Central muscarinic receptor subtypes involved in pilocarpine-induced salivation, hypertension and water intake

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fruit and seed ontogeny related to the seed behaviour of two tropical species of Caesalpinia (Leguminosae)

Caesalpinia echinata and C ferrea var. ferrea have different seed behaviours and seed and fruit types. Comparison of the seed ontogeny and anatomy partly explained the differences in seed behaviour between these two species of Brazilian legumes; some differences were also related to fruit development. The seed coat in C. ferrea consisted of two layers of osteosclereids, as well as macrosclereids and fibres, to form a typical legume seed coat, whereas C. echinata had only macrosclereids and fibres. In C. echinata, the developing seed coat had paracytic stomata, a feature rarely found in legume seeds. These seed coat features may account for the low longevity of C. echinata seeds. The embryogeny was similar in both species, with no differences in the relationship between embryo growth and seed growth. The seeds of both species behaved as typical endospermic seeds, despite their different morphological classification (exendospermic orthodox seeds were described for C. echinata and endospermic orthodox seeds for C. ferrea). Embryo growth in C. ferrea accelerated when the sclerenchyma of the pericarp was developing, whereas embryonic growth in C. echinata was associated with the conclusion of spine and secretory reservoir development in the pericarp. Other features observed included an endothelial layer that secreted mucilage in both species, a nucellar summit, which grew up into the micropyle, and a placental obturator that connected the ovarian tissue to the ovule in C. ferrea. (C) 2004 the Linnean Society of London.

Applied models to biodegradation kinetics of lubricant and vegetable oils in wastewater

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Central serotonergic and adrenergic/imidazoline inhibitory mechanisms on sodium and water intake

Recent studies have shown the existence of two important inhibitory mechanisms for the control of NaCl and water intake: one mechanism involves serotonin in the lateral parabrachial nucleus (LPBN) and the other depends on alpha(2)-adrenergic/imidazoline receptors probably in the forebrain areas. In the present study we investigated if alpha(2)-adrenergic/imidazoline and serotonergic inhibitory mechanisms interact to control NaCl and water intake. Male Holtzman rats with cannulas implanted simultaneously into the lateral ventricle (LV) and bilaterally into the LPBN were used. The ingestion of 0.3 M NaCl and water was induced by treatment with the diuretic furosemide (10 mg/kg of body weight)+the angiotensin converting enzyme inhibitor captopril (5 mg/kg) injected subcutaneously 1 h before the access of rats to water and 0.3 M NaCl. Intracerebroventricular (i.c.v.) injection of the alpha(1)-adrenergic/imidazoline agonist clonidine (20 nmol/l RI) almost abolished water (1.6 +/- 1.2, vs. vehicle: 7.5 +/- 2.2 ml/2 h) and 0.3 M NaCl intake (0.5 +/- 0.3, vs. vehicle: 2.2 0.8 ml/2 h). Similar effects were produced by bilateral injections of the 5HT(2a/2b) serotonergic agonist 2,5-dimetoxy-4-iodoamphetamine (DOI, 5 mug/0.2 mul each site) into the LPBN on water (3.6 +/- 0.9 ml/2 h) and 0.3 M NaCl intake (0.4 +/- 0.2 m1/2 h). Injection of the (alpha(2)-adrenergic/imidazoline antagonist idazoxan (320 nmol) i.c.v. completely blocked the effects of clonidine on water (8.4 +/- 1.5 ml/2 h) and NaCl intake (4.0 +/- 1.2 ml/2 h), but did not change the effects of LPBN injections of DOI on water (4.2 +/- 1.0 ml/2 h) and NaCl intake (0.7 +/- 0.2 ml/2 h). Bilateral injections of methysergide (4 mug/0.2 mul each site) into the LPBN increased 0.3 M NaCl intake (6.4 +/- 1.9 ml/2 h), not water intake. The inhibitory effect of i.c.v. clonidine on water and 0.3 M NaCl was still present after injections of methysergide into the LPBN (1.5 +/- 0.8 and 1.7 +/- 1.4 ml/2 h, respectively). The results show that the inhibitory effects of the activation of a,-adrenergic/imidazoline receptors in the forebrain are still present after blockade of the LPBN serotonergic mechanisms and vice versa for the activation of serotonergic mechanisms of the LPBN. Therefore, each system may act independently to inhibit NaCl and water intake. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Effects of central alpha-adrenergic agonists on hormone-induced 3% NaCl and water intake

