RESUMEN
Down syndrome (DS) is associated with congenital heart defects at birth, but cardiac function has not been assessed at older ages. We used the Ts65Dn mouse, a model of DS, to quantify heart structure and function with echocardiography in 18-mo male Ts65Dn and wild-type (WT) mice. Heart weight, nicotinamide adenine dinucleotide (NAD) signaling, and mitochondrial (citrate synthase) activity were investigated, as these pathways may be implicated in the cardiac pathology of DS. The left ventricle was smaller in Ts65Dn versus WT, as well as the anterior wall thickness of the left ventricle during both diastole (LVAW_d; mm) and systole (LVAW_s; mm) as assessed by echocardiography. Other functional metrics were similar between groups including left ventricular area end systole (mm2), left ventricular area end diastole (mm2), left ventricular diameter end systole (mm), left ventricular diameter end diastole (mm), isovolumetric relaxation time (ms), mitral valve atrial peak velocity (mm/s), mitral valve early peak velocity (mm/s), ratio of atrial and early peak velocities (E/A), heart rate (beats/min), ejection fraction (%), and fractional shortening (%). Nicotinamide phosphoribosyltransferase (NAMPT) protein expression, NAD concentration, and tissue weight were lower in the left ventricle of Ts65Dn versus WT mice. Sirtuin 3 (SIRT3) protein expression and citrate synthase activity were not different between groups. Although cardiac function was generally preserved in male Ts65Dn, the altered heart size and bioenergetic disturbances may contribute to differences in aging for DS.
Asunto(s)
NAD , Función Ventricular Izquierda , Masculino , Ratones , Animales , Función Ventricular Izquierda/fisiología , Citrato (si)-Sintasa , Diástole/fisiología , EcocardiografíaRESUMEN
Individuals with Down syndrome (Ds) are at increased risk of respiratory infection, aspiration pneumonia, and apnea. The Ts65Dn mouse is a commonly used model of Ds, but there have been no formal investigations of awake breathing and respiratory muscle function in these mice. We hypothesized that breathing would be impaired in Ts65Dn vs. wild-type (WT), and would be mediated by both neural and muscular inputs. Baseline minute ventilation was not different at 3, 6, or 12 mo of age. However, VT/Ti, a marker of the neural drive to breathe, was lower in Ts65Dn vs. WT and central apneas were more prevalent. The response to breathing hypoxia was not different, but the response to hypercapnia was attenuated, revealing a difference in carbon dioxide sensing, and/or motor output in Ts65Dn. Oxygen desaturations were present in room air, demonstrating that ventilation may not be sufficient to maintain adequate oxygen saturation in Ts65Dn. We observed no differences in arterial PO2 or PCO2, but Ts65Dn had lower hemoglobin and hematocrit. A retrospective medical record review of 52,346 Ds and 52,346 controls confirmed an elevated relative risk of anemia in Ds. We also performed eupneic in-vivo electromyography and in-vitro muscle function and histological fiber typing of the diaphragm, and found no difference between strains. Overall, conscious respiration is impaired in Ts65Dn, is mediated by neural mechanisms, and results in reduced hemoglobin saturation. Oxygen carrying capacity is reduced in Ts65Dn vs. WT, and we demonstrate that individuals with Ds are also at increased risk of anemia.
