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Prolonged bed rest impairs standing balance but the underlying mechanisms are uncertain. Previous research suggests strength loss is not the cause, leaving impaired sensorimotor control as an alternative. Here we examine vestibular control of posture in 18 male volunteers before and after 60 days of bed rest. Stochastic vestibular stimulation (SVS) was used to evoke sway responses before, 1 and 6 days after bed rest under different head yaw orientations. The directional accuracy and precision of these responses were calculated from ground reaction force vectors. Bed rest caused up to 63% increases in spontaneous standing sway and 31% reductions in leg strength, changes which were uncorrelated. The increase in sway was exacerbated when the eyes were closed. Mean directions of SVS-evoked sway responses were unaffected, being directed towards the anodal ear and rotating in line with head orientation in the same way before and after bed rest. However, individual trial analysis revealed 25%-30% increases in directional variability, which were significantly correlated with the increase in spontaneous sway (r = 0.48-0.71; P ≤ 0.044) and were still elevated on day 6 post-bed rest. This reveals that individual sway responses may be inappropriately oriented, a finding masked by the averaging process. Our results confirm that impaired balance following prolonged bedrest is not related to loss of strength. Rather, they demonstrate that the sensorimotor transformation process which converts vestibular feedback into appropriately directed balance responses is impaired. KEY POINTS: Prolonged inactivity impairs balance but previous research suggests this is not caused by loss of strength. Here we investigated vestibular control of balance before and after 60 days of bed rest using electrical vestibular stimulation (EVS) to evoke sway responses. Spontaneous sway significantly increased and muscle strength reduced following bed rest, but, in keeping with previous research, these two effects were not correlated. While the overall accuracy of EVS-evoked sway responses was unaffected, their directional variability significantly increased following bed rest, and this was correlated with the increases in spontaneous sway. We have shown that the ability to transform head-centred vestibular feedback into an appropriately directed body sway response is negatively affected by prolonged inactivity; this may contribute to the impaired balance commonly observed following bed rest.
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Reposo en Cama , Equilibrio Postural , Vestíbulo del Laberinto , Humanos , Masculino , Equilibrio Postural/fisiología , Adulto , Vestíbulo del Laberinto/fisiología , Adulto JovenRESUMEN
Receptor-mediated endocytosis is responsible for reabsorption of transferrin (Tf) in renal proximal tubules (PTs). Although the role of the megalin-cubilin receptor complex (MCRC) in this process is unequivocal, modalities independent of this complex are evident but as yet undefined. Here, using immunostaining and Tf-flux assays, FACS analysis, and fluorescence imaging, we report localization of Tf receptor 1 (TfR1), the cognate Tf receptor mediating cellular holo-Tf (hTf) acquisition, to the apical brush border of the PT, with expression gradually declining along the PT in mouse and rat kidneys. In functional studies, hTf uptake across the apical membrane of cultured PT epithelial cell (PTEC) monolayers increased in response to decreased cellular iron after desferrioxamine (DFO) treatment. We also found that apical hTf uptake under basal conditions is receptor-associated protein (RAP)-sensitive and therefore mediated by the MCRC but becomes RAP-insensitive under DFO treatment, with concomitantly decreased megalin and cubilin expression levels and increased TfR1 expression. Thus, as well as the MCRC, TfR1 mediates hTf uptake across the PT apical brush border, but in conditions of decreased cellular iron, hTf uptake is predominated by augmented apical TfR1. In conclusion, both the MCRC and TfR1 mediate hTf uptake across apical brush border membranes of PTECs and reciprocally respond to decreased cellular iron. Our findings have implications for renal health, whole-body iron homeostasis, and pathologies arising from disrupted iron balance.
