RESUMEN
It is now established that adverse conditions during pregnancy can trigger a fetal origin of cardiovascular dysfunction and/or increase the risk of heart disease in later life. Suboptimal environmental conditions during early life that may promote the development of cardiovascular dysfunction in the offspring include alterations in fetal oxygenation and nutrition as well as fetal exposure to stress hormones, such as glucocorticoids. There has been growing interest in identifying the partial contributions of each of these stressors to programming of cardiovascular dysfunction. However, in humans and in many animal models this is difficult, as the challenges cannot be disentangled. By using the chicken embryo as an animal model, science has been able to circumvent a number of problems. In contrast to mammals, in the chicken embryo the effects on the developing cardiovascular system of changes in oxygenation, nutrition or stress hormones can be isolated and determined directly, independent of changes in the maternal or placental physiology. In this review, we summarise studies that have exploited the chicken embryo model to determine the effects on prenatal growth, cardiovascular development and pituitary-adrenal function of isolated chronic developmental hypoxia.
Asunto(s)
Enfermedades Cardiovasculares/embriología , Hipoxia/embriología , Animales , Enfermedades Cardiovasculares/fisiopatología , Embrión de Pollo , Desarrollo Fetal , Humanos , Sistema Hipotálamo-Hipofisario/fisiología , Hipoxia/fisiopatología , Sistema Hipófiso-Suprarrenal/fisiologíaRESUMEN
Prenatal treatment with magnesium sulfate (MgSO4) has neuroprotective effects in very preterm infants but its use has been associated with an increased rate of patent ductus arteriosus (DA). MgSO4 is a vasodilator and thus may exert a direct relaxant effect in the DA. We aimed to investigate the vasoactive effects of MgSO4 in the DA using the chicken embryo as experimental model. DA rings from 15-d (E15), 17-d (E17) and 19-d (E19) chicken embryos (total incubation: 21-d) were mounted in a wire myograph for isometric tension recordings. Exposure of DA rings to 21% O2 induced a tonic contraction which was relaxed by MgSO4 (2.4 - 7.2 mmol L-1) in a concentration-dependent manner (mean maximal relaxation E19: 51.4%, SE 6.3; EC50: 3.5 mmol L-1, SE 0.7). The relaxation evoked by MgSO4 was not significantly different between E15, E17 and E19 DA and was not affected by removal of the endothelium or by the presence of the nitric oxide synthase inhibitor L-NAME, the soluble guanylate cyclase inhibitor ODQ, or the cyclooxygenase inhibitor indomethacin. In contrast, when the DA rings were incubated in Ca2+-free solution, or in the presence of inhibitors of L-type Ca2+ channels (nifedipine), or large-conductance Ca2+-activated K+ (BKCa) channels (iberiotoxin), MgSO4-induced relaxation was impaired. Preincubation of the DA rings with MgSO4 concentrations ranging from 0 to 6.0 mmol L-1 did not significantly affect O2-induced contraction that was only impaired by a concentration of 7.2 mmol L-1. In conclusion, MgSO4 induced endothelium-independent relaxation of chicken DA and this relaxation appeared to be mediated through stimulation of BKCa channels and blockade of transmembrane flux of extracellular Ca2+. However, O2-induced DA contraction was only impaired by suprapharmacological concentrations of MgSO4 (> 6.0 mmol L-1). Therefore, our data suggest that the higher incidence of patent DA in preterm infants exposed to MgSO4 is unlikely to be due to a direct pharmacological effect of the drug on the DA.
Asunto(s)
Conducto Arterial/efectos de los fármacos , Sulfato de Magnesio/farmacología , Animales , Calcio/metabolismo , Embrión de Pollo , Dinoprostona/fisiología , Conducto Arterial/fisiología , Conducto Arterioso Permeable/inducido químicamente , Óxido Nítrico/fisiología , Vasodilatación/efectos de los fármacosRESUMEN
Nonadrenergic noncholinergic (NANC) vasodilator mechanisms may contribute to the maintenance of adult pulmonary and systemic vascular tone. However, their actions in the neonatal circulation have not been studied. We aimed to investigate NANC vasorelaxation in neonatal and 2-week-old piglet pulmonary and mesenteric arteries and to examine the potential role of nitric oxide (NO) in this phenomenon. Responses to electric field stimulation (EFS, 50V, 0.25-32 Hz) were investigated in pulmonary and mesenteric artery rings (external diameter 150-200 microm) precontracted with the thromboxane A2 mimetic U46619, in the presence of guanethidine (10 microM) and atropine (10 microM). Under these conditions, EFS resulted in a frequency dependent relaxation of newborn pulmonary (maximal relaxation of 53+/-9.1%), mesenteric (68.8.2+/-7.1%) and 2-wk-old mesenteric (46 6.3%) arteries but this relaxation was significantly reduced (4.5+/-2.2%) in 2-week-old pulmonary arteries. In neonatal pulmonary arteries, the neurotoxin tetrodotoxin (0.3 muM), the NO synthase inhibitor L-NAME (0.1 mM), and the guanylyl cyclase inhibitor ODQ (10 microM) abolished EFS-induced relaxations, suggesting that NANC relaxation of porcine neonatal pulmonary arteries is mediated by NO, which is probably neuronal in origin. However, The expression in pulmonary arteries of the neuronal NO synthase (nNOS), as determined by Western-blot analysis, increased with postnatal age whereas the expression of the endothelial NOS (eNOS) did not change. In conclusion, NANC relaxation is present in neonatal pulmonary and mesenteric arteries and it is, at least partially, mediated through NO. NANC relaxation of porcine pulmonary and mesenteric arteries decreases with postnatal maturation.