Asunto(s)
Anemia , Síndrome de Down , Ratones , Animales , Oxígeno , Síndrome de Down/genética , Estudios Retrospectivos , Conservación de los Recursos Naturales , Respiración , HemoglobinasRESUMEN
This meta-analysis, which consisted of a scoping review and retrospective medical record review, is focused on potential sex differences in cardiovascular diseases in patients with Down syndrome. We limited our review to peer-reviewed, primary articles in the English language, in the PubMed and Web of Science databases from 1965 to 2021. Guidelines for scoping reviews were followed throughout the process. Four categorical domains were identified and searched using additional keywords: 1) congenital heart disease, 2) baseline physiology and risk factors, 3) heart disease and hypertension, and 4) stroke and cerebrovascular disease. Articles were included if they reported male and female distinct data, participants with Down syndrome, and one of our keywords. The retrospective medical record review was completed using 75 participating health care organizations to identify the incidence of congenital and cardiovascular diseases and to quantify cardiovascular risk factors in male and female patients. Female patients with Down syndrome are at higher risk of hypertension, ischemic heart disease, and cerebrovascular disease. The risk of congenital heart disease is higher in males with Down syndrome at all ages included in our analyses. Some of the male-to-female sex differences in cardiovascular disease risk in the general patient population are not present, or reversed in the Down syndrome population. This information should be considered for future investigations and ongoing patient care.NEW & NOTEWORTHY In patients with Down syndrome (DS), CHD is the leading cause of death <20 yr old and cardiovascular disease is a leading cause of death in individuals >20 yr old. Men with DS live longer than women. It is unknown if sex differences are present in cardiovascular disease and dysregulation in DS across the lifespan. We observed higher risk of hypertension, ischemic heart disease, and cerebrovascular disease in females and a higher risk of CHD in males with DS.
Asunto(s)
Enfermedades Cardiovasculares , Síndrome de Down , Cardiopatías Congénitas , Hipertensión , Isquemia Miocárdica , Accidente Cerebrovascular , Humanos , Masculino , Femenino , Síndrome de Down/epidemiología , Enfermedades Cardiovasculares/epidemiología , Estudios Retrospectivos , Caracteres Sexuales , Hipertensión/epidemiologíaRESUMEN
Coronavirus 2019 (COVID-19) disease has been a public health emergency of international concern with millions of confirmed cases globally. Closed environments with reduced ventilation contribute to the spread of COVID-19, including superspreading events. Exercising in closed places further increases the risk for transmission. Therefore, many fitness facilities were closed as part of mandated shutdowns early in the pandemic. Evidence-based safety protocols have now emerged and substantially reduce the risk of transmission. We report three positive cases of SARS-CoV-2 identified at a Dojo exercise facility in Manlius, NY, at three distinct time points. All cases were present in the Dojo 2 days prior to symptoms, a time period considered to be highly infectious. The safety protocols included universal mask wearing (no valves), multiple high-efficiency particulate air (HEPA) filters, and reduced capacity which resulted in no known spread of COVID-19.
Asunto(s)
COVID-19/transmisión , Centros de Acondicionamiento , Seguridad , Filtros de Aire , Desinfección , Medicina Basada en la Evidencia , Ejercicio Físico , Humanos , Máscaras , Pandemias , Equipo de Protección Personal , Riesgo , VentilaciónRESUMEN
Assessment methods vary widely across undergraduate physiology courses. Here, a cumulative oral examination was administered in two sections of a 300-level undergraduate physiology course. Student performance was quantified via instructor grading using a rubric, and self-perceptions (n = 55) were collected via survey. Overall, students affirmed that the oral examination assisted in their learning, specifically by leading them to begin preparation for their final written exam earlier than they otherwise would. The instructor considered the oral exam useful for student learning by providing a scaffold to the written final exam and a way to connect with students before a high-stakes final exam. Specific details of the examination format and suggestions and considerations for those considering this assessment approach are provided.
Asunto(s)
Evaluación Educacional , Fisiología , Diagnóstico Bucal , Humanos , Aprendizaje , Percepción , Fisiología/educación , EstudiantesRESUMEN
BACKGROUND: Recent advances in medical care have increased life expectancy and improved the quality of life for people with Down syndrome (DS). These advances are the result of both pre-clinical and clinical research but much about DS is still poorly understood. In 2020, the NIH announced their plan to update their DS research plan and requested input from the scientific and advocacy community. OBJECTIVE: The National Down Syndrome Society (NDSS) and the LuMind IDSC Foundation worked together with scientific and medical experts to develop recommendations for the NIH research plan. METHODS: NDSS and LuMind IDSC assembled over 50 experts across multiple disciplines and organized them in eleven working groups focused on specific issues for people with DS. RESULTS: This review article summarizes the research gaps and recommendations that have the potential to improve the health and quality of life for people with DS within the next decade. CONCLUSIONS: This review highlights many of the scientific gaps that exist in DS research. Based on these gaps, a multidisciplinary group of DS experts has made recommendations to advance DS research. This paper may also aid policymakers and the DS community to build a comprehensive national DS research strategy.
