Inflation reflex in the rat.

The Breuer-Hering inflation reflex [BHIR] was elicited in conscious and anaesthetized rats by inflating the lungs with constant pressures of 5--20 cm H2O. The reflex was elicited well in conscious animals, but even with the maximum stimulus [inflation of 20 cm H2O, corresponding to about 4.5-fold the tidal volume] the duration of apnoea did not exceed 4 control respiration cycles. In anaesthetized animals, the same stimulus let to apnoea lasting 180--400 control respiration cycles on the average, according to the type and depth of general anaesthesia. The duration of apnoea in occlusion of the air passages in the expiratory position increased with the depth of anaesthesia, while in occlusion of the air passages at the peak of inspiration it was shortened. Stimulation of chemoreceptors [inhalation of a mixture 4% CO2 in O2 or of 8% O2 in N2] did not influence the elicitability or duration of the BHIR, nor did cooling the rats to 28 degrees C or heating them to 38 degrees C. The mean respiration frequency was 98 c/min in unanaesthetized rats, 96 c/min in urethane anaesthesis and 79--48 c/min in halothane anaesthesia, according to the depth of anaesthesia. Bilateral cervical vagotomy reduced mean respiration frequency to 35.6 c/min in conscious rats and to 31 c/min in urethane-anaesthetized animals. The results indicate the existence of species-related differences between basic regulatory mechanisms in the rat and certain other mammals.     

Effect of lung inflation on the respiratory frequency and heart rate of premature neonates

Changes in the respiratory frequency and heart rate in response to 10 seconds' inflation of the lungs with oxygen by the CPAP method were studied in 32 premature neonates. Elevation of the pressure in the airways and lungs of 0.25, 0.49, 0.73 and 0.98 kPa led to a slower respiration rate or to apnoea. The incidence of apnoea rose in proportion to the pressure. When inflation was started, forced inspiratory efforts (a gasp reflex) occurred. The incidence of the efforts was higher at higher inflation pressures, but their intensity was not correlated to the size of the pressure. They were manifested in a drop in oesophageal pressure to a mean -2.0 +/- 0.2 kPa and their mean duration was 169 +/- 8 ms. The administration of oxygen without an increase in pressure produced neither apnoea nor forced inspiratory reactions. Neither an increase in the pressure in the airways and lungs to the above values, nor the plain administration of oxygen, produced any significant changes in the instantaneous heart rate.     

Breathing pattern of anesthetized humans during pancuronium-induced partial paralysis

We investigated the breathing patterns of 17 subjects anesthetized with enflurane before and after partial muscle paralysis produced by pancuronium bromide. In the face of significant muscle weakness produced by pancuronium, breathing patterns are characterized by decreases in both tidal volume and respiratory frequency. The decreased tidal volume corresponded to the decrease in occlusion pressure, indicating that the decreased tidal volume results solely from a decreased contractile force of the respiratory muscles. The decreased respiratory frequency was due to prolongation of both inspiratory and expiratory time without changing the ratio of the inspiratory time to the total breath time. Withdrawal of phasic vagal influence by airway occlusion before partial muscle paralysis revealed that an active Breuer-Hering inflation reflex was operative in only 8 of all 17 subjects. Since the contribution of the Breuer-Hering inflation reflex alone does not seem to account for the consistent decrease in respiratory frequency, some other mechanisms modulating respiratory frequency might be involved in the characteristic breathing patterns during partial muscle paralysis under enflurane anesthesia.     

The expiration reflex during ontogenesis in the rat.

The authors studied the elicitability of the expiration and aspiration reflex and of the respiratory reaction from the tracheobronchial area in 131 anaesthetized rats (aged 1-15 days, adult and biologically old). They found that the expiration reflex could be elicited, in the rat, from the first day of life, at a time when other respiratory reflexes were not yet stable. In young rats, the expiration reflex was often followed by a cough reaction which was absent in adult animals. The findings indicate that the expiration reflex is one of the most important respiratory reflexes of the early postnatal period in the rat, because the aspiration reflex and the respiratory reaction from the bronchi were not stable until the 15th day of life. In biological old rats, the expiration reflex is less frequently elicited and its intensity attains about half the value found in adult animals. The aspiration reflex and the respiratory reaction from the bronchi are likewise less readily elicited than in adult animals, but when the intensity of their maximum expiratory effort is increased, it is far greater.     

Respiratory mechanics and lung development in the rat from early age to adulthood.