Male rats received intracerebroventricular (ICV) renin (600 ng) or daily subcutaneous injections of deoxycorticosterone (5 mg) to induce 3% NaCl and water intake. Noradrenaline (NOR; 40-160 nmol) and clonidine (CLO; 5-20 nmol) injected ICV. induced 70 to 100% inhibition of the intakes. Phenylephrine (PHE; 40-160 nmol) injected ICV induced 60 to 95% inhibition of the intakes. NOR and PHE induced a stronger inhibition on the 3% NaCl intake induced by renin than on the intake induced by deoxycorticosterone (DOC), and CLO did the opposite. CLO was always more effective than PHE to induce inhibition of the intakes. The results suggest that NOR inhibits hormone (angiotensin II, aldosterone)-induced NaCl intake by acting mainly on alpha(2)-adrenergic receptors.

Receptor-mediated effects of clonidine on need-induced 3% NaCl and water intake

Clonidine combined with adrenergic antagonists were injected in the medial septal area in order to characterize the type of receptors involved with its inhibitory effect on 3% NaCl and water intake of sodium-depleted (furosemide + 24 h of removal of ambient sodium) and 30-h water-deprived rats, respectively. The inhibitory effect of clonidine (20 nmol) on need-induced water intake was reduced 50% by an 80-nmol dose of either idazoxan, yohimbine or prazosin. The inhibitory effect of clonidine (30 nmol) on need-induced 3% NaCl intake was completely antagonized by idazoxan (80, 160 nmol), not altered by yohimbine (40-160 nmol), and partially potentiated (40 nmol) or inhibited (160 nmol) by prazosin. Propranolol did not alter the effects of clonidine on either water (80 nmol) or 3% NaCl (40-160 nmol) intake. The results suggest that the inhibitory effects of clonidine on 3% NaCl and water intake are mediated by different types of alpha2-adrenergic receptors. Copyright (C) 1997 Elsevier B.V.

Paraventricular nucleus administration of DuP753 or PD123319 inhibits the effects of angiotensin on water and sodium intake

We determined the effects of DuP753 and PD123319 (both nonpeptides and selective antagonists of the AT(1) and AT(2) angiotensin receptors, respectively), and [Sar(1), Ala(8)]ANG II (a non-selective peptide antagonist of angiotensin receptors) on water and 3%NaCl intake induced by administration of angiotensin II (ANG II) into the paraventricular nucleus (PVN) of sodium-depleted Holtzman rats weighing 250-300 g. Twenty hours before the experiments, the rats were depleted of sodium using furosemide (10 ng/rat, sc). The volume of drug solution injected was 0.5 mu l over a period of 10-15 sec. Water and sodium intake were measured at 0.25, 0.5, 1.0 and 2.0 h. Pre-treatment with DuP753 (14 rats) at a dose of 60 ng completely abolished the water intake induced by injection of 12 ng of ANG II (15 rats) (6.4 +/- 0.6 vs 1.4 +/- 0.3 ml/2 h), where [Sar(1), Ala(8)]ANG II (12 rats) and PD123319 (10 rats) at the doses of 60 ng partially blocked water intake (6.4 +/- 0.6 vs 2.9 +/- 0.5 and 2.7 +/- 0.2 ml/2 h, respectively). In the same animals, [Sar(1), Ala(8)]ANG II, DuP753, and PD123319 blocked the sodium intake induced by ANG II (9.2 +/- 1.6 vs 3.3 +/- 0.6, 1.8 +/- 0.3, and 1.4 +/- 0.2 ml/2 h, respectively). These results indicate that both DuP753 and PD123319, administered into the PVN, blocked the water and sodium intake induced by administration of ANG II into the same site.

