Chronic Guanethidine Treatment Research Articles

Acute and chronic role of nitric oxide, renin-angiotensin system and sympathetic nervous system in the modulation of calcium sensitization in Wistar rats.

Principal vasoactive systems - renin-angiotensin system (RAS), sympathetic nervous system (SNS), nitric oxide (NO) and prostanoids - exert their vascular effects through the changes in calcium levels and/or calcium sensitization. To estimate a possible modulation of calcium sensitization by the above vasoactive systems, we studied the influence of acute and chronic blockade of particular vasoactive systems on blood pressure (BP) changes elicited in conscious normotensive rats by acute dose-dependent administration of Rho-kinase inhibitor fasudil. Adult male chronically cannulated Wistar rats were used throughout this study. The acute inhibition of NO synthase (NOS) by L-NAME enhanced BP response to fasudil, the effect being considerably augmented in rats deprived of endogenous SNS. The acute inhibition of prostanoid synthesis by indomethacin modified BP response to fasudil less than the acute NOS inhibition. The chronic NOS inhibition caused moderate BP elevation and a more pronounced augmentation of fasudil-induced BP changes compared to the effect of acute NOS inhibition. This indicates both short-term and long-term NO-dependent attenuation of calcium sensitization. Long-term inhibition of RAS by captopril caused a significant attenuation of BP changes elicited by fasudil. In contrast, a long-term attenuation of SNS by chronic guanethidine treatment (in youth or adulthood) had no effect on BP response to fasudil, suggesting the absence of SNS does not affect calcium sensitization in vascular smooth muscle of normotensive rats. In conclusion, renin-angiotensin system contributes to the long-term increase of calcium sensitization and its effect is counterbalanced by nitric oxide which decreases calcium sensitization in Wistar rats.

Role of catecholamines in glucagon-induced thermogenesis.

The present work was undertaken in order to investigate whether the observed thermogenesis following glucagon injection requires the participation of catecholamines. Our experiments aim at studying the effects of intraperitoneal injection of glucagon on metabolic rates, plasma catecholamine and fuel metabolites in guanethidine-treated ducklings reared at thermoneutrality (25 degrees C). The chronic guanethidine treatment induced a marked decrease in catecholamines levels in peripheral tissues (heart, muscle and intestine) but not in adrenals. At thermoneutrality, intraperitoneal injection of glucagon had lower thermogenic effects in guanethidine-treated compared to control ducklings. Glucagon injection elicited a concomitant increase of plasma norepinephrine, metabolic rate and energy metabolites in control ducklings, whereas in guanethidine-treated ducklings, the plasma catecholamines and metabolic rate did not undergo any consistent change. The thermogenic action of glucagon in birds involves at least the mobilization of lipids and catecholaminergic system stimulation.

Effect of Chronic Guanethidine Treatment on Amylobarbitone—Induced Anaesthesia

Elsa, A.T., Osunkwo, U.A. and Onyeyili, P.A. 2003. Effect of chronic guanethidine treatment of amylobarbitone—induced anaesthesia. J. Appl. Anim. Res., 24: 101–104. The effect of chronic treatment with guanethidine on the pattern of amylobarbitone induced anaesthesia in adult rabbits was studied to find out whether chemical sympathectomy influences barbiturate anaesthesia. The onset and duration of amylobarbitone induced anaesthesia decreased linearly (P<0.05), when treated with 2.5 or 5.0 mg/kg body weight guanethidine. Thus, guanethidine treatment may enhance induction but decrease duration of amylobarbitone anaesthesia in rabbits.