The purpose of the present study was to establish how the dependence of respiratory mechanics on lung inflation changes during development. We studied seven groups of rats from 10 days to 3 mo of age at five levels of positive end-expiratory pressure (PEEP) from 0 to 7 hPa (1 hPa = 0.1 kPa approximately 1 cmH(2)O). At each PEEP level, we measured respiratory system resistance and elastance at both 0.9 and 4.8 Hz to partition the mechanical properties into its airway and tissue components. Elastance increased more rapidly with PEEP in the younger animals, which we interpret as reflecting a more pronounced strain stiffening of the younger parenchyma. However, the decrease in airway resistance with PEEP was more pronounced in the older animals. Morphometric analysis showed that mean tissue density decreased and total alveolar surface area increased with age. Our data suggest that the mechanical interdependence between airways and parenchyma is weaker in very young animals compared with mature animals. This may play a role in the hyperresponsiveness of immaturity.     

Interrupter technique for measurement of respiratory mechanics in anesthetized cats

In six spontaneously breathing anesthetized cats (pentobarbital sodium, 35 mg/kg ip), airflow, changes in lung volume, and tracheal and esophageal pressures were measured. Airflow was interrupted by brief airway occlusions during relaxed expirations (elicited via the Breuer-Hering inflation reflex) and throughout spontaneous breaths. A plateau in tracheal pressure occurred throughout relaxed expirations and the latter part of spontaneous expirations indicating respiratory muscle relaxation. Measurement of tracheal pressure, immediately preceding airflow, and corresponding volume enabled determination of respiratory system elastance and flow resistance. These were partitioned into lung and chest wall components using esophageal pressure. Respiratory system elastance was constant over the tidal volume range, divided approximately equally between the lung and chest wall. While the passive pressure-flow relationship for the respiratory system was linear, those for the lung and chest wall were curvilinear. Volume dependence of chest wall flow resistance was demonstrated. During inspiratory interruptions, tracheal pressure increased progressively; initial tracheal pressure was estimated by backward extrapolation. Inspiratory flow resistance of the lung and total respiratory system were constant. Force-velocity properties of the contracting inspiratory muscles contributed little to overall active resistance.     

Ontogenetic development of etomidate action in rats

Action of etomidate was studied in 219 albino rats 7, 12, 18, 25 days old and adult. Etomidate at the dose of 20 mg/kg i.p. led to a loss of the righting reflex in all age groups. Sleeping time increased with decreasing age up to the 12th day. Seven-day-old rat pups exhibited short sleeping time, approximately at the level of adult animals. Etomidate can be used for general anaesthesia even in young animals but the quantitative differences in its efficacy have to be taken into account.     

Recovery of breathing pattern after 15 min of cerebral ischemia in rabbits

The study was undertaken to ascertain the neural control of breathing and vagal reflexes during and after cerebral ischemia. The experiments were performed on anesthetized, paralyzed, and artificially ventilated rabbits. Cerebral ischemia was induced by reversible intrathoracic occlusion of the brachiocephalic trunk and the left subclavian and both internal thoracic arteries for 15 min. The effect of cerebral ischemia on breathing pattern was assessed by monitoring the integrated activities of phrenic and recurrent laryngeal nerves. Ischemia produced enhancement of breathing followed by apnea and gasping. During enhanced breathing as well as during gasping, the inspiratory-inhibiting effect of lung inflation (Breuer-Hering reflex) was abolished. When brain circulation was restored, respiratory activity started with gasps, which later were intermingled with eupneic type of inspirations. During the onset of a eupneic breath, lung inflation produced inspiratory facilitation but never an inhibition. However, after 30 min of recovery from cerebral ischemia, the Breuer-Hering reflex was restored. Results show that precise analysis of vagal reflexes and respiratory pattern during ischemia and resuscitation may be used as an indicator of resumption of autonomic activity in the brain stem.     

The effect of tachykinins microinjections into the solitary tract nucleus on respiration and blood circulation in rats

Administration of substance P and kassinin into the solitary tract nucleus of anesthetized rats induced a dose-dependent increase in ventilation, tidal volume, inspiratory muscle activity, and a decrease in the mean blood pressure and heart rate. Microinjections of peptides caused a decrease in ventilatory response to hypoxia and an inhibition of the Breuer-Hering reflex. The data obtained suggest involvement of tachykinins in the respiratory and circulatory control via the solitary tract nucleus.     

