Biomedical subjects

A Arce

Publications and source records attributed to A Arce.

At least 37 records · Page 2Linked to original sources

PubMed1998

Development of 24-hour rhythms in serum prolactin and luteinizing hormone levels in rats neonatally administered melatonin.

To obtain information on the chronobiological consequences of neonatal melatonin administration to rats, we examined the 24h variations of serum prolactin and luteinizing hormone (LH) levels in 21-, 25-, and 60-day-old male rats injected with 100 microg of melatonin or vehicle on the fifth day of life and kept under light from 08:00 to 20:00 daily. In 21-day-old rats, circulating prolactin varied diurnally in melatonin-treated rats only, showing a maximum at 16:00 and a secondary peak at 08:00. In 25-day-old rats, both vehicle- and melatonin-injected rats exhibited significant time-of-day-related changes in serum prolactin, with 77% of the variation in melatonin-injected rats explained by a single 24h component. Acrophase for prolactin rhythm of melatonin-treated rats occurred at 19:51. At 60 days of age, rats receiving vehicle or melatonin showed similar, significant 24h variations in serum prolactin, with acrophases at 17:31-18:15. Serum LH levels attained a maximum at midnight in 21-day-old rats treated with vehicle and at noon in those injected with melatonin. After a cosinor analysis, 71% of the variation in melatonin-injected rats was explained by a single 24h component, with the acrophase at 11:08. In 25-day-old rats, a complex pattern of serum LH concentration was found, with two maxima in vehicle-injected controls, at noon and midnight, and a maximum at noon together with a plateau from 20:00 to 04:00 in melatonin-treated rats. In 60-day-old rats, the maximum of serum LH was found at noon, with a similar extent showing in vehicle- and melatonin-treated animals. The acrophases were at 10:20 (vehicle) and 10:57 (melatonin) and did not differ significantly from that found in 21-day-old melatonin-treated rats. The results suggest that exposure to melatonin early in life may accelerate maturation of 24h prolactin and LH profiles toward an adult form.

Animals↗

PubMed1998

Effects of immunoneutralization of substance P on hypothalamic neurotransmitters in normal mice and in transgenic mice expressing bovine growth hormone.

It is well known that transgenic mice expressing bovine growth hormone have altered neuroendocrine functions. Substance P was shown to influence the secretion of gonadotropins. In this investigation, the effect of a single injection of an antiserum to substance P was investigated in intact and castrated transgenic (MT-bGH) mice and in their normal litter mates. In the median eminence, the administration of antisubstance P serum resulted in a decreased dihydroxyphenyl acetic acid/dopamine index in intact and castrated normal mice but was without effect in transgenics. The homovanillic/dopamine index was decreased in normal mice (intact or castrated) but unchanged in transgenics. Norepinephrine and epinephrine were increased in normal mice (intact and castrated) treated with the anti-SP serum, but in transgenic mice, the anti-SP serum induced significant changes of norepinephrine only in intact animals, with no modifications in epinephrine levels. In the whole hypothalamus (minus the median eminence), the injection of antisubstance P serum resulted in an increased dihydroxyphenyl acetic acid/dopamine index in castrated, but not in intact, normal mice. In transgenic mice, this index was increased in intact but decreased in castrated animals. The homovanillic/dopamine index was decreased in normal intact mice treated with the antiserum but increased in intact transgenic mice. Norepinephrine and epinephrine were decreased by the antiserum treatment in normal intact mice but were unchanged in transgenics, except for norepinephrine in castrated transgenics, in which it was found increased. The administration of the antiserum did not affect plasma LH, FSH, or prolactin in normal mice but it reduced LH levels in intact transgenic mice. These results indicate that the response to the treatment with the antiserum to substance P shows considerable alterations in transgenic mice as compared with their litter-mate, normal controls, producing divergent effects on hypothalamic catecholamine metabolism. The present findings confirm that transgenic mice overexpressing the bGH gene have marked neuroendocrine alterations as compared with their normal litter mates.

Animals↗

PubMed1998

Effects of acute and chronic administration of cyclosporine on dopamine metabolism in the rat cochlea.