RESUMEN
Unrestrained barometric plethysmography (UBP) is a method for quantifying the pattern of breathing in mice, where breathing frequency, tidal volume, and minute ventilation are routinely reported. Moreover, information can be collected regarding the neural output of breathing, including the existence of central apneas and augmented breaths. An important consideration for UBP is obtaining a breathing segment with a minimal impact of anxious or active behaviors, to elucidate the response to breathing challenges. Here, we present a protocol that allows for short, quiet baselines to be obtained in aged mice, comparable to waiting for longer bouts of quiet breathing. The use of shorter time segments is valuable, as some strains of mice may be increasingly excitable or anxious, and longer periods of quiet breathing may not be achieved within a reasonable timeframe. We placed 22 month-old mice in a UBP chamber and compared four 15 s quiet breathing segments between minutes 60-120 to a longer 10 min quiet breathing period that took 2-3 h to acquire. We also obtained counts of central apneas and augmented breaths prior to the quiet breathing segments, following a 30 min familiarization period. We show that 10 min of quiet breathing is comparable to using a much shorter 15 s duration. Additionally, the time leading up to these 15 s quiet breathing segments can be used to gather data regarding apneas of central origin. This protocol allows investigators to collect pattern-of-breathing data in a set amount of time and makes quiet baseline measures feasible for mice that may exhibit increased amounts of excitable behavior. The UBP methodology itself provides a useful and noninvasive way to collect pattern-of-breathing data and allows for mice to be tested over several time points.
Asunto(s)
Pletismografía/métodos , Respiración/inmunología , Animales , Masculino , RatonesRESUMEN
Down syndrome (Ds) is the most common chromosomal cause of intellectual disability that results from triplication of chromosome 21 genes. Lower blood pressure (BP) and heart rate (HR) in response to exercise and other stressors are prevalent in Ds, and are mediated by autonomic dysfunction. The Ts65Dn mouse is a model of Ds that is commonly used in preclinical studies, but has not been formally investigated for cardiovascular responses in conscious mice. Based on human studies of Ds, we hypothesized Ts65Dn would have lower BP and HR, but similar arterial stiffness. BP was quantified in conscious wild-type (WT) and Ts65Dn. A main effect for strain was observed for all BP measures (systolic, diastolic, mean, pulse pressure), with WT higher than Ts65Dn. Pulse wave velocity was similar between WT and Ts65Dn. High-frequency power spectra was higher in WT suggesting autonomic differences between strains. Freely moving HR was higher in WT versus Ts65Dn in both the dark and light cycles, although a main effect of circadian cycle was also present (dark> light). Similar to what is observed in humans, Ts65Dn has a lower BP which may be attributed to autonomic differences and result in preservation of arterial function with advancing age. Ts65Dn thus appears to capture the Ds cardiovascular phenotype across the lifespan. These data support further use of Ts65Dn to investigate mechanisms that may lead to altered BP and HR responses in Ds.
