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Air-breathing vertebrates exhibit cardiovascular responses to diving including heart rate reduction (diving bradycardia). Field studies on aquatic mammals and birds have shown that the intensity of bradycardia can vary depending on diving behaviour, such as the depth of dives and dive duration. However, in aquatic reptiles, the variation in heart rate during deep dives under natural conditions has not been fully investigated. In this study, we released five loggerhead sea turtles (Caretta caretta) outfitted with recorders into the sea and recorded their electrocardiogram, depth, water temperature and longitudinal acceleration. After 3 days, the recorders automatically detached from the turtles. The heart rate signals were detected from the electrodes placed on the surface of the plastron. The mean (±s.d.) heart rate of 12.8±4.1 beats min-1 during dives was significantly lower than that of 20.9±4.1 beats min-1 during surface periods. Heart rate during dives varied with dive depth, although it remained lower than that at the surface. When the turtle dived deeper than 140 m, despite the relatively high flipper stroke rate (approximately 19 strokes min-1), the heart rate dropped rapidly to approximately 2 beats min-1 temporarily. The minimum instantaneous heart rate during dives was lower at deeper dive depths. Our results indicate that loggerhead sea turtles show variations in the intensity of diving bradycardia depending on their diving behaviour, similar to that shown by marine mammals and birds.
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Caniformia , Tortugas , Animales , Bradicardia , Frecuencia Cardíaca , Aceleración , CetáceosRESUMEN
The relationship between the environment and marine animal small-scale behavior is not fully understood. This is largely due to the difficulty in obtaining environmental datasets with a high spatiotemporal precision. The problem is particularly pertinent in assessing the influence of environmental factors in rapid, high energy-consuming behavior such as seabird take-off. To fill the gaps in the existing environmental datasets, we employed novel techniques using animal-borne sensors with motion records to estimate wind and ocean wave parameters and evaluated their influence on wandering albatross take-off patterns. Measurements revealed that wind speed and wave heights experienced by wandering albatrosses during take-off ranged from 0.7 to 15.4 m/s and 1.6 to 6.4 m, respectively. The four indices measured (flapping number, frequency, sea surface running speed, and duration) also varied with the environmental conditions (e.g., flapping number varied from 0 to over 20). Importantly, take-off was easier under higher wave conditions than under lower wave conditions at a constant wind speed, and take-off effort increased only when both wind and waves were gentle. Our data suggest that both ocean waves and winds play important roles for albatross take-off and advances our current understanding of albatross flight mechanisms.
Wandering albatrosses are large seabirds with one of the most impressive wingspans found in the animal kingdom. While they spend most of their time efficiently gliding above the waves, they do have to regularly land on sea to snatch their prey. To resume flight, the birds turn into the wind and flap their wings as they run on the surface of the ocean; this causes their heart to beat three to four times faster than normal. In contrast, flying barely leads to a change in pulse rate compared to rest. As for many other marine birds, sea take-offs therefore represent one of the major energy costs that albatrosses face when out foraging. Scientists have long assumed that the amount of effort required for this manoeuvre depends on factors such as wind speed and, potentially, the height of the waves. However, this is difficult to establish for sure because direct information about the environment that a bird faces as it takes off is rarely available. Often, the best that researchers can do is to reconstruct this data based on global weather patterns, ocean climatic models or evidence collected from nearby locations. To address this problem, Uesaka et al. devised innovative ways to use data from animal-borne sensors. They equipped 44 albatrosses with these instruments and recorded over 1,500 hours of foraging sea trips. Wind parameters such as speed and direction were estimated based on the animals' flying paths, and wave height calculated from their floating motion. Sensor data also gave an insight into the energy cost of each take-off, which was estimated based on four parameters (running duration, running speed, number of wing flaps, and flapping frequency). The analyses confirmed that albatrosses take off into a headwind, with stronger winds reducing the amount of effort required. However, wave height also had a profound impact, suggesting that this parameter should be included in future studies. Overall, the birds flapped their wings less and ran on the surface of the water for shorter amounts of time when the wind was strong, or the waves were high. Even with weak winds, take offs were easier when waves were taller, and they were most costly when both the sea and wind were calm. The work by Uesaka et al. helps to capture how environmental factors influence the energy balance of albatrosses and other marine birds. As ocean weather patterns become more volatile and extreme climate events more frequent, such knowledge is acutely needed to understand how these creatures may respond to their changing world.