Alveolar liquid pressures in newborn and adult rabbit lungs

To study the effects of lung maturation and inflation on alveolar liquid pressures, we isolated lungs from adult and newborn rabbit pups (1-11 days old). We used the micropuncture technique to measure alveolar liquid pressure at several transpulmonary pressures on lung deflation. Alveolar liquid pressure was greater than pleural pressure but less than airway pressure at all transpulmonary pressures. Alveolar liquid pressure decreased further below airway pressure with lung inflation. At high transpulmonary pressure, alveolar liquid pressure was less in newborn than in adult lungs. To study the effects of edema, we measured alveolar liquid pressures in newborn lungs with different wet-to-dry weight ratios. Alveolar liquid pressure increased with progressive edema. In addition, we compared alveolar liquid and perivenular interstitial pressures in perfused newborn lungs and found that they were similar. Thus alveolar liquid pressure can be used to estimate perivenular interstitial pressure. We conclude that the transvascular pressure gradient for fluid flux into the interstitium might increase with lung inflation and decrease with progressive edema. Furthermore, this gradient might be greater in newborn than adult lungs at high inflation pressures.     

Compliance of the respiratory system in newborn and adult rats after gestation in hypoxia

We hypothesized that hypoxia during gestation modifies the compliance of the respiratory system of newborn and adult rats. Pregnant rats were placed in a hypobaric chamber at an inspired oxygen pressure of 86 mmHg (equivalent to 12% O₂ in normobaria) from day 4 of gestation until day 2 post-partum. Three- day-old rat pups were smaller than controls, with higher hematocrit; the lungs were also small, with less protein and DNA content. The pressure (x-axis)-volume (y-axis) curve of the respiratory system was displaced to the right of the control curve, and the compliance of the respiratory system, measured on the inflation or deflation limb of the pressure-volume curve, was decreased by ∼20–25%, depending upon the normalization procedure (per body mass or per dry lung weight). In 50-day-old rats exposed to hypoxia during gestation, body weight, hematocrit, lung mass and DNA content were normal; the compliance of the respiratory system, measured at ventilation frequencies between 20 cpm and 100 cpm, was higher than in controls by ∼20%. It is concluded that the effects of prenatal hypoxia on the compliance of the respiratory system can vary with age. In the rat the process of alveolar formation initiates postnatally. Hence, in the newborn the effects of the prenatal hypoxia on the compliance of the respiratory system are likely to be dominated by the hypoxic pulmonary hypoplasia and hypertension, which decrease the compliance of the respiratory system. In the adult, the effects of the decreased alveolar formation are the prevailing ones, increasing the compliance of the respiratory system. Accepted: 3 January 2000     

Participation of the Breuer-Hering inflation reflex in regulation of respiration frequency in anaesthetized rats.

The respiration frequency and Breuer-Hering inflation reflex (BHIR) values were compared in anaesthetized rats after unilateral vagotomy. Nos significant differences were found in the decrease in respiration frequency after transection of the right of left vagus and there were no differences in BHIR values in correlation to conduction by the right or left vagus. With unilateral vagotomy and an inflation pressure of 20 cm H20 the BHIR values fell to about one fourth of the control values. Progressive cooling of the contralateral vagus led at 8-10 degrees C to a non-significant decrease in respiration frequency to 87% of the initial value; in this state the BHIR could not be elicited. Further cooling of the vagus to 0 degrees C reduced respiration frequency to the same values as in bilateral vagotomy. The discrepancy between the mild decrease in respiration frequency and simultaneous disappearance of the BHIR shows that changes in respiration frequency in anaesthetized rats are not directly correlated to the presence of the BHIR.     

bFGF increases collateral blood flow in aged rats with femoral artery ligation

We tested the hypothesis that aged animals are as responsive as the young adult animals in expanding collateral vasculature under a similar treatment of basic fibroblast growth factor (bFGF). Two age groups of male Fischer 344 rats (11 mo old; n = 32, 23 mo old; n = 43) weighing approximately 385 g were subdivided into normal, acute ligation [femoral artery (FA) ligated 3 days before blood flow (BF) measurement] or ligated groups for 16 days and received recombinant human bFGF intra-arterial infusion at doses of 0, 0.5, 5, and 50 microg x kg(-1) x day(-1). BF was determined with (85)Sr- and (141)Ce-labeled microspheres during treadmill running at 15 and 20 m/min at 15% grade. Blood pressure (BP) values were approximately 149 and approximately 163 mmHg (p < 0.05); heart rates were approximately 496 and approximately 512 beats/min in the aged and young adult groups during running, respectively. Maximal collateral BF values were confirmed by no additional BF increase in the calf muscle at the higher speed. Ligation of the FA for 3 days reduced the BF reserve to the calf muscle by approximately 90%. Calf muscle BF was modestly greater (10 ml x min(-1) x 100 g(-1)) by 16 days in the carrier group. bFGF infusion expanded collateral BF in a dose-dependent manner with an increase of 33 and 42 ml x min(-1) x 100 g(-1) (P < 0.001) in the 5 and 50 microg x kg(-1) x day(-1) bFGF groups, respectively. Aged animals showed similar BF improvements as observed with the adult groups in response to ligation surgery and bFGF treatment. Our data indicate that the aged rats (approximately 23 mo old) remain responsive to exogenous bFGF induced in developing collateral-dependent BF as the young adult (approximately 11 mo old) controls. This suggests that the influence of bFGF in expanding collateral BF should not be preempted in the aged group, the population most affected by peripheral arterial insufficiency.     