Chemical sympathectomy and maternal separation affect neonatal stress responses and adrenal sensitivity to ACTH

The participation of sympathetic adrenal innervation in the control of the neonatal adrenocortical system and in changes in adrenal sensitivity after maternal separation for 24 h was tested in 10- and 23-day-old pups. Chemical sympathectomy by guanethidine (20 mg/kg body wt) decreased basal and stimulated corticosterone compound B (B) secretion without affecting adrenocorticotropic hormone (ACTH) release, abolished the enhanced adrenal sensitivity to ACTH induced by maternal separation in 10-day-old pups, but did not modify adrenal sensitivity following ether stress in 23-day-old pups. Guanethidine treatment did not affect body and adrenal weight or adrenal choline acetyltransferase activity, but it increased tyrosine hydroxylase activity at both ages. Both chronic guanethidine treatment and acute corticotropin-releasing factor immunoneutralization reduced plasma B levels after maternal separation without affecting plasma ACTH levels. Maternal separation in 10-day-old pups enhanced basal and stimulated ACTH and B secretion after exposure to ether vapors and insulin-induced hypoglycemia (IIH). In nonseparated pups, IIH did not stimulate ACTH secretion and caused small increases in B secretion; however, the enhanced response of separated pups to IIH was due to the effects of intraperitoneal injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Different reinnervation patterns in the celiac/mesenteric and superior cervical ganglia following guanethidine sympathectomy in adult rats

We sought to determine whether chronic guanethidine (Gu) treatment in adult rats produces depletion of sympathetic neurons and hyperinnervation by sensory neuropeptides in the celiac/superior mesenteric (C/SMG) ganglion. Rats received Gu 40 mg/kg per day i.p or saline for 5 weeks. Upon completion of treatment, the C/SMG and the superior cervical ganglion (SCG) were examined for neuropeptide Y (NPY), substance P (SP), calcitonin gene-related peptide (CGRP) and vasoactive intestinal polypeptide (VIP), both by immunocytochemistry (ICC) and radioimmunoassay (RIA). Gu produced marked depletion of NPY-containing neurons and NPY content in the C/SMG, similar to that in the SCG (−89 ± 2vs.−92 ± 4%, respectively). SP and CGRP immunoreactivities were significantly higher iontrol C/SMG as compared with SCG; after Gu treatment, there was no significant increase in either SP or CGRP in the C/SMG, however, both increased in the SCG. In contrast, VIP levels were similar in the SCG and C/SMG in controls and increased in the C/SMG but not in the SCG after Gu treatment. Thus, in adult rats, the C/SMG is as susceptible as the SCG to Gu treatment; the different pattern of hyperinnervation by SP, CGRP and VIP of the C/SMG as compared with the SCG may reflect the different sources for these neuropeptides in prevertebral as compared with paravertebral ganglia.

Effect of chemical sympathectomy on scintigraphic gastric and small bowel transit in the rat

Our aims were to measure gastric and small bowel transit of radiolabeled chow or liquids in rats and to assess the effect of pharmacologically induced chronic sympathectomy of at least 5 weeks duration. Three series of experiments were performed. In series I, four rats in each group underwent i.p. chloral hydrate anesthesia, tracheostomy with intubation and mechanical ventilation: they received by gavage 1.0 g rat chow dissolved in 1.5 ml saline labeled with 1.0 mCi 99mTc-DTPA In series II, four rats in each group were anesthetized with only i.p. chloral hydrate and received 1.5 ml labeled saline. Rats were placed on a gamma camera for dynamic acquisition of 1-min images for 7 h in series I, and 4 h in series II. Activity was quantitated in stomach and cecum; gastric emptying was analyzed by the power exponential model. In series III, four rats in each group were studied for 1 h on gamma camera and sacrificed after 5 h. At the end of each study, the stomach, small bowel, cecum and colon were removed and radioactivity counted in each organ in a dose calibrator. Quantitation of radioactivity by gamma camera was highly accurate: labeled chow in the stomach at 7 h was identical by external camera and counting of the excised organ (linear regression slope = 1.01, r = 0.97, P < 0.0001). Similar precision was observed with labeled liquids (slope = 0.93, r = 0.93, P = 0.001). Chow emptied slower than liquid in both groups ( P = 0.048). Ileocecal movement of isotope occurred by bolus transfers. Chemical sympathectomy did not alter gastric emptying or small bowel transit. In conclusion, gastric emptying and cecal filling in rats can be accurately quantitated by dynamic scintigraphy. The rat stomach discriminates between solids and liquids; cecal filling occurs by bolus transfers. Chronic guanethidine treatment, as a model of sympathectomy, does not alter upper gastrointestinal transit.

