Serotonin agonists and antagonists in obstructive sleep apnea: therapeutic potential

Obstructive sleep apnea hypopnea syndrome (OSAHS) is a prevalent disorder associated with substantial cardiovascular and neurobehavioral morbidity. Yet this is a disorder for which there are no widely effective pharmacotherapies. The pathophysiology of obstructive sleep apnea namely, normal respiration in waking with disordered breathing only in sleep, suggests that this disorder should be readily amenable to drug therapy. Over the past 10 years, we have gained tremendous insight into the neurochemical mechanisms involved in state-dependent control of respiration. It is apparent from this work that there are many potential avenues for pharmacotherapies, including several seemingly conflicting directions for serotonergic therapies. Serotonin delivery is reduced to upper airway dilator motor neurons in sleep, and this contributes, at least in part, to sleep-related reductions in dilator muscle activity and upper airway obstruction. The dilator motor neuron post-synaptic serotonin receptors are 5-HT(2A) and 5-HT(2C) subtypes, and in adults the presynaptic 5-HT receptor in motor nuclei is 5-HT(1B), an inhibitory receptor. Serotonin receptors are also found within central respiratory neuronal groups, and these receptor subtypes include 5-HT(1A) (inhibitory) and 5-HT(2) receptors. Peripherally, stimulation of 5-HT(2A), 5-HT(2C) and 5-HT(3) receptor subtypes have an inhibitory effect on respiration via action at the nodose ganglion. Many of these receptor subtypes and their signal transduction pathways may be affected by oxidative stress in obstructive sleep apnea. These alterations will make finding drug therapies for sleep apnea more challenging, but not insurmountable. Future directions are suggested for elucidating safe, well-tolerated serotonergic drugs for this disorder. Tryptophan was one of the first serotonergic drugs tested for OSAHS. This drug was withdrawn from the market as a result of reports linking tryptophan use with eosinophilic myalgia syndrome and life-threatening pulmonary hypertension. Newer drugs with serotonergic activity tested in persons with sleep-disordered breathing include buspirone, fluoxetine and paroxetine. Trials are presently being conducted to evaluate the effects of 5-HT(2A) and 5-HT(3) antagonists on OSAHS. Many of the drugs tested have not shown significant improvement in sleep apnea. However, with continued effort to elucidate the pharmacology of neurochemical control of state-dependent changes in respiratory control, the availability of pharmacological therapy for this disorder is not too far away.     

Pharmacotherapies for obstructive sleep apnea: how close are we?

Obstructive sleep apnea is a prevalent disorder associated with significant neurobehavioral and cardiovascular morbidities. At present, however, there are no widely effective, well-tolerated pharmacotherapies for obstructive sleep apnea. The pathogenesis of this disorder predicts that sleep apnea should respond to drug therapies. Specifically, respiration during waking in persons with sleep apnea is normal, while collapse of the upper airway occurs exclusively in sleep. This state-dependency in upper airway patency suggests state-dependent changes in neurochemical control of the upper airway dilator motoneurons, and this, in turn, suggests that appropriate medications would maintain upper airway dilator function in sleep and prevent sleep related collapse of the upper airway. The past few years have brought significant insight into the neural mechanisms governing upper airway dilator muscle function. This article provides updates on neurochemical mechanisms, emphasizing a role for serotonergic control, and reviews recent drug therapy trials for sleep apnea. We are currently well poised to develop effective pharmacotherapies for obstructive sleep apnea, with opportunities to target several regions involved in respiratory control.     

Elevated anxiety and antidepressant-like responses in serotonin 5-HT1A receptor mutant mice

The brain serotonin (5-hydroxytryptamine; 5-HT) system is a powerful modulator of emotional processes and a target of medications used in the treatment of psychiatric disorders. To evaluate the contribution of serotonin 5-HT_1A receptors to the regulation of these processes, we have used gene-targeting technology to generate 5-HT_1A receptor-mutant mice. These animals lack functional 5-HT_1A receptors as indicated by receptor autoradiography and by resistance to the hypothermic effects of the 5-HT_1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). Homozygous mutants display a consistent pattern of responses indicative of elevated anxiety levels in open-field, elevated-zero maze, and novel-object assays. Moreover, they exhibit antidepressant-like responses in a tail-suspension assay. These results indicate that the targeted disruption of the 5-HT_1A receptor gene leads to heritable perturbations in the serotonergic regulation of emotional state. 5-HT_1A receptor-null mutant mice have potential as a model for investigating mechanisms through which serotonergic systems modulate affective state and mediate the actions of psychiatric drugs.     

