David Perrin

Behavioral and neurochemical effects induced by subchronic exposure to 40 ppm toluene in rats.

Chronic toluene inhalation at concentrations above occupational exposure limits (e.g., 100 ppm; NIOSH) has been repeatedly shown to induce neurotoxic effects. In contrast, although few clinical and experimental data are available on the effects of toluene exposure at concentrations below occupational exposure standards, some of these data may support adverse effects of long-term exposure to low toluene concentrations. To test this hypothesis, we investigated the neurobehavioral and neurochemical effects of 40 ppm inhaled toluene in a rat model of 16-week subchronic exposure, examining locomotor and rearing activities; adaptation/sensitization to narcosis produced by acute exposure to toluene at high concentration; and tyrosine hydroxylase and tryptophan hydroxylase activities, and dopamine (DA) and serotonin (5-HT) turnovers in the caudate-putamen, nucleus accumbens, hippocampus, prefrontal cortex, and cerebellum. Our results mainly show that subchronic exposure to 40 ppm toluene significantly resulted in a sensitization to toluene-induced narcosis, a decrease in rearing activity, and alterations in DA and 5-HT transmissions. This demonstrates that subchronic toluene exposure at a low concentration may lead to adverse changes in neurobehavioral and neurochemical functioning, and further questions in a public health perspective the actual neurotoxic potential of toluene and other organic compounds, because deficits in functioning are generally viewed as precursors of more serious adverse effects.

Long-term prenatal hypoxia alters maturation of adrenal medulla in rat.

Catecholamine release from the adrenal medulla glands plays a vital role in postnatal adaptation. A number of pathologic situations are characterized by oxygen deficiency. The objective of the present study was to determine the influence of long-term prenatal hypoxia on maturation of the adrenal medulla. Pregnant rats were subjected to hypoxia (10% O2) from the fifth to the 20th d of gestation. The offspring were examined on the 19th d of gestation (E19), the day of birth (P0), and at postnatal (P) day of life P3, P7, P14, P21, and P68. The catecholamine content and activity of tyrosine hydroxylase (TH) in vivo were assayed by HPLC with electrochemical detection. Cellular expression of TH and phenylethanolamine N-methyl transferase was evaluated by protein immunohistochemistry and in situ hybridization of the corresponding mRNA species. Exposure to prenatal hypoxia reduced the epinephrine content of the adrenal medulla on E19, P0, P3, and P7 while increasing the norepinephrine content on E19, P0, and P14. Furthermore, the peak epinephrine to norepinephrine ratio appearing between P7 and P10 in the normoxic offspring was absent in the hypoxic offspring. The in vivo TH activity was increased on P3 and P14 and decreased on P68. The percentage of chromaffin cells in the medulla expressing TH and phenylethanolamine N-methyl transferase was lowered on E19, P0, and P7. TH and phenylethanolamine N-methyl transferase mRNA levels were reduced on P7. Clearly prenatal hypoxia results in major changes in adrenal catecholamine stores and synthesis during the perinatal period, which persist into adulthood. The capacity to cope with postnatal stress might be disturbed as a consequence of prenatal hypoxia.

Central and peripheral changes in catecholamine biosynthesis and turnover in rats after a short period of ozone exposure.

We investigated in rat the effects of ozone exposure (0.7 ppm) for 5 h on the catecholamine biosynthesis and turnover in sympathetic efferents and various brain areas. For this purpose, the activity of tyrosine hydroxylase, the rate-limiting enzyme in catecholamine biosynthesis, was assessed in superior cervical ganglia and in two major noradrenergic cell groups, A2 and A6 (locus coeruleus). Tyrosine hydroxylase activity was estimated in vivo by measuring the accumulation of l-dihydroxyphenylalanine after pharmacological blockade of L-aromatic acid decarboxylases by NSD-1015 (100 mg/kg i.p.). The catecholamine turnover rate was measured after inhibition of tyrosine hydroxylase by alpha-methyl-para-tyrosine (AMPT, 250 mg/kg, i.p., 2.5 h) in peripheral sympathetic target organ (heart and lungs) as well as in some brain catecholamine terminal areas (cerebral cortex, hypothalamus and striatum). Ozone caused differential effects according to the structure. Catecholamine biosynthesis was stimulated in superior cervical ganglia (+44%, P < 0.05) and caudal A2 subset (+126%, P < 0.01), whereas catecholamine turnover was increased in heart (+183%, P < 0.01) and cortex (+22%, P < 0.05). On the other hand, catecholamine turnover was inhibited in lungs (-53%, P < 0.05) and striatum (-24%, P < 0.05). A brief exposure to ozone, at a concentration chosen to mimic pollution level encountered in urban areas, can modulate catecholamine biosynthesis and utilization rate in the sympathetic and central neurones.

Neurochemical development of the brainstem catecholaminergic cell groups in rat.