The effect of cyclosporine (CyA) on dopamine (DA) metabolism at the cochlear level was analyzed. Adult male rats were submitted to CyA treatment at the doses of 1, 5 or 20 mg/Kg/day, for 1 day (acute) or 8 days (chronic). Cochlear contents of DA and its metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were measured by using high performance liquid chromatography (HPLC-ED). Either dose of acutely administered CyA did not modify cochlear DA content and markedly reduced that of DOPAC, in a non dose-dependent way. Acute administration of 5 mg/Kg of CyA decreased HVA content while the highest dose increased it. DOPAC/DA index was significantly reduced with either CyA dose, although HVA/DA index was not modified. Chronic treatment with CyA markedly reduced cochlear DA and DOPAC contents in a non dose-dependent way. However, HVA content decreased after the highest administration dose of the drug. DOPAC/DA index was further reduced after the drug chronic administration. An increased HVA/DA index was surprisingly observed, after chronic administration of either dose of the drug, the response being dose- dependent. These data show that acute treatment with CyA mainly affects the DA reuptake, while chronic treatment affected both DA reuptake and metabolism at the cochlear level.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1998

Circadian rhythms in adenohypophysial hormone levels and hypothalamic monoamine turnover in mycobacterial-adjuvant-injected rats

The effect of Freund's adjuvant injection on 24-hour variation in circulating ACTH, prolactin, growth hormone (GH) and thyroid-stimulating hormone (TSH) levels, and of hypothalamic norepinephrine (NE) content and dopamine (DA) and serotonin (5HT) turnover was examined in adult rats. In control rats, serum ACTH and prolactin exhibited peak values at the light-dark transition while the maximum in TSH was found in the late afternoon. GH levels did not vary on a 24-hour basis. In Freund's-adjuvant-injected rats, 24-hour variations in TSH levels became blunted while 24-hour variations in prolactin and ACTH persisted. Freund's adjuvant treatment augmented serum ACTH and prolactin levels, and decreased GH and TSH levels. Hypothalamic NE content, and turnover of DA and 5HT varied on a 24-hour basis in rats receiving adjuvant's vehicle. The NE content of the anterior, medial and posterior hypothalamus peaked at 04.00 h, while that of the median eminence attained its maximum at 16.00-20.00 h. Maxima in hypothalamic DA and 5HT turnover ocurred at 04.00 h regardless of the region examined. In Freund's-adjuvant-injected rats, reduced amplitude of daily variations of NE content in the median eminence and anterior and medial hypothalamus, as well as a phase advance in the 24-hour rhythm of the posterior hypothalamic NE content were seen. Mycobacterial adjuvant injection also reduced the amplitude of circadian rhythm in hypothalamic 5HT turnover, shifted the maximum in median eminence DA turnover towards light-dark transition, and decreased the amplitude of DA turnover rhythm in the anterior, medial and posterior hypothalamus. Administration of the immunosuppressant drug cyclosporine restored the augmented ACTH and prolactin levels and the depressed GH and TSH levels found in Freund's-adjuvant-injected rats. Cyclosporine was also effective to restore 24-hour rhythmicity of serum ACTH and TSH, but not of prolactin levels. Immunosuppression restored rhythmicity of NE content and of DA and 5HT turnover in anterior, medial and posterior hypothalamic regions. Cyclosporine did not modify the effect of Freund's adjuvant on median eminence but in was able to counteract the changes in the DA and 5HT turnover in the median eminence found after immunization. The results are in accord with a significant effect of immune-mediated inflammatory response at an early phase after Freund's adjuvant injection on ACTH, GH, prolactin and TSH release mechanisms, which was partially sensitive to immunosuppression induced by cyclosporine.