Pharmacological characterization of serotonergic receptor activity in the hypoglossal nucleus

State-dependent reductions in serotonin delivery to upper airway dilator motoneuron activity may contribute to sleep apnea. The functional significance of serotonin receptor subtypes implicated in excitation of dilator motor neurons was evaluated in anesthetized, paralyzed, mechanically ventilated adult rats (n = 108). The effects of antagonists selective for serotonin receptor subtypes 2A, 2C, or 7 on intrinsic hypoglossal activity and on serotonin agonist (serotonin, 5-carboxamidotryptamine maleate, and RO-600175) dose responses were characterized. All drugs were injected unilaterally into the hypoglossal nucleus. The 2A antagonist, MDL-100907, dropped intrinsic hypoglossal nerve respiratory activity by 61 +/- 6% (p < 0.001) and suppressed serotonin excitation of hypoglossal nerve activity (p < 0.05). The 2C antagonist, SB-242084, dropped hypoglossal nerve activity 17 +/- 6% (p < 0.05) and suppressed the dose-response curve for the 2C agonist. Rapid desensitization occurred with the 2C agonist only (p < 0.05). The 7 antagonist, SB-269970, had no effect on either intrinsic activity or agonist responses. We conclude that serotonin 2A is the predominant excitatory serotonin receptor subtype at hypoglossal motor neurons. The serotonin 2C excitatory effects are of lower magnitude and are associated with rapid desensitization. There is no evidence for serotonin 7 activity in the hypoglossal nucleus. This characterization of serotonin receptor subtypes in the hypoglossal nucleus provides a focus for the development of pharmacotherapies for sleep apnea.     

Structural requirements for 5-HT2A and 5-HT1A serotonin receptor potentiation by the biologically active lipid oleamide

Oleamide is an endogenous fatty acid primary amide that possesses sleep-inducing properties in animals and that has been shown to effect serotonergic receptor responses and block gap junction communication. Herein, the potentiation of the 5-HT_1A receptor response is disclosed, and a study of the structural features of oleamide required for potentiation of the 5-HT_2A and 5-HT_1A response to serotonin (5-HT) is described. Of the naturally occurring fatty acids, the primary amide of oleic acid (oleamide) is the most effective at potentiating the 5-HT_2A receptor response. The structural features required for activity were found to be highly selective. The presence, position, and stereochemistry of the Δ⁹-cis double bond is required, and even subtle structural variations reduce or eliminate activity. Secondary or tertiary amides may replace the primary amide but follow a well defined relationship requiring small amide substituents, suggesting that the carboxamide serves as a hydrogen bond acceptor but not donor. Alternative modifications at the carboxamide as well as modifications of the methyl terminus or the hydrocarbon region spanning the carboxamide and double bond typically eliminate activity. A less extensive study of the 5-HT_1A potentiation revealed that it is more tolerant and accommodates a wider range of structural modifications. An interesting set of analogs was identified that inhibit rather than potentiate the 5-HT_2A, but not the 5-HT_1A, receptor response, further suggesting that such analogs may permit the selective modulation of serotonin receptor subtypes and even have opposing effects on the different subtypes.     