PROGESTERONE ADMINISTRATION TO OVARIECTOMIZED RATS REDUCES WATER AND SALT INTAKE INDUCED BY CENTRAL ADMINISTRATION OF ANGIOTENSIN-II

We tested the effects of estradiol, progesterone and testosterone on water and salt intake induced by angiotensin II (ANG II) injected into the third ventricle of female Holtzman rats weighing 250-300 g. The water and salt ingestion observed after 120 min in the control experiments (injection of 0.5 mu l of 0.15 M NaCl into the third ventricle) was 1.6 +/- 0.3 ml (N = 10) and 0.3 +/- 0.1 ml (N = 8) in intact rats, respectively, and 1.4 +/- 0.3 ml (N = 10) and 0.2 +/- 0.1 (N = 8) in ovariectomized rats, respectively. ANG II injected in intact rats (4, 6, 12, 25, and 50 ng, icv, in 0.5 mu l saline) induced an increase in water intake (4.3 +/- 0.6, 5.4 +/- 0.7. 7.8 +/- 0.8, 10.4 +/- 1.2, 11.2 +/- 1.4 ml/120 min, respectively) (N = 43). The same doses of icv ANG II in intact rats increased the 3% NaCl intake (0.9 +/- 0.2; 1.4 +/- 0.3, 2.3 +/- 0.4, 2.2 +/- 0.3. and 2.5 +/- 0.4 ml/120 min, respectively) (N = 42). When administered to ovariectomized rats ANG II induced comparable amounts of water intake (4.0 +/- 0.5, 4.8 +/- 0.6, 6.9 +/- 0.7. 9.6 +/- 0.8, and 10.9 +/- 1.2 ml/120 min, respectively) (N = 43) but there was a significant decrease of 3% NaCl solution ingestion (0.3 +/- 0.1, 0.4 +/- 0.1, 0.8 +/- 0.2, 0.7 +/- 0.2, and 0.6 +/- 0.2 ml/120 min, respectively) (N = 44). Estrogen (50 mu g), progesterone (25 ng), and testosterone (300 mu g) were injected daily into ovariectomized rats for 21 days. Treatment with estrogen decreased the water intake and abolished the saline ingestion induced by icy injection of ANG II (12 ng (2.8 +/- 1.2 and 0.3 +/- 0.1 ml/120 min, respectively) (N = 8). Treatment with progesterone also reduced the water intake (3.3 +/- 0.6 ml/120 min) (N = 8) and abolished the ANG II-induced saline ingestion (0.4 +/- 0.1 ml/120 min) (N = 8), but these effects were not observed with testosterone (6.4 +/- 0.8 and 2.2 +/- 0.3 ml/120 min, respectively) (N = 8). These results indicate that ANG II induces a greater increase in sodium intake in intact female rats than in ovariectomized rats and that estrogen and progesterone impair water and sodium intake in ovariectomized rats.

Effect of ibotenate lesions of the ventromedial hypothalamus on the water and salt intake induced by activation of the median preoptic nucleus in sodium-depleted rats

In this study we investigated the influence of a ventromedial hypothalamus (VMH) lesion with ibotenic acid on water and sodium intake and presser responses induced by combined treatment of the median preoptic nucleus (MnPO) with angiotensin Il (ANG II) and adrenergic agonists (phenylephrine, norepinephrine, isoproterenol and clonidine). Male Holtzman rats with a stainless steel cannula implanted into the MnPO and bilateral sham (vehicle) or VMH lesions with ibotenic acid were used. The ingestion of water and sodium and mean arterial pressure (MAP) were determined in separate groups submitted to sodium depletion with the diuretic furosemide (20 mg/rat). ANG II (10 pmol) injection into the MnPO of sham-lesioned rats induced water and sodium intake and presser responses. VMH-lesion reduced ANG II-induced water intake and increased saline intake, In sham rats phenylephrine (80 nmol) into MnPO increased, whereas norepinephrine (80 nmol) and clonidine (40 nmol) reduced ANG II-induced water intake while sodium intake was reduced only by clonidine into MnPO. In VMH-lesioned rats, phenylephrine reduced, noradrenaline increased and clonidine produced no effect on ANG II-induced water intake. In lesioned rats ANG II-induced sodium intake was reduced by phenylephrine and noradrenaline, whereas clonidine produced no change. ANG II-induced presser response was reduced in VMH-lesioned rats, but the presser response combining ANG II and phenylephrine or noradrenaline in VMH-lesioned rats was bigger than sham rats. These results show that the VMH is important for the changes in water and sodium intake and cardiovascular responses induced by angiotensinergic and adrenergic activation of the MnPO. (C) 1997 Elsevier B.V. B.V.

