Role of spinal alpha1-adrenoceptor subtypes in the bladder reflex in anesthetized rats

The contribution of different subtypes of alpha1-adrenoceptors in the lumbosacral spinal cord to the control of the urinary bladder was examined in urethane-anesthetized rats. Bladder pressure was recorded via a transurethral catheter under isovolumetric conditions. Drugs were administered intrathecally at the L6-S1 segmental level of spinal cord. RS-100329 (an alpha1A-antagonist) in doses of 25, 50, and 100 nmol significantly decreased bladder-contraction amplitude by 38%, 52%, and 95%, respectively, whereas (+)-cyclazosin (an alpha1B-antagonist) significantly decreased bladder-contraction amplitude (48% reduction) only in a 50-nmol but not a 100-nmol dose. Fifty nanomoles of RS-100329 and (+)-cyclazosin increased bladder-contraction frequency by 54% and 44%, respectively. BMY7378 (an alpha1D-antagonist), in doses of 25, 50, and 100 nmol, did not change bladder activity. These studies suggest that reflex-bladder activity is modulated by two types of spinal alpha1-adrenergic mechanisms: 1) alpha1A- or alpha1B-inhibitory control of the frequency of voiding reflexes presumably mediated by an alteration in the processing of bladder afferent input and 2) alpha(1A)-facilitatory modulation of the descending efferent limb of the micturition-reflex pathway. Spinal alpha1D-adrenoceptors do not appear to have a significant role at either site.     

A glycoprotein hormone expressed in corticotrophs exhibits unique binding properties on thyroid-stimulating hormone receptor

Corticotroph-derived glycoprotein hormone (CGH), also referred to as thyrostimulin, is a noncovalent heterodimer of glycoprotein hormone alpha 2 (GPHA2) and glycoprotein hormone beta 5 (GPHB5). Here, we demonstrate that both subunits of CGH are expressed in the corticotroph cells of the human anterior pituitary, as well as in skin, retina, and testis. CGH activates the TSH receptor (TSHR); (125)I-CGH binding to cells expressing TSHR is saturable, specific, and of high affinity. In competition studies, unlabeled CGH is a potent competitor for (125)I-TSH binding, whereas unlabeled TSH does not compete for (125)I-CGH binding. Binding and competition analyses are consistent with the presence of two binding sites on the TSHR transfected baby hamster kidney cells, one that can interact with either TSH or CGH, and another that binds CGH alone. Transgenic overexpression of GPHB5 in mice produces elevations in serum T(4) levels, reductions in body weight, and proptosis. However, neither transgenic overexpression of GPHA2 nor deletion of GPHB5 produces an overt phenotype in mice. In vivo administration of CGH to mice produces a dose-dependent hyperthyroid phenotype including elevation of T(4) and hypertrophy of cells within the inner adrenal cortex. However, the distinctive expression patterns and binding characteristics of CGH suggest that it has endogenous biological roles that are discrete from those of TSH.     

The nociceptin/orphanin FQ receptor ligand acetyl-RYYRIK-amide exhibits antagonistic and agonistic properties

The hexapeptide acetyl-RYYRIK-amide (Ac-RYYRIK-NH(2)) has recently been reported to act as partial agonist of the nociceptin/orphanin FQ (noc/OFQ) receptor expressed in CHO cells. In addition, this peptide acts as a competitive antagonist of noc/OFQ-stimulated GTPgamma(35)S binding in rat brain membranes as well as of the noc/OFQ-evoked chronotropic effect in rat cardiomyocytes. In contrast to this antagonism, in the present study, Ac-RYYRIK-NH(2) was found to behave as an agonist at noc/OFQ receptors, affecting spontaneous locomotor activity. When administered intracerebroventricularly (i.c.v.), noc/OFQ and Ac-RYYRIK-NH(2) inhibited spontaneous locomotor activity in mice with ID(50) of 1.1 and 0.07 nmol, respectively. Co-administration of both peptides lead to additive effects. The higher potency of Ac-RYYRIK-NH(2) could not be clearly explained by differential metabolism, because in vivo microdialysis in rat striatum and in vitro metabolic inactivation by rat and mouse brain membranes revealed extensive inactivation of both peptides. Similar to Ac-RYYRIK-NH(2), [Phe(1)psi(CH(2)-NH)Gly(2)]noc/OFQ(1-13)-NH(2) ([F/G]NC(1-13)NH(2)) inhibited the noc/OFQ-stimulated GTPgamma(35)S binding in rat brain membranes (Schild constant 3.83 nM) and mouse brain sections, although several reports have shown that this peptide exhibits agonist activity of noc/OFQ in the CNS. Changes in the optimum conditions of the in vitro assay for GTP binding increased low partial agonism of Ac-RYYRIK-NH(2) in GTP binding response. To explain the discrepancy between the in vitro antagonism of G protein coupling of the noc/OFQ receptor and in vivo agonism of Ac-RYYRIK-NH(2) and of [F/G]NC(1-13)NH(2), it is suggested that low partial agonism of receptor/G protein coupling in native systems may be sufficient to evoke full biologic responses. The extent of partial agonism for GTP binding and of coupling reserve may vary in different systems, thus explaining why [F/G]NC(1-13)NH(2) and Ac-RYYRIK-NH(2) were reported to exhibit antagonist, partial agonist, or even full agonist properties, depending on the system studied.     