Drugs affecting serotonin receptors

A greater understanding of the various serotonin receptor subtypes has led to a clearer appreciation of the role of serotonin in gastrointestinal motility, sensation and secretion. Serotonin is definitely involved in the aetiopathogenesis of cisplatin-induced emesis and carcinoid diarrhoea. The application of serotonergic drugs in clinical therapeutics for gut disturbances is presently dominated by the use of 5-HT₃ antagonists for acute chemotherapy-induced nausea and vomiting, and the use of substituted benzamides which are 5-HT₄ agonists stimulating gut motor function through 5-HT₄ neuronal receptors. The best-studied 5-HT₄ agonist is cisapride, which has been shown to stimulate motility at several levels of the gut. Cisapride is approved for healing and maintenance treatment of reflux oesophagitis and is used in several countries for the alleviation of symptoms consistent with regional stasis, from dyspepsia to constipation.Carcinoid diarrhoea is a prototypic disease associated with deranged serotonin metabolism, and a rationale for using 5-HT₃ or 5-HT₄ antagonists is based on the recent appreciation of the important role of impaired gut motor function in carcinoid diarrhoea. In the future, greater understanding of the serotonin receptor subtypes and their role in gut disorders may lead to novel approaches to alleviate increased visceral perception of functional gastrointestinal disorders, to correct changes in colonic capacitance, or to alter gastrointestinal motility that contributes to diarrhoea or constipation. However, at the present time, it must be stressed that these uses are still at an experimental stage and that careful validation and proper controlled studies are still required.     

Perturbed dentate gyrus function in serotonin 5-HT2C receptor mutant mice

Serotonin systems have been implicated in the regulation of hippocampal function. Serotonin 5-HT_2C receptors are widely expressed throughout the hippocampal formation, and these receptors have been proposed to modulate synaptic plasticity in the visual cortex. To assess the contribution of 5-HT_2C receptors to the serotonergic regulation of hippocampal function, mice with a targeted 5-HT_2C-receptor gene mutation were examined. An examination of long-term potentiation at each of four principal regions of the hippocampal formation revealed a selective impairment restricted to medial perforant path–dentate gyrus synapses of mutant mice. This deficit was accompanied by abnormal performance in behavioral assays associated with dentate gyrus function. 5-HT_2C receptor mutants exhibited abnormal performance in the Morris water maze assay of spatial learning and reduced aversion to a novel environment. These deficits were selective and were not associated with a generalized learning deficit or with an impairment in the discrimination of spatial context. These results indicate that a genetic perturbation of serotonin receptor function can modulate dentate gyrus plasticity and that plasticity in this structure may contribute to neural mechanisms underlying hippocampus-dependent behaviors.     

阻塞性睡眠呼吸暂停低通气综合征患者睡眠结构紊乱与肝酶水平升高的相关性研究

目的 研究阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者的睡眠结构紊乱及其对肝酶水平的影响。方法 研究2010-2013年苏州大学附属第二医院神经内科收治的多导睡眠监测(PSG)确诊的OSAHS组和对照组的睡眠参数,且根据睡眠呼吸暂停和(或)低通气指数(AHI)将OSAHS组分为轻、中、重度三组,比较各组间肝酶及睡眠结构的变化,并将肝酶水平与睡眠结构指标进行相关性分析。结果 与对照组比较,OSAHS组患者的AHI、呼吸相关微觉醒指数、最长呼吸暂停时间、睡眠时间1+2期比例及血清丙氨酸转氨酶(ATL)、天冬氨酸转氨酶(ASL)、血清γ-谷氨酰转肽酶(γ-GGT)水平显著增高,最低血氧饱和度(L-SaO2)及慢波睡眠比例显著降低(均P<0.05)。OSAHS组各亚组间比较,AHI差异有统计学意义;与重度亚组比较,轻、中度亚组呼吸相关微觉醒指数、最长呼吸暂停时间、睡眠时间1+2期比例、慢波睡眠比例及血清ATL、ASL的变化均有统计学意义;与重度亚组比较,轻度亚组的血清γ-GGT显著升高(P<0.05)。偏相关分析显示,血清ALT、AST与睡眠1+2期比例呈正相关,血清ALT与慢波睡眠比例呈负相关,与呼吸睡眠微觉醒呈正相关(均P<0.05)。结论 OSAHS患者存在睡眠结构紊乱及肝脏损伤,其肝酶水平的升高与睡眠结构紊乱有关。     