Myelin sheath survival after guanethidine-induced axonal degeneration.

Membrane-membrane interactions between axons and Schwann cells are required for initial myelin formation in the peripheral nervous system. However, recent studies of double myelination in sympathetic nerve have indicated that myelin sheaths continue to exist after complete loss of axonal contact (Kidd, G. J., and J. W. Heath. 1988. J. Neurocytol. 17:245-261). This suggests that myelin maintenance may be regulated either by diffusible axonal factors or by nonaxonal mechanisms. To test these hypotheses, axons involved in double myelination in the rat superior cervical ganglion were destroyed by chronic guanethidine treatment. Guanethidine-induced sympathectomy resulted in a Wallerian-like pattern of myelin degeneration within 10 d. In doubly myelinated configurations the axon, inner myelin sheath (which lies in contact with the axon), and approximately 75% of outer myelin sheaths broke down by this time. Degenerating outer sheaths were not found at later periods. It is probably that outer sheaths that degenerated were only partially displaced from the axon at the commencement of guanethidine treatment. In contrast, analysis of serial sections showed that completely displaced outer internodes remained ultrastructurally intact. These internodes survived degeneration of the axon and inner sheath, and during the later time points (2-6 wk) they enclosed only connective tissue elements and reorganized Schwann cells/processes. Axonal regeneration was not observed within surviving outer internodes. We therefore conclude that myelin maintenance in the superior cervical ganglion is not dependent on direct axonal contact or diffusible axonal factors. In addition, physical association of Schwann cells with the degenerating axon may be an important factor in precipitating myelin breakdown during Wallerian degeneration.

Source of Stress‐Induced Increase in Plasma Met‐Enkephalin in Rats: Contribution of Adrenal Medulla and/or Sympathetic Nerves*

Abstract The contribution of the adrenal medulla and/or the sympathetic nerves to the plasma levels of Met-enkephalin was investigated. Rats were divided into four groups: sham-operated/saline, sham-operated/guanethidine, adrenal-demedullated/saline, demedulla-ted/guanethidine. After 4 weeks of injection with either saline or guanethidine (25 mg/kg/day), animals were cannulated in the left carotid artery for blood sampling. Three days later, blood samples were taken before and at 2 and 30 min of restraint stress. Adrenal demedullation lowered basal plasma epinephrine levels markedly and prevented entirely the increase induced by restraint stress. Chronic guanethidine treatment lowered basal plasma norepinephrine levels and decreased the response to stress. Guanethidine treatment increased the basal plasma epinephrine level without affecting the response to stress. The combination of guanethidine plus adrenal demedullation lowered basal plasma concentrations of all three catecholamines and further attenuated the norepinephrine response. Restraint stress increased plasma native and peptidase-derivable Met-enkephalin. Adrenal demedullation, resulting in greater than 95% depletion of adrenal catecholamines and significant depletion of adrenal Met-enkephalin, did not inhibit the stress-induced increase in plasma Met-enkephalin, and in fact, was associated with a potentiated response to stress. Guanethidine treatment with or without demedullation increased baseline plasma native Met-enkephalin and abolished the stress-induced increase in plasma native and peptidase-derivable Met-enkephalin. Thus, the stress-induced increase in plasma Metenkephalin results from release from sympathetic nerves, rather than adrenal medulla. However, the sympathetic nerves and adrenal medulla together do not appear to account entirely for basal concentrations of circulating Met-enkephalin. Hepatic portal vein plasma concentration of native Met-enkephalin was greater than that in the carotid artery, suggesting contribution from the gastrointestinal tract; however, evisceration did not decrease plasma native Met-enkephalin. Some compensatory mechanism results in elevation of basal plasma native Met-enkephalin in sympathectomized rats. Also, in the absence of the adrenal medulla there is a compensatory increase in the amount of Met-enkephalin released into the circulation in response to stress.