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Hierro/metabolismo , Túbulos Renales Proximales/metabolismo , Proteína 2 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , Receptores de Superficie Celular/metabolismo , Receptores de Transferrina/metabolismo , Transferrina/metabolismo , Animales , Línea Celular Transformada , Masculino , Ratones , Ratones Endogámicos C57BL , Ratas , Ratas Endogámicas WKYRESUMEN
KEY POINTS: Reaching movements can be perturbed by vestibular input, but the function of this response is unclear. Here, we applied galvanic vestibular stimulation concurrently with real body movement while subjects maintained arm position either fixed in space or fixed with respect to their body. During the fixed-in-space conditions, galvanic vestibular stimulation caused large changes in arm trajectory consistent with a compensatory response to maintain upper-limb accuracy in the face of body movement. Galvanic vestibular stimulation responses were absent during the body-fixed task, demonstrating task dependency in vestibular control of the upper limb. The results suggest that the function of vestibular-evoked arm movements is to maintain the accuracy of the upper limb during unpredictable body movement, but only when reaching in an earth-fixed reference frame. ABSTRACT: When using our arms to interact with the world, unintended body motion can introduce movement error. A mechanism that could detect and compensate for such motion would be beneficial. Observations of arm movements evoked by vestibular stimulation provide some support for this mechanism. However, the physiological function underlying these artificially evoked movements is unclear from previous research. For such a mechanism to be functional, it should operate only when the arm is being controlled in an earth-fixed rather than a body-fixed reference frame. In the latter case, compensation would be unnecessary and even deleterious. To test this hypothesis, subjects were gently rotated in a chair while being asked to maintain their outstretched arm pointing towards either earth-fixed or body-fixed memorized targets. Galvanic vestibular stimulation was applied concurrently during rotation to isolate the influence of vestibular input, uncontaminated by inertial factors. During the earth-fixed task, galvanic vestibular stimulation produced large polarity-dependent corrections in arm position. These corrections mimicked those evoked when chair velocity was altered without any galvanic vestibular stimulation, indicating a compensatory arm response to a sensation of altered body motion. In stark contrast, corrections were completely absent during the body-fixed task, despite the same chair movement profile and arm posture. These effects persisted when we controlled for differences in limb kinematics between the two tasks. Our results demonstrate that vestibular control of the upper limb maintains reaching accuracy during unpredictable body motion. The observation that such responses occurred only when reaching within an earth-fixed reference frame confirms the functional nature of vestibular-evoked arm movement.
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Retroalimentación Fisiológica , Movimiento , Potenciales Vestibulares Miogénicos Evocados , Vestíbulo del Laberinto/fisiología , Adulto , Brazo/fisiología , Femenino , Humanos , MasculinoRESUMEN
KEY POINTS: When standing and holding an earth-fixed object, galvanic vestibular stimulation (GVS) can evoke upper limb responses to maintain balance. In the present study, we determined how these responses are affected by grip context (no contact, light grip and firm grip), as well as how they are co-ordinated with the lower limbs to maintain balance. When GVS was applied during firm grip, hand and ground reaction forces were generated. The directions of these force vectors were co-ordinated such that the overall body sway response was always aligned with the inter-aural axis (i.e. craniocentric). When GVS was applied during light grip (< 1 N), hand forces were secondary to body movement, suggesting that the arm performed a mostly passive role. These results demonstrate that a minimum level of grip is required before the upper limb becomes active in balance control and also that the upper and lower limbs co-ordinate for an appropriate whole-body sway response. ABSTRACT: Vestibular stimulation can evoke responses in the arm when it is used for balance. In the present study, we determined how these responses are affected by grip context, as well as how they are co-ordinated with the rest of the body. Galvanic vestibular stimulation (GVS) was used to evoke balance responses under three conditions of manual contact with an earth-fixed object: no contact, light grip (< 1 N) (LG) and firm grip (FG). As grip progressed along this continuum, we observed an increase in GVS-evoked hand force, with a simultaneous reduction in ground reaction force (GRF) through the feet. During LG, hand force was secondary to the GVS-evoked body sway response, indicating that the arm performed a mostly passive role. By contrast, during FG, the arm became actively involved in driving body sway, as revealed by an early force impulse in the opposite direction to that seen in LG. We then examined how the direction of this active hand vector was co-ordinated with the lower limbs. Consistent with previous findings on sway anisotropy, FG skewed the direction of the GVS-evoked GRF vector towards the axis of baseline postural instability. However, this was effectively cancelled by the hand force vector, such that the whole-body sway response remained aligned with the inter-aural axis, maintaining the craniocentric principle. These results show that a minimum level of grip is necessary before the upper limb plays an active role in vestibular-evoked balance responses. Furthermore, they demonstrate that upper and lower-limb forces are co-ordinated to produce an appropriate whole-body sway response.