Asunto(s)
Neuronas Nitrérgicas/metabolismo , Óxido Nítrico/metabolismo , Arteria Pulmonar/metabolismo , Vasodilatación , Animales , Animales Recién Nacidos , Western Blotting , Estimulación Eléctrica , Inhibidores Enzimáticos/farmacología , Guanilato Ciclasa/antagonistas & inhibidores , Técnicas In Vitro , Arterias Mesentéricas/inervación , Arterias Mesentéricas/metabolismo , Miografía , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa de Tipo I/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo I/metabolismo , Óxido Nítrico Sintasa de Tipo III/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo III/metabolismo , Oxadiazoles/farmacología , Arteria Pulmonar/efectos de los fármacos , Arteria Pulmonar/inervación , Quinoxalinas/farmacología , Porcinos , Tetrodotoxina/farmacología , Vasodilatación/efectos de los fármacosRESUMEN
Encephalopathy due to perinatal asphyxia (PA) is a major cause of neonatal morbidity and mortality in the period around birth. Preterm infants are especially at risk for cognitive, attention and motor impairments. Therapy for this subgroup is limited to supportive care, and new targets are thus urgently needed. Post-asphyxic excitotoxicity is partially mediated by excessive nitric oxide (NO) release. The aims of this study were to determine the timing and distribution of nitric oxide (NO) production after global PA in brain areas involved in motor regulation and coordination. This study focused on the rat striatum and cerebellum, as these areas also affect cognition or attention, in addition to their central role in motor control. NO/peroxynitrite levels were determined empirically with a fluorescent marker on postnatal days P5, P8 and P12. The distributions of neuronal NO synthase (nNOS), cyclic guanosine monophosphate (cGMP), astroglia and caspase-3 were determined with immunohistochemistry. Apoptosis was additionally assessed by measuring caspase-3-like activity from P2-P15. On P5 and P8, increased intensity of NO-associated fluorescence and cGMP immunoreactivity after PA was apparent in the striatum, but not in the cerebellum. No changes in nNOS immunoreactivity or astrocytes were observed. Modest changes in caspase-3-activity were observed between groups, but the overall time course of apoptosis over the first 11 days of life was similar between PA and controls. Altogether, these data suggest that PA increases NO/peroxynitrite levels during the first week after birth within the striatum, but not within the cerebellum, without marked astrogliosis. Therapeutic benefits of interventions that reduce endogenous NO production would likely be greater during this time frame.
Asunto(s)
Asfixia Neonatal/metabolismo , Cerebelo/metabolismo , Cuerpo Estriado/metabolismo , Óxido Nítrico/metabolismo , Animales , Apoptosis/efectos de los fármacos , Astrocitos/metabolismo , Caspasa 3/metabolismo , GMP Cíclico/metabolismo , Femenino , Masculino , Óxido Nítrico Sintasa de Tipo I/metabolismo , Ácido Peroxinitroso/metabolismo , Periodo Posparto/metabolismo , Embarazo , RatasRESUMEN
Chronic or repeated stress during human fetal brain development has been associated with various learning, behavioral, and/or mood disorders, including depression in later life. The mechanisms accounting for these effects of prenatal stress are not fully understood. The aim of this study was to investigate the effects of prenatal stress on early postnatal brain development, a disturbance of which may contribute to this increased vulnerability to psychopathology. We studied the effects of prenatal stress on fetal growth, stress-induced corticosterone secretion, brain cell proliferation, caspase-3-like activity and brain-derived neurotrophic factor protein content in newborn Fischer 344 rats. In addition to a slight reduction in birth weight, prenatal stress was associated with elevated corticosterone levels (33.8%) after 1 h of maternal deprivation on postnatal day 1, whereas by postnatal day 8 this pattern was reversed (-46.5%). Further, prenatal stress resulted in an approximately 50% decrease in brain cell proliferation just after birth in both genders with a concomitant increase in caspase-3-like activity within the hippocampus at postnatal day 1 (36.1%) and at postnatal day 5 (females only; 20.1%). Finally, brain-derived neurotrophic factor protein content was reduced in both the olfactory bulbs (-24.6%) and hippocampus (-28.2%) of prenatally stressed male offspring at postnatal days 1 and 5, respectively. These detrimental central changes observed may partly explain the increased susceptibility of prenatally stressed subjects to mood disorders including depression in later life.
