The neonatal treatment with clomipramine decreases sexual motivation and increases estrogen receptors expression in the septum of male rats: Effects of the apomorphine.

Administering clomipramine during the early days of life induced several behavioral and neurochemical alterations in adult male rats, which resemble major depression disorder. The alterations included poor sexual performance, which is considered a reward-seeking behavior regulated by dopaminergic system. Given that estrogen receptors are expressed in different areas of the brain involved in regulating reproductive behavior, motivation and mood. The objective of this study was to analyze the effect of a non-selective dopamine agonist (apomorphine) on sexual incentive motivation in rats exposed to clomipramine (CMI) in the neonatal period. In addition, we evaluated the expression of mRNA ERα and ERß in nucleus accumbens (NAcc) and septum of CMI rats. We found that only a few rats subjected to neonatal CMI treatment performed mounts, intromissions and ejaculations. Also, those rats spent less time exploring the sexual incentive zone and had lower preference scores; this effect was reverted by administering 0.1 mg/kg of apomorphine. Finally, the CMI rats presented higher levels of mRNA ERα and ERß, only in septum area. These data indicate that neonatal treatment with CMI altered the expression of mRNA ERα and ERß in the septum, which participates in regulating the motivational component of sexual behavior.

Widespread blunting of hypothalamic and amygdala-septal activity and behavior in rats with long-term hyperglycemia.

Anxiety and depression in diabetic patients contributes to a poor prognosis, but possible causal relationships have been controversial. Anxiety, fear, and anhedonia are mediated by interactions between different deep structures of the temporal lobe (e.g., amygdala complex and hippocampus) and other forebrain-related structures (e.g., lateral septal nucleus). Connections between these structures and the hypothalamic orexinergic system are necessary for the maintenance of energy and wakefulness. However, few studies have explored the impact of long-term hyperglycemia in these structures on anxiety. We induced long-term hyperglycemia (glucose levels of ∼500mg/dl) in Wistar rats by injecting them with alloxan and simultaneously protecting them from hyperglycemia by injecting them daily with a low dose of insulin (i.e., just enough insulin to avoid death), thus maintaining hyperglycemia and ketonuria for as long as 6 weeks. Compared with controls, long-term hyperglycemic rats exhibited a significant reduction of Fos expression in the lateral septal nucleus and basolateral amygdala, but no differences were found in cerebellar regions. Orexin-A cells appeared to be inactive in the lateral hypothalamus. No differences were found in sucrose consumption or behavior in the elevated plus maze compared with the control group, but a decrease in general locomotion was observed. These data indicate a generalized blunting of the metabolic brain response, accompanied by a decrease in locomotion but no changes in hedonic- or anxiety-like behavior.

The recombinant C-terminal fragment of tetanus toxin protects against cholinotoxicity by intraseptal injection of ß-amyloid peptide (25-35) in rats.

The recombinant C-terminal domain of tetanus toxin (Hc-TeTx) is a new non-toxic peptide of the tetanus toxin that exerts a protective action against glutamate excitotoxicity in motoneurons. Moreover, its efficacy as a neuroprotective agent has been demonstrated in several animal models of neurodegeneration. The eleven amino acids in the ß amyloid peptide (Aß25-35) mimic the toxic effects of the full ß amyloid peptide (Aß1-42), causing the impairment of the cholinergic system in the medial septum (MS) which, in turn, alters the septo-hippocampal pathway and leads to learning and memory impairments. The aim of this study was to examine the neuroprotective effects of the Hc-TeTx fragment against cholinotoxicity. The Hc-TeTx fragment (100 ng) was injected into the rats intercranially, with the Aß(25-35) (2 µg) then injected into their MS. The animals were tested for spatial learning and memory in the eight-arm radial maze. The brains were removed to assess cholinergic markers, such as choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), and to explore neurodegeneration in the MS and hippocampus, using amino-cupric silver and H&E staining. Finally, capase-3, a marker of apoptosis, was examined in the MS. Our results clearly demonstrate that the application of Hc-TeTx prevents the loss of cholinergic markers (ChAT and AChE), the activation of capase-3, and neurodegeneration in the MS and the CA1 and CA3 subfields of the hippocampus. All these improvements were reflected in spatial learning and memory performance, and were significantly higher compared with animals treated with Aß(25-35). Interestingly, the single administration of Hc-TeTx into the MS modified the ChAT and AChE expression that affect cognitive processes, without inducing neurodegeneration or an increase in capase-3 expression in the MS and hippocampus. In summary, our findings suggest that the recombinant Hc-TeTx fragment offers effective protection for the septo-hippocampal pathway, given that it reduces the neurodegeneration caused by Aß(25-35) and improves learning and memory processes.