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Extremidad Inferior/fisiología , Equilibrio Postural , Extremidad Superior/fisiología , Vestíbulo del Laberinto/fisiología , Adulto , Femenino , Fuerza de la Mano , Humanos , MasculinoRESUMEN
This study aims to illustrate potential transport mechanisms behind the divergent approaches to nitrogen excretion seen in the ureotelic toadfish (Opsanus beta) and the ammoniotelic plainfin midshipman (Porichthys notatus). Specifically, we wish to confirm the expression of a urea transporter (UT), which is found in the gill of the toadfish and which is responsible for the unique "pulsing" nature of urea excretion and to localize the transporter within specific gill cells and at specific cellular locations. Additionally, the localization of ammonia transporters (Rhesus glycoproteins; Rhs) within the gill of both the toadfish and midshipman was explored. Toadfish UT (tUT) was found within Na(+)-K(+)-ATPase (NKA)-enriched cells, i.e., ionocytes (probably mitochondria-rich cells), especially along the basolateral membrane and potentially on the apical membrane. In contrast, midshipman UT (pnUT) immunoreactivity did not colocalize with NKA immunoreactivity and was not found along the filaments but instead within the lamellae. The cellular location of Rh proteins was also dissimilar between the two fish species. In toadfish gills, the Rh isoform Rhcg1 was expressed in both NKA-reactive cells and non-reactive cells, whereas Rhbg and Rhcg2 were only expressed in the latter. In contrast, Rhbg, Rhcg1 and Rhcg2 were expressed in both NKA-reactive and non-reactive cells of midshipman gills. In an additional transport epithelium, namely the intestine, the expression of both UTs and Rhs was similar between the two species, with only subtle differences being observed.
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Amoníaco/metabolismo , Batrachoidiformes/metabolismo , Proteínas de Peces/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Urea/metabolismo , Animales , Anticuerpos/metabolismo , Western Blotting , Perros , Glicoproteínas/metabolismo , Inmunohistoquímica , Células de Riñón Canino Madin Darby , Reproducibilidad de los Resultados , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Transformación Genética , Transportadores de UreaRESUMEN
The divalent metal transporter 1 (DMT1) is the best characterized Fe²âº transporter involved in cellular iron uptake in mammals. Four possible isoforms have been identified as a result of alternative promoter (DMT1-1A and DMT1-1B) and alternative splicing involving the C-terminus and producing transcripts with or without an iron responsive element [DMT1-IRE⺠and DMT1-IREâ», respectively]. Despite the general importance of DMT1 in controlling iron homeostasis, the distribution and the role of the transporter in the CNS is still controversial. In this study, we characterize the expression of DMT1 in hippocampal neurons and astrocytes. We found that the main isoform endogenously expressed is DMT1-1B/IREâº, which shows cytoplasmic distribution, colocalization with late endosome/lysosome markers and iron regulation, as expected from the presence of an iron responsive element. Our results also show that DMT1-1B/IRE⺠isoform does not sustain iron entry, even after its neuronal over-expression. Overall, our results argue against a physiological role of the endogenous DMT1 in neuronal iron uptake but do not exclude that, under pathological conditions, the expression of other DMT1 isoforms might contribute to iron overload.