Asunto(s)
Encéfalo/embriología , Desarrollo Fetal/fisiología , Efectos Tardíos de la Exposición Prenatal , Estrés Psicológico/fisiopatología , Animales , Encéfalo/metabolismo , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Caspasa 3 , Caspasas/metabolismo , Proliferación Celular , Corticosterona/sangre , Femenino , Masculino , Embarazo , Ratas , Ratas Endogámicas F344 , Restricción Física/efectos adversosRESUMEN
Exposure of a pregnant woman to physical and/or psychological stress might affect her offspring by promoting the development of various learning, behavioral and/or mood disorders in later life. The 5-HT1A and 5-HT2A receptors are prominently implicated in the modulation of anxiety and mood-related behaviors. Using a semi-quantitative radiolabel immunocytochemical analysis (immunobinding), we studied the effect of prenatal stress on binding of these two receptor subtypes in the hippocampus of 4-week-old male and female Fischer 344 rats. Levels of 5-HT1A immunobinding in the ventral hippocampus, which is primarily implicated in emotional processing, were significantly decreased in male offspring after prenatal stress. A trend towards a decrease was observed in the ventral hippocampus of females. In contrast, 5-HT1A immunobinding within the dorsal hippocampus, which is mainly related to learning and memory, was not affected by prenatal stress in offspring of either gender. Likewise, no significant differences between control and prenatally stressed rats were observed for levels of 5-HT2A immunobinding in either part of the hippocampus or gender. The observed reduction in hippocampal 5-HT1A receptor binding in male offspring after prenatal stress may have important consequences for adult anxiety- and depressive-like behavior.
Asunto(s)
Hipocampo/fisiopatología , Trastornos del Humor/etiología , Efectos Tardíos de la Exposición Prenatal/metabolismo , Receptor de Serotonina 5-HT1A/metabolismo , Serotonina/deficiencia , Estrés Fisiológico/metabolismo , Animales , Trastornos de Ansiedad/etiología , Trastornos de Ansiedad/metabolismo , Trastornos de Ansiedad/fisiopatología , Sitios de Unión/fisiología , Unión Competitiva/fisiología , Trastorno Depresivo/etiología , Trastorno Depresivo/metabolismo , Trastorno Depresivo/fisiopatología , Modelos Animales de Enfermedad , Femenino , Hipocampo/metabolismo , Aprendizaje/fisiología , Masculino , Trastornos del Humor/metabolismo , Trastornos del Humor/fisiopatología , Embarazo , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Ratas , Ratas Endogámicas F344 , Receptor de Serotonina 5-HT2A/metabolismo , Estrés Fisiológico/fisiopatología , Transmisión Sináptica/fisiologíaRESUMEN
BACKGROUND: Epidemiological findings suggest an association between low-for-age birth weight and the risk to develop coronary heart diseases in adulthood. During pregnancy, an imbalance between fetal demands and supply may result in permanent alterations of neuroendocrine development in the fetus. We evaluated whether chronic prenatal hypoxia increases arterial sympathetic innervation. METHODS AND RESULTS: Chicken embryos were maintained from 0.3 to 0.9 of the 21-day incubation period under normoxic (21% O(2)) or hypoxic conditions (15% O(2)). At 0.9 incubation, the degree of sympathetic innervation of the embryonic femoral artery was determined by biochemical, histological, and functional (in vitro contractile reactivity) techniques. Chronic hypoxia increased embryonic mortality (32% versus 13%), reduced body weight (21.9+/-0.4 versus 25.4+/-0.6 g), increased femoral artery norepinephrine (NE) content (78.4+/-9.4 versus 57.5+/-5.0 pg/mm vessel length), and increased the density of periarterial sympathetic nerve fibers (14.4+/-0.7 versus 12.5+/-0.6 counts/10(4) microm(2)). Arteries from hypoxic embryos were less sensitive to NE (pD(2), 5.99+/-0.04 versus 6. 21+/-0.10). In the presence of cocaine, however, differences in sensitivity were no longer present. In the embryonic heart, NE content (156.9+/-11.0 versus 108.1+/-14.7 pg/mg wet wt) was also increased after chronic hypoxia. CONCLUSIONS: In the chicken embryo, chronic moderate hypoxia leads to sympathetic hyperinnervation of the arterial system. In humans, an analogous mechanism may increase the risk for cardiovascular disease in adult life.