Glucuronoxylomannan exhibits potent immunosuppressive properties

Cryptococcus neoformans is an opportunistic fungus responsible for life-threatening infections in immunocompromised and occasionally in immunocompetent hosts. The fungus is endowed with several virulence factors such as its capsular polysaccharide, which plays a key role in virulence. The major component of capsular material of C. neoformans is glucuronoxylomannan, a polysaccharide that exhibits potent immunosuppressive properties in vitro and in vivo. Here we describe a new aspect relative to glucuronoxylomannan-induced immunosuppression. In this review we analyze the suppressive effects of glucuronoxylomannan and the mechanisms leading to induction of apoptosis in T cells.     

Peroxisome proliferator-activated receptor alpha agonist-induced down-regulation of hepatic glucocorticoid receptor expression in SD rats

It was reported that glucocorticoid production was inhibited by fenofibrate through suppression of type-1 11β-hydroxysteroid dehydrogenase gene expression in liver. The inhibition might be a negative-feedback regulation of glucocorticoid receptor (GR) activity by peroxisome proliferator-activated receptor alpha (PPARα), which is quickly induced by glucocorticoid in the liver. However, it is not clear if GR expression is changed by fenofibrate-induced PPARα activation. In this study, we tested this possibility in the liver of Sprague-Dawley rats. GR expression was reduced by fenofibrate in a time- and does-dependent manner. The inhibition was observed in liver, but not in fat and muscle. The corticosterone level in the blood was increased significantly by fenofibrate. These effects of fenofibrate were abolished by PPARα inhibitor MK886, suggesting that fenofibrate activated through PPARα. In conclusion, inhibition of GR expression may represent a new molecular mechanism for the negative feedback regulation of GR activity by PPARα.     

The nuclear receptor coactivator PGC-1alpha exhibits modes of interaction with the estrogen receptor distinct from those of SRC-1

Estrogen receptor (ER) function is mediated by multi-domain co-regulator proteins. A fluorescently labelled fragment of the human PGC-1alpha co-regulator (residues 91-408) bearing the two motifs most strongly implicated in interactions with nuclear receptors (NR box2 and NR box3), was used to characterize in vitro binding of PGC-1alpha to ER. Anisotropy measurements revealed that the affinity of this PGC-1alpha fragment for human ERalpha and beta was fairly strong in the presence of estradiol (approximately 5 nM), and that unlike a similar fragment of SRC-1 (570-780), PGC-191-408 exhibited ligand-independent interactions with ER, particularly with ERbeta (Kd approximately 30 nM). Competition experiments of the complex between ERalpha and fluorescently labelled PGC-1 91-408 with unlabelled SRC-1 570-780 showed that PGC-1 91-408 was an efficient competitor of SRC-1 570-780, while the inverse was not true, underscoring their distinct modes of binding. The anisotropy data provide strong evidence for a ternary complex between ERalpha, SRC-1 570-780 and PGC-1 91-408. GST-pull-down experiments with deletion mutants of ERalpha revealed that the constitutive binding of PGC-1 91-408 requires the presence of the linker domain between the DNA binding and ligand binding domains (DBD and LBD). Homology modeling studies of the different regions of full length PGC-1alpha confirmed the lack of compact tertiary structure of the N-terminal region bearing the NR box motifs, and suggested a slightly different mode of interaction compared to the NR box motifs of SRC-1. They also provided reasonable structural models for the coiled-coil dimerization motif at residues 633-675, as well as the C-terminal putative RNA binding domain, raising important questions concerning the stoichiometry of its complex with the nuclear receptors.     