Asunto(s)
Presión Sanguínea , Síndrome de Down/fisiopatología , Frecuencia Cardíaca , Animales , Sistema Nervioso Autónomo/fisiopatología , Ritmo Circadiano , Síndrome de Down/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Rigidez VascularRESUMEN
Unrestrained barometric plethysmography is a common method used for characterizing breathing patterns in small animals. One source of variation between unrestrained barometric plethysmography studies is the segment of baseline. Baseline may be analyzed as a predetermined time-point, or using tailored segments when each animal is visually calm. We compared a quiet, minimally active (no sniffing/grooming) breathing segment to a predetermined time-point at 1 h for baseline measurements in young and middle-aged mice during the dark and light cycles. Additionally, we evaluated the magnitude of change for gas challenges based on these two baseline segments. C57BL/6JEiJ x C3Sn.BliA-Pde6b+ /DnJ male mice underwent unrestrained barometric plethysmography with the following baselines used to determine breathing frequency, tidal volume (VT) and minute ventilation (VE): (1) 30-sec of quiet breathing and (2) a 10-min period from 50 to 60 min. Animals were also exposed to 10 min of hypoxic (10% O2 , balanced N2 ), hypercapnic (5% CO2 , balanced air) and hypoxic hypercapnic (10% O2 , 5% CO2 , balanced N2 ) gas. Both frequency and VE were higher during the predetermined 10-min baseline versus the 30-sec baseline, while VT was lower (P < 0.05). However, VE/VO2 was similar between the baseline time segments (P > 0.05) in an analysis of one cohort. During baseline, dark cycle testing had increased VT values versus those in the light (P < 0.05). For gas challenges, both frequency and VE showed higher percent change from the 30-sec baseline compared to the predetermined 10-min baseline (P < 0.05), while VT showed a greater change from the 10-min baseline (P < 0.05). Dark cycle hypoxic exposure resulted in larger percent change in breathing frequency versus the light cycle (P < 0.05). Overall, light and dark cycle pattern of breathing differences emerged along with differences between the 30-sec behavior observational method versus a predetermined time segment for baseline.
Asunto(s)
Envejecimiento/fisiología , Ritmo Circadiano , Hipercapnia/fisiopatología , Hipoxia/fisiopatología , Respiración , Animales , Masculino , Ratones , Ratones Endogámicos C57BLRESUMEN
Capsaicin is an agonist for transient receptor potential vanilloid 1 (TRPV1), and acute injection results in an increased frequency and tidal volume in young rats. It is unknown how capsaicin influences breathing in aged mice. We tested the hypothesis that capsaicin supplementation would elicit an augmented pattern of breathing in old mice compared to controls. Male 22-month old C57BL/6 J mice consumed a diet containing capsaicin (50 ppm) or lecithin control for one month. Breathing patterns were obtained prior to/following the dietary supplementation period using unrestrained barometric plethysmography. Frequency, tidal volume (VT), minute ventilation (VE), VE to expelled carbon dioxide ratio (VE/VCO2) and VT divided by inspiratory time (VT/Ti) were analyzed at baseline and during a 15-minute hypoxic exposure (10% O2). Capsaicin supplemented mice showed greater VE, VE/VCO2 and TV/Ti during hypoxic exposure compared to controls, with no change at baseline. Overall, these findings suggest an acute augmented response to hypoxia following capsaicin administration in older mice.
Asunto(s)
Capsaicina/farmacología , Hipoxia/fisiopatología , Frecuencia Respiratoria/efectos de los fármacos , Frecuencia Respiratoria/fisiología , Canales Catiónicos TRPV/agonistas , Animales , Capsaicina/administración & dosificación , Masculino , Ratones , Ratones Endogámicos C57BL , Pletismografía , Volumen de Ventilación Pulmonar/efectos de los fármacos , Volumen de Ventilación Pulmonar/fisiologíaRESUMEN
Isoflurane (ISO) is a commonly used anesthetic that offers rapid recovery for laboratory animal research. Initial studies indicated no difference in arterial Pco2 ( PaCO2 ) or pH between conscious (NO ISO) and 1% ISO-exposed CD-1 mice. Our laboratory investigated whether arterial blood sampling with 1% ISO is a suitable alternative to NO ISO sampling for monitoring ventilation in a commonly studied mouse strain. We hypothesized similar blood chemistry, breathing patterns, and cardiovascular responses with NO ISO and 1% ISO. C57BL/6J mice underwent unrestrained barometric plethysmography to quantify the pattern of breathing. Mice exposed to hypoxic and hypercapnic gas under 1% ISO displayed blunted responses; with air, there were no breathing differences. Blood pressure and heart rate were not different between NO ISO and 1% ISO-exposed mice breathing air. Oxygen saturation was not different between groups receiving 2% ISO, 1% ISO, or air. Breathing frequency stabilized at ~11 min of 1% ISO following 2% ISO exposure, suggesting that 11 min is the optimal time for a sample in C57BL/6J mice. Blood samples at 1% ISO and NO ISO revealed no differences in blood pH and PaCO2 in C57BL/6J mice. Overall, this method reveals similar arterial blood sampling values in awake and 1% ISO CD-1 and C57BL/6J mice exposed to air. Although this protocol may be appropriate in other mouse strains when a conscious sample is not feasible, caution is warranted first to identify breathing frequency responses at 1% ISO to tailor the protocol. NEW & NOTEWORTHY Conscious arterial blood sampling is influenced by extraneous factors and is a challenging method due to the small size of mice. Through a series of experiments, we show that arterial blood sampling with 1% isoflurane (ISO) is an alternative to awake sampling in C57BL/6J and CD-1 male mice breathing air. Monitoring breathing frequency during 1% ISO is important to the protocol and should be closely followed to confirm adequate recovery after the catheter implantation.