Asunto(s)
Arterias/inervación , Hipoxia Fetal/fisiopatología , Sistema Nervioso Simpático/crecimiento & desarrollo , Arterias/crecimiento & desarrollo , Enfermedades Cardiovasculares/etiología , Modelos Animales de Enfermedad , Humanos , Hipoxia , Factores de RiesgoRESUMEN
OBJECTIVE: The distribution of cardiac output to different organs is well described in the mammalian fetus. Chick embryos are not often used in perinatal cardiovascular research and therefore it is not known whether they can serve as an animal model for this purpose. In this study we documented cardiac output distribution in chick embryos at increasing incubation time. METHODS: Fertilized eggs from day 10 to 19 with an incubation time of 21 days were studied in 3 increasing incubation time groups (10-13, 14-16 and 17-19 days). For the experiment, the egg was placed in a holder in an incubator. The egg was opened at the air cell and a small vein of the chorioallantoic membrane was catheterized. Twenty thousand fluorescent 15 microns microspheres in 0.2 ml were injected. After 5 min, the embryo was sacrificed and the different organs were dissected and digested for microsphere isolation and subsequent fluorescence analysis. RESULTS: The chorioallantoic membrane, which is the placenta equivalent of the chick embryo, received a relatively large fraction of the combined cardiac output: 52.08% (interquartile range [IQR] 12.67%) on days 10-13 and 40.95% (IQR 27.24%) on days 17-19. Relatively small fractions were distributed: to the heart 2.03% (IQR 1.58) on days 10-13 and 3.18% (IQR 1.95) on days 17-19, and to the brain 3.20% (IQR 1.80) on days 10-13 and 5.02% (IQR 3.39) on days 17-19. As incubation time advanced, the fraction of the combined cardiac output to the chorioallantoic membrane and yolk-sac decreased significantly in favor of the heart and brain. CONCLUSION: This distribution shows great similarity to the one found in the mammalian fetus. The chick embryo is an attractive model for perinatal cardiovascular research.
Asunto(s)
Gasto Cardíaco/fisiología , Embrión de Pollo/fisiología , Alantoides/fisiología , Animales , Encéfalo/fisiología , Corion/fisiología , Edad Gestacional , Corazón/fisiología , Microscopía Fluorescente , Microesferas , Modelos Biológicos , Saco Vitelino/fisiologíaRESUMEN
Recent work has implicated plasminogen activator released from macrophages as a possible mediator of the demyelinating process in experimental allergic encephalomyelitis and multiple sclerosis (MS). We have studied the capacity of white matter and plaques from MS patients to break down fibrin clots, using a histochemical technique. Fibrinolytic activity was localized exclusively to areas around blood vessels and capillaries in both patients and controls. While there was marked variation between individuals, the unaffected white matter from MS patients was, on the average, not more active than that of controls, but plaques tended to show more numerous foci of lysis, often also more intense, than adjacent white matter; there was no correlation with disease activity or age of the plaques as determined by histological criteria. The localization and degree of fibrinolysis observed were not related to the presence of lymphocytic infiltrates, gliosis, or macrophages. However, the findings do not exclude an involvement of fibrinolytic enzymes (although originating from vascular endothelium rather than macrophages) in the genesis of the MS plaque, which commonly starts around a small vein.
Asunto(s)
Encéfalo/fisiopatología , Fibrinólisis , Esclerosis Múltiple/fisiopatología , Humanos , Factores de TiempoRESUMEN
The effect of chronic supraphysiological doses of anabolic-androgenic steroids, such as those illegally used by recreational, amateur and professional athletes to increase muscle mass and strength, on motoneurons has not been established. The choline acetyltransferase activity levels of perineal muscles in the male rat are modulated by plasma testosterone levels. These muscles are innervated by the sexually dimorphic motoneurons of the spinal nucleus of the bulbocavernosus. Since the primary source of choline acetyltransferase in muscle is motoneuronal, testosterone may modulate perineal muscle choline acetyltransferase activity by regulating choline acetyltransferase messenger RNA levels in motoneurons. The purpose of this study was to determine if choline acetyltransferase messenger RNA levels in cervical and lumbar spinal motoneurons are affected by chronic (four weeks) changes of plasma testosterone levels in the adult male rat. Using in situ hybridization, choline acetyltransferase messenger RNA levels were analysed in four motor columns: the spinal nucleus of the bulbocavernosus, the retrodorsal lateral nucleus of the lumbar spinal cord, and the lateral motor columns of the cervical and lumbar spinal cords. Chronic exposure to supraphysiological levels of testosterone (five- to ten-times physiologic levels) significantly increased choline acetyltransferase messenger RNA in all four motor columns. Subsequent to castration, choline acetyltransferase messenger RNA levels decreased in motoneurons of the spinal nucleus of the bulbocavernosus and the retrodorsal lateral nucleus. This observation suggests that the decrease in choline acetyltransferase activity levels of muscles innervated by spinal nucleus of the bulbocavernosus motoneurons may be due to changes in choline acetyltransferase protein levels. Indeed, testosterone replacement therapy of castrated males prevented the decline of choline acetyltransferase messenger RNA levels in motoneurons. The results of this study demonstrate that anabolic-androgenic steroids can affect the levels of specific messenger RNAs in motoneuron populations throughout the spinal cord suggesting that motoneuronal characteristics are modulated by circulating anabolic-androgenic steroid levels regardless of the purported "androgen sensitivity" of the specific neuromuscular system.