Septal serotonin depletion in rats facilitates working memory in the radial arm maze and increases hippocampal high-frequency theta activity.

Hippocampal theta activity, which is strongly modulated by the septal medial/Broca׳s diagonal band neurons, has been linked to information processing of the hippocampus. Serotonin from the medial raphe nuclei desynchronises hippocampal theta activity, whereas inactivation or a lesion of this nucleus induces continuous and persistent theta activity in the hippocampus. Hippocampal serotonin depletion produces an increased expression of high-frequency theta activity concurrent with the facilitation of place learning in the Morris maze. The medial septum-diagonal band of Broca complex (MS/DBB) has been proposed as a key structure in the serotonin modulation of theta activity. We addressed whether serotonin depletion of the MS/DBB induces changes in the characteristics of hippocampal theta activity and whether the depletion is associated with learning in a working memory spatial task in the radial arm maze. Sprague Dawley rats were depleted of 5HT with the infusion of 5,7-dihydroxytriptamine (5,7-DHT) in MS/DBB and were subsequently trained in the standard test (win-shift) in the radial arm, while the CA1 EEG activity was simultaneously recorded through telemetry. The MS/DBB serotonin depletion induced a low level of expression of low-frequency (4.5-6.5Hz) and a higher expression of high-frequency (6.5-9.5Hz) theta activity concomitant to a minor number of errors committed by rats on the working memory test. Thus, the depletion of serotonin in the MS/DBB caused a facilitator effect on working memory and a predominance of high-frequency theta activity.

Blockade of 5-Ht3 receptors in the septal area increases Fos expression in selected brain areas.

Serotonin is widely distributed throughout the brain and is involved in a multiplicity of visceral, cognitive and behavioral responses. It has been previously shown that injections of different doses of ondansetron, a 5-HT3 receptor antagonist, into the medial septum/vertical limb of the diagonal band complex (MS/vDB) induce a hypertensive response in rats. On the other hand, administration of m-CPBG, a 5-HT3 agonist, into the MS/vDB inhibits the increase of blood pressure during restraint stress. However, it is unclear which neuronal circuitry is involved in these responses. The present study investigated Fos immunoreactive nuclei (Fos-IR) in different brain areas following the blockade of 5-HT3 receptors located in the MS/vDB in sham and in sinoaortic denervated (SAD) rats. Ondansetron injection into the MS/vDB increases Fos-IR in different brain areas including the limbic system (central amygdala and ventral part of the bed nucleus of the stria terminalis), hypothalamus (medial parvocellular parts of the paraventricular nucleus, anterodorsal preoptic area, dorsomedial hypothalamic nucleus), mesencephalon (ventrolateral periaqueductal gray region) and rhombencephalon (lateral parabrachial nucleus) in sham rats. Barodenervation results in higher Fos expression at the parvocellular and magnocellular part of the paraventricular nucleus, the lateral parabrachial nucleus, the central nucleus of amygdala, the locus coeruleus, the medial part of the nucleus of the solitary tract, the rostral ventrolateral medulla and the caudal ventrolateral medulla following 5-HT3receptor blockade in the MS/vDB. Based on the present results and previous data showing a hypertensive response to ondansetron injected into the MS/vDB, it is reasonable to suggest that 5-HT3receptors in the MS/vDB exert an inhibitory drive that may oscillate as a functional regulatory part of the complex central neuronal network participating in the control of blood pressure.

5-HT(2A) receptor activation in the dorsolateral septum facilitates inhibitory avoidance in the elevated T-maze.