Journal Article↗

PubMed1998

Acute and chronic effects of superior cervical ganglionectomy on in vitro mitogenic responses of lymphocytes from submaxillary lymph nodes of pituitary-grafted rats

Male rats were grafted an anterior pituitary within breast muscles or received a sham operation on day 5 of life. At the 60th day of life, the sympathetic denervation of rat submaxillary lymph nodes was achieved by a bilateral sympathetic superior cervical ganglionectomy (SCGx; at 15.00 h). Rats were killed either 18 h later (acute SCGx) or after 12 days (chronic SCGx) to measure lipopolysaccharide (LPS)- and concanavalin A (ConA)-induced cell proliferation in submaxillary lymph nodes, submaxillary lymph node cellularity and serum prolactin levels. In control rats, acute SCGx significantly augmented LPS or ConA activity on lymph cells while chronic SCGx had no effect. In pituitary-grafted rats, acute SCGx depressed the mitogenic effect of LPS or ConA whereas chronic SCGx augmented it. A global inhibitory effect of surgical stress on submaxillary lymph node cellularity was found in rats subjected to SCGx or its sham operation 18 h earlier. Serum prolactin levels increased significantly in pituitary-grafted rats, particularly in those subjected to chronic SCGx. In pituitary-grafted rats, a significant effect of acute SCGx was apparent, with serum prolactin levels augmenting about twice in sham-SCGx rats, and to a significantly less extent in acute SCGx rats. The results provide further evidence of the immunomodulatory role of local sympathetic nerves in submaxillary lymph nodes.

Journal Article↗

PubMed1997

Learning and novelty induced increase of central benzodiazepine receptors from chick forebrain, in a food discrimination task.

Young chicks were trained to discriminate food grains from inedible pebbles. On Day 1 and Day 2 of the task, latency to peck, and number of pecks were scored and the forebrain [3H]flunitrazepam receptor binding was also determined at 0 and 30 min after an 8-min training session. Compared with quiet controls, the receptor density increased 46%, 30 min after the training session on Day 1. Compared with chicks that had learned the discrimination and were merely repeating already learned behavior on Day 2, the receptor increased more than 46%. Since chicks that had learned the discrimination had a higher behavioral activity, we interpret that the learning of a new task is itself responsible in addition to stress for the receptor density increase. Stressful factors accompanying the learning task as handling and novelty increased 17% the receptor density, 30 min after a training session without food, compared with quiet controls. However, receptor density did not increase in chicks repeating the same housing conditions, suggesting that chicks were habituated to handling and novelty on Day 2. Differences in receptor density were not observed between quiet controls and experimental groups, at 0 min after the training session, indicating that changes were time dependent. In all cases the affinity remained unchanged. Our results suggest that, the GABA(A) receptor (i) is involved in early stages of memory formation and in stress adaptive responses, and (ii) is modulated by new non-repetitive environmental conditions.

Animals↗

PubMed1997

Effect of local autonomic denervation on in vitro responsiveness of lymphocytes from rat submaxillary lymph nodes.

The local autonomic denervation of rat submaxillary lymph nodes was achieved by a unilateral sympathetic superior cervical ganglionectomy and/or the unilateral section of chorda tympani (that resulted in ipsilateral parasympathetic decentralization of the submandibular territory). This study was performed to determine: (1) whether local sympathetic and/or parasympathetic denervation of rat submaxillary lymph nodes brought about changes in lymph node cellularity, natural killer activity and lipopolysaaccharide (LPS)- and concanavalin A (Con A)-induced cell proliferation in Freund's adjuvant-injected rats; (2) whether the effect of the immunosuppressive drug cyclosporine in rat submaxillary lymph nodes was affected by a single or combined unilateral ganglionectomy plus decentralization. A unilateral ganglionectomy, or the combination of ganglionectomy plus decentralization, performed 7 days earlier, decreased significantly cellularity in ipsilateral submaxillary lymph nodes, while a unilateral decentralization failed to affect it. Natural killer activity increased ipsilaterally after ganglionectomy or decentralization, and decreased after the combined surgical procedure. LPS-induced cell proliferation augmented significantly after ganglionectomy or decentralization, while Con A-induced T lymphocyte proliferation remained unaffected. In the sham-operated side, cyclosporine decreased submaxillary lymph node cell number and natural killer activity, while it increased the proliferative response to LPS. The depressive effect of cyclosporine on lymph node cellularity was no longer observed in ganglionectomized or decentralized lymph nodes, but was found after the combined surgical denervation. Decentralization, or decentralization plus ganglionectomy, blunted the depressive effect of cyclosporine on natural killer activity. The stimulatory effect of cyclosporine on lymphocyte proliferation induced by LPS was reversed both by ganglionectomy or by decentralization and was suppressed by the combined surgical procedure. Neither treatment affected Con A-induced proliferation of T lymphocytes. The results further indicate that an appropriate sympathetic and parasympathetic local environment may be needed for immunomodulation, as well as for cyclosporine activity in lymphoid tissue.