Asunto(s)
Anestésicos por Inhalación , Recolección de Muestras de Sangre/métodos , Arteria Femoral/cirugía , Isoflurano , Vigilia , Animales , Presión Sanguínea , Frecuencia Cardíaca , Masculino , Ratones Endogámicos C57BL , Oximetría , RespiraciónRESUMEN
The hindlimb casting model was developed to study skeletal muscle reloading following a period of unloading. It is unknown if ventilation parameters of mice are affected by the casting model. We tested the hypothesis that hindlimb casted mice have similar ventilatory patterns compared to mice with the casts removed. Male CD-1 mice underwent 14 days of hindlimb immobilization via plaster casting. Breathing parameters were obtained utilizing unrestrained barometric plethysmography (UBP). Breathing traces were analyzed with Ponemah software for breathing frequency, tidal volume (TV), and minute ventilation (MV). Frequency, TV and MV did not show any differences in quiet breathing patterns during or post-casting in mice. Thus, the hindlimb casting model does not complicate breathing during and after casting and should not interfere with the unloading and reloading of skeletal muscle.
Asunto(s)
Suspensión Trasera/efectos adversos , Atrofia Muscular/complicaciones , Atrofia Muscular/etiología , Respiración , Animales , Peso Corporal , Modelos Animales de Enfermedad , Masculino , Ratones , Pletismografía , Volumen de Ventilación Pulmonar/fisiologíaRESUMEN
Saline (0.9% NaCl) is used in clinical and research settings as a vehicle for intravenous drug administration. While saline is a standard control in mouse studies, there are reports of hyperchloremic metabolic acidosis in high doses. It remains unknown if metabolic acidosis occurs in mice and/or if compensatory increases in breathing frequency and tidal volume accompany saline administration. It was hypothesized that saline administration alters blood pH and the pattern of breathing in conscious CD-1 male mice exposed to air or hypoxia (10% O2 , balanced N2 ). Unrestrained barometric plethysmography was used to quantify breathing frequency (breaths/min; bpm), tidal volume (VT; mL/breath/10 g body weight (BW)), and minute ventilation (VE; mL/min/10 g BW) in two designs: (1) 11-week-old mice with no saline exposure (n = 11) compared to mice with 7 days of 0.9% saline administration (intraperitoneal, i.p.; 10 mL/kg body mass; n = 6). and (2) 17-week-old mice tested before (PRE) and after 1 day (POST1, n = 6) or 7 days (POST7, n = 5) of saline (i.p.; 10 mL/kg body mass). There were no differences when comparing frequency, VT, or VE between groups for either design with room air or hypoxia exposures. Hypoxia increased frequency, VT, and VE compared to room air. Moreover, conscious blood sampling showed no differences in pH, paCO2 , paO2 , or HCO3- in mice without or with 7 days of saline. These findings reveal no differences in ventilation following 1 and/or 7 days of saline administration in mice. Therefore, the use of 0.9% saline as a control is supported for studies evaluating the control of breathing in mice.