Serotonin in the lateral septum has been implicated in the modulation of defense and hence in anxiety. However, it deserves investigation how changes in 5-HT-mechanisms in this area modulate defensive responses associated with specific subtypes of anxiety disorders. We evaluated the effects of intra-dorsolateral septum (DLS) injections of the preferential 5-HT(2A) receptor agonist DOI (8 and 16nmol), the 5-HT(2C) selective agonist MK-212 (0.1 and 1nmol) and the preferential 5-HT(2A) antagonist ketanserin (10 and 20nmol) in rats exposed to the elevated T-maze (ETM), a model which allows the measurement of two defensive responses: inhibitory avoidance and escape. These responses have been respectively related to generalized anxiety and panic disorder. All animals were tested in an open-field after the ETM for locomotor activity assessments. Results showed that intra-DLS DOI increased avoidance latencies, an anxiogenic effect. MK and ketanserin were without effect. Also, none of the drugs administered affected the escape performance. Ketanserin blocked the anxiogenic effect caused by DOI. No changes to locomotion were observed. The data suggests that DLS 5-HT(2A) receptors are involved in the control of inhibitory avoidance and that a failure in this mechanism may be of importance to the physiopathology of generalized anxiety.

Antidepressant-like effects of mineralocorticoid but not glucocorticoid antagonists in the lateral septum: interactions with the serotonergic system.

The lateral septum (LS) is a limbic brain region that receives serotonergic projections from raphe neurons and participates in the modulation of stress responses and affective states. The present study determined whether mineralocorticoid receptors (MRs) and/or glucocorticoid receptors (GRs) located in the LS interact with the serotonergic system in the regulation of depressive-like behavior of rats subjected to the forced swimming test (FST). We also studied the effect of corticosterone release induced by the FST on MR- and GR-mRNA expression in the LS. Specifically, we studied the antidepressant-like effects of spironolactone (a MR antagonist), mifepristone (a GR antagonist), and the antidepressant clomipramine (CMI) administered directly into the LS. In addition, spironolactone and CMI actions were studied in animals with serotonergic depletion induced by dl-p-chlorophenylalanine (pCPA). Finally, adrenalectomized and Sham-operated rats were subjected to the FST to determine MR- and GR-mRNA expression in the LS at different post-FST intervals. The results showed that intraseptal injection of spironolactone, but not mifepristone induced antidepressant-like actions in the FST; this effect was blocked by pCPA treatment. CMI and spironolactone increased 5-HT concentrations in the LS of rats subjected to the FST. Increases in corticosterone release, induced by the FST, correlated with a decrease in MR-mRNA expression in the LS; no correlation was found with GR-mRNA expression. In conclusion, MRs in the lateral septum, but not GRs, participate in the regulation of depressive-like behavior of animals subjected to the FST. Both serotonin and corticosterone play an important role in MR actions in the LS.

Medial septal area ANG II receptor subtypes in the regulation of urine and sodium excretion induced by vasopressin.

INTRODUCTION: The present study was designed to determine the effects of selective antagonists of angiotensin II receptor types AT(1) and AT(2) on the flow of urine and sodium excretion induced by arginine vasopressin (AVP). MATERIALS AND METHODS: To this end, the drugs were microinjected into the medial septal area (MSA) of the brains of male Holtzman rats. Intravenous infusion of hypotonic saline was used to promote urinary flow, which was collected for one hour. RESULTS: MSA microinjections of AVP decreased the urinary flow and increased sodium excretion in a dose-dependent manner. Microinjection into MSA of an AT(2) antagonist (PD-123319) had a significantly greater effect than with an AT(1) antagonist (losartan) in increasing urinary flow and decreasing sodium excretion. These effects were more pronounced when both antagonists were injected together, before the AVP. CONCLUSIONS: These results indicate that MSA AT(1) and AT(2) receptors act synergistically in the regulation of urine and sodium excretion induced by AVP.

Activation of GABA-B receptors induced by systemic amphetamine abolishes dopamine release in the rat lateral septum.

The lateral septum is a brain nucleus involved in various mental disorders such as anxiety and drug addiction. In the present study, we investigated whether systemic amphetamine, known to release dopamine (DA) in nucleus accumbens, will also release DA in lateral septum. Our results show that systemic amphetamine administration (2 mg/kg i.p.) induced a significant increase in DA extracellular levels in nucleus accumbens but not in lateral septum. Interestingly, intralateral septum perfusion of amphetamine through the microdialysis probe induced a significant increase in DA extracellular levels. To test if GABAergic neurotransmission in lateral septum was responsible for inhibiting the release of DA when amphetamine was administered systemically, we perfused a GABA-B selective antagonist (CGP-52432) intra lateral septum. Systemic amphetamine administration induced a significant increase in lateral septum DA release when CGP-52432 was concomitantly superfused. Our results indicate that the systemic administration of amphetamine induces an increase in lateral septum GABA release and the consequent activation of GABA-B receptors counteracting the direct effect of amphetamine on lateral septum DA release.