Asunto(s)
Anabolizantes/farmacología , Colina O-Acetiltransferasa/biosíntesis , Neuronas Motoras/enzimología , ARN Mensajero/biosíntesis , Médula Espinal/metabolismo , Animales , Autorradiografía , Hibridación in Situ , Masculino , Neuronas Motoras/efectos de los fármacos , Ratas , Médula Espinal/citología , Médula Espinal/efectos de los fármacos , Testosterona/sangre , Testosterona/farmacologíaRESUMEN
It has previously been suggested that anabolic-androgenic steroids may affect neuromuscular function through their potential action as glucocorticoid receptor antagonists. Alternatively, androgens may regulate the sensitivity of neuromuscular systems to glucocorticoids by modulating GR levels. The purpose of this study was to determine the effects of chronic pharmacologic testosterone treatment of gonadally intact male rats on glucocorticoid receptor alpha immunoreactivity (GRalpha-IR) of motor neurons in the lumbosacral spinal cord. Circulating testosterone levels were chronically increased by subcutaneous Silastic capsules containing crystalline testosterone propionate (TP) for 7, 14, and 28 days. Age-matched sham-operated gonadally intact males served as controls. Relative cytoplasmic and nuclear GRalpha-IR of motor neurons located in the lateral motor column of spinal cord segments L(3) and L(4) (L(Lat); innervating rat hindlimb muscles) and the spinal nucleus of the bulbocavernosus (SNB; innervating the external anal sphincter, bulbocavernosus and levator ani muscles) was measured densitometrically. TP treatment duration had a significant impact on the mean GRalpha levels of both cellular compartments regardless of motor column (two-way ANOVA, P<0.001). The mean nuclear GRalpha-IR of lumbar motor neurons was significantly reduced after 7 days (OD: 0.239+/-0.013 S.E.M.; P<0.016) and 14 days (OD: 0.196+/-0.013; P<0.001) from the GRalpha-IR levels observed among the control group (OD: 0.296+/-0.012) by 20 and 40%, respectively. Interestingly, nuclear GRalpha-IR levels were similar to control levels after 28 days of TP treatment (OD: 0.307+/-0.010). Treatment-dependent changes in cytoplasmic GRalpha-IR paralleled the observed changes in nuclear GRalpha-IR. These data suggest that pharmacologic testosterone treatment effects on motor neuron gene expression may be mediated by testosterone-induced temporal fluctuations of GRalpha-dependent gene regulation.
Asunto(s)
Andrógenos/farmacología , Glucocorticoides/metabolismo , Neuronas Motoras/efectos de los fármacos , Músculo Esquelético/inervación , Receptores de Glucocorticoides/efectos de los fármacos , Médula Espinal/efectos de los fármacos , Andrógenos/metabolismo , Animales , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Citoplasma/efectos de los fármacos , Citoplasma/metabolismo , Miembro Posterior/inervación , Miembro Posterior/fisiología , Hipocampo/citología , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Inmunohistoquímica , Vértebras Lumbares , Masculino , Neuronas Motoras/citología , Neuronas Motoras/metabolismo , Músculo Esquelético/fisiología , Diafragma Pélvico/inervación , Diafragma Pélvico/fisiología , Ratas , Ratas Sprague-Dawley , Receptores de Glucocorticoides/metabolismo , Médula Espinal/citología , Médula Espinal/metabolismoRESUMEN
The aim of our study was to investigate the effect of perinatal asphyxia on developmental apoptosis in the cervical and lumbar spinal cord in the neonatal rat. Perinatal asphyxia was induced by keeping pups at term in utero in a water bath at 37 degrees C for 20 min, followed by resuscitation. Effects of this treatment on developmental apoptosis were studied on postnatal days 2, 5 and 8 using terminal deoxynucleotidyl transferase (TdT)-dUTP-biotin nick end labelling (TUNEL) and caspase-3 staining. TUNEL positive cells were identified using double immunostaining. On postnatal day 2 an increase of 215% in TUNEL positive cells was detected (P=0.005) in laminae IV-VII of the lumbar spinal cord of rats which underwent perinatal asphyxia compared to controls. An increase of 55% compared to controls (P=0.03) was seen in laminae I-III of the lumbar spinal cord at postnatal day 8. TUNEL positive cells could be partly identified as microglia cells (ED1 positive) and oligodendrocytes (O4 positive). The effect of perinatal asphyxia on programmed cell death in the neonatal rat spinal cord was mainly observed in the intermediate zone and dorsal horn of the lumbar spinal cord. We conclude that perinatal asphyxia has a pronounced effect on the survival of cells in a specific region of the spinal cord and thus may have a profound effect on the development of motor networks.