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Vuelo Animal , Viento , Animales , Aves , Conducta Animal , Movimiento (Física)RESUMEN
The authors investigated the relationship between frame rates and subpopulation structure of bovine sperm divided by their motility analyzed by a computer-assisted sperm analysis (CASA). Kinematic parameters of bovine sperm incubated in Brackett & Oliphant medium with and without calcium ionophore for 0, 1, 5, 10, 15, 20, 25, and 30 min were evaluated by a CASA at 150 frames per second (fps) and analyzed structure of sperm motility subpopulation by cluster analysis. Then, we converted CASA data at 150 fps to 75, 50, and 30 fps and evaluated the structures of sperm motility subpopulation at different fps in each sperm by a discriminant analysis. As the results, the structure of sperm motility subpopulation was affected by frame rate. Sperm were divided into six clusters at 150, 75, and 50 fps; on the other hand, there were five clusters at 30 fps. Straight-line velocity was similar at all frame rates. However, as the frame rate became higher, curvilinear velocity and beat cross frequency of sperm head increased significantly, whereas lateral sperm head displacement decreased significantly. In conclusion, higher frame rate at 150 fps is recommended to capture the trajectory of sperm accurately by CASA in the present study.
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Semen , Motilidad Espermática , Masculino , Bovinos , Animales , Análisis de Semen/veterinaria , Espermatozoides , ComputadoresRESUMEN
Diving bradycardia is a reduction in the heart rate mediated by the parasympathetic system during diving. Although diving bradycardia is pronounced in aquatic mammals and birds, the existence of this response in aquatic reptiles, including sea turtles, remains under debate. Using the parasympathetic blocker atropine, we evaluated the involvement of the parasympathetic nervous system in heart rate reduction of loggerhead sea turtles (Caretta caretta) during voluntary diving in tanks. The heart rate of the control group dropped by 40-60% from the pre-dive value at the onset of diving; however, administration of atropine significantly inhibited heart rate reduction (P<0.001). Our results indicate that, similar to mammals and birds, the heart rate reduction in sea turtles while diving is primarily mediated by the parasympathetic nervous system. In conclusion, we suggest that diving bradycardia exists not only in aquatic mammals and birds but also in aquatic reptiles.
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Tortugas , Animales , Atropina/farmacología , Bradicardia , Frecuencia Cardíaca/fisiología , Mamíferos , Tortugas/fisiologíaRESUMEN
Heart rate measurement is an essential method for evaluating the physiological status of air-breathing diving animals. However, owing to technical difficulties, many marine animals require an invasive approach to record an electrocardiogram (ECG) in water, limiting the application of this approach in a wide range of marine animals. Recently, a non-invasive system was reported to measure the ECG of hard-shelled sea turtles by pasting the electrodes on the dorsal side of the shell, although the ECG obtained from the moving turtle contains noise produced by muscle contraction. Here, we report that clear ECGs can be obtained by placing the electrodes on the ventral side rather than the dorsal side in loggerhead sea turtles. Using our method, clearer ECG signals were obtained with less electrical noise, even when turtles are swimming. According to the anatomical features, the electrode position on the ventral side is closer to the heart than the dorsal side, minimizing the effects of noise generated by the skeletal muscle. This new biologging technique will elucidate the functioning of the circulatory system of sea turtles during swimming and their adaptabilities to marine environments. This article is part of the theme issue "Methods and Applications in Physio-logging."
RESUMEN
Heart rates of air-breathing diving animals can change on a short time scale due to the diving response during submergence. Heart rate is used frequently as a proxy for indirectly estimating metabolic rates on a fine time scale. However, most studies to date have been conducted on endothermic diving animals, and the relationships between metabolic rates and heart rates in ectothermic diving animals have not been well studied. Sea turtles are unique model organisms of diving ectotherms because they spend most of their life in the ocean and perform deep and/or long dives. In this study, we examined the relationship between heart rates and metabolic rates in captive loggerhead turtles, Caretta caretta, to estimate oxygen consumption rates during each dive based on heart rates. The oxygen consumption rates (VÌO2: mlO2 min-1 kg-1) and average heart rates (fH: beats min-1) were measured simultaneously in indoor tanks at water temperatures of 15-25°C. Our results showed that oxygen consumption rate was affected by heart rate and water temperature in loggerhead turtles. Based on the collected data, we formulated the model equation as VÌO2=0.0124fH+0.0047Tw - 0.0791. The equation can be used for estimating fine-scaled field metabolic rates in free-ranging loggerhead turtles. The results of this study will contribute to future comparative studies of the physiological states of ectothermic diving animals.
