Katarzyna Kaczyńska

Peripheral 5-HT1A receptors are not essential for increased ventilation evoked by systemic 8-OH-DPAT challenge in anaesthetized rats.

The respiratory effects resulting from stimulation of 5‐HT1A receptors were studied in spontaneously breathing rats that were: (i) neurally intact and subsequently bilaterally vagotomized; (ii) subjected to bilateral midcervical vagotomy followed by supranodosal vagotomy; (iii) midcervically vagotomized and treated by carotid sinus/body denervation; or (iv) subjected to infra‐ and supranodosal vagotomy followed by pharmacological blockade of 5‐HT1A receptors. An intravenous bolus of the 5‐HT1A receptor agonist 8‐hydroxy‐dipropylaminotetralin (8‐OH‐DPAT, 10 μg kg−1) evoked increases in both breathing rate and tidal volume. After section of the midcervical and supranodosal vagi, 8‐OH‐DPAT challenge still increased the respiratory rate and tidal volume. Carotid sinus/body denervation did not reduce the augmentation of the tidal volume, but prevented the increase in breathing rate. Blockade of 5‐HT1A receptors with intravenous doses of 1‐(2‐metoxyphenyl)‐4‐[4‐(2‐phthalimido) butyl] piperazine (NAN 190; 20 μg kg−1) abolished all respiratory effects of 8‐OH‐DPAT challenge. In all the neural states, 8‐OH‐DPAT evoked a significant fall in mean arterial blood pressure. Pretreatment with NAN 190 reduced baseline values of mean arterial pressure and prevented 8‐OH‐DPAT‐induced hypotension. These results indicate that: (i) 8‐OH‐DPAT‐evoked activation of 5‐HT1A receptors increases breathing rate and tidal volume, which persists after section of the lung vagi and the nodose ganglia, but only the increase in breathing rate was abolished by carotid sinus/body denervation; and (ii) 8‐OH‐DPAT hyperventilatory and hypotensive responses result from the excitation of presumed 5‐HT1A carotid receptors and the central 5‐HT1A‐expressing neurones.

Clonidine-evoked respiratory effects in anaesthetized rats

The respiratory effects of stimulation of α2‐adrenergic receptors were studied in spontaneously breathing anaesthetized rats that were neurally intact, or bilaterally vagotomized, or subjected to bilateral combined midcervical vagotomy and section of the carotid sinus nerves. An intravenous clonidine bolus (15 μg kg−1) evoked a prolonged slowing of the respiratory rate in all the neural states explored. Vagotomy reduced the early clonidine‐evoked decline, but not the augmentation of tidal volume that followed the decline. After section of the carotid sinus nerves, clonidine challenge continued to decrease the respiratory rate, but not the tidal volume. Blockade of α2‐adrenergic receptors with intravenous doses of SKF 86466 (200 μg kg−1) abolished all respiratory effects of the clonidine challenge. In all the neural states studied, clonidine evoked a significant short‐lived rise in mean arterial blood pressure followed by a decrease below the respective prechallenge value. The SKF 86466 pretreatment lowered mean arterial blood pressure control values and reduced the magnitude of postclonidine changes. These results indicate that: (i) clonidine‐evoked activation of α2‐adrenergic receptors affects the two components of the breathing pattern differently, and this occurs beyond the lung vagi; and (ii) changes in tidal volume result from excitation of the carotid bodies and are coupled with centrally mediated slowing of the respiratory rhythm.

NPY Y1 receptors are involved in cardio-respiratory responses to intravenous injection of neuropeptide Y in anaesthetized rats

The respiratory effects evoked by systemic injection of neuropeptide Y (NPY) were studied in anaesthetized, spontaneously breathing rats that were (i) neurally intact; (ii) subjected to bilateral midcervical vagotomy (MC vagi cut); (iii) midcervically vagotomized and treated by supranodosal denervation (NG vagi cut); (iv) neurally intact, before and after pharmacological blockade of the NPY Y(1) and NPY Y(2) receptors. An intravenous (iv) bolus of NPY (100 μg/kg) induced slowing down of the respiratory rate, decreased tidal volume and heart rate, and increased mean arterial blood pressure. After section of midcervical vagi, NPY still evoked the cardio-respiratory changes. Supranodose vagotomy abolished the fall in respiratory rate and reduced significantly the decreases in tidal volume and minute ventilation. This level of vagotomy did not affect vasopressor and bradycardic effects of NPY. Blockade of NPY Y(1) receptors with an intravenous dose of 5 mg/kg of BMS 193885, reduced significantly the cardio-respiratory effects of NPY injection. Pre-treatment with BIIE 0246, NPY (2) receptor antagonist at a dose of 1-2.5 mg/kg was not effective in blocking the response to NPY. The results of this study indicate that NPY-evoked activation of NPY Y(1) receptors decreases both components of the breathing pattern, and this response is primarily mediated central to the cervical vagi. Bradycardia and hypertensive effect of NPY are attributed to the excitation of peripheral and central NPY Y(1) receptors and occur outside of the vagal pathways.