Dynamics and Patterning of 5-Hydroxytryptamine 2 Subtype Receptors in JC Polyomavirus Entry

The organization and dynamics of plasma membrane receptors are a critical link in virus-receptor interactions, which finetune signaling efficiency and determine cellular responses during infection. Characterizing the mechanisms responsible for the active rearrangement and clustering of receptors may aid in developing novel strategies for the therapeutic treatment of viruses. Virus-receptor interactions are poorly understood at the nanoscale, yet they present an attractive target for the design of drugs and for the illumination of viral infection and pathogenesis. This study utilizes super-resolution microscopy and related techniques, which surpass traditional microscopy resolution limitations, to provide both a spatial and temporal assessment of the interactions of human JC polyomavirus (JCPyV) with 5-hydroxytrypamine 2 receptors (5-HT₂Rs) subtypes during viral entry. JCPyV causes asymptomatic kidney infection in the majority of the population and can cause fatal brain disease, and progressive multifocal leukoencephalopathy (PML), in immunocompromised individuals. Using Fluorescence Photoactivation Localization Microscopy (FPALM), the colocalization of JCPyV with 5-HT₂ receptor subtypes (5-HT_2A, 5-HT_2B, and 5-HT_2C) during viral attachment and viral entry was analyzed. JCPyV was found to significantly enhance the clustering of 5-HT₂ receptors during entry. Cluster analysis of infected cells reveals changes in 5-HT₂ receptor cluster attributes, and radial distribution function (RDF) analyses suggest a significant increase in the aggregation of JCPyV particles colocalized with 5-HT₂ receptor clusters in JCPyV-infected samples. These findings provide novel insights into receptor patterning during viral entry and highlight improved technologies for the future development of therapies for JCPyV infection as well as therapies for diseases involving 5-HT₂ receptors.     

Serotonin receptor subtypes influence prolactin secretion in the turkey

Serotonin (5-HT) stimulation of prolactin (PRL) secretion is mediated through the dopaminergic (DAergic) system, with 5-HT ligands having no direct effect on pituitary PRL release. Infusion of 5-HT into the third ventricle (ICV) or electrical stimulation (ES) of the medial preoptic area (POM) or the ventromedial nucleus (VMN) induces an increase in circulating PRL in the turkey. These increases in PRL do not occur when a selective antagonist blocks the D₁ dopamine (DA) receptors in the infundibular area (INF). In this study, the ICV infusion of (R)(−)-DOI hydrochloride (DOI), a selective 5-HT_2A eceptor agonist, caused PRL to increase. Pretreatment with Ketanserin tartrate salt (KETAN), a selective 5-HT_2A receptor antagonist, blocked DOI-induced PRL secretion, attesting to the specificity of the response. DOI-induced PRL secretion was prevented when the D₁ DA receptors in the INF were blocked by the D₁ DA receptor antagonist, R(+)-SCH-23390 hydrochloride microinjection, suggesting that the DAergic activation of the vasoactive intestinal peptide (VIP)/PRL system is mediated by a stimulatory 5-HT_2A receptor subtype. The DOI-induced PRL increase did not occur when (±)-8-OH-DPAT (DPAT) was concurrently infused. DPAT is a 5-T_1A receptor agonist which appears to mediate the inhibitory influence of 5-HT on PRL secretion. When DPAT was microinjected directly into the VMN, it blocked the PRL release affected by ES in the POM. These data suggested that when 5-HT_2A receptors are activated, they influence the release of DA to the INF. When 5-HT_1A receptors are stimulated, they somehow inhibit the PRL-releasing actions of 5-HT_2A receptors. This inhibition could take place centrally, or it could occur postsynaptically at the pituitary level. It is known that D₂ DA receptors in the pituitary antagonize PRL-releasing effect of VIP. A release of DA to the pituitary, initiated by 5-HT_1A receptors, could effectively inhibit PRL secretion.     