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Buceo , Tortugas , Animales , Buceo/fisiología , Frecuencia Cardíaca/fisiología , Consumo de Oxígeno/fisiología , Temperatura , Tortugas/fisiologíaRESUMEN
The study of seabird behaviour has largely relied on animal-borne tags to gather information, requiring interpretation to estimate at-sea behaviours. Details of shallow-diving birds' foraging are less known than deep-diving species due to difficulty in identifying shallow dives from biologging devices. Development of smaller video loggers allow a direct view of these birds' behaviours, at the cost of short battery capacity. However, recordings from video loggers combined with relatively low power usage accelerometers give a means to develop a reliable foraging detection method. Combined video and acceleration loggers were attached to streaked shearwaters in Funakoshi-Ohshima Island (39°24'N,141°59'E) during the breeding season in 2018. Video recordings were classified into behavioural categories (rest, transit, and foraging) and a detection method was generated from the acceleration signals. Two foraging behaviours, surface seizing and foraging dives, are reported with video recordings. Surface seizing was comprised of successive take-offs and landings (mean duration 0.6 and 1.5s, respectively), while foraging dives were shallow subsurface dives (3.2s mean duration) from the air and water surface. Birds were observed foraging close to marine predators, including dolphins and large fish. Results of the behaviour detection method were validated against video recordings, with mean true and false positive rates of 90% and 0%, 79% and 5%, and 66% and <1%, for flight, surface seizing, and foraging dives, respectively. The detection method was applied to longer duration acceleration and GPS datasets collected during the 2018 and 2019 breeding seasons. Foraging trips lasted between 1 - 8 days, with birds performing on average 16 surface seizing events and 43 foraging dives per day, comprising <1% of daily activity, while transit and rest took up 55 and 40%, respectively. This foraging detection method can address the difficulties of recording shallow-diving foraging behaviour and provides a means to measure activity budgets across shallow diving seabird species.
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Aves , Conducta Alimentaria , Animales , Estaciones del AñoRESUMEN
To measure the heart rate of unrestrained sea turtles, it has been believed that a probe must be inserted inside the body owing to the presence of the shell. However, inserting the probe is invasive and difficult to apply to animals in the field. Here, we have developed a non-invasive heart rate measurement method for some species of sea turtles. In our approach, an electrocardiogram (ECG) was performed using an animal-borne ECG recorder and two electrodes-which were electrically insulated from seawater-pasted on the carapace. Based on the measured ECG, the heartbeat signals were identified with an algorithm using a band-pass filter. We implemented this algorithm in a user-friendly program package, ECGtoHR. In experiments conducted in a water tank and in a lagoon, we successfully measured the heart rate of loggerhead, olive ridley and black turtles, but not green and hawksbill turtles. The average heart rate of turtles when resting underwater was 6.2 ± 1.9 beats min-1 and that when moving at the surface was 14.0 ± 2.4 beats min-1. Our approach is particularly suitable for endangered species such as sea turtles, and has the potential to be extended to a variety of other free-ranging species. This article is part of the theme issue 'Measuring physiology in free-living animals (Part I)'.
