin ^treatmentsturtiy showsha decreased serotonergic receptor kinetic parameters to control level. receptor function. These changes in adrenergic and serotonergic receptor function were suggested to be important in insulin function during STZ diabetes.

Enhancement of [m-methoxy 3H]MDL100907 binding to 5HT 2A receptors in cerebral cortex and brain stem of streptozotocin induced diabetic rats

5-Hydroxytryptamine2A (5-HT2A) receptor kinetics was studied in cerebral cortex and brain stem of streptozotocin (STZ) induced diabetic rats. Scatchard analysis with [3H] (±) 2,3dimethoxyphenyl-l-[2-(4-piperidine)-methanol] ([3H]MDL100907) in cerebral cortex showed no significant change in maximal binding (Bmax) in diabetic rats compared to controls. Dissociation constant (K) of diabetic rats showed a significant decrease (p < 0.05) in cerebral cortex, which was reversed to normal by insulin treatment. Competition studies of [3H]MDL100907 binding in cerebral cortex with ketanserin showed the appearance of an additional low affinity site for 5-HT2A receptors in diabetic state, which was reversed to control pattern by insulin treatment. In brain stem, scatchard analysis showed a significant increase (p < 0.05) in Bmax accompanied by a significant increase (p < 0.05) in Kd. Competition analysis in brain stem also showed a shift in affinity towards a low affinity State for 5-HT2A receptors. All these parameters were reversed to control level by insulin treatment. These results show that in cerebral cortex there is an increase in affinity of 5-HT2A receptors without any change in its number and in the case of brain stem there is an increase in number of 5HT2A receptors accompanied by a decrease in its affinity during diabetes. Thus, from the results we suggest that the increase in affinity of 5-HT2A receptors in cerebral cortex and upregulation of 5-HT2A receptors in brain stem may lead to altered neuronal function in diabetes.