EOG responses in anesthetized freely breathing rats

In mammals, access of odor molecules to the olfactory receptor neurons is controlled by respiratory activity. Thus, anesthetized, freely breathing rats were used to record from the olfactory mucosa in the intact nasal cavity (electroolfactogram or EOG) so as to study global response characteristics to odor stimuli. During alternation of the inspiratory phases of odor sampling and expiratory phases, the response was a succession of individual EOG events synchronized with respiration. These were characterized by a steep decrease that started approximately 100-150 ms after the beginning of inhalation, reached its maximum at the transition between inspiration and expiration and was followed by a slower rise until the next inhalation. They were greater during the first respiratory cycles following odor stimulation onset. Thereafter their amplitudes decreased throughout odor delivery, but a significant EOG signal was still present at the end of short (10 s) and long (60 s) odor presentations. Amplitude increased with odor concentration, but much less than expected from concentration changes. Lastly, for some odors EOG responses persisted well beyond the end of stimulation. These results are in agreement with the respiratory synchronization of mitral cell activities observed during short odor presentations and long duration odor exposures. They underline again the importance of taking into account the respiratory activity in studies on the functioning of the olfactory system.     

Increased estrogen receptor alpha immunoreactivity in the forebrain of sexually satiated rats

Estrogen receptor alpha (ERalpha) participates in the neuroendocrine regulation of male sexual behavior, primarily in brain areas located in the limbic system. Males of many species present a long-term inhibition of sexual behavior after several ejaculations, known as sexual satiety. It has been shown that androgen receptor density is reduced 24 h after a single ejaculation or mating to satiety, in the medial preoptic area, nucleus accumbens and ventromedial hypothalamus. The aim of this study was to analyze if the density of ERalpha was also modified 24 h after a single ejaculation or mating to satiety. Sexual satiety was associated with an increased ERalpha density in the anteromedial bed nucleus of the stria terminalis (BSTMA), ventrolateral septum (LSV), posterodorsal medial amygdala (MePD), medial preoptic area (MPA) and nucleus accumbens core (NAc). A single ejaculation was related to an increase in ERalpha density in the BSTMA and MePD. ERalpha density in the arcuate (Arc) and ventromedial hypothalamic nuclei (VMN), and serum estradiol levels remained unchanged 24 h after one ejaculation or mating to satiety. These data suggest a relationship between sexual activity and an increase in the expression of ERalpha in specific brain areas, independently of estradiol levels in systemic circulation.     

Downregulation of alpha7 nicotinic acetylcholine receptor in two-kidney one-clip hypertensive rats

ABSTRACT: BACKGROUND: Inflammation processes are important participants in the pathophysiology of hypertension and cardiovascular diseases. The role of the alpha7 nicotinic acetylcholine receptor (alpha7nAChR) in inflammation has recently been identified. Our previous study has demonstrated that the alpha7nAChR-mediated cholinergic anti-inflammatory pathway is impaired systemically in the genetic model of hypertension. In this work, we investigated the changes of alpha7nAChR expression in a model of secondary hypertension. METHODS: The 2-kidney 1-clip (2K1C) hypertensive rat model was used. Blood pressure, vagus nerve function, serum tumor necrosis factor-alpha (TNF-alpha) and both the mRNA and protein levels of alpha7nAChR in tissues from heart, kidney and aorta were measured at 4, 8 and 20 weeks after surgery. RESULTS: Compared with age-matched control, it was found that vagus nerve function was significantly decreased in 2K1C rats with the development of hypertension. Serum levels of TNF-alpha were greater in 2K1C rats than in age-matched control at 4, 8 and 20 weeks. alpha7nAChR mRNA in the heart was not altered in 2K1C rats. In the kidney of 2K1C rats, alpha7nAChR expression was significantly decreased at 8 and 20 weeks, but markedly increased at 4 weeks. alpha7nAChR mRNA was less in aorta of 2K1C rats than in age-matched control at 4, 8 and 20 weeks. These findings were confirmed at the protein levels of alpha7nAChR. CONCLUSIONS: Our results suggested that secondary hypertension may induce alpha7nAChR downregulation, and the decreased expression of alpha7nAChR may contribute to inflammation in 2K1C hypertension.     