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Organismos Acuáticos/fisiología , Frecuencia Cardíaca/fisiología , Fisiología/instrumentación , Tortugas/fisiología , Exoesqueleto , Animales , Agua de MarRESUMEN
Heart rate monitoring in free-ranging cetaceans to understand their behavioural ecology and diving physiology is challenging. Here, we developed a simple, non-invasive method to monitor the heart rate of cetaceans in the field using an electrocardiogram-measuring device and a single suction cup equipped with an electrode. The unipolar suction cup was placed on the left lateral body surface behind the pectoral fin of Risso's dolphins (Grampus griseus) and a false killer whale (Pseudorca crassidens) in captivity; their heart rate was successfully monitored. We observed large heart rate oscillations corresponding to respiration in the motionless whales during surfacing (a false killer whale, mean 47 bpm, range 20-75 bpm; Risso's dolphins, mean ± s.d. 61 ± 15 bpm, range 28-120 bpm, n = 4 individuals), which was consistent with the sinus arrhythmia pattern (eupneic tachycardia and apneic bradycardia) observed in other cetaceans. Immediately after respiration, the heart rate rapidly increased to approximately twice that observed prior to the breath. Heart rate then gradually decreased at around 20-50 s and remained relatively constant until the next breath. Furthermore, we successfully monitored the heart rate of a free-swimming Risso's dolphin. The all-in-one suction cup device is feasible for field use without restraining animals and is helpful in further understanding the diving physiology of free-ranging cetaceans. This article is part of the theme issue 'Measuring physiology in free-living animals (Part II)'.
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Delfines/fisiología , Pruebas de Función Cardíaca/veterinaria , Frecuencia Cardíaca , Fisiología/instrumentación , Animales , Femenino , Pruebas de Función Cardíaca/instrumentación , MasculinoRESUMEN
In the present study, bull sperm in the first and second ejaculates were divided into subpopulations based on their motility characteristics using a cluster analysis of data from computer-assisted sperm motility analysis (CASA). Semen samples were collected from 4 Japanese black bulls. Data from 9,228 motile sperm were classified into 4 clusters; 1) very rapid and progressively motile sperm, 2) rapid and circularly motile sperm with widely moving heads, 3) moderately motile sperm with heads moving frequently in a short length, and 4) poorly motile sperm. The percentage of cluster 1 varied between bulls. The first ejaculates had a higher proportion of cluster 2 and lower proportion of cluster 3 than the second ejaculates.
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Bovinos/fisiología , Motilidad Espermática/fisiología , Espermatozoides/clasificación , Espermatozoides/fisiología , Animales , Análisis por Conglomerados , Eyaculación/fisiología , Masculino , Análisis de Semen/veterinariaRESUMEN
Inbreeding avoidance is essential to providing offspring with genetic diversity. Females' mate choice is more crucial than males' for successful reproduction because of the high cost of producing gametes and limited chances to mate. However, the mechanism of female inbreeding avoidance is still unclear. To elucidate the mechanism underlying inbreeding avoidance by females, we conducted Y-maze behavioral assays using BALB/c and C57BL/6 female mice. In both strains, the avoidance of male urine from the same strain was lower in the low estrogen phase than in the high estrogen phase. The estrous cycle-dependent avoidance was completely prevented by vomeronasal organ (VNO) removal. To assess the regulation of the vomeronasal system by estrogen, the neural excitability was evaluated by immunohistochemistry of the immediate early gene products. Although estrogen did not affect neural excitability in the VNO, estrogen enhanced the neural excitability of the mitral cell layer in the AOB induced by urine from the cognate males. These results suggest that female mice avoid odor from genetically similar males in an estrogen-dependent manner via the vomeronasal system and the excitability of the mitral cells in the AOB is presumed to be regulated by estrogen.
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Odorantes , Órgano Vomeronasal/metabolismo , Animales , Conducta Animal/fisiología , Estradiol/administración & dosificación , Femenino , Inmunohistoquímica , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Bulbo Olfatorio/metabolismo , Feromonas/orina , Proteínas Proto-Oncogénicas c-fos/metabolismoRESUMEN
We tagged 82 lactating northern fur seals (Callorhinus ursinus) with tri-axial accelerometers and magnetometers on two eastern Bering Sea islands (Bogoslof and St. Paul) with contrasting population trajectories. Using depth data, accelerometer data and spectral analysis we classified time spent diving (30%), resting (~7%), shaking and grooming their pelage (9%), swimming in the prone position (~10%) and two types of previously undocumented rolling behavior (29%), with the remaining time (~15%) unspecified. The reason for the extensive rolling behavior is not known. We ground-truthed the accelerometry signals for shaking and grooming and rolling behaviors--and identified the acceleration signal for porpoising--by filming tagged northern fur seals in captivity. Speeds from GPS interpolated data indicated that animals traveled fastest while in the prone position, suggesting that this behavior is indicative of destination-based swimming. Very little difference was found in the percentages of time spent in the categorical behaviors with respect to breeding islands (Bogoslof or St. Paul Island), forager type (cathemeral or nocturnal), and the region where the animals foraged (primarily on-shelf <200 m, or off-shelf > 200 m). The lack of significant differences between islands, regions and forager type may indicate that behaviors summarized over a trip are somewhat hardwired even though foraging trip length and when and where animals dive are known to vary with island, forager type and region.