Asunto(s)
Apoptosis , Asfixia/complicaciones , Enfermedades de la Médula Espinal/etiología , Médula Espinal/crecimiento & desarrollo , Médula Espinal/patología , Animales , Animales Recién Nacidos , Asfixia/patología , Vértebras Cervicales , Femenino , Etiquetado Corte-Fin in Situ , Vértebras Lumbares , Masculino , Ratas , Ratas Wistar , Enfermedades de la Médula Espinal/patologíaRESUMEN
Perinatal asphyxia can cause neuronal loss and depletion of neurotransmitters within the striatum. The striatum plays an important role in motor control, sensorimotor integration and learning. In the present study we investigated whether perinatal asphyxia leads to motor deficits related to striatal damage, and in particular to the loss of GABAergic neurons. Perinatal asphyxia was induced in time-pregnant Wistar rats on the day of delivery by placing the uterus horns, containing the pups, in a 37 degrees C water bath for 20 min. Three motor performance tasks (open field, grip test and walking pattern) were performed at 3 and 6 weeks of age. Antibodies against calbindin and parvalbumin were used to stain GABAergic striatal projection neurons and interneurons, respectively. The motor tests revealed subtle effects of perinatal asphyxia, i.e. small decrease in motor activity. Analysis of the walking pattern revealed an increase in stride width at 6 weeks of age after perinatal asphyxia. Furthermore, a substantial loss of calbindin-immunoreactive (-22%) and parvalbumin-immunoreactive (-43%) cells was found in the striatum following perinatal asphyxia at two months of age. GABA(A) receptor autoradiography revealed no changes in GABA binding activity within the striatum, globus pallidus or substantia nigra. We conclude that perinatal asphyxia resulted in a loss of GABAergic projection neurons and interneurons in the striatum without alteration of GABA(A) receptor affinity. Despite a considerable loss of striatal neurons, only minor deficits in motor performance were found after perinatal asphyxia.
Asunto(s)
Asfixia/metabolismo , Asfixia/fisiopatología , Actividad Motora/fisiología , Destreza Motora/fisiología , Receptores de GABA-A/metabolismo , Animales , Animales Recién Nacidos , Cuerpo Estriado/química , Cuerpo Estriado/metabolismo , Femenino , Masculino , Embarazo , Ratas , Ratas Wistar , Receptores de GABA-A/análisis , Ácido gamma-Aminobutírico/análisis , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Along with acetylcholine, motoneurons express several neuromodulatory peptides. The most extensively studied of these peptides is calcitonin gene-related peptide (CGRP). CGRP modulates the biochemical, physiological and metabolic properties of skeletal muscle primarily through activation of membrane receptors. Virtually all motor pool contain motoneurons that are immunoreactive for CGRP. The purpose of this study was to determine the proportions of motoneurons that express alpha-CGRP in motor pools innervating muscles with different motor unit compositions. These include the soleus, extensor digitorum longus, tensor fascia latae and the diaphragm muscles as well as the spinal nucleus of the bulbocavernosus. The spinal nucleus of the bulbocavernosus provides innervation to the bulbocavernosus/levator ani muscle complex and the external anal sphincter muscle. The spinal nucleus of the bulbocavernosus contained the greatest proportion of alpha-CGRP mRNA-positive motoneurons, followed in descending rank order by the tensor fascia latae, the extensor digitorum longus, the soleus and the diaphragm motor pools. In addition, significant differences between motor pools were observed in the mean relative alpha-CGRP mRNA level among those motoneurons expressing alpha-CGRP. The highest mean relative alpha-CGRP mRNA level was observed in soleus and the extensor digitorum longus motor pools; followed in descending rank order by the tensor fascia latae, the diaphragm and the spinal nucleus of the bulbocavernosus. We have previously shown that muscle contractile inactivity increases the number of motoneurons that express alpha-CGRP and in the relative mRNA levels. The results of the present study suggest that the proportion of motoneurons within a motor pool that express alpha-CGRP may be closely related to the contractile activity (i.e. activation history) of the target muscle.