缓激肽抑制麻醉大鼠颈动脉窦压力感受器反射

在36只隔离灌流左侧颈动脉窦区的麻醉大鼠上观察了缓激肽(bradykinin,BK)对颈动脉窦压力感受器反射的影响.结果如下:(1)以BK (1.0μmol/L)隔离灌流大鼠颈动脉窦区时,压力感受器机能曲线向右上方移位,曲线的最大斜率(peak slope,PS)由0.44±0.14降至0.31±0.01(P<0.01),反射性血压下降幅度(reflex decrease,RD)由6.85±0.18 kPa降至4.46±0.16 kPa (P<0.01),阈压(TP)由7.76±0.20增至10.04±0.09kPa (P<0.05),其中RD,PS和TP的变化呈明显的剂量依赖性.(2)用环氧酶抑制剂消炎痛(indomethacin,10 μmol/L)预处理后,对BK (1.0μmol/L)抑制压力感受器的作用无影响; (3)预先灌流NO合酶阻断剂(L-NAME,100μmol/L),则可完全消除BK (1.0μmol/L)对压力感受器反射的抑制效应; (4)预先给予转换酶抑制剂(captopril,20μmol/L),可加强BK对压力感受器反射的抑制作用.以上结果表明:BK对大鼠颈动脉窦压力感受器反射有抑制作用,此作用系BK引起血管内皮细胞生成NO所致.     

Renal effects of SK&F 82526 in anesthetized rats

Dopamine affects renal hemodynamics, renal tubular functions, and the secretion of renin. We have studied the renal effects of SK&F 82526 (an agonist which is selective for the DA1 subclass of dopamine receptors) in anesthetized rats. Infused intravenously at 0.005 mumol/min/kg, this drug increased renal plasma flow and the clearances of PAH and insulin, effects which are consistent with decreased renovascular resistance. Concomitantly, urine flow and K excretion increased, and Na excretion tended to increase. All these effects of SK&F 82526 were antagonized by intravenous metoclopramide (1 mumol/min/kg). Despite its diuretic effect and despite its lack of effect on arterial blood pressure, SK&F 82526 increased arterial plasma renin concentration, suggesting a stimulatory effect on renin secretory rate. Taken together, our results demonstrate that the renal effects of SK&F 82526 mimic those of dopamine.     

辣椒素对颈动脉窦压力感受性反射的易化作用

在30只隔离灌流颈动脉窦区的麻醉大鼠,观察了辣椒素(capsaicin,CAP)对颈动脉窦压力感受性反射的影响.结果显示(1)以CAP(1 μmol/L)隔离灌流大鼠左侧颈动脉窦区时,颈动脉窦压力感受性机能曲线向左下方移位,曲线最大斜率(peak slope,PS)由0.34±0.01增至0.42±0.01(P＜0.01),反射性血压下降幅度(reflex decrease,RD)由36.51±1.26增至45,01±0.71 mmHg(P＜0.01).阈压、平衡压和饱和压分别从70.43±2.09、95.5±1.71和177.60±1.37 mmHg下降至52.86±2.80、87.00±1.58、163.55±2.12 mmHg(P＜0.01).其中PS和RD的变化呈明显的剂量依赖性.(2)用香草酸受体亚型(vanilloid receptor subtypel,VRl)阻断剂钌红(ruthenium red,100 μmo1/L)预处理后,CAP的上述反射效应即被阻断.(3)先给予KArp通道阻断剂格列苯脲(glibenclamide,20 μmo1/L)也取消了CAP对压力感受性反射的影响.结果表明,CAP对大鼠颈动脉窦压力感受性反射有易化作用,此作用似与VR1介导的KATP通道开放有关.     

Regulation of estrogen receptor alpha by estradiol in pregnant and estradiol treated rats

Estrogens play an important role in tissue metabolism through specific regulation of several intracellular pathways. We studied ERalpha regulation in muscle and adipose tissue from pregnant and estradiol treated rats. In both groups, we identified three different ERalpha inmunoreactive proteins (80, 67 and 46 kDa) using total protein extracts. Because it has been showed that estrogens are able to promote rapid effects in several cellular models, we looked for three ERalpha-related proteins at plasma membrane. In skeletal muscle of both groups, we positively identified the three ERalpha-related isoforms in plasma membrane, but in adipose tissue from pregnant we were not able to identify ERalpha67, and in estradiol treated animals ERalpha80 was absent. Taking together, our results showed a tissue-specific regulation of whole-cell ERalpha-related proteins and ERalpha located at plasma membrane, which should be involved in non-genomic actions of 17beta-estradiol. The role of the three ERalpha inmunoreactive proteins is unknown, however, seems probably related to rapid activation of signalling pathways.     