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Conducta Animal/fisiología , Lobos Marinos/fisiología , Lactancia/fisiología , Actividad Motora/fisiología , Natación/fisiología , Acelerometría , Animales , Ecosistema , Femenino , IslasRESUMEN
Contamination levels of coplanar polychlorinated biphenyls (Co-PCBs), polycyclic aromatic hydrocarbons (PAHs), and dichlorodiphenyltrichloroethane (DDTs) were measured in the entire body of the large Japanese field mouse (Apodemus speciosus) collected from Hokkaido (Ishikari and Rankoshi) and Aomori prefecture (Takko) in Japan. Higher concentrations of PCBs including Co-PCBs, were observed in the mice collected from Ishikari than those from Rankoshi. The concentration of PAHs in the soil from Ishikari was also higher than that in the other sampling sites. The findings suggest that Ishikari is the most polluted area, probably because of human activities, depending on the population distribution. However, the observed contaminant levels were extremely lower compared to those in previous studies. The ratio of testis weight to body weight (TW/BW) was the lowest in the mice collected from Ishikari, which is the area contaminated with PAHs and p,p'-dichlorodiphenyldichloroethylene (DDE). However, the serum testosterone levels of mice from the Ishikari area were higher than those from the non-contaminated other areas although no significant differences. Previous studies have shown that a low-level exposure to dioxin related compounds (DRCs) disturbances in sexual function, resulting in the production of testosterone. This study showed that POPs exposure is one of the possibility of the high testosterone concentration in the mice of the Ishikari area in addition to a cause of biological and environmental factors such as habitat density, age, temperatures and/or food riches.
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Diclorodifenildicloroetano/toxicidad , Contaminantes Ambientales/toxicidad , Murinae , Bifenilos Policlorados/toxicidad , Hidrocarburos Policíclicos Aromáticos/toxicidad , Animales , Peso Corporal , Diclorodifenildicloroetano/química , Diclorodifenildicloroetano/metabolismo , Monitoreo del Ambiente , Contaminantes Ambientales/química , Contaminantes Ambientales/metabolismo , Japón , Masculino , Tamaño de los Órganos , Bifenilos Policlorados/química , Bifenilos Policlorados/metabolismo , Hidrocarburos Policíclicos Aromáticos/química , Hidrocarburos Policíclicos Aromáticos/metabolismo , Testículo/anatomía & histologíaRESUMEN
Albatrosses are known to expend only a small amount of energy during flight. The low energy cost of albatross flight has been attributed to energy-efficient gliding (soaring) with sporadic flapping, although little is known about how much time and energy albatrosses expend in flapping versus gliding during cruising flight. Here, we examined the heart rates (used as an instantaneous index of energy expenditure) and flapping activities of free-ranging black-browed albatrosses (Thalassarche melanophrys) to estimate the energy cost of flapping as well as time spent in flapping activities. The heart rate of albatrosses during flight (144 beats min(-1)) was similar to that while sitting on the water (150 beats min(-1)). In contrast, heart rate was much higher during takeoff and landing (ca. 200 beats min(-1)). Heart rate during cruising flight was linearly correlated with the number of wing flaps per minute, suggesting an extra energy burden of flapping. Albatrosses spend only 4.6±1.4% of their time flapping during cruising flight, which was significantly lower than during and shortly after takeoff (9.8±3.5%). Flapping activity, which amounted to just 4.6% of the time in flight, accounted for 13.3% of the total energy expenditure during cruising flight. These results support the idea that albatrosses achieve energy-efficient flight by reducing the time spent in flapping activity, which is associated with high energy expenditure.