Sympathetic regulation of adrenal medullary function during aging

Our recent studies on changes in sympathoadrenal medullary function with age in anesthetized Wistar rats were reviewed. Although secretion rates of adrenaline and noradrenaline from the adrenal gland under resting conditions varied among animals, they gradually increased after 300 days and reached a level 2-4 times higher at 800-900 days compared with that of 100 days. Spontaneous activity of a single sympathetic nerve fiber under resting conditions also increased during aging in a manner similar to the catecholamine secretion rates. Reflex responses of mass activity of adrenal sympathetic nerve fibers to stimulation of baroreceptor and cutaneous mechanoreceptors were compared in young adult (4 months old) and aged (26 months old) Wistar rats under strictly controlled conditions for anesthesia, respiration and body temperature. Under these conditions the reflex depression in response to baroreceptor stimulation and cutaneous brushing as well as reflex excitation in response to cutaneous pinching were quite well maintained in the aged rats.     

Vagal afferent activity and body temperature in 3 to 10-day-old and adult rats

This study examined the possible contribution of vagal stretch receptor activity to the increased power of the Hering-Breuer reflex in hyperthermia in rats during the early postnatal period. Experiments were performed on 10 anesthetized (pentobarbital 40 mg/kg, i.p.) 3 to 10-day-old (body weight of 16 +/- 1 g; SE) and, for comparison, 18 adult Sprague-Dawley rats (body weight of 336 +/- 35 g). Animals were tracheostomized and artificially ventilated with oxygen. The left vagus nerve was cut. In adult animals, single receptor fibers or a bundle of a few fibers were recorded using a bipolar stainless-steel electrode under mineral oil. In the young rats, a suction electrode filled with normal saline was used. Positive pressure of either 5 or 10 cmH2O was applied to the trachea when the respirator was turned off. The vagal activity was amplified and monitored on a storage oscilloscope for calculation of the frequency of vagal afferent activity during a given pressure application at different rectal temperatures (T(R); range 28 to 42 degrees C). In total, 30 and 31 sets of vagal activity in the young and adult rats, respectively, were analyzed. In all cases, an increase in tracheal pressure (P(TR)) from 5 to 10 cmH2O increased the frequency of vagal firing. The increase was greater in the adult versus the young animals; at 36 degrees C the increase was 49 +/- 11% and 16 +/- 3% in the adult and young rats, respectively (P < 0.01). In all animals, vagal receptors showed temperature-sensitivity, but less so in the young than in the adult rats (P < 0.0004 and P < 0.003; for P(TR) of 5 and 10 cmH2O, respectively). In addition, the relationship between temperature-sensitivity and T(R) had significant slopes (P < 0.001 for both inflation pressures) in the adults but not in the young rats, indicating that in the latter the temperature-sensitivity of vagal receptors is independent of TR. These results imply that temperature-sensitivity of vagal receptors could have contributed to the increased power of the Hering-Breuer reflex in rats during the early postnatal period in the warmer environment.     

Adaptive responses of 25-hydroxyvitamin D3 1-alpha hydroxylase expression to dietary phosphate restriction in young and adult rats

Regulation of vitamin D metabolism alters with age. The present study is undertaken to investigate if the loss of renal 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) production in response to dietary phosphate (P) restriction in adult rats is due to an alteration in the renal expression of 25-hydroxyvitamin D(3) 1-alpha hydroxylase (1-OHase). Young (4-6 weeks old) and adult (12-14 weeks old) male Sprague Dawley rats were fed either normal P (NPD) or low P diet (LPD) for 0-5 days. Basal expression of 1-OHase protein was higher in adult rats. Young rats, but not adult rats, significantly increased 1-OHase protein and mRNA expressions in response to LPD in a time-dependent manner. To determine if the stability of renal 1-OHase protein changes with LPD feeding, young and adult rats fed either NPD or LPD for 5 days were injected intravenously with cycloheximide (CHX), a protein synthesis inhibitor. CHX decreased 1-OHase protein expression in young rats fed NPD. However, CHX did not alter 1-OHase protein expression in young rats fed LPD nor in adult rats fed either diet. The results indicate that the stability of renal 1-OHase protein increased with age and that LPD increased its stability only in young rats.     