Asunto(s)
Péptido Relacionado con Gen de Calcitonina/genética , Neuronas Motoras/metabolismo , Músculo Esquelético/inervación , ARN Mensajero/metabolismo , Análisis de Varianza , Animales , Masculino , Contracción Muscular/fisiología , RatasRESUMEN
Deficits in cognitive function have been related to quantitative changes in synaptic population, particularly in the cerebral cortex. Here, we used an established model of perinatal asphyxia that induces morphological changes, i.e. neuron loss in the cerebral cortex and striatum, as well as behavioural deficits. We hypothesized that perinatal asphyxia may lead to a neurodegenerative process resulting in cognitive impairment and altered presynaptic bouton numbers in adult rats. We studied cognitive performance at 18 months and presynaptic bouton numbers at 22 months following perinatal asphyxia. Data of the spatial Morris water escape task did not reveal clear memory or learning deficits in aged asphyctic rats compared to aged control rats. However, a memory impairment in aged rats versus young rats was observed, which was more pronounced in asphyctic rats. We found an increase in presynaptic bouton density in the parietal cortex, whereas no changes were found in striatum and frontal cortex in asphyctic rats. An increase of striatal volume was observed in asphyctic rats, leading to an increase in presynaptic bouton numbers in this area. These findings stress the issue that volume measurements have to be taken into account when determining presynaptic bouton density. Furthermore, perinatal asphyxia led to region-specific changes in presynaptic bouton numbers and it worsened the age-related cognitive impairment. These results suggest that perinatal asphyxia induced neuronal loss, which is compensated for by an increase in presynaptic bouton numbers.
Asunto(s)
Asfixia/patología , Asfixia/psicología , Conducta Animal/fisiología , Corteza Cerebral/patología , Neostriado/patología , Terminales Presinápticos/patología , Adulto , Animales , Cognición/fisiología , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Imagenología Tridimensional , Inmunohistoquímica , Aprendizaje por Laberinto/fisiología , Perfusión , Embarazo , Terminales Presinápticos/fisiología , Terminales Presinápticos/ultraestructura , Ratas , Ratas Wistar , Reproducibilidad de los ResultadosRESUMEN
Changes in the contractile and fatigue properties of the cat diaphragm muscle were examined during the first 6 wk of postnatal development. Both twitch contraction time and half-relaxation time decreased progressively with age. Correspondingly, the force-frequency curve was shifted to the left early in development compared with adults. The ratio of peak twitch force to maximum tetanic force decreased with age. Fatigue resistance of the diaphragm was highest at birth and then progressively decreased with age. At birth, most diaphragm muscle fibers stained darkly for myofibrillar adenosinetriphosphatase after alkaline preincubation and thus would be classified histochemically as type II. During subsequent postnatal development, the proportion of type I fibers (lightly stained for adenosinetriphosphatase) increased while the number of type II fibers declined. At birth, type I fibers were larger than type II fibers. The size of both fiber types increased with age, but the increase in cross-sectional area was greater for type II fibers. On the basis of fiber type proportions and mean cross-sectional areas, type I fibers contributed 15% of total muscle mass at birth and 25% in adults. Thus postnatal changes in diaphragm contractile and fatigue properties cannot be attributed to changes in the relative contribution of histochemically classified type I and II fibers. However, the possibility that these developmental changes in diaphragm contractile and fatigue properties correlated with the varying contractile protein composition of muscle fibers was discussed.
Asunto(s)
Animales Recién Nacidos/fisiología , Diafragma/fisiología , Fatiga/fisiopatología , Músculos Respiratorios/fisiología , Adenosina Trifosfatasas/metabolismo , Envejecimiento/metabolismo , Envejecimiento/fisiología , Animales , Peso Corporal/fisiología , Gatos , Diafragma/crecimiento & desarrollo , Estimulación Eléctrica , Fatiga/enzimología , Femenino , Histocitoquímica , Técnicas In Vitro , Masculino , Contracción Muscular/fisiología , Desarrollo de Músculos , Oxidación-Reducción , Músculos Respiratorios/enzimología , Músculos Respiratorios/crecimiento & desarrollo , Succinato Deshidrogenasa/metabolismoRESUMEN
The influence of prolonged nutritional deprivation on the succinate dehydrogenase (SDH) activity and cross-sectional areas of individual fibers in the rat diaphragm and deep portion of the medial gastrocnemius (MGr) muscles was determined. Fatigue resistance of the diaphragm was measured by means of an in vitro nerve-muscle strip preparation. Fiber SDH activity and cross-sectional area were quantified by means of an image processing system. Diaphragm fatigue resistance was significantly improved in the nutritionally deprived (ND) group. In both muscles, nutritional deprivation resulted in a significant decrease in fiber cross-sectional area (both type I and II), type II fibers showing greater atrophy. The SDH activities of type I and II fibers in the diaphragm were not affected by nutritional deprivation. This contrasted with a significant decrease in the SDH activity of both type I and II fibers in the MGr of ND animals. An assessment of the interrelationships between fiber atrophy and fiber SDH activity revealed a greater effect of malnutrition on those diaphragm type II fibers that had the lowest relative SDH activities and the largest cross-sectional areas. By comparison, the effect of malnutrition on type I and II fibers in the MGr was nonselective with regard to fiber SDH activity. We conclude that the enhanced diaphragm fatigue resistance in the ND animals does not result from an increase in the oxidative capacity of muscle fibers and is best explained by the pattern of diaphragm muscle fiber atrophy.