Papel dos receptores serotoninérgicos do tipo 5-HT3 localizados no núcleo septal medial/banda diagonal de broca sobre o controle da pressão arterial em ratos / Role of serotonin receptor type 5-HT3 receptors located in the medial septal nucleus / diagonal band drill over the control of blood pressure in rats

Diversos estudos têm mostrado a participação das vias serotoninérgicas tanto centrais, quanto periféricas no controle cardiovascular. Os principais receptores serotoninérgicos envolvidos na regulação cardiovascular são do tipo 5-HT1A, 5-HT2, 5-HT3 e 5-HT7. A ativação serotoninérgica em diferentes áreas cerebrais resulta tanto em aumento, quanto diminuição da pressão arterial. As principais áreas cerebrais relacionadas com os efeitos cardiovasculares induzidos pela estimulação serotoninérgica são: região rostro ventrolateral do bulbo, núcleo paraventricular do hipotálamo, núcleo dorsal motor do vago, núcleo do trato solitário. Entre as estruturas prosencefálicas, tem sido mostrado que o complexo núcleo septal mediallbanda diagonal de Broca (SM/vDB) desempenha importante papel no controle do equilíbrio hidrossalino e cardiovascular. Desde que esta área apresenta receptores serotoninérgicos 5-HT3 e recebe aferências serotoninérgicas do núcleo mediano da rafe, os objetivos do presente estudo foram: a) investigar o papel dos receptores 5-HT3 localizados no SM/vDB sobre o controle cardiovascular b) analisar o padrão de expressão de c-Fos no cérebro de ratos induzido pelo bloqueio farmacológico dos receptores 5-HT3 presentes no SM/vDB. Ratos Wistar (280-300g) anestesiados com cetamina/xilazina (80/11,5 mg/kg; i.p.) foram submetidos a implante de cânulas no SM/vDB. Os animais foram novamente anestesiados 48 h antes dos experimentos para cateterização da carótida e veia femoral. Na sessão experimental a pressão pulsátil (PAP) foi registrada durante 30 minutos para cálculo dos valores basais de pressão arterial média (PAM) e da freqüência cardíaca (FC). A seguir os animais receberam no SM/vDB microinjeçães de m-CPBG (160 nmol), ondansetrona (160, 80 e 40 nmol) ou NaCI 0,9%, em um volume de 0,2 /-lI e a PAP foi registrada durante mais 100 minutos...

Participação dos receptores serotoninérgicos do subtipo 5-HT³ localizados na área septal medial sobre o controle da ingestão de água e sal em ratos sódio-depletados / Participation of serotonin receptor subtype 5-HT ³ located in the medial septal area on the control of water intake and salt in sodium-depleted rats

A regulação do equilíbrio hidroelétrico é desempenhada por um conjunto de mecanismos viscerais e comportamentais, nos quais se incluem a ingestão de água e sal controlados por diversos neurotransmissores e áreas cerebrais. Estudos sugerem a participação de mecanismos Serotoninérgicos na indução das respostas fisiológicas relacionadas com a regulação da ingestão de água e sal. No presente estudo analisamos o envolvimento dos receptores 5-HT³ Serotoninérgicos localizados na ASM sobre a ingestão de água e sal em ratos sódio-depletados. Verificamos que a estimulação farmacológica específica dos receptores Serotoninérgicos 5-HT³ localizados na ASM, pelo agonista 5-HT³ seletivo m-CPBG, aumenta a ingestão de sal em ratos sódio-depletados. A especificidade deste efeito parece assegurada pelo fato de que o pré-tratamento com Ondansetrona, antagonista dos receptores Serotoninérgicos 5-HT³, aboliu completamente a elevação na ingestão de sal produzida pela administração do m-CPBG na ASM. Esse efeito Natriorexigênio parece não ser dependente de alterações na pressão arterial, visto que a micro-injeção na ASM do m-CPBG não foi capaz de modificar este parâmetro. Além disso, constatamos que a administração isolada do antagonista Ondasterona é incapaz de produzir efeito significativo sobre a ingestão de água e sal em ratos sódio-depletados. Tomando em conjunto os achados acima mencionados sugere-se que os receptores Serotoninérgicos do subtipo 5-HT³ localizados na ASM quando ativados farmacologicamente são capazes de ativar mecanismos centrais envolvidos com a procura e a ingestão de sódio.