Summary. The postnatal development of tyrosine hydroxylase activity has been studied in the brainstem catecholaminergic cell groups (A1C1, A2C2, A5, A6, A7), involved in cardiorespiratory control. In rat, at birth and at postnatal days P3, P7, P14, P21 ant P68, we used a microdissection technique followed by in vivo measurement of the tyrosine hydroxylase (TH) activity, the rate-limiting enzyme in catecholamine synthesis. There is two successive marked increases in TH activity: at P3 in every catecholaminergic cell groups (A1C1, +225%; A2C2, +300%; A5, +190%; A6, +205% compared to birth) and during the third postnatal week with a peak of TH activity at P14 (A6, +90% above the P7 level) or at P21 (A1C1, +715%; caudal A2C2, +585%; rostral A2C2, +15%; A5, +445%; A7, +180% compared to P7). The data suggest the existence of two temporal windows during the neurochemical development of the catecholaminergic cell groups, which correspond to two metabolic transitions. The first one could be related to the intra-, extrauterine transition and the second one, to a deep energetic phase of maturation in the rat brain, closely related to the maturation of cardiorespiratory processes.

Sympathetic and brain monoaminergic regulation of energy balance in obesity-resistant rats (Lou/C)

In contrast to the Wistar rat, the Lou/C rat does not develop obesity with age. To determine the role of sympathetic output and brain monoamines in the different energy balance of Lou/C rats, the monoamine contents and activity of rate-limiting enzymes in catecholamine and serotonin biosynthesis were assessed in brain structures involved in energy balance regulation, i.e., brainstem noradrenergic (A6, A5, A2) and serotonergic cell groups (dorsal raphe, and median raphe), and two hypothalamic nuclei (ventromedial nucleus and paraventricular nucleus). In vivo tyrosine hydroxylase activity and noradrenaline content were measured in sympathetic target organs storing fuel substrates, the liver, white adipose and brown adipose tissues in the Lou/C rat and compared to the Wistar rat. In Lou/C rats, indirect calorimetric measurements showed a higher energy expenditure despite a reduced food intake. The Lou/C rat displayed selective monoamine features. The catecholaminergic activity was higher in the white adipose tissue and interscapular brown adipose tissue but lower in the liver and adrenal gland of Lou/C rats. The noradrenergic activity in A2, A6 and ventromedial nucleus, and the serotonergic pattern in A6, dorsal raphe and median raphe were lower in Lou/C. The metabolic particularities of Lou/C rats are associated with (i) a selectively enhanced sympathetic activity restricted to the white adipose tissue and brown adipose tissue, (ii) a reduced noradrenergic activity in selective brainstem and hypothalamic areas, which control the energy expenditure and food intake.

Prenatal nicotine alters maturation of breathing and neural circuits regulating respiratory control.

While perinatal nicotine effects on ventilation have been widely investigated, the prenatal impact of nicotine treatment during gestation on both breathing and neural circuits involved in respiratory control remains unknown. We examined the effects of nicotine, from embryonic day 5 (E5) to E20, on baseline ventilation, the two hypoxic ventilatory response components and in vivo tyrosine hydroxylase (TH) activity in carotid bodies and brainstem areas, assessed at postnatal day 7 (P7), P11 and P21. In pups prenatally exposed to nicotine, baseline ventilation and hypoxic ventilatory response were increased at P7 (+48%) and P11 (+46%), with increased tidal volume (p<0.05). Hypoxia blunted frequency response at P7 and revealed unstable ventilation at P11. In carotid bodies, TH activity increased by 20% at P7 and decreased by 48% at P11 (p<0.05). In most brainstem areas it was reduced by 20-33% until P11. Changes were resolved by P21. Prenatal nicotine led to postnatal ventilatory sequelae, partly resulting from impaired maturation of peripheral chemoreceptors and brainstem integrative sites.

A6 Noradrenergic Cell Group Modulates the Hypoxic Ventilatory Response

A6 noradrenergic cell groups or locus cœruleus (LC) are a pair of nuclei located in the peri-ventricular grey-matter of the rostral pons(Dalhstrom & Fuxe, 1964). A6 possesses a high concentration of noradrenergic perikarya accounting for nearly 45 % of the whole brain noradrenergic cell bodies. It extensively projects in the brain and provides noradrenergic innervation to many brain areas and to the spinal cord (Fritschy & Grzanna, 1990). Kaehler et al.(1999) demonstrate an activation of A6 cell group under hypoxia. Indeed, LC neurones exhibit an increased firing rate (Elam et al., 1981, Guyenet et al., 1993) and an immediate early gene induction (c-fos) under exposure to systemic hypoxia (Breen et al., 1996). Several studies (Li et al, 1992, Guyenet et al, 1993) strongly suggest that A6 is involved in central modulation of breathing, however its role in the ventilatory response to hypoxia remains to be defined. The aim of the present study was to define in adult rat the contribution of the LC noradrenergic neurons for the establishment of the ventilatory response to short-term hypoxia. Hence, we developped a selective lesion of the LC noradrenergic component using 6-hydroxydopamine (6-OHDA). The ventilatory effect of this lesion was tested in awake and unrestained animals by analyzing the hypoxic ventilatory response using whole body plethysmography.