5-HT4 and 5-HT2 receptors antagonistically influence gap junctional coupling between rat auricular myocytes

5-hydroxytryptamine-4 (5-HT₄) receptors have been proposed to contribute to the generation of atrial fibrillation in human atrial myocytes, but it is unclear if these receptors are present in the hearts of small laboratory animals (e.g. rat). In this study, we examined presence and functionality of 5-HT₄ receptors in auricular myocytes of newborn rats and their possible involvement in regulation of gap junctional intercellular communication (GJIC, responsible for the cell-to-cell propagation of the cardiac excitation). Western-blotting assays showed that 5-HT₄ receptors were present and real-time RT-PCR analysis revealed that 5-HT_4b was the predominant isoform. Serotonin (1 μM) significantly reduced cAMP concentration unless a selective 5-HT₄ inhibitor (GR113808 or ML10375, both 1 μM) was present. Serotonin also reduced the amplitude of L-type calcium currents and influenced the strength of GJIC without modifying the phosphorylation profiles of the different channel-forming proteins or connexins (Cxs), namely Cx40, Cx43 and Cx45. GJIC was markedly increased when serotonin exposure occurred in presence of a 5-HT₄ inhibitor but strongly reduced when 5-HT_2A and 5-HT_2B receptors were inhibited, showing that activation of these receptors antagonistically regulated GJIC. The serotoninergic response was completely abolished when 5-HT₄, 5-HT_2A and 5-HT_2B were simultaneously inhibited. A 24 h serotonin exposure strongly reduced Cx40 expression whereas Cx45 was less affected and Cx43 still less. In conclusion, this study revealed that 5-HT₄ (mainly 5-HT_4b), 5-HT_2A and 5-HT_2B receptors coexisted in auricular myocytes of newborn rat, that 5-HT₄ activation reduced cAMP concentration, I_CaL and intercellular coupling whereas 5-HT_2A or 5-HT_2B activation conversely enhanced GJIC.     

Increased anxiety of mice lacking the serotonin1A receptor

Brain serotonin (5-HT) has been implicated in a number of physiological processes and pathological conditions. These effects are mediated by at least 14 different 5-HT receptors. We have inactivated the gene encoding the 5-HT_1A receptor in mice and found that receptor-deficient animals have an increased tendency to avoid a novel and fearful environment and to escape a stressful situation, behaviors consistent with an increased anxiety and stress response. Based on the role of the 5-HT_1A receptor in the feedback regulation of the 5-HT system, we hypothesize that an increased serotonergic neurotransmission is responsible for the anxiety-like behavior of receptor-deficient animals. This view is consistent with earlier studies showing that pharmacological activation of the 5-HT system is anxiogenic in animal models and also in humans.     

Treatment of Pregnant Alcohol-Consuming Rats with Buspirone: Effects on Serotonin and 5-Hydroxyindoleacetic Acid Content in Offspring

This laboratory previously demonstrated that in utero ethanol exposure markedly impairs the development of the serotonergic system in rat brain. Developmental abnormalities could be detected as early as G15 in the brainstem and G19 in the cortex. Because of the importance of fetal serotonin (5-HT) and 5-HT_1A receptors for the normal development of 5-HT containing neurons, we initiated studies to determine whether administration of a 5-HT_1A agonist, buspirone, to pregnant rats could overcome the adverse effects of in utero ethanol exposure on the developing serotonergic system in offspring. Female, Sprague-Dawley rats were given daily subcutaneous injections of buspirone (4.5 mg/kg) from gestational day 13 (G13) to G20. 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) content were determined in the cortex and cortical regions. These experiments demonstrated that the ethanol-associated abnormalities in the development of the serotonergic system can be partially overcome by in utero exposure to buspirone. Specifically, whereas untreated ethanol rats had a deficiency of 5-HT and/or 5-HIAA in whole cortex on PN5, and in the motor cortex on PN19 and 35, no significant differences were detected in these regions of the age-matched offspring of buspirone-treated, ethanol-fed rats. In contrast, the 5-HT and 5-HIAA deficiency in the somatosensory cortex of 19-dayold offspring of ethanol-fed rats was not corrected by in utero buspirone treatment. These results suggest that the abnormal development of cortical projections of serotonergic neurons may be due in part to the low fetal 5-HT content in ethanol-exposed rats and may potentially be overcome by in utero treatment with a 5-HT_1A agonist. However, additional studies are needed to elucidate the importance of dose and timing of buspirone administration, and the effects of in utero buspirone exposure on other components of serotonergic and nonserotonergic neurons.     