Vasopressin and angiotensin receptors of the medial septal area of the brain in the control of thirst and salt appetite induced by vasopressin in water-deprived and sodium-depleted rats.

In this study we investigated the influence of d(CH(2))(5)-Tyr (Me)-AVP (A(1)AVP) and [Adamanteanacatyl(1),D-ET-D-Tyr(2), Val(4), aminobutyril(6),A(8,9)]-AVP (A(2)AVP), antagonists of V(1) and V(2) arginine(8)-vasopressin (AVP) receptors, respectively, as well as the effects of losartan and CGP42112A, antagonists of angiotensin II (ANGII) AT(1) and AT(2,) receptors, respectively, on water and 0.3 M sodium intake induced by water deprivation or sodium depletion (furosemide treatment) and enhanced by AVP injected into the medial septal area (MSA). A stainless steel cannula was implanted into the medial septal area (MSA) of male Holtzman rats AVP injection enhanced water and sodium intake in a dose-dependent manner. Pretreatment with V(1) antagonist injected into the MSA produced a dose-dependent reduction, whereas prior injection of V(2) antagonist increased, in a dose-dependent manner, the water and sodium responses elicited by the administration of AVP. Both AT(1) and AT(2) antagonists administered into the MSA elicited a concentration-dependent decrease in water and sodium intake induced by AVP, while simultaneous injection of the two antagonists was more effective in decreasing AVP responses. These results also indicate that the increase in water and sodium intake induced by AVP was mediated primarily by MSA AT(1) receptors.

c-Fos expression related to sexual satiety in the male rat forebrain.

The long term inhibition of masculine sexual behavior after repeated ejaculations is known as sexual satiety. To investigate the brain areas that may regulate sexual satiety, c-Fos expression was studied in different groups of sexually experienced male rats: controls not allowed to copulate, males allowed two or four ejaculations and animals allowed to reach sexual satiety. Interestingly, males that ejaculated two or four times had similar c-Fos densities in all the evaluated brain regions, except for the suprachiasmatic nucleus. Similarly, sexually satiated males had analogous c-Fos densities in all the evaluated brain areas independently of the number of ejaculations required to reach satiety. Sexual activity (evidenced in males that ejaculated two or four times) increased c-Fos levels in the anteromedial bed nucleus of the stria terminalis, claustrum, entorhinal cortex, medial preoptic area, nucleus accumbens core, suprachiasmatic nucleus and supraoptic nucleus; however, sexual satiety did not modify c-Fos expression in these regions. Sexually satiated males had increased c-Fos densities in the ventrolateral septum and the anterodorsal and posteroventral medial amygdala, compared with animals allowed to copulate but that did not reach sexual satiety, and decreased c-Fos density in the piriform cortex. These results suggest that the network that underlies sexual satiety is different from that which regulates copulation.

Memory consolidation and amnesia modify 5-HT6 receptors expression in rat brain: an autoradiographic study.