Effects of Adrenalectomy and Chronic Guanethidine Treatment on Tissue Adrenaline Concentrations in Swimming-Exposed Rats

Adrenaline is known to increase in extra-adrenal peripheral organs as well as urine and plasma in the rat after swimming stress. In order to clarify the physiological significance of adrenaline in stress responses, the origin and location of adrenaline in the peripheral organs were examined by measuring adrenaline in the heart, spleen, submaxillary gland and some other organs after adrenalectomy or guanethidine treatment. After bilateral adrenalectomy, adrenaline in the peripheral organs mostly disappeared. Swimming stress caused no increase in the adrenaline content of the organs in the adrenalectomized rats, while a marked increase was observed in the sham-operated animals. In guanethidine-treated rats, the adrenaline content of the peripheral organs was extremely low as compared to that in the saline-treated animals, and the swimming-induced increase in adrenaline level in the organs was greatly suppressed. These observations suggest that the organ adrenaline, which exists in sympathetic nerve endings, is mostly derived from the adrenal gland. It is considered that adrenaline would be involved in sympathetic neurotransmission, especially under stressful conditions.

Effects of adrenalectomy and chronic guanethidine treatment on tissue adrenaline concentrations in swimming-exposed rats.

Adrenaline is known to increase in extra-adrenal peripheral organs as well as urine and plasma in the rat after swimming stress. In order to clarify the physiological significance of adrenaline in stress responses, the origin and location of adrenaline in the peripheral organs were examined by measuring adrenaline in the heart, spleen, submaxillary gland and some other organs after adrenalectomy or guanethidine treatment. After bilateral adrenalectomy, adrenaline in the peripheral organs mostly disappeared. Swimming stress caused no increase in the adrenaline content of the organs in the adrenalectomized rats, while a marked increase was observed in the sham-operated animals. In guanethidine-treated rats, the adrenaline content of the peripheral organs was extremely low as compared to that in the saline-treated animals, and the swimming-induced increase in adrenaline level in the organs was greatly suppressed. These observations suggest that the organ adrenaline, which exists in sympathetic nerve endings, is mostly derived from the adrenal gland. It is considered that adrenaline would be involved in sympathetic neurotransmission, especially under stressful conditions.

Degeneration of myelinated sympathetic nerve fibres following treatment with guanethidine.

The specificity and characteristics of the degeneration of myelinated axons after chronic guanethidine treatment have been investigated in sympathetic and non-sympathetic nerves. Adult male Sprague-Dawley rats aged approximately 43 weeks were treated with guanethidine sulphate (50 mg per kg body weight per day) for between ten days and six weeks. Tissues were examined by qualitative and quantitative light and electron microscopy. In the superior cervical (sympathetic) ganglion (SCG), guanethidine treatment produced a 78% decrease (P = 0.009) in the mean number of myelinated fibres at a standard level of section, compared to the contralateral control ganglion which was removed surgically prior to drug treatment. This reduction in the treated SCG was apparent after 10 days, though complete degeneration of nerve cell bodies was not widespread at this stage. Degeneration of unmyelinated axons was extensive. Degenerating myelinated fibres were consistently small in diameter (up to approximately 3 microns). In individual myelinated fibres the earliest signs of degeneration involved disruption of axonal organelles, particularly the cytoskeleton, and focal widening of the periaxonal space. Myelin breakdown followed these events; degeneration of myelin still associated with a structurally intact axon was not observed. Myelin breakdown appeared to take place initially within the Schwann cell, at least to the stage of 'loosened' membranes. However, infiltrating cells were also involved in myelin phagocytosis. At all stages of treatment some small diameter myelinated fibres remained intact, and there was no evidence of degeneration of the larger diameter fibres (up to approximately 15 microns) which are consistently present in small numbers in the SCG. In the cervical sympathetic trunk, which carries preganglionic axons to the SCG and the vagus and sciatic nerves, degeneration only of unmyelinated axons was detected. These results indicate that guanethidine does not exert a primary degenerative influence on myelin or myelinating Schwann cells and that the myelin degeneration observed in the SCG is a secondary result of the previously documented selectively destructive effect of guanethidine on postganglionic sympathetic neurons. Surviving, small diameter myelinated fibres in the SCG could be either preganglionic or processes of resistant postganglionic neurons, while the larger diameter fibres are likely to be somatic. While the cervical sympathetic trunk, vagus and sciatic nerves all contain postganglionic sympathetic fibres it appears that few of these are myelinated, at least at the levels sampled in this study.