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Proteínas de Transporte de Catión/metabolismo , Regulación de la Expresión Génica/fisiología , Hipocampo/citología , Hierro/metabolismo , Neuronas/metabolismo , Análisis de Varianza , Animales , Animales Recién Nacidos , Astrocitos/metabolismo , Fraccionamiento Celular/métodos , Células Cultivadas , Deferoxamina/farmacología , Duodeno/citología , Regulación de la Expresión Génica/efectos de los fármacos , Proteínas Fluorescentes Verdes/genética , Hierro/farmacología , Proteínas de Membrana de los Lisosomas/metabolismo , Microscopía Confocal , ARN Mensajero , Ratas , Ratas Sprague-Dawley , Receptores de Transferrina/metabolismo , Sideróforos/farmacología , Transfección/métodos , Transferrina/metabolismo , Proteínas de Transporte Vesicular/metabolismo , Grabación en VideoRESUMEN
Wearable devices are a popular training tool to measure biomechanical performance indicators during running, including vertical oscillation (VO). VO is a contributing factor in running economy and injury risk, therefore VO feedback can have a positive impact on running performance. The validity and reliability of the VO measurements from wearable devices is crucial for them to be an effective training tool. The aims of this study were to test the validity and reliability of VO measurements from wearable devices against video analysis of a single trunk marker. Four wearable devices were compared: the INCUS NOVA, Garmin Heart Rate Monitor-Pro (HRM), Garmin Running Dynamics Pod (RDP), and Stryd Running Power Meter Footpod (Footpod). Fifteen participants completed treadmill running at five different self-selected speeds for one minute at each speed. Each speed interval was completed twice. VO was recorded simultaneously by video and the wearables devices. There was significant effect of measurement method on VO (p < 0.001), with the NOVA and Footpod underestimating VO compared to video analysis, while the HRM and RDP overestimated. Although there were significant differences in the average VO values, all devices were significantly correlated with the video analysis (R > = 0.51, p < 0.001). Significant agreement between repeated VO measurements for all devices, revealed the devices to be reliable (ICC > = 0.948, p < 0.001). There was also significant agreement for VO measurements between each device and the video analysis (ICC > = 0.731, p < = 0.001), therefore validating the devices for VO measurement during running. These results demonstrate that wearable devices are valid and reliable tools to detect changes in VO during running. However, VO measurements varied significantly between the different wearables tested and this should be considered when comparing VO values between devices.
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Carrera , Dispositivos Electrónicos Vestibles , Humanos , Reproducibilidad de los Resultados , Carrera/fisiología , Prueba de Esfuerzo , Modalidades de FisioterapiaRESUMEN
Myocardial inflammation contributes to cardiomyopathy in diabetic patients through incompletely defined underlying mechanisms. In both human and time-course experimental samples, diabetic hearts exhibited abnormal ER, with a maladaptive shift over time in rodents. Furthermore, as a cardiac ER dysfunction model, mice with cardiac-specific p21-activated kinase 2 (PAK2) deletion exhibited heightened myocardial inflammatory response in diabetes. Mechanistically, maladaptive ER stress-induced CCAAT/enhancer-binding protein homologous protein (CHOP) is a novel transcriptional regulator of cardiac high-mobility group box-1 (HMGB1). Cardiac stress-induced release of HMGB1 facilitates M1 macrophage polarization, aggravating myocardial inflammation. Therapeutically, sequestering the extracellular HMGB1 using glycyrrhizin conferred cardioprotection through its anti-inflammatory action. Our findings also indicated that an intact cardiac ER function and protective effects of the antidiabetic drug interdependently attenuated the cardiac inflammation-induced dysfunction. Collectively, we introduce an ER stress-mediated cardiomyocyte-macrophage link, altering the macrophage response, thereby providing insight into therapeutic prospects for diabetes-associated cardiac dysfunction.