EFFECT OF LATERAL HYPOTHALAMUS LESIONS ON THE WATER AND SALT INTAKE, AND SODIUM AND URINE EXCRETION INDUCED BY ACTIVATION OF THE MEDIAN PREOPTIC NUCLEUS IN CONSCIOUS RATS

In this-study we investigated the influence of electrolytic lesion of the lateral hypothalamus (LH) on the water and salt appetite, and the natriuretic, diuretic and cardiovascular effects induced by angiotensinergic, cholinergic and noradrenergic stimulation of the median preoptic nucleus (MnPO) in rats. Male Holtzman rats were implanted with a cannula into the MnPO. Other groups of sham- and LH-lesioned rats received a stainless steel cannula implanted into the MnPO. ANGII injection into the MnPO induced water and sodium intake, and natriuretic, diuretic, presser and tachycardic responses. Carbachol induced water intake, and natriuretic, presser and bradycardic responses, whereas noradrenaline increased urine, sodium excretion and blood pressure, and induced bradycardia. In rats submitted to LH-lesion only, water and sodium intake was reduced compared with sham rats. LH lesion also reduced the sodium ingestion induced by ANGII (12 ng) into the MnPO. In LH-lesioned rats, the dipsogenic, diuretic and presser responses induced by ANGII (12 ng), carbachol (2 nmol) and noradrenaline (20 nmol) injection into the MnPO were reduced. The same occurred with sodium excretion when carbachol (2 nmol) and noradrenaline (20 nmol) were injected into the MnPO of LH-lesioned rats, whereas ANGII(12 ng) induced an increase in sodium excretion. These data show that electrolytic lesion of the LH reduces fluid and sodium intake, and presser responses to angiotensinergic, cholinergic and noradrenergic activation of the MnPO. LH involvement with MnPO excitatory and inhibitory mechanisms related to water and sodium intake, sodium excretion and cardiovascular control is suggested.

Central alpha-adrenergic agonists and need-induced 3% NaCl and water intake

In the present study, noradrenaline (NOR, alpha-non-specific adrenergic agonist), clonidine (CLO, alpha(2)), phenylephrine (PHE, alpha(1)) or isoproterenol (ISO, beta-agonist) was injected in the medial septal area (MSA) of water-deprived, sodium-deplete or food-deprived rats. NOR (80, 160 nmol) inhibited the intake of 3% NaCl, water deprivation-induced and meal-associated water intake. Food deprivation-induced food intake and 10% sucrose intake were not altered by NOR. CLO (10, 20, 30, 40 nmol) inhibited (80-100% inhibition compared to control during 60 min) the intake of 3% NaCl, water deprivation-induced and meal-associated water intake. CLO had a weaker inhibition on food and 10% sucrose intake (30-50% less than the control during 60 and 15 min, respectively). PHE (160 nmol) inhibited 3% NaCl intake and 10% sucrose intake (30% less than the control for 15-30 min). ISO (160 nmol) did not after water or 3% NaCl intake. NOR induced an increase, CLO and ISO induced a decrease, and PHE no alteration in mean arterial pressure. NOR did not alter water or 3% NaCl intake when injected unilaterally into the caudate nucleus. The results suggest that NOR injected in the MSA acts on alpha(2)-adrenergic receptors inducing a specific inhibition of 3% NaCl and water intake. (C) 1997 Elsevier B.V.

Effects of subtypes alpha- and beta-adrenoceptors of the lateral hypothalamus on the water and sodium intake induced by angiotensin II injected into the subfornical organ

The present experiments were conducted to investigate the role of the alpha (1A)-, alpha (1B), beta (1),- and beta (2)-adrenoceptors of the lateral hypothalamus (LH) on the water and salt intake responses elicited by subfornical organ (SFO) injection of angiotensin II (ANG II) in rats. 5-methylurapidil (an alpha (1A)-adrenergic antagonist), cyclazosin (an alpha (1B)-adrenergic antagonist) and ICI-118,551 (a beta (2)-adrenergic antagonist) injected into the LH produced a dose-dependent reduction, whereas efaroxan (an alpha (2)-antagonist) increased the water intake induced by administration of ANG II into the SFO. These data show that injection of 5-methylurapidil into the LH prior to ANG II into the SFO increased the water and sodium intake induced by the injection of ANG II. The present data also show that atenolol (a beta (1)-adrenergic antagonist), ICI-118,551, cyclazosin, or efaroxan injected into the LH reduced in a dose-dependent manner the water and sodium intake to angiotensinergic activation of SFO. Thus, the alpha (1)- and beta -adrenoceptors of the LH are possibly involved with central mechanisms dependent on ANG II and SFO that control water and sodium intake. (C) 2000 Elsevier B.V. B.V. All rights reserved.