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Aves/fisiología , Metabolismo Energético/fisiología , Vuelo Animal/fisiología , Frecuencia Cardíaca/fisiología , Alas de Animales/fisiología , Aceleración , Animales , Conducta Animal/fisiología , Peso Corporal/fisiología , Electrocardiografía , Océanos y MaresRESUMEN
Corticosterone has received considerable attention as the principal hormonal mediator of allostasis or physiological stress in wild animals. More recently, it has also been implicated in the regulation of parental care in breeding birds, particularly with respect to individual variation in foraging behavior and provisioning effort. There is also evidence that prolactin can work either inversely or additively with corticosterone to achieve this. Here we test the hypothesis that endogenous corticosterone plays a key physiological role in the control of foraging behavior and parental care, using a combination of exogenous corticosterone treatment, time-depth telemetry, and physiological sampling of female macaroni penguins (Eudyptes chrysolophus) during the brood-guard period of chick rearing, while simultaneously monitoring patterns of prolactin secretion. Plasma corticosterone levels were significantly higher in females given exogenous implants relative to those receiving sham implants. Increased corticosterone levels were associated with significantly higher levels of foraging and diving activity and greater mass gain in implanted females. Elevated plasma corticosterone was also associated with an apparent fitness benefit in the form of increased chick mass. Plasma prolactin levels did not correlate with corticosterone levels at any time, nor was prolactin correlated with any measure of foraging behavior or parental care. Our results provide support for the corticosterone-adaptation hypothesis, which predicts that higher corticosterone levels support increased foraging activity and parental effort.
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Corticosterona/sangre , Conducta Alimentaria/fisiología , Conducta Materna/fisiología , Spheniscidae/fisiología , Animales , Femenino , Prolactina/sangreRESUMEN
The odor preference of female mice for male odor is reported to have cyclical variations in relation to the estrus cycle. Females prefer the odor of genetically dissimilar males to that of genetically similar ones, but the causal relation between this preference and the estrus cycle has scarcely been investigated. The Y-maze test demonstrated that BALB/c females stayed for a longer duration near the urine of C57BL/6 males than that of BALB/c males when they were in metestrus, diestrus and proestrus, but not in estrus. The prolonged stay disappeared after ovariectomy, and administration of estradiol-17ß restored the tendency. The present results suggest that the odor preference of BALB/c females for C57BL/6 over BALB/c males temporally changes according to the estrus cycle and that estrogen can be one of endogenous factors regulating this phenomenon.
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Estradiol/farmacología , Ciclo Estral/fisiología , Ratones Endogámicos BALB C/fisiología , Conducta Sexual Animal/fisiología , Animales , Femenino , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Aprendizaje por Laberinto/fisiología , Ratones , Ratones Endogámicos C57BL , Odorantes , Conducta Sexual Animal/efectos de los fármacosRESUMEN
In an attempt to explore the functioning of nitric oxide (NO) in pancreatic exocrine cells, we have recently obtained several lines of circumstantial evidence indicating that one of molecular targets of NO is phospholipase C (PLC), the activation of which leads to an increase in the cytosolic Ca2+ concentration ([Ca2+]i) via inositol 1, 4, 5-trisphosphate, IP3. However, whether IP3 is actually produced by NO has not yet been substantiated. The present study was therefore designed to directly measure the intracellular IP3, concentration ([IP3]i) for better understanding of the underlying mechanisms with the help of pharmacological tools. [IP3]i was measured using a fluorescence polarization technique (HitHunter). We obtained the following results: 1) varying concentrations of an NO donor, sodium nitroprusside (SNP), elevated [IP3]i, 2) this elevation was completely inhibited in the presence of the soluble guanylyl cyclase (sGC) inhibitor, 1H-[1, 2, 4] oxadiazolo [4, 3-a] quinoxalin-1-one (ODQ), 3) varying concentrations of the cGMP analogue, 8-Br-cGMP, also increased [IP3]i, 4) the cGMP analogue-induced IP3 production was abolished by pretreatment with either a PLC inhibitor, U73122, or a G-protein inhibitor, GP2A, and 5) KT5823, a potent and highly selective inhibitor of cGMP-dependent protein kinase G (PKG), also abolished the IP3 production induced by 8-Br-cGMP. These results suggest that the NO-induced [Ca2+]i increase is triggered by an increase in [IP3]i located downstream from intracellular cGMP elevation. In this intracellular pathway, each sGC, cGMP-dependent PKG, G-protein and PLC were suggested to be involved. The present work provides new insights into the intracellular signaling accelerated by NO. NO triggers a [Ca2+]I increase via cGMP and IP3 in pancreatic acinar cells.