Cardio-respiratory effects of systemic neurotensin injection are mediated through activation of neurotensin NTS1 receptors

The purpose of our study was to determine the cardio-respiratory pattern exerted by the systemic injection of neurotensin, contribution of neurotensin NTS(1) receptors and the neural pathways mediating the responses. The effects of an intravenous injection (i.v.) of neurotensin were investigated in anaesthetized, spontaneously breathing rats in following experimental schemes: (i) control animals before and after midcervical vagotomy; (ii) in three separate subgroups of rats: neurally intact, vagotomized at supranodosal level and initially midcervically vagotomized exposed to section of the carotid sinus nerves (CSNs); (iii) in the intact rats 2 minutes after blockade of neurotensin NTS(1) receptors with SR 142948. Intravenous injection of 10 μg/kg of neurotensin in the intact rats evoked prompt increase in the respiratory rate followed by a prolonged slowing down coupled with augmented tidal volume. Midcervical vagotomy precluded the effects of neurotensin on the frequency of breathing, while CSNs section reduced the increase in tidal volume. In all the neural states neurotensin caused significant fall in mean arterial blood pressure preceded by prompt hypertensive response. The cardio-respiratory effects of neurotensin were blocked by pre-treatment with NTS(1) receptor antagonist. The results of this study showed that neurotensin acting through NTS(1) receptors augments the tidal component of the breathing pattern in a large portion via carotid body afferentation whereas the respiratory timing response to neurotensin depends entirely on the intact midcervical vagi. Blood pressure effects evoked by an intravenous neurotensin occur outside vagal and CSNs pathways and might result from activation of the peripheral vascular NTS(1) receptors.

Peripheral cardiorespiratory effects of bombesin in anaesthetized rats

The respiratory effects evoked by systemic injection of bombesin were studied in spontaneously breathing rats that were (i) neurally intact and subsequently bilaterally vagotomized, (ii) intact, before and after pharmacological blockade of the bombesin BB(1) and BB(2) receptors. An intravenous bolus of bombesin (10 microg/kg) evoked sighs, decrease in the breathing rate, augmentation of tidal volume and an increase in mean arterial blood pressure. Midcervical vagotomy abolished all respiratory changes evoked by bombesin challenge, but did not prevent the increase in blood pressure. Blockade of BB(1) and BB(2) receptors with an intravenous dose of 50 microg/kg of [D-Phe](12)-bombesin, reduced significantly the cardio-respiratory effects due to bombesin administration. The BB(1) receptors antagonist, BIM 23127, at a dose of 100 microg/kg did not block the response to bombesin. These results indicate that bombesin given systemically stimulates ventilation by activation of BB(2) receptors affecting mainly the tidal component of the breathing pattern, and that the response is mediated by the lung vagi. The hypertensive effect of bombesin resulted from the excitation of BB(2) receptors, but occurred outside vagal afferentation from the lungs.

Long-Term Ultrastructural Indices of Lead Intoxication in Pulmonary Tissue of the Rat

In the present research long-term pulmonary toxicity of lead was investigated in rats treated by intraperitoneal administration of lead acetate for three consecutive days (25 mg/kg per day). Five weeks after treatment average lead content in the whole blood was 0.41 μg/dL ± 0.05, in the lung homogenates it measured 3.35 μg/g ± 0.54, as compared to the control values of 0.13 ± 0.07 μg/dL and 1.03 μg/g ± 0.59, respectively. X-ray microanalysis of lung specimens displayed lead localized mainly within type II pneumocytes and macrophages. At the ultrastructural level the effects of lead toxicity were found in lung capillaries, interstitium, epithelial cells, and alveolar lining. Alveolar septa showed intense fibrosis, consisting of collagen, elastin, and fibroblasts. Thinned alveolar septa had emphysematous tissue with some revealing signs of angiogenesis. Type II pneumocytes contained lamellar bodies with features of laminar destruction. Fragments of the surfactant layer were often detached from the alveolar epithelium. These findings indicate that 5 weeks after exposure, lead provokes reconstruction of the alveolar septa including fibrosis and emphysematous changes in the lung tissue.

Depressive cardio-respiratory effects of somatostatin in anaesthetized rats.

Cardio-respiratory effects of intravenous injection of somatostatin were investigated in anaesthetized, spontaneously breathing rats that were: (i) neurally intact and subsequently bilaterally vagotomized in the neck, or (ii) midcervically vagotomized with later vagotomized at the supranodosal level, or (iii) midcervically vagotomized and subjected to section of the carotid sinus nerves (CSNs). Intravenous injection of 100mug/kg of somatostatin before and after midcervical vagotomy-induced immediate slowing down of the respiratory rate and decreased tidal volume, mean arterial blood pressure (MAP) and heart rate. Supranodose vagotomy did not abolish somatostatin-induced respiratory depression. CSNs section eliminated all respiratory effects of somatostatin challenge. In all the neural states, somatostatin caused significant falls in mean arterial blood pressure and heart rate. The results of this study suggest that hypoventilation induced by intravenous somatostatin administration occurs outside vagal afferentation to the medulla and is mediated via carotid body afferents. Sino-aortic chemoreceptors and baroreceptors do not contribute to bradycardia and a fall in blood pressure. These cardiovascular effects are presumably mediated due to excitation of somatostatin receptors within the heart and/or in the brain.