Effect of chronic guanethidine treatment on vascular norepinephrine content in the neonatal domestic pig

Vinblastine, 6-hydroxydopamine, and antibody to nerve growth factor have been shown to destroy sympathetic neurons in neonatal rats. High doses of guanethidine produce permanent sympathectomies in the adult or newborn rat. Similar studies in the cat, rabbit, and hamster demonstrate that the sympathetic nervous systems are not destroyed with guanethidine. To produce permanent sympathectomy in a large animal model, guanethidine was administered to neonatal pigs. As with the cat, rabbit, and hamster, the sympathetic nervous system in the piglets was not destroyed by guanethidine.

KCl contractions in the rat intact and bisected vas deferens: contribution of endogenous noradrenaline release.

1. KCl produced a biphasic contraction in the intact rat vas deferens. Both components were larger and the initial rapid phasic component was faster in the prostatic portion than the epididymal portion. In some experiments the epididymal phasic response was a single slow contraction, while in others it had a mixture of fast and slow responses. 2. Phentolamine reduced the phasic response but not the tonic response of the intact vas deferens. This effect was not observed after denervation produced by chronic guanethidine treatment. 3. Both phases of the response to KCl 160 mmol/l were substantially reduced by phentolamine in the epididymal portion. In the prostatic portion phentolamine produced only slight inhibition of the phasic component and had no effect on the tonic component. 4. Isoprenaline had no effect on the response to KCl 160 mmol/l but reduced both phases of the response to KCl 50 mmol/l. This effect was antagonized by propranolol. 5. It is concluded that part of the phasic component of the response to KCl in the rat vas deferens is due to the release of noradrenaline from intramural nerves.

The effect of chronic neonatal guanethidine treatment on body composition in pigs.

The effect of neonatal guanethidine administration (30 mg/kg/day) during the first 32 days of life on body composition at 20 weeks of age, was assessed in two breeds of pig which were known to differ in their sympathetic responsiveness. Guanethidine administration increased the proportion of fat and decreased the proportion and absolute weight of lean in the Pietrain breed; and it had no effect in Gloucester Old Spots. Thus it can be inferred that endogenous noradrenaline is responsible for promoting leanness in a pig breed with known sympathetic overactivity.

Role of the autonomic nervous system in the acute responses to renal artery pressure reduction in conscious dogs.

1. Plasma renin and arterial pressure responses to acute renal artery pressure reduction were compared in intact dogs and "autonomically-blocked" dogs subjected to adrenalectomy, chronic guanethidine treatment and acute vagal block (methscopolamine). 2. Following reduction of renal artery pressure plasma renin activity and concentration rose more in the autonomically blocked dogs than in the intact dogs. When renal artery pressure was held at 30 mmHg for 1 h, plasma renin activity rose by 19.1 ng/ml per h (range 11.6-28.7) in autonomically blocked dogs and 3.65 ng/ml per h (range 1.54-5.89) in intact dogs. When renal artery pressure was held at 60 mmHg plasma renin activity rose 3.28 ng/ml per h (range 2.4-4.7) and 1.90 ng/ml per h (range 1.30-3.56), respectively. 3. Arterial blood pressure also rose more in autonomically blocked dogs in accord with the greater rise in plasma renin activity. The relationships between the increases in arterial pressure and plasma renin were closely similar in the two groups. 4. We conclude that the release of renin and increase in arterial blood pressure in response to renal artery stenosis is normally inhibited by arterial baroreflexes.