Traxoprodil,a selective antagonist of the NR2B subunit of the NMDA receptor,potentiates the antidepressant-like effects of certain antidepressant drugs in the forced swim test in mice

One of the newest substances, whose antidepressant activity was shown is traxoprodil, which is a selective antagonist of the NR2B subunit of the NMDA receptor. The main goal of the present study was to evaluate the effect of traxoprodil on animals’ behavior using the forced swim test (FST), as well as the effect of traxoprodil (10 mg/kg) on the activity of antidepressants, such as imipramine (15 mg/kg), fluoxetine (5 mg/kg), escitalopram (2 mg/kg) and reboxetine (2.5 mg/kg). Serotonergic lesion and experiment using the selective agonists of serotonin receptors 5-HT_1A and 5-HT₂ was conducted to evaluate the role of the serotonergic system in the antidepressant action of traxoprodil. Brain concentrations of tested agents were determined using HPLC. The results showed that traxoprodil at a dose of 20 and 40 mg/kg exhibited antidepressant activity in the FST and it was not related to changes in animals’ locomotor activity. Co-administration of traxoprodil with imipramine, fluoxetine or escitalopram, each in subtherapeutic doses, significantly affected the animals’ behavior in the FST and, what is important, these changes were not due to the severity of locomotor activity. The observed effect of traxoprodil is only partially associated with serotonergic system and is independent of the effect on the 5-HT_1A and 5-HT₂ serotonin receptors. The results of an attempt to assess the nature of the interaction between traxoprodil and the tested drugs show that in the case of joint administration of traxoprodil and fluoxetine, imipramine or escitalopram, there were interactions in the pharmacokinetic phase.     

Serotonin responsiveness through 5-HT2A and 5-HT4 receptors is differentially regulated in hypertrophic and failing rat cardiac ventricle

Cardiac ventricular responsiveness to serotonin appears in rat postinfarction congestive heart failure (CHF), mainly mediated by 5-HT₄ receptors in chronic dilated CHF and 5-HT_2A receptors in acute CHF. To differentiate between the effects of left ventricular (LV) hypertrophy and failure on 5-HT_2A- and 5-HT₄-mediated inotropic serotonin response, male Wistar rats with increasing LV hypertrophy (AB1-3) and failure (ABHF) 6 weeks after banding of the ascending aorta were screened for contractile function in vivo (echocardiography) and ex vivo in LV papillary muscles, and mRNA expression level determined by RT-PCR. Both AB1-3 and ABHF displayed LV hypertrophy and remodelling. In ABHF, systolic LV and left atrial diameter increased and cardiac output decreased compared to AB3. Serotonin induced a positive inotropic response (PIR) in papillary muscles correlated with the degree of hypertrophy reaching a maximum in ABHF. Both 5-HT_2A and 5-HT₄ receptors contributed to the PIR. The 5-HT_2A contribution increased with increasing hypertrophy, and the 5-HT₄ contribution increased upon transition to heart failure. No 5-HT_2B-mediated PIR was observed, consistent with increased 5-HT_2B mRNA only in non-cardiomyocytes. The 5-HT_2A, 5-HT_2B and 5-HT₄ mRNA levels increased in AB1-3 and increased further in ABHF compared to AB3, but did not correlate with degree of hypertrophy. 5-HT_2A mRNA was also increased in LV of terminally failing human hearts. In conclusion, functional 5-HT_2A and 5-HT₄ receptors are differentially induced in LV hypertrophy and failure. While the 5-HT_2A-mediated PIR is linearly correlated with the degree of hypertrophy, the 5-HT₄-mediated PIR seems to increase with LV dilatation, as also seen in postinfarction CHF.     