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Ferroportin 1 (FPN1) is an iron export protein expressed in liver and duodenum, as well as in reticuloendothelial macrophages. Previously, we have shown that divalent metal transporter 1 (DMT1) is expressed in late endosomes and lysosomes of the kidney proximal tubule (PT), the nephron segment responsible for the majority of solute reabsorption. We suggested that following receptor mediated endocytosis of transferrin filtered by the glomerulus, DMT1 exports iron liberated from transferrin into the cytosol. FPN1 is also expressed in the kidney yet its role remains obscure. As a first step towards determining the role of renal FPN1, we localized FPN1 in the PT. FPN1 was found to be located in association with the basolateral PT membrane and within the cytosolic compartment. FPN1 was not expressed on the apical brush-border membrane of PT cells. These data support a role for FPN1 in vectorial export of iron out of PT cells. Furthermore, under conditions of iron loading of cultured PT cells, FPN1 was trafficked to the plasma membrane suggesting a coordinated cellular response to export excess iron and limit cellular iron concentrations.
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Proteínas de Transporte de Catión/metabolismo , Sobrecarga de Hierro/metabolismo , Hierro/metabolismo , Túbulos Renales Proximales/metabolismo , Animales , Proteínas de Transporte de Catión/biosíntesis , Proteínas de Transporte de Catión/genética , Línea Celular , Membrana Celular/metabolismo , Masculino , Transporte de Proteínas , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Ratas , Ratas Sprague-DawleyRESUMEN
The aim of this study was to determine the iron (Fe) concentration profile within the lumen of the S2 renal proximal convoluted tubule (PCT) and to resolve whether this nephron segment transported Fe. To do this, we performed in vivo renal micropuncture on Wistar rats, collected PCT tubular fluid from superficial nephrons, and measured Fe concentration. The Fe concentration profile along the S2 PCT suggested significant Fe reabsorption. Proximal tubules were also microperfused in vivo with physiological solutions containing Fe and Zn, Cu, Mn, or Cd. PCTs perfused with 12µmol.l-1 55FeCl3 reabsorbed 105.2±12.7 fmol.mm-1.min-1 Fe, 435±52pmol.mm-1.min-1 Na, and 2.7±0.2nl.mm-1.min-1 water (mean ± SEM; n=19). Addition of ascorbate (1mmol.l-1) to the perfusate did not significantly alter Fe, Na, or water reabsorption. Supplementing the control perfusate with 60µmol.l-1 FeSO4 significantly decreased 55Fe uptake. Recalculating for the altered molar activity following addition of unlabeled Fe revealed a three-fold increase in Fe flux. Addition to the perfusate 12µmol.l-1 CuSO4, MnSO4, CdSO4, or ZnSO4 did not affect Fe, Na, or water flux. In conclusion, (1) in vivo, S2 PCTs of rat reabsorb Fe and (2) Fe is reabsorbed along the PCT via a pathway that is insensitive to Cu, Mn, Cd, or Zn. Together, these data demonstrate for the first time the hitherto speculated process of renal Fe filtration and subsequent tubular Fe reabsorption in a living mammal.
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Gut microbiota has been implicated as a modifier of childhood growth. Here, 16S rRNA sequencing-based fecal microbiota profiles of 18-24 month old Indian children were evaluated (n = 41), in relation to their anthropometric parameters, intestinal permeability, body composition and total energy expenditure. Pathway analyses were conducted to assess microbial functions related to stunting, underweight and wasting. The fecal microbiota was enriched in Prevotella 9, Bifidobacterium and Escherichia-Shigella. Weight, weight-for-age Z-scores (WAZ) and weight-for-length Z-scores (WLZ), along with age, acted as covariates of microbiota variation specifically in boys (n = 23). Bifidobacterium longum subsp longum abundance was positively associated with WAZ while Bifidobacterium bifidum and Bifidobacterium breve abundances were negatively associated with age. The lipopolysaccharide biosynthesis pathway was upregulated in stunted (n = 16) and wasted (n = 8) children. Findings from this study indicate that child sex may be a critical modifier of the role of gut microbiota on childhood growth.