Developmental plasticity of the carotid chemoafferent pathway in rats that are hypoxic during the prenatal period

The chemoreflex pathway undergoes postnatal maturation, and the perinatal environment plays a critical role in shaping respiratory control system. We investigated the role of prenatal hypoxia on the maturation of the chemoreflex neural circuits regulating ventilation in rat. Effects of hypoxia (10% O2) from the 5th to the 20th day of gestation were studied on male offspring at birth and on postnatal days 3, 7, 21 and 68. Maturation of the respiratory control system was assessed by in vivo tyrosine hydroxylase (TH) activity measurement in peripheral chemoreceptors (carotid bodies, petrosal ganglia), and in brainstem catecholaminergic cell groups (A2C2c and A1C1 areas in the medulla, A5 and A6 areas in the pons). Resting ventilation and ventilatory response to hypoxia were evaluated as functional sequelae. In peripheral structures, prenatal hypoxia reduced TH activity within the first postnatal week and enhanced it later. In contrast, in central areas, prenatal hypoxia upregulated TH activity within the first postnatal week and downregulated it later. The in vivo TH activity impairment is therefore tissue specific, with an opposite effect on the peripheral and central neural circuits. A shift of the effect of prenatal hypoxia occurred between 1 and 3 weeks, indicating a postnatal temporal effect of prenatal hypoxia. An important period in the development of the chemoafferent pathway occurred between the first and the third postnatal week. Functionally, prenatal hypoxia impaired resting ventilation and ventilatory response to hypoxia. The alterations of the catecholaminergic components of the chemoafferent pathway resulting from prenatal hypoxia might contribute to impair postnatal respiratory behaviour.

Deployment of stent grafts in curved aneurysmal arteries: toward a predictive numerical tool

The mechanical behavior of aortic stent grafts plays an important role in the success of endovascular surgery for aneurysms. In this study, finite element analysis was carried out to simulate the expansion of five marketed stent graft iliac limbs and to evaluate quantitatively their mechanical performances. The deployment was modeled in a simplified manner according to the following steps: (i) stent graft crimping and insertion in the delivery sheath, (ii) removal of the sheath and stent graft deployment in the aneurysm, and (iii) application of arterial pressure. In the most curved aneurysm and for some devices, a decrease of stent graft cross-sectional area up to 57% was found at the location of some kinks. Apposition defects onto the arterial wall were also clearly evidenced and quantified. Aneurysm inner curve presented significantly more apposition defects than outer curve. The feasibility of finite element analysis to simulate deployment of marketed stent grafts in curved aneurysm models was demonstrated. The study of the influence of aneurysm tortuosity on stent graft mechanical behavior shows that increasing vessel curvature leads to stent graft kinks and inadequate apposition against the arterial wall. Such simulation approach opens a very promising way toward surgical planning tools able to predict intra and/or post-operative short-term stent graft complications.

Patient-specific simulation of endovascular repair surgery with tortuous aneurysms requiring flexible stent-grafts

The rate of post-operative complications is the main drawback of endovascular repair, a technique used to treat abdominal aortic aneurysms. Complex anatomies, featuring short aortic necks and high vessel tortuosity for instance, have been proved likely prone to these complications. In this context, practitioners could benefit, at the preoperative planning stage, from a tool able to predict the post-operative position of the stent-graft, to validate their stent-graft sizing and anticipate potential complications. In consequence, the aim of this work is to prove the ability of a numerical simulation methodology to reproduce accurately the shapes of stent-grafts, with a challenging design, deployed inside tortuous aortic aneurysms. Stent-graft module samples were scanned by X-ray microtomography and subjected to mechanical tests to generate finite-element models. Two EVAR clinical cases were numerically reproduced by simulating stent-graft models deployment inside the tortuous arterial model generated from patient pre-operative scan. In the same manner, an in vitro stent-graft deployment in a rigid polymer phantom, generated by extracting the arterial geometry from the preoperative scan of a patient, was simulated to assess the influence of biomechanical environment unknowns in the in vivo case. Results were validated by comparing stent positions on simulations and post-operative scans. In all cases, simulation predicted stents deployed locations and shapes with an accuracy of a few millimetres. The good results obtained in the in vitro case validated the ability of the methodology to simulate stent-graft deployment in very tortuous arteries and led to think proper modelling of biomechanical environment could reduce the few local discrepancies found in the in vivo case. In conclusion, this study proved that our methodology can achieve accurate simulation of stent-graft deployed shape even in tortuous patient specific aortic aneurysms and may be potentially helpful to help practitioners plan their intervention.