Animals↗

PubMed1997

Effect of diazepam and A beta-carboline on open-field and T-maze behaviors in 2-day-old chicks.

The effects of diazepam and the beta-carboline FG 7142 in chicks were examined on several behavioral measures in open-field and T-maze tasks. In the open field, only the higher doses of diazepam affected behavior, suggesting a sedative-like effect, while FG 7142 influenced behavior as would a fear-inducing manipulation. After a low dose of either drug was injected, testing in a T-maze showed that diazepam improved and FG 7142 impaired the escape performance from the isolation chamber, without affecting the time spent in the T-corridor. In three groups of chicks selected on the basis of their first escape performance, only lower performance chicks were affected by an anxiolytic dose of diazepam. T-maze results suggest that: (a) T-maze is more sensitive than open-field test to behavior changes induced by anxiolytic doses of diazepam; (b) isolation chamber behavior could be an index of general emotionality in young chicks; (c) diazepam and FG 7142 do not modify the social motivation to escape the maze; (d) higher performance chicks present an escape behavior of a less anxious type than lower performance chicks. The results suggest that the GABAergic system is involved in the behavioral expression of fear and anxiety in young chicks.

Animals↗

PubMed1997

Twenty-four hour rhythms of serum prolactin, growth hormone and luteinizing hormone levels, and of medial basal hypothalamic corticotropin-releasing hormone levels and dopamine and serotonin metabolism in rats neonatally administered melatonin.

To obtain information on long-term circadian consequences of administering melatonin neonatally to rats, we assessed the 24-hour rhythms of 1) serum prolactin (PRL), luteinizing hormone (LH), and growth hormone (GH), and 2) medial basal hypothalamic dopamine (DA) and serotonin (5-HT) metabolism and corticotropin-releasing hormone (CRH) content in 60-day old male rats injected with 100 micrograms of melatonin on the 5th day of life. Controls receiving vehicle alone showed serum PRL concentration (when 60 days of age) attaining its maximum at the end of the light period (i.e., at 2000 hr), while in melatonin-injected rats high PRL values were found between 1200 and 2000 hr. Twenty-four hour changes in serum LH levels exhibited a maximum at noon, and to a similar extent in vehicle- and melatonin-treated rats. Neonatal melatonin injection did not affect serum GH concentration when the rats were adult. In the medial basal hypothalamus (MBH), the dihydroxyphenyl acetic acid (DOPAC)/ DA ratio attained a maximum at midnight, its amplitude being significantly higher in melatonin- than in vehicle-treated rats. Neonatally melatonin-injected rats also exhibited a second maximum in DOPAC/DA ratio at noon, coinciding with a minimum in DA levels of MBH. The 5-hydroxyindoleacetic acid (5-HIAA)/5-HT ratio in MBH showed significant diurnal variations in vehicle-injected controls with maxima at 1200 and 0400 hr, while in melatonin-treated rats a single maximum occurred at 2400 hr. These maximum correlated with minima in 5-HT content of MBH. Neonatal melatonin injection brought about a significant increase in the CRH content of MBH, as well as distortion of its diurnal rhythmicity, a maximum being found at noon in controls and at 1800 hr in melatonin-treated rats. The results indicate that exposure to melatonin early in life affects subsequent diurnal rhythmicity of PRL release, and of DA and 5-HT turnover and CRH content in the MBH of rats.

Analysis of Variance↗

PubMed1997

Twenty-four-hour rhythms of serum ACTH, prolactin, growth hormone, and thyroid-stimulating hormone, and of median-eminence norepinephrine, dopamine, and serotonin, in rats injected with Freund's adjuvant.