Central moxonidine on water and NaCl intake

In this study we investigated: (a) the effects of intracerebroventricular (i.c.v.) injections of moxonidine (an alpha(2)-adrenergic and imidazoline receptor agonist) on the ingestion of water and NaCl induced by 24 h of water deprivation; (b) the effects of i.c.v. injection of moxonidine on central angiotensin II (ANG II)- and carbachol-induced water intake; (c) the effects of the pre-treatment with i.c.v, idazoxan (an alpha(2)-adrenergic and imidazoline receptor antagonist) and RX 821002 (a selective alpha(2)-adrenergic antagonist) on the antidipsogenic action of central moxonidine. Male Holtzman rats had stainless steel cannulas implanted in the lateral cerebral ventricle. Intracerebroventricular injection of moxonidine (5 and 20 nmol/1 mu l) reduced the ingestion of 1.5% NaCl solution (4.1 +/- 1.1 and 2.9 +/- 2.5 ml/2 h, respectively vs. control = 7.4 +/- 2.1 ml/2 h) and water intake (2.0 +/- 0.6 and 0.3 +/- 0.2 ml/h, respectively vs. control = 13.0 +/- 1.4 ml/h) induced by water deprivation, Intracerebroventricular moxonidine (5 nmol/1 mu l) also reduced i.c.v. ANG Ii-induced water intake (2.8 +/- 0.9 vs. control = 7.9 +/- 1.7 ml/1 h) and i.c.v. moxonidine (10 and 20 nmol/1 mu l) reduced i.c.v. carbachol-induced water intake (4.3 +/- 1.7 and 2.1 +/- 0.9, respectively vs. control = 9.2 +/- 1.0 ml/1 h). The pre-treatment with i.c.v. idazoxan (40 to 320 nmol/1 mu l) abolished the inhibitory effect of i.c.v, moxonidine on carbachol-induced water intake. Intracerebroventricular idazoxan (320 nmol/1 mu l) partially reduced the inhibitory effect of moxonidine on water deprivation-induced water intake and produced only a tendency to reduce the antidipsogenic effect of moxonidine on ANG Ii-induced water intake. RX 821002 (80 and 160 nmol/1 mu l) completely abolished the antidipsogenic action of moxonidine on ANG Ii-induced water intake. The results show that central injections c: moxonidine strongly inhibit water and NaCl ingestion. They also suggest the involvement of central alpha(2)-adrenergic receptors in the antidipsogenic action of moxonidine. (C) 1999 Elsevier B.V.

EFFECT OF CHOLINERGIC STIMULATION OF THE AMYGDALOID COMPLEX ON WATER AND SALT INTAKE

The effect of carbachol (80 nmol/mul) injection into the amygdaloid nuclear complex (AMG) on sodium appetite and water intake was studied in male Holtzman rats weighing 240-270 g. Twenty-five satiated rats and 38 water-deprived rats were used in the experiment on water intake. In the experiment on sodium intake, 19 rats were injected with atropine + carbachol and 9 rats with hexamethonium + carbachol. After carbachol injection into the AMG, water intake decreased in rats submitted to 30 h of water deprivation (10.28 +/- 1.04 ml/120 min vs 0.69 +/- 0.22 ml/120 min). The decrease in water intake was blocked by prior local injection of a tropine (20 nmol/1 mul)(11.66 +/- 1.46 ml/120 min vs 0.69 +/- 0.22 ml/120 min), but not of hexamethonium (30 nmol/1 mul), into the AMG. In water-deprived animals, carbachol injection into the AMG caused a decrease in sodium chloride intake (6.16 +/- 1.82 ml/h vs 0.88 +/- 0.54 ml/h) which was blocked by previous injection of hexamethonium but not of a tropine. These results suggest that the cholinergic system of the AMG plays a role in the control of water and salt intake.