Asunto(s)
Diafragma/análisis , Contracción Muscular , Músculos/anatomía & histología , Trastornos Nutricionales/fisiopatología , Succinato Deshidrogenasa/metabolismo , Animales , Atrofia , Peso Corporal , Diafragma/enzimología , Diafragma/fisiopatología , Técnicas In Vitro , Masculino , Músculos/enzimología , Músculos/fisiopatología , Ratas , Ratas Endogámicas , Valores de ReferenciaRESUMEN
The oxidative capacity and cross-sectional area of muscle fibers were compared between the costal and crural regions of the cat diaphragm and across the abdominal-thoracic extent of the muscle. Succinate dehydrogenase (SDH) activity of individual fibers was quantified using a microphotometric procedure implemented on an image-processing system. In both costal and crural regions, population distributions of SDH activities were unimodal for both type I and II fibers. The continuous distribution of SDH activities for type II fibers indicated that no clear threshold exists for the subclassification of fibers based on differences in oxidative capacity (e.g., the classification of fast-twitch glycolytic and fast-twitch oxidative glycolytic fiber types). No differences in either SDH activity or cross-sectional area were noted between fiber populations of the costal and crural regions. Differences in SDH activity and cross-sectional area were noted, however, between fiber populations located on the abdominal and thoracic sides of the costal region. Both type I and II fibers on the abdominal side of the costal diaphragm were larger and more oxidative than comparable fibers on the thoracic side.
Asunto(s)
Músculos/metabolismo , Consumo de Oxígeno , Succinato Deshidrogenasa/metabolismo , Animales , Gatos , Diafragma/anatomía & histología , Diafragma/metabolismo , Músculos/anatomía & histología , Especificidad de ÓrganosRESUMEN
In the cat diaphragm, fiber capillarity, cross-sectional area, and succinate dehydrogenase (SDH) activity were measured across the first 6 wk of postnatal development. Fibers were classified as type I, IIa, IIb, or IIc on the basis of staining for myofibrillar adenosinetriphosphatase (ATPase). Capillaries were identified in sections stained for ATPase at pH 4.2. Fiber cross-sectional areas and SDH activities were quantified using an image-processing system. During postnatal development, the proportions of type I fibers increased while type II fibers decreased. At birth, all type II fibers were IIc. From the 1st to the 2nd postnatal wk, the proportion of type IIc fibers decreased while the numbers of IIa and IIb increased. Thereafter the proportion of type IIb fibers continued to increase while the number of IIa steadily declined. At birth, capillarity, cross-sectional areas, and SDH activities of type I and II fibers were low compared with other postnatal age groups. Fiber cross-sectional areas increased progressively with age. The number of capillaries surrounding type I and II fibers increased markedly by the 2nd wk and then continued to increase at a slower rate. The number of capillaries per fiber area reached a peak by the 2nd wk and then declined as fiber cross-sectional area increased. Postnatal changes in capillarity depended on fiber type, being greatest in IIb. SDH activities of type I and II fibers were initially low during the first 2 postnatal wk and then peaked by the 3rd wk. After the 6th wk, fiber SDH activities decreased to adult values. Among the type II fibers, IIb showed the greatest change in SDH activity during early postnatal development.
Asunto(s)
Diafragma/irrigación sanguínea , Adenosina Trifosfatasas/metabolismo , Factores de Edad , Animales , Capilares/anatomía & histología , Capilares/crecimiento & desarrollo , Gatos , Diafragma/crecimiento & desarrollo , Diafragma/metabolismo , Femenino , Masculino , Desarrollo de Músculos , Oxidación-Reducción , Succinato Deshidrogenasa/metabolismoRESUMEN
Motor units in cat diaphragm and tibialis posterior muscles were classified physiologically as slow-twitch, fast-twitch, fatigue-resistant, fast-twitch fatigue-intermediate, or fast-twitch fatigable. Motor unit fibers were then identified by glycogen depletion and classified as type I, IIa, IIb, or IIx on the basis of myofibrillar adenosinetriphosphatase-staining profiles and immunoreactivity for myosin heavy-chain (MHC) isoforms. In both muscles, slow-twitch and fast-twitch fatigue-resistant units comprised type I and IIa fibers expressing MHC-slow and MHC-2A isoforms, respectively. Fast-twitch fatigue-intermediate and fast-twitch fatigable units comprised type IIx fibers expressing the MHC-2X isoform. Some fast-twitch fatigue-intermediate units had a mixed composition with a few fibers (approximately 10%) expressing the MHC-2A isoform. Motor unit fiber succinate dehydrogenase (SDH) activity was quantified, and variability was estimated by the interquartile range, which was lower among motor unit fibers than in adjacent fibers of the same histochemical type but comparable to that along the length of individual fibers. We conclude that, despite the mixed-MHC phenotype of some diaphragm and tibialis posterior motor units, SDH activity is relatively uniform. This supports the hypothesis that motoneurons exert a predominant influence on muscle fiber SDH activity.