Traditionally, the search for memory circuits has been centered on examinations of amnesic and AD patients, cerebral lesions and, neuroimaging. A complementary alternative might be the use of autoradiography with radioligands. Indeed, ex vivo autoradiographic studies offer the advantage to detect functionally active receptors altered by pharmacological tools and memory formation. Hence, herein the 5-HT(6) receptor antagonist SB-399885 and the amnesic drugs scopolamine or dizocilpine were used to manipulate memory consolidation and 5-HT(6) receptors expression was determined by using [(3)H]-SB-258585. Thus, memory consolidation was impaired in scopolamine and dizocilpine treated groups relative to control vehicle but improved it in SB-399885-treated animals. SB-399885 improved memory consolidation seems to be associated with decreased 5-HT(6) receptors expression in 15 out 17 brain areas. Scopolamine or dizocilpine decreased 5-HT(6) receptors expression in nine different brain areas and increased it in CA3 hippocampus or other eight areas, respectively. In brain areas thought to be in charge of procedural memory such basal ganglia (i.e., nucleus accumbens, caudate putamen, and fundus striate) data showed that relative to control animals amnesic groups showed diminished (scopolamine) or augmented (dizocilpine) 5-HT(6) receptor expression. SB-399885 showing improved memory displayed an intermediate expression in these same brain regions. A similar intermediate expression occurs with regard to amygdala, septum, and some cortical areas in charge of explicit memory storage. However, relative to control group amnesic and SB-399885 rats in the hippocampus, region where explicit memory is formed, showed a complex 5-HT(6) receptors expression. In conclusion, these results indicate neural circuits underlying the effects of 5-HT(6) receptor antagonists in autoshaping task and offer some general clues about cognitive processes in general.

Acute morphine administration increases extracellular DA levels in the rat lateral septum by decreasing the GABAergic inhibitory tone in the ventral tegmental area.

We studied the effect of an acute systemic administration of morphine and of a local intra-ventral tegmental area (VTA) infusion of the same drug on extracellular levels of dopamine (DA) in the lateral septum (LS) by in vivo microdialysis in anesthetized rats. The extracellular levels of 5-hydroxytryptamine (5-HT) were also measured in all dialysate samples. The acute systemic administration of morphine dose-dependently increased extracellular levels of DA but not of 5-HT in the LS, in the absence or presence of fluoxetine. This morphine effect was antagonized by the previous administration of naloxone, a specific opioid antagonist. The local infusion of morphine in the VTA also induced a significant increase of the extracellular levels of DA in the LS, concomitantly with a decrease of gamma-aminobutyric acid (GABA) extracellular levels in the VTA itself. Intriguingly, the LS extracellular levels of DA returned to basal values before the VTA GABA extracellular levels recovered. Our results show for the first time that an acute administration of morphine increases DA extracellular levels in the LS. The results also suggest that DA cells in the VTA and innervating the LS are under an inhibitory GABAergic tone sensitive to morphine. Taken together, our neurochemical data and previous studies involving LS DA in stress-related behavior support the hypothesis that DA in the LS plays a significant role in addictive behavior. The participation of LS DA and 5-HT systems in stress-induced relapse to drug seeking should be studied further.

Corticotropin-releasing hormone and urocortin: redundant or distinctive functions?

Neuropeptides play important roles in synaptic transmission. Among them, the peptides of the corticotropin-releasing hormone (CRH) family present interesting features. The two main mammalian peptides of this family, CRH and urocortin (UCN), signal through the same receptors, CRH-R1 and CRH-R2. The question arises as to whether these peptides have redundant or distinctive functions. The fact that CRH and UCN have high affinity for both receptors has hampered the possibility to define the functional contribution of each peptide. Recent studies conducted on mice deficient in CRH, CRH-R1, CRH-R2 and CRH-R1/CRH-R2, as well as in two different UCN-deficient mice, have added relevant information towards the understanding of the role of this peptide family in the CNS. Our new anatomical evidence of UCN expression in the septum will be discussed in this context.

Effects of V1 and angiotensin receptor subtypes of the paraventricular nucleus on the water intake induced by vasopressin injected into the lateral septal area.

In this study, we investigated the influence of d(CH(2))(5)-Tyr (Me)-AVP (AAVP) an antagonist of V(1) receptors of arginine(8)-vasopressin (AVP) and the effects of losartan and CGP42112A (selective ligands of the AT(1) and AT(2) angiotensin receptors, respectively) injections into the paraventricular nucleus (PVN) on the thirst effects of AVP stimulation of the lateral septal area (LSA). AVP injection into the LSA increased the water intake in a dose-dependent manner. AAVP injected into the PVN produced a dose-dependent reduction of the drinking responses elicited by LSA administration of AVP. Both the AT(1) and AT(2) ligands administered into the PVN elicited a concentration-dependent inhibition in the water intake induced by AVP injected into the LSA, but losartan was more effective than CGP42112A the increase in the AVP response. These results indicate that LSA dipsogenic effects induced by AVP are mediated primarily by PVN AT(1) receptors. However, doses of losartan were more effective when combined with CGP42112A than when given alone, suggesting that the thirst induced by AVP injections into LSA may involve activation of multiple angiotensin II (ANG II) receptor subtypes. These results also suggests that facilitatory effects of AVP on water intake into the LSA are mediated through the activation of V(1)-receptors and that the inhibitory effect requires V(2)-receptors. Based on the present findings, we suggest that the administration of AVP into the LSA may play a role in the PVN control of water control.