The effect of Freund's adjuvant injection on 24-h variation of circulating ACTH, prolactin, growth hormone (GH), and thyroid-stimulating hormone (TSH) levels, and of norepinephrine (NE) content, and dopamine (DA) and serotonin (5HT) turnover in median eminence, was examined in adult rats kept under light between 0800 and 2000 h daily. Groups of 6-10 animals received Freund's complete adjuvant or its vehicle at 1100 h 3 days before sacrifice and were killed by decapitation at six different time intervals throughout a 24-h cycle. In rats injected with adjuvant's vehicle, serum ACTH and prolactin exhibited peak values around the light-dark transition (p < 0.0001 and < 0.04, respectively), while the maximum in TSH was found in the late afternoon (p < 0.0001, one-way ANOVA). GH levels did not vary on a 24-h basis. In Freund's adjuvant-injected rats, 24-h variations of TSH levels became blunted, while 24-h variations of prolactin and ACTH persisted. Freund's adjuvant augmented serum ACTH and prolactin levels, and decreased GH and TSH levels (p < 0.0007, factorial ANOVA). Median-eminence NE content, and turnover of DA, assessed by measuring dihydroxyphenylacetic acid, DOPAC/DA ratio, and of 5HT, assessed by measuring 5-hydroxyindoleacetic acid, HIAA/5HT ratio, varied on a 24-h basis in rats receiving adjuvant's vehicle (p < 0.02). Median-eminence NE content attained its maximum at 1600-2000 h, while maxima in DOPA/DA and HIAA/5HT ratios occurred at 0400 h. Injection with Freund's adjuvant reduced the amplitude of the daily variation of NE content, shifted the maximum of DOPAC/DA ratio toward the light-dark transition, and blunted the daily variation in HIAA/5HT ratio in median eminence. The administration at 1200 of the immunosuppressant drug cyclosporine (5 mg/kg, 5 days) restored the augmented ACTH and prolactin levels (p < 0.0001, factorial ANOVA) and depressed GH and TSH levels (p < 0.02) found in Freund's adjuvant-injected rats. Cyclosporine was also effective in restoring 24-h rhythmicity of serum ACTH and TSH, but not of prolactin, levels. Cyclosporine did not modify the effect of Freund's adjuvant on time-of-day changes of median-eminence NE content, but it was effective in counteracting the changes of DA and 5HT turnover found after immunization. The results are compatible with a significant effect of immune-mediated inflammatory response at an early phase after Freund's adjuvant injection on ACTH, GH, prolactin, and TSH release, which is partially sensitive to immunosuppression by cyclosporine.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1996

Twenty-four-hour rhythms in immune responses in rat submaxillary lymph nodes and spleen: effect of cyclosporine.

Twenty-four-hour variations in cellularity, lipopolysaccharide (LPS)- and concanavalin A (Con A)-induced cell proliferation, and natural killer (NK) activity were examined in submaxillary lymph nodes and spleen of rats injected with Freund's complete adjuvant or its vehicle and kept under light from 08:00 to 20:00 h daily. A significant daily variation in cellularity was detected, exhibiting maxima at 09:00 h in submaxillary lymph nodes (nonimmunized and immunized rats) and at 13:00 h in spleen (immunized rats only). Submaxillary lymph node LPS- and Con A-mitogenic effect displayed maximal activity during daytime (peak at 13:00-17:00 h). In spleen, the maxima for 24-h rhythm in LPS-induced cell proliferation and NK activity occurred at midnight and at early morning (09:00 h), respectively. Con A-induced spleen cell proliferation peaked at midday in nonimmunized rats only. Injection of the immunosuppressive drug cyclosporine decreased Freund's adjuvant-induced augmentation of LPS and Con A mitogenic effect in both tissues and diminished spleen cell number. Cyclosporine blunted circadian rhythms in submaxillary lymph node Con A response and cell number, while it shifted the maximum in LPS effect to peak at 01:00 h. Cyclosporine also suppressed the circadian changes in LPS- and Con A-induced spleen cell proliferation, but not those found in NK activity. The results indicate the existence of 24-h rhythms in immune responses of rat submaxillary lymph nodes and spleen with maxima at different times of the day and that were significantly affected by cyclosporine injection.