Chronic guanethidine treatment of female rats including effects on the fetus.

Adult virgin female rats were injected daily with low doses (5 or 10 mg/kg) or a high dose (30 mg/kg) of guanethidine for 12 or 18 weeks respectively. 'Short' and 'long' noradrenergic neurones were unaffected by low doeses. This contrasts markedly to earlier findings in male rats in which long-term damage of 'short' noradrenergic neurones occurred, and indicates a basic difference between 'short' noradrenergic neurones in male and female rats. Widespread degeneration of both types of neurones followed treatment with high doses and little reinnervation was observed 8 weeks after cessation of treatment. Fertility, pregnancy and litter size were apparently unaffected. Some teratogenic effects were observed in the offspring of female rats treated with guanethidine (10 or 25 mg/kg/day) before and throughout pregnancy. However, these effects had largely disappeared by the time the offspring were 10 weeks old. Since noradrenergic neurones of newborn rats are particularly sensitive to damage by guanethidine it would appear that either very little guanethidine crosses the placental barrier or that noradrenergic neurones are not susceptible during prenatal development to the cytotoxic effects of guanethidine.

Reinnervation following guanethidine-induced sympathectomy of adult rats

The reinnervation of various tissues by autonomic neurons has been studied in adult rats which had been sympathectomized by chronic guanethidine treatment (30 or 60 mg/kg/day for 8--15 weeks). In the heart, iris, ileum, mesentery, vas deferens and epididymis, fluorescence histochemistry reveals an almost complete disappearance of adrenergic fibres which is very longlasting. Even after 63 weeks few fluorescent fibres can be seen in these tissues and the density of innervation is not increased by incubation in alpha-methylnoradrenaline. The superior cervical ganglion represents an exception; large numbers of fluorescent fibres but few fluorescent ganglion cells were apparent during recovery. Axon counts carried out by electron microscopy on the vas deferens showed that after recovery for one year the number of axons was similar to that of controls; however, the pattern of innervation was abnormal, small granular vesicles were rarely seen and there was little uptake of 5-hydroxydopamine. On the basis of this evidence and of some pharmacological data it is suggested that there is a limited reinnervation by adrenergic axons accompanied by a great increase in the number of non-adrenergic, possibly cholinergic, axons.

Chronic guanethidine treatment increases cardiac β-adrenergic receptors

VARIOUS pharmacologically active agonists capable of altering cell function have been shown to interact primarily with specific receptors on the outer cell membrane1. Evidence has accumulated from several systems to suggest that there may be a dynamic and inverse relationship between the number or density of receptors on the membrane surface and the degree of receptor occupancy by agonist2,3. We present here experiments which demonstrate in rat hearts that chronic withdrawal of adrenergic stimulation produced by guanethidine treatment in vivo results in increased cardiac β-adrenergic receptor density. Further, in isolated, perfused hearts obtained from guanethidine-treated animals, there is an increased cyclic AMP (cAMP) accumulation in response to the β-adrenergic agonist isoprenaline, suggesting that the increased numbers of receptors may be related to the increased biochemical response.

Selectivity of neuronal degeneration produced by chronic guanethidine treatment.

Chronic guanethidine treatment of rats produced extensive damage to sympathetic neurons of the superior cervical ganglion and pelvic plexus. No ultrastuctural changes were observed in parasympathetic cholinergic neurons in the ciliary ganglion and pelvic plexus, nor in sensory neurons in nodose and dorsal root ganglia. A total of only six nerve cell bodies free of degenerative changes were observed in sections of superior cervical ganglia from 20 rats. This suggests either that the earlier estimates of 5% cholinergic neurons in the superior cervical ganglion based on acetylcholinesterase staining are too high, or implies that sympathetic cholinergic neurons, unlike parasympathetic neurons, are damaged by chronic guanethidine treatment.

Prolonged effects of chronic guanethidine treatment on the sympathetic innervation of the genitalia of male rats.

Prolonged effects of chronic guanethidine treatment on the sympathetic innervation of the genitalia of male rats.