Characterization of proliferation and differentiation of EGF-responsive striatal and septal precursor cells.

The epigenetic manipulation of precursors may provide data to elucidate the potential interactions among these cells in different brain regions. However, the response to epigenetic signals is modulated by the environment in which the cells are manipulated. Therefore, data regarding the action of a particular factor must be considered in the light of a specific system. To compare septal and striatal precursors, we have tested the effect of nerve growth factor (NGF) on the proliferation and neuronal differentiation of epidermal growth factor (EGF)-responsive cells from these brain regions. Precursors were cultivated as 'neuropheres' in serum free medium (SFM) to which NGF was added. NGF did not support the proliferation of EGF-generated precursors so that no differences in the cell magnitude with respect to control cultures were observed. Differentiation of precursors in SFM plus 1% fetal bovine serum (FBS) on poly-D-lysine showed that the neuron number was increased two-fold in septal cultures treated with NGF but not in those from striatum. A quantitative evaluation of the soma surface and the number of primary neurites showed differences between both populations of precursor-generated neurons. In addition, we also observed no influence of NGF on these parameters of cellular morphology. Thus, taken together these results seem to indicate that at this developmental stage in which these populations of precursors were isolated, heterogeneities exist between them, which is probably related to their origin and/or functional roles in vivo.

Dissociation of memory and anxiety in a repeated elevated plus maze paradigm: forebrain cholinergic mechanisms.

The effect of intraseptal injection of the cholinergic immunotoxin 192-IgG-saporin on behavior in the elevated plus maze was investigated. A 5-min test-retest paradigm, with minute-by-minute analysis of the first session, was used to evaluate both anxiety and memory in this task. Biochemical analyses revealed a decrease in acetylcholinesterase (AChE) activity in the hippocampus (HPC), septum, and frontal cortex of animals injected with IgG-192 saporin (237.5 ng) when compared with controls. No statistical differences were found between groups in terms of behaviors associated with locomotor activity, conventional measures of anxiety, or ethological behaviors during either session 1 or 2. During test session 2 the controls exhibited decreased exploratory activity and increased indices of anxiety. In contrast, the saporin-treated rats did not exhibit these experience-dependent behavioral changes from session 1 to 2. The minute-by-minute analysis showed a significant decrease in exploratory as well in anxiety associated behaviors during the first session for the control group, but not for the saporin-treated group. These results suggest that the cholinergic innervation of the HPC, the frontal cortex, or both forebrain structures, modulate the initiation of exploratory activity which, results in the acquisition and retention of spatial information, but does not affect the expression of anxiety in the elevated plus-maze.

Expression of the vesicular acetylcholine transporter, proteins involved in exocytosis, and functional calcium signaling in varicosities and soma of a murine septal cell line.

The expression and localization of the vesicular acetylcholine transporter in a septal cell line, SN56, were investigated. Immunoprecipitation and immunoblot analysis of postnuclear supernatants indicated that this cell line expresses reasonable amounts of the transporter. Immunofluorescence and confocal microscopy experiments showed that the vesicular transporter is present in varicosities and also in the cell body of differentiated cells. Varicosities have the potential to be functional sites of transmitter release because they responded to depolarization with calcium influx through voltage-gated calcium channels and expressed the synaptic proteins synaptotagmin, SV2, synaptophysin, and a subunit of P/Q calcium channels. In the soma of SN56 cells, the transporter immunoreactivity was similar to that for synaptotagmin, and it colocalized with synaptophysin, but it did not colocalize with SV2. Labeling for SV2 appeared prominently in a defined perinuclear structure, whereas the two former proteins were widely distributed in the soma, where several endocytic compartments could be identified with the vital dye FM4-64. These data suggest that distinct synaptic vesicle proteins exist in different subcellular compartments, and consequently they may follow distinct pathways in neurites before reaching sites of transmitter storage and release in SN56 cells.