Animals↗

PubMed1996

Changes in mediobasal hypothalamic dopamine and indoleamine metabolism after superior cervical ganglionectomy of rats.

Eight days after bilateral superior cervical ganglionectomy (Gx) of rats, norepinephrine content of medial basal hypothalamus (MBH) decreased significantly by 44-50%. To obtain information on other possible neurochemical sequela of Gx in MBH, we examined the metabolism of dopamine and serotonin in MBH of Gx rats by employing a high pressure liquid chromatography procedure. Eight days after Gx, MBH dopamine levels augmented significantly. Assessment of dopamine metabolism by measuring dihydroxyphenylacetic acid (DOPAC)/dopamine and homovanillic acid (HVA)/dopamine indexes indicated a significant decrease of MBH DOPAC/dopamine ratio after Gx. MBH serotonin levels increased, and 5-hydroxyindoleacetic acid (5-HIAA)/serotonin index decreased significantly in Gx rats. To examine the interaction Gx-induced changes on MBH dopamine and serotonin with the modified hormonal milieu produced by an ectopic pituitary transplant, adult male rats bearing an ectopic pituitary within the pectoral muscles from day 5 of life were submitted to Gx on day 60 of life and were studied 8 days later. MBH dopamine content increased significantly after pituitary grafting, an effect counteracted by a subsequent Gx, while Gx alone augmented MBH dopamine levels. DOPAC and HVA contents augmented in pituitary-grafted animals, an effect counteracted by Gx. Gx increased MBH serotonin content in control but not in pituitary-grafted rats. After pituitary grafting a decrease in MBH 5-HIAA levels was found, an effect reversed by Gx. Pituitary transplants brought about a significant increase of MBH DOPAC/dopamine index, and a significant decrease in 5-HIAA/serotonin index, both effects being counteracted by Gx. Gx of control rats resulted in a significant decrease of MBH 5-HIAA/serotonin index. Analyzed as a main effect in a factorial analysis of variance, Gx decreased MBH DOPAC/dopamine and HVA/dopamine indexes significantly. Plasma prolactin increased in pituitary-grafted rats, an effect further increased by a subsequent Gx. In pituitary-grafted, Gx rats plasma GH levels augmented significantly. The data suggest that superior cervical ganglion removal affects differentially dopamine and indoleamine metabolism in MBH of control and pituitary-grafted rats.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1996

Benzodiazepine receptors increase induced by stress and maze-learning performance in chick forebrain.

Two-day-old chicks were selected on their second escape performance in a one-trial, maze-learning task, and termed high-performance (H-P), moderate-performance (M-P), and low-performance (L-P) chicks. The learning degree was expressed by the escape time improvement being respectively the 64, 46, and 24%. Then, the three selected groups were maintained to reach 15 days of age and then submitted to acute swimming stress, and [3H]flunitrazepam and [3H]Ro 5-4864 receptor bindings were performed on synaptosomal/mitochondrial membranes from forebrain. The receptor number for both radioligands in stressed high-performance chicks was significantly higher than in stressed low-performance chicks. The results suggest that higher performance chicks were more susceptible than lower performance chicks to acute stress associated to increase of both central and peripheral type benzodiazepine receptors, probably due to differences in the degree of endogenous emotionality.

Animals↗

PubMed1996

Recruitment of peripheral-type benzodiazepine receptors after acute stress in chick forebrain membranes: action of Triton X-100.

A significant increase in the number of measurable [3H]Ro 5-4864 receptors was found in forebrain membranes of chicks submitted to 15 min of acute swim stress compared to non-stressed chicks. In addition, low subsolubilizing concentrations of Triton X-100 caused a significant increase in the measurable [3H]Ro 5-4864 receptor number in forebrain membranes from non-stressed chicks. However, this increase caused by Triton X-100 was not observed when tested in forebrain membranes from stressed chicks. In all cases the affinity remained unchanged. These results suggest that: (1) acute stress and Triton X-100 induce receptor increase by enhancing [3H]Ro 5-4864 accessibility to a pool of receptors not detected before stress or in the absence of detergent; (2) the pool of non-measured receptors represents about a third of the total in control chicks; (3) the increments are not additive and could involve receptors coming from the same non-measured pool; (4) the receptor increase during a short time of stress could be explained by recruitment of receptors but not by an increase in the receptor protein biosynthesis; (5) stress induces a maximal recruitment of measurable [-3H]Ro 5-4864 receptors.