Asunto(s)
Ventilación Pulmonar/efectos de los fármacos , Respiración/efectos de los fármacos , Solución Salina/administración & dosificación , Acidosis/sangre , Acidosis/inducido químicamente , Administración Intravenosa/efectos adversos , Animales , Peso Corporal/efectos de los fármacos , Masculino , Ratones , Volumen de Ventilación Pulmonar/efectos de los fármacosRESUMEN
Down syndrome (DS) is a genetic condition caused by the triplication of chromosome 21. Persons with DS exhibit pronounced muscle weakness, which also occurs in the Ts65Dn mouse model of DS. Oxidative stress is thought to be an underlying factor in the development of DS-related pathologies including muscle dysfunction. High-levels of oxidative stress have been attributed to triplication and elevated expression of superoxide dismutase 1 (SOD1); a gene located on chromosome 21. The elevated expression of SOD1 is postulated to increase production of hydrogen peroxide and cause oxidative injury and cell death. However, it is unknown whether SOD1 protein expression is associated with greater oxidant production in skeletal muscle from Ts65Dn mice. Thus, our objective was to assess levels of SOD1 expression and oxidant production in skeletal myofibers from the flexor digitorum brevis obtained from Ts65Dn and control mice. Measurements of oxidant production were obtained from myofibers loaded with 2',7'-dichlorodihydrofluorescein diacetate (DCFH2-DA) in the basal state and following 15min of stimulated unloaded contraction. Ts65Dn myofibers exhibited a significant decrease in basal DCF emissions (p < 0.05) that was associated with an approximate 3-fold increase in SOD1 (p < 0.05). DCF emissions were not affected by stimulating contraction of Ts65Dn or wild-type myofibers (p > 0.05). Myofibers from Ts65Dn mice tended to be smaller and myonuclear domain was lower (p < 0.05). In summary, myofibers from Ts65Dn mice exhibited decreased basal DCF emissions that were coupled with elevated protein expression of SOD1. Stimulated contraction in isolated myofibers did not affect DCF emissions in either group. These findings suggest the skeletal muscle dysfunction in the adult Ts65Dn mouse is not associated with skeletal muscle oxidative stress.
Asunto(s)
Síndrome de Down/metabolismo , Peróxido de Hidrógeno/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Superóxido Dismutasa-1/metabolismo , Animales , Células Cultivadas , Masculino , Ratones , Contracción Muscular , Fibras Musculares Esqueléticas/fisiología , Oxidantes/metabolismo , Estrés Oxidativo , Superóxido Dismutasa-1/genéticaRESUMEN
The flipped classroom was utilized in a two-semester, high-content science course that enrolled between 50 and 80 students at a small liberal arts college. With the flipped model, students watched ~20-min lectures 2 days/wk outside of class. These videos were recorded via screen capture and included a detailed note outline, PowerPoint slides, and review questions. The traditional format included the same materials, except that lectures were delivered in class each week and spanned the entire period. During the flipped course, the instructor reviewed common misconceptions and asked questions requiring higher-order thinking, and five graded case studies were performed each semester. To determine whether assessments included additional higher-order thinking skills in the flipped vs. traditional model, questions across course formats were compared via Blooms Taxonomy. Application-level questions that required prediction of an outcome in a new scenario comprised 38 ± 3 vs. 12 ± 1% of summative assessment questions (<0.01): flipped vs. traditional. Final letter grades in both formats of the course were compared with major GPA. Students in the flipped model performed better than their GPA predicted, as 85.5% earned a higher grade (vs. 42.2% in the traditional classroom) compared with their major GPA. These data demonstrate that assessments transitioned to more application-level compared with factual knowledge-based questions with this particular flipped model, and students performed better in their final letter grade compared with the traditional lecture format. Although the benefits to a flipped classroom are highlighted, student evaluations did suffer. More detailed studies comparing the traditional and flipped formats are warranted.