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Bacterias , Desarrollo Infantil , Microbioma Gastrointestinal , Caracteres Sexuales , Bacterias/clasificación , Bacterias/genética , Bacterias/crecimiento & desarrollo , Preescolar , Femenino , Humanos , Lactante , MasculinoRESUMEN
Over the last decade there has been an explosion in our understanding of the proteins that modulate iron homeostasis. Much research has focused on the tissues classically associated with iron absorption and metabolism, namely the duodenum, the liver and the reticulo-endothelial system. Expression profiling has highlighted that many of the components associated with iron homeostasis, are also expressed in tissues which hitherto have received relatively little attention in terms of iron research. These include, testis, lung and, the subject of this review, the kidney. The latter is of great interest because other than a source of erythropoietin, a function that is of course of utmost importance for iron homeostasis, the kidney is regarded as more or less irrelevant in terms of iron handling. However, the fact that the kidneys of our favourite subjects, namely rats, mice and humans, contain many if not all of the proteins that are central to iron balance, that in some cases are expressed in considerable amounts, implies that the kidney handles iron in some way that has demanded evolutionary conservation and therefore is likely to be of importance. This review will document the evidence of iron transporter expression in the kidney, detail data showing the expression of other proteins associated with iron homeostasis and discuss the relevance of renal iron transport to pathophysiological states. Based on these data, a hypothetical model of renal iron handling will be presented.
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Hierro/metabolismo , Riñón/metabolismo , Animales , Proteínas de Transporte de Catión/metabolismo , Proteínas de Transporte de Catión/fisiología , Hemocromatosis/complicaciones , Homeostasis/fisiología , Humanos , Riñón/patología , Enfermedades Renales/etiología , Proteínas de la Membrana/fisiología , Modelos Biológicos , Siderosis/etiologíaRESUMEN
Ankle proprioception is crucial for balance and relies upon accurate input from calf muscle spindles. Spindle input, in turn, depends upon the physiological and mechanical properties of surrounding muscle tissue. Altering these properties could affect ankle proprioception, with potential consequences for balance. Here we determine the effects of prior muscle cooling, stretch and contraction upon performance of a contralateral ankle joint matching task. Participants stood passively leaning against a board oriented 22° rearward from vertical. Their right ankle was rotated to a randomised position between ± 6° plantar/dorsiflexion. The task was to align the left ankle to the same position, without vision. In the first experiment, immediately prior to each testing session, participants either produced a strong calf muscle contraction in a fully plantarflexed (tiptoe) posture or underwent 15° dorsiflexion stretch. Contraction had no effect on task performance, whereas stretch produced a significant bias in ankle placement of 0.89 ± 0.6°, indicating that participants perceived their foot to be more plantarflexed compared to a control condition. In the second experiment, the right lower leg was cooled in iced water (≤ 5°C) for 10 minutes. Cooling increased joint matching error by ~0.4°, through a combination of increased bias and variability. These results confirm that conditioning the triceps surae muscles can alter perception of ankle joint position. Since body movement during quiet stance is in the order of 1°, the magnitude of these changes are relevant for balance.
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Tobillo/fisiología , Músculo Esquelético/fisiología , Propiocepción , Adulto , Retroalimentación Fisiológica , Femenino , Humanos , Masculino , Contracción Muscular , Temperatura Cutánea , Adulto JovenRESUMEN
Urea transporters (UTs) encoded by the Slc14a1 (UT-B) and Slc14a2 (UT-A) genes mediate urea flux across cellular membranes. Considerable research has accrued detailing the function and distribution of members of both subfamilies. Much research effort has focused on the kidney, where UTs are highly expressed and function to promote urine concentration. Interestingly, UTs are also expressed in several other tissues that are historically not primarily associated with urea metabolism. In this review, I describe the phenotypes of UT knockout and transgenic mice and highlight the major advances made possible by use of these animal models. Where pertinent, I contrast these findings with known human phenotypes associated with UT mutations.