Animals↗

PubMed1996

Estimation of the binding affinity constants of soluble ligand-receptor complexes by a rapid filtration technique: [3H]-flunitrazepam-bovine serum albumin as an example.

A method for determining the equilibrium dissociation constant (KA) of a soluble ligand (L) from a soluble receptor (A) in the presence of another solid phase receptor (R) for the same ligand was developed. The total and nonspecific binding of L to R was measured in the presence and in the absence of A. The separation of bound and free L was done by a rapid filtration technique so that only the complex RL, but not AL, was recovered. An apparent dissociation constant (KR,app) was calculated from the saturation curve obtained in the presence of A. The magnitude of KA could be determined from this KR,app and the value of the equilibrium dissociation constant of the complex R-L (KR) calculated from the saturation curve in the absence of A. The equality of the Bmax values obtained in the presence and in the absence of A assured the accuracy in the determination of KA so that the fulfillment of this condition could be used as an internal control. For the correct definition of nonspecific binding, the displacement agent (L1) should be used at concentrations within the range 10(2).KR < L1 < 10. K4. This fact constraints the applicability of the method to systems where KA/KR > 10(3). The highest sensitivity of the method can be attained when 0.33 < [At]/KA < 3. The equilibrium binding constant of [3H]-flunitrazepam to non-delipidized bovine serum albumin determined by the present approach (31 +/- 7 mumol/L) did not differ significantly from the literature.

Binding Sites↗

PubMed1995

Effect of stress and cyclosporine on ornithine decarboxylase activity in rat submaxillary lymph nodes.

This study was performed: (1) to assess whether the stress produced in rats by daily turpentine oil injections for 5 days, or by restraining the animals for 30 min during 5 days, affected basal and Freund's adjuvant-stimulated submaxillary lymph node ornithine decarboxylase activity, an indicator of cell proliferation; (2) to analyze whether the activity of the immunosuppressive drug cyclosporine on submaxillary lymph node ornithine decarboxylase activation after Freund's adjuvant injection was modified in stressed rats; (3) to examine the mediation of stress effects on submaxillary lymph node ornithine decarboxylase activation by regional sympathetic or parasympathetic nerves. Animals subjected to a unilateral superior cervical ganglionectomy, or to a unilateral chorda tympani section, together with a contralateral sham-operation were employed. After turpentine oil or restraint stress, a significant decrease in submaxillary lymph node ornithine decarboxylase was found. A unilateral sympathetic denervation of submaxillary lymph nodes counteracted in part the inhibitory effect of stress on ornithine decarboxylase activation, as well as augmented the enzyme response in innervated submaxillary lymph nodes. Ornithine decarboxylase activation attained similar values in parasympathetic decentralized or intact submaxillary lymph nodes and the unilateral parasympathetic decentralization did not interfere with the inhibition of enzyme activity found in turpentine oil-stressed rats. Cyclosporine administration (5 or 20 mg/kg) significantly decreased Freund's adjuvant-induced ornithine decarboxylase activity in the submaxillary lymph nodes of control rats, but failed to modify it in turpentine oil-stressed animals. In this latter group, a higher (40 mg/kg) dose of cyclosporine decreased ornithine decarboxylase activity on the innervated side only. A diminished inhibitory response to cyclosporine was found in the parasympathetic decentralized submaxillary lymph nodes of unstressed rats. The results support the view that the immunosuppressive effects of cyclosporine may diminish during stress, in part due to changes in the traffic of neural signals in local sympathetic nerves.

Animals↗

PubMed1995

Cyclosporine effects on in vitro responsiveness of anterior pituitary hormone release to dopamine and thyrotropin-releasing hormone in young female rats.