Asunto(s)
Educación de Pregrado en Medicina/métodos , Evaluación Educacional/métodos , Aprendizaje Basado en Problemas/métodos , Pensamiento , Femenino , Humanos , MasculinoRESUMEN
The rise in non-heme iron (NHI) concentration observed in skeletal muscle of aging rodents is thought to contribute to the development of sarcopenia. The source of the NHI has not been identified, nor have the physiological ramifications of elevated iron status in aged muscle been directly examined. Therefore, we assessed plantaris NHI and heme iron (HI) levels in addition to expression of proteins involved in iron uptake (transferrin receptor-1; TfR1), storage (ferritin), export (ferroportin; FPN), and regulation (iron regulatory protein-1 (IRP1) and -2 (IRP2)) of male F344xBN F1 rats (n=10/group) of various ages (8, 18, 28, 32, and 36 months) to further understand iron regulation in aging muscle. In a separate experiment, iron chelator (pyridoxal isonicotinoyl hydrazone; PIH) or vehicle was administered to male F344xBN F1 rats (n=8/group) beginning at 30 months of age to assess the impact on plantaris muscle mass and function at ~36 months of age. Principle findings revealed the increased NHI concentration in old age was consistent with concentrating effects of muscle atrophy and reduction in HI levels, with no change in the total iron content of the muscle. The greatest increase in muscle iron content occurred during the period of animal growth and was associated with downregulation of TfR1 and IRP2 expression. Ferritin upregulation did not occur until senescence and the protein remained undetectable during the period of muscle iron content elevation. Lastly, administration of PIH did not significantly (p>0.05) impact NHI or measures of muscle atrophy or contractile function. In summary, this study confirms that the elevated NHI concentration in old age is largely due to the loss in muscle mass. The increased muscle iron content during aging does not appear to associate with cytosolic ferritin storage, but the functional consequences of elevated iron status in old age remains to be determined.
Asunto(s)
Envejecimiento/metabolismo , Hierro/metabolismo , Músculo Esquelético/metabolismo , Envejecimiento/genética , Envejecimiento/patología , Animales , Proteínas de Transporte de Catión/genética , Proteínas de Transporte de Catión/metabolismo , Ferritinas/genética , Ferritinas/metabolismo , Expresión Génica , Hemo/metabolismo , Quelantes del Hierro/farmacología , Proteína 1 Reguladora de Hierro/genética , Proteína 1 Reguladora de Hierro/metabolismo , Proteína 2 Reguladora de Hierro/genética , Proteína 2 Reguladora de Hierro/metabolismo , Isoniazida/análogos & derivados , Isoniazida/farmacología , Masculino , Contracción Muscular/efectos de los fármacos , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/patología , Atrofia Muscular/metabolismo , Atrofia Muscular/patología , Piridoxal/análogos & derivados , Piridoxal/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas BN , Ratas Endogámicas F344 , Receptores de Transferrina/genética , Receptores de Transferrina/metabolismo , Sarcopenia/metabolismo , Sarcopenia/patologíaRESUMEN
Persons with Down syndrome (DS) exhibit low muscle strength that significantly impairs their physical functioning. The Ts65Dn mouse model of DS also exhibits muscle weakness in vivo and may be a useful model to examine DS-associated muscle dysfunction. Therefore, the purpose of this experiment was to directly assess skeletal muscle function in the Ts65Dn mouse and to reveal potential mechanisms of DS-associated muscle weakness. Soleus muscles were harvested from anesthetized male Ts65Dn and wild-type (WT) colony controls. In vitro muscle contractile experiments revealed normal force generation of nonfatigued Ts65Dn soleus, but a 12% reduction in force was observed during recovery from fatiguing contractions compared with WT muscle (P < 0.05). Indicators of oxidative stress and mitochondrial oxidative capacity were assessed to reveal potential mechanisms of DS-associated muscle weakness. Protein expression of copper-zinc superoxide dismutase (SOD1), a triplicated gene in persons with DS and Ts65Dn mice, was increased 25% (P < 0.05) in Ts65Dn soleus. Nontriplicated antioxidant protein expression was similar between groups. Lipid peroxidation was unaltered in Ts65Dn animals, but protein oxidation was 20% greater compared with controls (P < 0.05). Cytochrome-c oxidase expression was 22% lower in Ts65Dn muscle (P < 0.05), while expression of citrate synthase was similar between groups. Microarray analysis revealed alteration of numerous pathways in Ts65Dn muscle, including proteolysis, glucose and fat metabolism, neuromuscular transmission, and ATP biosynthesis. In summary, despite biochemical and gene expression differences in soleus muscle of Ts65Dn animals, the functional properties of skeletal muscle likely contribute a minor part to the in vivo muscle weakness.