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Membrana Celular/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Urea/metabolismo , Animales , Transporte Biológico/fisiología , Modelos Animales de Enfermedad , Proteínas de Transporte de Membrana/genética , Ratones , Ratones Noqueados , Ratones Transgénicos , Transportadores de UreaRESUMEN
In upright stance, light touch of a space-stationary touch reference reduces spontaneous sway. Moving the reference evokes sway responses which exhibit non-linear behavior that has been attributed to sensory reweighting. Reweighting refers to a change in the relative contribution of sensory cues signaling body sway in space and light touch cues signaling finger position with respect to the body. Here we test the hypothesis that the sensory fusion process involves a transformation of light touch signals into the same reference frame as other sensory inputs encoding body sway in space, or vice versa. Eight subjects lightly gripped a robotic manipulandum which moved in a circular arc around the ankle joint. A pseudo-randomized motion sequence with broad spectral characteristics was applied at three amplitudes. The stimulus was presented at two different heights and therefore different radial distances, which were matched in terms of angular motion. However, the higher stimulus evoked a significantly larger sway response, indicating that the response was not matched to stimulus angular motion. Instead, the body sway response was strongly related to the horizontal translation of the manipulandum. The results suggest that light touch is integrated as the horizontal distance between body COM and the finger. The data were well explained by a model with one feedback loop minimizing changes in horizontal COM-finger distance. The model further includes a second feedback loop estimating the horizontal finger motion and correcting the first loop when the touch reference is moving. The second loop includes the predicted transformation of sensory signals into the same reference frame and a non-linear threshold element that reproduces the non-linear sway responses, thus providing a mechanism that can explain reweighting.
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Dedos , Modelos Biológicos , Equilibrio Postural/fisiología , Percepción del Tacto/fisiología , Adulto , Femenino , Humanos , MasculinoRESUMEN
Cholecystokinin (CCK) is an archetypal incretin hormone secreted by intestinal enteroendocrine cells (EEC) in response to ingested nutrients. The aim of this study was to determine whether CCK modulates enterocyte fatty acid uptake by primary mouse duodenal cells. Exposure of primary mouse duodenal cells to 10 pM sulfated CCK-8 caused a two fold increase in dodecanoic acid fatty acid (FA) uptake. The selective CCK A receptor antagonist loxiglumide (100 µM) completely abolished the CCK-8 induced FA uptake. The CD36 fatty acid translocase-specific inhibitor sulfo-N-succinimidyl oleate (1 µM) also completely inhibited CCK-8 induced FA uptake, as did treatment with 200 µM phloretin. Together these data show CCK induces FA uptake into duodenal enterocytes; this action involves the CCK-RA receptor and is carrier mediated by CD36.
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Maintaining a correct balance of N is essential for life. In mammals, the major sources of N in the diet are amino acids and peptides derived from ingested proteins. The immediate endproduct of mammalian protein catabolism is ammonia, which is toxic to cells if allowed to accumulate. Therefore, amino acids are broken down in the liver as part of the ornithine-urea cycle, which results in the formation of urea - a highly soluble, biochemically benign molecule. Mammals cannot break down urea, which is traditionally viewed as a simple waste product passed out in the urine. However, urea from the bloodstream can pass into the gastrointestinal tract, where bacteria expressing urease cleave urea into ammonia and carbon dioxide. The bacteria utilise the ammonia as an N source, producing amino acids and peptides necessary for growth. Interestingly, these microbial products can be reabsorbed back into the host mammalian circulation and used for synthetic processes. This entire process is known as 'urea nitrogen salvaging' (UNS). In this review we present evidence supporting a role for this process in mammals - including ruminants, non-ruminants and man. We also explore the possible mechanisms involved in UNS, including the role of specialised urea transporters.