Serotonin and the GI tract

Serotonin (5-hydroxytryptamine, 5-HT) participates in several functions of the gastrointestinal tract. Receptors in seven families (5-HT₁-5-HT₇) were identified, many of which are present on enterocytes, intrinsic and extrinsic neurons, interstitial cells, and gut myocytes. Most 5-HT is released from enterochromaffin cells in response to physiologic and pathologic stimuli. Roles of 5-HT in health include control of normal gut motor activity, secretion, and sensation, and regulation of food intake and cell growth. Abnormalities of serotonergic function contribute to symptom genesis in functional bowel disorders, inflammatory and infectious diseases of the gut, emetic responses to varied stimuli, obesity, and dysregulation of cell growth. Therapies acting as agonists or antagonists of 5-HT receptors or that modulate 5-HT reuptake play prominent roles in managing these conditions, although use of many agents is hampered by cardiopulmonary complications. Novel agents are in testing, which may exhibit efficacy without significant toxicity.     

Adrenergic,dopaminergic and serotonergic gene expression in low dose,long time insulin and somatotropin treatment to ageing rats: rejuvenation of brain function

Somatotropin (GH) and insulin (INS) low dose, long-term brain rejuvenation effect was studied in the cerebral cortex using NE, EPI, DA and 5-HT receptor subtypes of young (group I—treatment started 4 weeks continued to 16 weeks) and old rats (group II—treatment started 60 weeks continued to 90 weeks). GH and INS treatment showed significant decrease in NE and EPI content in cerebral cortex of both young and old rats. α_2A-adrenergic receptors showed decreased expression whereas β₂-adrenergic receptors showed enhanced expression with age. GH and INS treatment significantly increased α_2A-adrenergic receptor protein in group I rats whereas INS treatment could increase β₂-adrenergic receptor protein expression in group II rats. DA and 5-HT content decrease with age. GH and INS administration showed increase in DA and 5-HT content in the brain regions-corpus striatum and brainstem of both young and old rats. Also, DA D₂ and 5-HT_2C receptor gene expression were increased significantly by GH and INS treatment in both young and old rats. In conclusion, low dose, long-term treatment of INS and GH to ageing rats improved the adrenergic, dopaminergic and serotonergic receptor subtypes activity and rejuvenation of brain function.     

The role in spawning of a putative serotonin receptor isolated from the germ and ciliary cells of the gonoduct in the gonad of the Japanese scallop, Patinopecten yessoensis

Serotonin (5-hydroxytriptamin, 5-HT) triggers germinal vesicle breakdown (GVBD) of oocytes and the transporting of the mature oocyte through the gonoduct via cilia motility in bivalves. The 5-HT receptor in the oocyte membrane of the Japanese scallop, Patinopecten yessoensis, has been pharmacologically characterized as a mixed profile of 5-HT₁/5-HT₂ and is induced by estradiol-17β (E₂). Here we report the isolation, cloning, and tissue expression of the 5-HT receptor from the gonad of the Japanese scallop. A full-length cDNA (1818 bp) encoding a putative 5-HT receptor (5-HT_py) of 454 amino acid residues was isolated from the ovary and shared 53.3% and 40.2% homology with the Aplysia 5-HT_1ap and mouse 5-HT_1A, respectively. The 5-HT_py sequence possessed typical characteristics of 5-HT₁, including seven transmembrane domains, a long third inner loop, and a short fourth inner terminal. Phylogenetic analysis suggested that 5-HT_py was classified into the 5-HT₁ subtype as well as other invertebrate 5-HT₁ receptors. Semi-quantitative RT-PCR showed the expression of the 5-HT_py gene in both the nervous system and peripheral tissues and the induction of expression by E₂ in the ovarian tissue. In situ hybridization revealed a strong 5-HT_py signal in the oocytes, spermatids, and ciliary epithelium of the gonoducts in the ovary and testis. These results suggest that the effects of 5-HT on the induction of oocyte maturation, sperm motility, and transport of mature oocytes and sperm through the ciliated epithelium of the gonoducts are mediated by 5-HT_py.