Endocrine side effects of the immunosuppressive drug cyclosporine (CyA) include changes in anterior pituitary hormone secretion. The aim of the present study was to examine the effects of CyA on the responsiveness of in situ and ectopic anterior pituitary prolactin (PRL), growth hormone (GH) and luteinizing hormone (LH) release response to dopamine (DA) and thyrotropin-releasing hormone (TRH) treatment in young female rats, and to evaluate the possible PRL participation in these effects. Thirty day old rats were rendered hyperprolactinemic by transplanting an anterior pituitary gland of a littermate donor, under the kidney capsule, and were then injected with CyA or vehicle for 2 or 8 days. Sham-operated rats were used as controls and treated in the same way. CyA treatment prevented the increase in plasma PRL levels which occurred in controls after pituitary grafting. In vitro basal PRL release of in situ pituitaries from either sham-operated and/or pituitary-grafted animals was decreased by CyA treatment at any point studied. Basal in vitro secretion of GH was only decreased in the in situ pituitaries from grafted animals after 2 days of CyA therapy. The presence of an ectopic pituitary lead to an increase in the in vitro basal LH secretion from in situ pituitaries as compared to those from sham-operated rats. Basal LH release rates were not changed by CyA treatment, although the LH release in vitro did increase in the in situ pituitaries from sham-operated animals treated with the drug for 2 days. DA addition to the incubation media decreased the in vitro release of PRL, GH and LH from the in situ pituitaries of sham-operated and pituitary-grafted animals treated with vehicle. In CyA treated animals, DA decreased in vitro PRL release from the in situ pituitaries of animals, independently of the presence or absence of an ectopic pituitary. Reductions of the in vitro GH and LH release release after DA treatment were higher in the in situ pituitaries from grafted animals on day 8 of CyA or vehicle treatment. TRH increased the in vitro release of the three hormones with differential effects related to the length of the treatment with CyA and/or the presence of an ectopic pituitary. In vitro release of PRL and GH by ectopic pituitaries was inhibited by previous treatment with CyA and this effect was decreased proportional to the duration of the treatment with the drug, while LH secretion was not modified. Addition of DA to the incubation media resulted in a marked reduction of in vitro PRL and GH release, but only at day 8 of vehicle treatment on GH release did DA addition to media further decrease the release of both hormones from ectopic pituitaries from animals treated for 2 or 8 days with the drug, whereas LH secretion was not modified. TRH addition to the incubation media of ectopic pituitaries surprisingly reduced PRL and GH secretion on day 8 of CyA treatment or after surgery. The results of these studies suggest that CyA can act directly at the hypophyseal level modifying pituitary responsiveness to external stimuli. CyA seems to exert its main effects on lactotroph activity while its effects on somatotrophs and gonadotrophs are less.

Animals↗

PubMed1995

Effects of cyclosporine at the hypothalamic-pituitary axis in pituitary-grafted young female rats.

This work was designed to investigate the effects of cyclosporine on prolactin secretion by an ectopically grafted heterologous pituitary gland, and on the hypothalamic content of norepinephrine, dopamine and serotonin. The administration of cyclosporine prevented the augmentation in plasma prolactin levels which occurred following an ectopic graft of a litter-mate pituitary gland. In contrast, in sham-operated rats, cyclosporine increased prolactin levels on day 8 of treatment. Both pituitary grafting and cyclosporine treatment in sham-operated rats decreased hypothalamic norepinephrine content. In grafted rats, cyclosporine returned hypothalamic norepinephrine to normal. Hypothalamic serotonin content decreased 8 days after pituitary grafting but increased to the values of control animals after cyclosporine administration. Cyclosporine treatment for 2 and 8 days increased serotonin content in sham-operated animals. As expected, the hypothalamic dihydroxphenylacetic acid/dopamine index increased after pituitary grafting and administration of cyclosporine for 8 days resulted in a further increase. Cyclosporine administration for 2 days, however, decreased this index to the values observed in control animals while drug treatment of control rats for 8 days decreased the dihydroxyphenylacetic acid/dopamine index. In vitro release of prolactin from the ectopic gland was markedly decreased in animals treated with cyclosporine for 2 days and this effect was less evident in 8-day treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)

3,4-Dihydroxyphenylacetic Acid↗