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Enteroendocrine (EEC) cells have a pivotal role in intestinal nutrient sensing and release hormones that orchestrate food digestion and regulate appetite. EEC cells are found scattered throughout the intestine and have typically been classified based on the primary hormone they contain. I cells represent a subset of EEC cells that secrete cholecystokinin (CCK) and are mainly localized to the duodenum. Recent studies have shown that I cells express mRNAs encoding several gut hormones. In this study, we investigated the hormonal profile of murine fluorescence-activated cell sorting-sorted duodenal I cells using semiquantitative RT-PCR, liquid chromatography tandem mass spectrometry, and immunostaining methods. We report that I cells are enriched in mRNA transcripts encoding CCK and also other key gut hormones, including neurotensin, glucose-dependent insulinotropic peptide (GIP), secretin, peptide YY, proglucagon, and ghrelin (Ghrl). Furthermore, liquid chromatography tandem mass spectrometry analysis of fluorescence-activated cell sorting-purified I cells and immunostaining confirmed the presence of these gut hormones in duodenal I cells. Immunostaining highlighted that subsets of I cells in both crypts and villi coexpress differential amounts of CCK, Ghrl, GIP, or peptide YY, indicating that a proportion of I cells contain several hormones during maturation and when fully differentiated. Our results reveal that although I cells express several key gut hormones, including GIP or proglucagon, and thus have a considerable overlap with classically defined K and L cells, approximately half express Ghrl, suggesting a potentially important subset of duodenal EEC cells that require further consideration.
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Colecistoquinina/metabolismo , Duodeno/metabolismo , Células Enteroendocrinas/metabolismo , Ghrelina/metabolismo , Animales , Células Cultivadas , Colecistoquinina/genética , Duodeno/citología , Células Enteroendocrinas/citología , Citometría de Flujo , Hormonas Gastrointestinales/genética , Hormonas Gastrointestinales/metabolismo , Ghrelina/genética , Masculino , RatonesRESUMEN
Enteroendocrine cells have a critical role in regulation of appetite and energy balance. I-cells are a subtype of enteroendocrine cells localized in duodenum that release cholecystokinin in response to ingested fat and amino-acids. Despite their potentially pivotal role in nutrient sensing and feeding behaviour, native I-cells have previously been difficult to isolate and study. Here we describe a robust protocol for the isolation and characterization of native duodenal I-cells and additionally, using semi-quantitative RT-PCR we determined that mouse duodenal I-cells contain mRNA transcripts encoding key fatty acid and endocannabinoid receptors including the long chain fatty acid receptors GPR40/FFAR1, GPR120/O3FAR1; short chain fatty acid receptors GPR41/FFAR3 and GPR43/FFAR2; the oleoylethanolamide receptor GPR119 and the classic endocannabinoid receptor CB1. These data suggest that I-cells sense a wide range of gut lumen nutrients and also have the capacity to respond to signals of fatty-acid derivatives or endocannabinoid peptides.
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Duodeno/metabolismo , Endocannabinoides/metabolismo , Células Enteroendocrinas/metabolismo , Ácidos Grasos/metabolismo , ARN Mensajero/genética , Receptores Acoplados a Proteínas G/genética , Animales , ADN Complementario , Duodeno/citología , Expresión Génica , Masculino , Ratones , Ratones Transgénicos , Receptores de Colecistoquinina/genética , Receptores de Colecistoquinina/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Reproducibilidad de los Resultados , Transcripción GenéticaRESUMEN
The ideal serum level of phosphate in patients on dialysis, and the benefits of controlling levels of phosphate in serum remain unclear despite observational studies that associate phosphate levels with mortality. In the absence of robust data from trials, current guidelines are necessarily based on opinion. Oral phosphate binders are required by the majority of patients on dialysis, and all of these binders can control serum levels of phosphate to similar degrees. Patient preference and adherence to prescribed therapy is at least as important as the efficacy of the prescribed binder. Avoidance of calcium-containing binders has become accepted practice where the alternatives are affordable, but incontrovertible evidence in favor of this approach is lacking. Use of sevelamer and lanthanum avoids calcium loading, but at considerable financial cost and with no reliable patient outcome data to prove their value. Additional approaches to aid control of serum levels of phosphate include blockade of gastrointestinal phosphate absorption and possibly binding of salivary phosphate. Importantly, the role of phosphate control in determining patient outcomes must be quantified, which is likely to require a large randomized, controlled study of two levels of phosphate control. Without such a study we will continue to rely on observational data with all its uncertainties and potential to mislead.