链霉素对颈动脉窦压力感受器反射的抑制作用

在 2 3只隔离灌流颈动脉窦区的麻醉大鼠 ,观察了链霉素对颈动脉窦压力感受器反射的影响。结果如下 :(1)以链霉素 (10 0 μmol/L)隔离灌流大鼠左侧颈动脉窦区时 ,压力感受器反射机能曲线向左下方移位 ,曲线最大斜率 (PS)由 0 40± 0 0 1kPa降至 0 33± 0 0 1kPa (P <0 0 0 1) ,血压反射性下降 (reflexdecrease ,RD)幅度由 6 2 2±0 13kPa降至 5 0 2± 0 11kPa (P <0 0 0 1) ,阈压 (TP)、平衡压 (EP)和饱和压 (SP)则分别从 8 2 7± 0 2 5 ,12 71± 0 2 1和 2 4 41± 0 14kPa增至 10 33± 0 32 (P <0 0 1) ,13 33± 0 30 (P <0 0 1)和 2 6 11± 0 2 8kPa (P <0 0 1)。其中RD ,PS和TP的变化呈明显的剂量依赖性。 (2 )应用腺苷隔离灌流大鼠颈动脉窦区 ,引起颈动脉窦压力感受器反射的易化 ;在用链霉素预处理后 ,此易化效应不仅完全被阻断 ,且可使反射效应小于应用腺苷前的对照值。以上结果表明 ,链霉素对大鼠颈动脉窦压力感受器反射有明显的抑制作用。     

Exercise pressor reflex in decerebrate and anesthetized rats

I investigated whether muscular contraction evokes cardiorespiratory increases (exercise pressor reflex) in alpha-chloralose- and chloral hydrate-anesthetized and precollicular, midcollicular, and postcollicular decerebrated rats. Mean arterial pressure (MAP), heart rate (HR), and minute ventilation (Ve) were recorded before and during 1-min sciatic nerve stimulation, which induced static contraction of the triceps surae muscles, and during 1-min stretch of the calcaneal tendon, which selectively stimulated mechanosensitive receptors in the muscles. Anesthetized rats showed various patterns of MAP response to both stimuli, i.e., biphasic, depressor, pressor, and no response. Sciatic nerve stimulation to muscle in precollicular decerebrated rats always evoked spontaneous running, so the exercise pressor reflex was not determined from these preparations. None of the postcollicular decerebrated rats showed a MAP response or spontaneous running. Midcollicular decerebrated rats consistently showed biphasic blood pressure response to both stimulations. The increases in MAP, HR, and Ve were related to the tension developed. The static contractions in midcollicular decerebrated rats (381 +/- 65 g developed tension) significantly increased MAP, HR, and Ve from 103 +/- 12 to 119 +/- 24 mmHg, from 386 +/- 30 to 406 +/- 83 beats/min, and from 122 +/- 7 to 133 +/- 25 ml/min, respectively. After paralysis, sciatic nerve stimulation had no effect on MAP, HR, or Ve. These results indicate that the midcollicular decerebrated rat can be a model for the study of the exercise pressor reflex.     

Spinal proerectile effect of oxytocin in anesthetized rats

The spinal cord contains the neural network that controls penile erection. This network is activated by information from peripheral and supraspinal origin. We tested the hypothesis that oxytocin (OT), released at the lumbosacral spinal cord level by descending projections from the paraventricular nucleus, regulated penile erection. In anesthetized male rats, blood pressure and intracavernous pressure (ICP) were monitored. Intrathecal (it) injection of cumulative doses of OT and the selective OT agonist [Thr(4),Gly(7)]OT at the lumbosacral level elicited ICP rises whose number, amplitude, and area were dose dependent. Thirty nanograms of OT and one-hundred nanograms of the agonist displayed the greatest proerectile effects. Single injections of OT also elicited ICP rises. Preliminary injection of a specific OT-receptor antagonist, hexamethonium, or bilateral pelvic nerve section impaired the effects of OT injected it. NaCl and vasopressin injected it at the lumbosacral level and OT injected it at the thoracolumbar level or intravenously had no effect on ICP. The results demonstrate that OT, acting at the lumbosacral spinal cord, elicits ICP rises in anesthetized rats. They suggest that OT, released on physiological activation of the PVN in a sexually relevant context, is a potent activator of spinal proerectile neurons.