Serotonergic system in hypoxic ventilatory response in unilateral rat model of Parkinson’s disease

BackgroundMalfunctioning of the serotonergic system in Parkinson’s disease may contribute to non-motor symptoms such as respiratory complications. Thus the aim of our study was to investigate the role of serotonin 5-HT2 receptors in the modulation of normoxic breathing and the hypoxic ventilatory response (HVR) in rat model of Parkinson’s disease.MethodsWistar rats were lesioned unilaterally with double 6-hydroxydopamine (6-OHDA) injection to the right medial forebrain bundle (MFB). Before lesion and two weeks later animals were put in whole body plethysmography chamber and exposed to hypoxia (8% O2). Before hypoxic tests animals received intraperitoneal injections of DOI and ketanserin. Efficacy of lesion was confirmed by cylinder test, assessing limb use asymmetry.ResultsDegeneration of the nigrostriatal pathway augmented response of tidal volume and minute ventilation to hypoxia. DOI administration in control and lesion state caused a significant rise in normoxic respiratory rate and minute ventilation. Yet, ventilatory response of these parameters to hypoxia was attenuated. Post-DOI magnitude of HVR in lesioned state was decreased in compare to pre-lesion control. Subsequent ketanserin injection reverted DOI-induced respiratory effects. We demonstrated that 6-OHDA treatment decreased the content of serotonin in the injured striatum and on both sides of the brainstem, leaving the concentration of noradrenaline on unchanged level.ConclusionsThese observations showed that damage of the nigrostriatal system initiates changes in the serotonergic system, confirmed by reduced concentration of serotonin in the striatum and brainstem, which affects the magnitude of respiratory response to hypoxia after activation of 5-HT2 receptors.

Hypoxic ventilatory response after dopamine D2 receptor blockade in unilateral rat model of Parkinson's disease.

Modified non-motor brainstem ventilatory control might be involved in Parkinsons disease. Our study was designed to investigate the impact of degeneration of the nigrostriatal dopaminergic pathway on resting breathing and hypoxic ventilatory response in conscious rats. The role of central and peripheral dopamine D2 receptors in the modulation of the hypoxic ventilatory response in conditions of dopamine shortage was examined. Adult Wistar rats received a unilateral double 6-hydroxydopamine lesion of the right medial forebrain bundle. After surgery, animals were placed in whole-body plethysmographic chamber and exposed to hypoxia (8% O2). One group of animals received inraperitoneal injections of either haloperidol or domperidone before hypoxia. Levels of dopamine and its metabolite in the brainstem and striatum were assessed. Neurotoxin treatment evoked limb use asymmetry. No effect on the resting normoxic respiration was observed. An increase in tidal volume and a decrease in respiratory rate during respiratory response to hypoxia with short magnification of minute ventilation were predominant effects. Domperidone treatment in intact animals evoked a significant increase in normoxic tidal volume, while haloperidol potentiated tidal volume increase in response to hypoxia. After the lesion, the effects of both antagonists were absent. In rats with Parkinsons, the content of dopamine and its metabolite decreased substantially in the injured striatum. Augmentation of a tidal volume response to hypoxia, and the absence of stimulatory effect of intraperitoneal domperidone on normoxic and haloperidol on hypoxic tidal volume, in lesioned rats indicated altered control of breathing. This could be the result of a dopamine deficiency in the striatum and an increased turnover of DOPAC/DA in the brainstem.

Vasopressor and heart rate responses to systemic administration of bombesin in anesthetized rats

The aim of the present study was to investigate the effects of aortic depressor nerve (ADN) transection, supranodosal vagi denervation (NG vagi cut) and adrenergic receptor blocker treatment on the cardiovascular responses evoked by systemic injection of bombesin. The cardiovascular effects were studied in spontaneously breathing rats that were (i) bilaterally, midcervically vagotomized (MC vagi cut) and subjected to section of the aortic depressor nerves, (ii) midcervically vagotomized and subsequently vagotomized at the supranodosal level or (iii) midcervically vagotomized before and after pharmacological blockade of α- or β-adrenergic receptors with phentolamine and propranolol, respectively. An intravenous bolus of bombesin (10 μg/kg) in midcervically vagotomized and ADN denervated animals increased mean arterial blood pressure (MAP) and heart rate (HR). An approximate 20% increase in blood pressure occurred immediately following bombesin injection and lasted for 2-3 min. Augmentation of the heart rate occurred 30-60 s after the bombesin challenge and persisted for more than 10 min. After section of the supranodosal vagi, bombesin failed to induce an increase in heart rate. Blockade of α-adrenergic receptors with an intravenous dose of phentolamine significantly reduced post-bombesin hypertension. These results indicate that bombesin-evoked increases in blood pressure do not require aortic depressor nerves and supranodosal vagi and are presumably mediated by the activation of peripheral α-adrenergic receptors. Bombesin-induced tachycardia was dependent on an intact supranodose pathway and was amplified by activation of β-adrenoceptors.