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Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Inositol 1,4,5-Trifosfato/biosíntesis , Óxido Nítrico/metabolismo , Páncreas Exocrino/metabolismo , Animales , Calcio/metabolismo , GMP Cíclico/análogos & derivados , GMP Cíclico/farmacología , GMP Cíclico/fisiología , Proteínas Quinasas Dependientes de GMP Cíclico/antagonistas & inhibidores , Donantes de Óxido Nítrico/farmacología , Nitroprusiato/farmacología , Páncreas Exocrino/efectos de los fármacos , Páncreas Exocrino/enzimología , Ratas , Ratas Wistar , Fosfolipasas de Tipo C/metabolismoRESUMEN
The aim of the present study was to investigate the possible interaction between intracellular Ca(2+) and nitric oxide (NO) in rat pancreatic acinar cells, especially intracellular signaling events. (1) Nitric oxide donors SNP (0.1-100 µM) and NOR-3 (50-400 µM) induced Ca(2+) oscillations in fluo-4-loaded acini, that appeared to be analogous to what we usually observe in acini stimulated with physiological secretagogues such as CCK-8 and this oscillations were abolished in the presence of carboxy-PTIO. (2) The NO donors-evoked Ca(2+) oscillations were not abolished even in the absence of extracellular Ca(2+) but totally disappeared when cells were pretreated with thapsigargin, a sarcoplasmic-endoplasmic reticulum Ca(2+) ATPase (SERCA) inhibitor. (3) Inhibition of guanylate cyclase with 1 H-[1,2,4] oxadiazolo [4,3-a] quinoxaline-1-one (ODQ) attenuated Ca(2+) oscillations evoked by SNP in the absence of extracellular Ca(2+). (4) Inhibitors of phospholipase C activity, U73122 and the IP(3)R blocker xestospongin C, both abolished the SNP-induced Ca(2+) response. (5) Furthermore, we found that both CCK-8 and carbachol (CCh) induced NO production in DAF-2-loaded acinar cells and that an inhibitor of NO synthase, N(G)-monomethyl-l-arginine (L-NMMA), significantly reduced CCK-8-induced Ca(2+) oscillation. These results indicate that NO mobilizes Ca(2+) from internal stores through activation of guanylate cyclase and resultant cGMP production. In addition, PLC activation of IP(3) production is also suggested to be involved in Ca(2+) mobilization via IP(3) receptors. This suggests the presence of cross-talk between Ca(2+) and NO in pancreatic acini and this cascade may, at least partially, participate in physiological secretagogue-evoked Ca(2+) dynamics in pancreatic acinar cells.
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Calcio/metabolismo , Carcinoma de Células Acinares/metabolismo , Óxido Nítrico/metabolismo , Neoplasias Pancreáticas/metabolismo , Fosfolipasas de Tipo C/metabolismo , Animales , Estrenos/farmacología , Inositol 1,4,5-Trifosfato/antagonistas & inhibidores , Inositol 1,4,5-Trifosfato/metabolismo , Compuestos Macrocíclicos/farmacología , Masculino , Donantes de Óxido Nítrico/farmacología , Nitroprusiato/farmacología , Oxadiazoles/farmacología , Oxazoles/farmacología , Inhibidores de Fosfodiesterasa/farmacología , Pirrolidinonas/farmacología , Quinoxalinas/farmacología , Ratas , Ratas Wistar , Transducción de Señal/efectos de los fármacos , Sincalida/metabolismo , Fosfolipasas de Tipo C/antagonistas & inhibidoresRESUMEN
The objective of this study was to investigate and characterize the metabolic activities of CYP1A in deer, cattle and horses in comparison to those of rats using ethoxyresorufin O-deethylation (EROD) and methoxyresorufin O-demethylation (MROD) assays. We performed an inhibition study for these activities using anti-rat CYP1A1 antibody and identified that these activities were due to the CYP1A subfamily. Interspecies differences in the CYP1A-dependent activities were highly observed in this study. In particular, we found that the horse had the highest EROD and MROD activities among the examined animal species. In the kinetic analysis, the horses showed the highest Vmax and catalytic efficiency (Vmax/Km), followed by the cattle, deer and rats.