Postnatal development of the scratch reflex in rabbits]

The development of the scratch reflex was studied in newborn (up to 2 months old) rabbits in norm and after elimination or activation of some parts of their nervous system (reticular formation, cerebellum, caudate nucleus, cerebral cortex, superior cervical sympathetic ganglia). The experiments with the section of the brain stem at the border between the medulla and the midbrain showed that in very young (5-10 days old) rabbits in norm the scratch reflex is controlled by the spinal cord with no influences of structures situated above the section's level. Later on the spinal mechanism of the scratch reflex becomes subject to supraspinal influences, among which in 2-3 weeks old animals facilitatory effects are predominant produced, in particular, by the reticular formation and the cerebellum, whereas in older age prevail inhibitory influences of the cerebral cortex, cerebellum, caudate nucleus and the sympathetic nervous system.     

Hering-Breuer reflex in conscious newborn rats: effects of changes in ambient temperature during hypoxia.

In a previous study in conscious normoxic newborn rats, we found that the strength of the Hering-Breuer reflex (HB reflex) was greater (188%) at high (36 degrees C) than at low (24 degrees C) ambient temperature (T(a); D. Merazzi and J. P. Mortola. Pediatr. Res. 45: 370-376, 1999). We now asked what the effect would be of changes in T(a) during hypoxia. Rat pups at 3-4 days of age were studied in a double-chamber airflow plethysmograph. The HB reflex was induced by negative body surface pressures of 5 or 10 cmH(2)O and quantified from the inhibition of breathing during maintained lung inflation. Rats were first studied at T(a) = 32 degrees C in normoxia, followed by hypoxia (10% O(2) breathing). During hypoxia, oxygen consumption (VO(2)) averaged 47%, and HB reflex 115%, of the corresponding normoxic values, confirming that in the newborn, differently from the adult, hypoxia does not decrease the strength of the HB reflex. As hypoxia was maintained, lowering T(a) to 24 degrees C or increasing it to 36 degrees C, on average, had no significant effects on VO(2) and the HB reflex. However, with 5-cmH(2)O inflations, the HB reflex during the combined hypoxia and hyperthermia was significantly stronger than in normoxia. We conclude that in conscious newborn rats during normoxia the T(a) sensitivity of the HB reflex is largely mediated by the effects of T(a) on thermogenesis and VO(2); in hypoxia, because thermogenesis is depressed and VO(2) varies little with T(a), the HB reflex is T(a) independent. The observation that the reflex response to lung inflations during hypoxic hyperthermia can be greater than in normoxia may be of importance in the pathophysiology of apneas during the neonatal period.     

Effect of age on passive elastic stiffness of rat heart muscle.

A thick-wall spherical model for the rat left ventricle was used to deduce passive wall stiffness from diastolic pressure-volume data. This was done for rats in three age classes: young (1 mo), adult (17 mo) and old (17 mo). The model was based on finite deformation elasticity theory consistent with the magnitude of observed deformation. A least-squares procedure was used to determine elastic constants in postulated nonlinear stress-stretch relations for the myocardium. It was found that at a given level of stress, wall stiffness for ventricles in the young age class was consistently greater than wall stiffness in the other two classes. In addition, the difference in wall stiffness between rats in the adult and old age classes was found to be approximately 10%.     

Age differences in responsiveness of brainstem chemosensitive neurons to extracellular pH changes

The sensitivity of neurons in the caudal chemosensitive area on the ventrolateral surface of the medulla oblongata (VMS) to extracellular pH changes was examined in newborn and young developing kittens and compared to that of adult cats. The pH was varied by superfusion of the VMS with mock cerebrospinal fluid (CSF) of pH 7.4 (control), 7.0 (acid) and 7.8 (alkaline). A total of 97 neuronal units in the three age groups changed their firing rates inversely in response to extracellular fluid (ECF) pH changes. The greatest sensitivity was found in the adult group where acid superfusion caused an increase in neuronal activity. The least sensitivity was observed in the newborn group (1-6 days old), whereas the young kitten group (4-6 weeks old) exhibited an intermediate sensitivity. Neurons of kittens older than 7 weeks of age demonstrated a response pattern characteristic of the adult group. Neurons of neonates older than seven days, exhibited a response pattern characteristic of the young kitten group.