Asunto(s)
Modelos Animales de Enfermedad , Síndrome de Down/fisiopatología , Debilidad Muscular/fisiopatología , Músculo Esquelético/fisiopatología , Animales , Síndrome de Down/metabolismo , Complejo IV de Transporte de Electrones/metabolismo , Técnicas In Vitro , Peroxidación de Lípido/fisiología , Masculino , Ratones , Ratones Mutantes , Contracción Muscular/fisiología , Fatiga Muscular/fisiología , Debilidad Muscular/metabolismo , Músculo Esquelético/metabolismo , Estrés Oxidativo/fisiología , Superóxido Dismutasa/metabolismoRESUMEN
Pompe disease is a muscular dystrophy that results in respiratory insufficiency. We characterized the outcomes of targeted delivery of recombinant adeno-associated virus serotype 1 (rAAV2/1) vector to diaphragms of Pompe mice with varying stages of disease progression. We observed significant improvement in diaphragm contractile strength in mice treated at 3 months of age that is sustained at least for 1 year and enhanced contractile strength in mice treated at 9 and 21 months of age, measured 3 months post-treatment. Ventilatory parameters including tidal volume/inspiratory time ratio, minute ventilation/expired CO2 ratio, and peak inspiratory airflow were significantly improved in mice treated at 3 months and tested at 6 months. Despite early improvement, mice treated at 3 months and tested at 1 year had diminished normoxic ventilation, potentially due to attenuation of correction over time or progressive degeneration of nontargeted accessory tissues. However, for all rAAV2/1-treated mice (treated at 3, 9, and 21 months, assayed 3 months later; treated at 3 months, assayed at 1 year), minute ventilation and peak inspiratory flows were significantly improved during respiratory challenge. These results demonstrate that gel-mediated delivery of rAAV2/1 vectors can significantly augment ventilatory function at initial and late phases of disease in a model of muscular dystrophy.
Asunto(s)
Dependovirus/genética , Técnicas de Transferencia de Gen , Terapia Genética/métodos , Enfermedad del Almacenamiento de Glucógeno Tipo II/genética , Enfermedad del Almacenamiento de Glucógeno Tipo II/terapia , Distrofias Musculares/terapia , Respiración , Animales , Dióxido de Carbono/química , Progresión de la Enfermedad , Geles , Vectores Genéticos , Ratones , Ratones Transgénicos , Contracción Muscular , Distrofias Musculares/genética , Factores de TiempoRESUMEN
Metallothionein (MT) is a small molecular weight protein possessing metal binding and free radical scavenging properties. We hypothesized that MT-1/MT-2 null (MT(-/-)) mice would display exacerbated soleus muscle atrophy, oxidative injury, and contractile dysfunction compared with the response of wild-type (WT) mice following acute spinal cord transection (SCT). Four groups of mice were studied: WT laminectomy, WT transection, MT(-/-) laminectomy (MT(-/-) lami), and MT(-/-) transection (MT(-/-) trans). Laminectomy animals served as surgical controls. Mice in SCT groups experienced similar percent body mass (BM) losses at 7 days postinjury. Soleus muscle mass (MM) and MM-to-BM ratio were lower at 7 days postinjury in SCT vs. laminectomy mice, with no differences observed between strains. However, soleus muscles from MT(-/-) trans mice showed reduced maximal specific tension compared with MT(-/-) lami animals. Mean cross-sectional area (microm(2)) of type I and type IIa fibers decreased similarly in SCT groups compared with laminectomy controls, and no difference in fiber distribution was observed. Lipid peroxidation (4-hydroxynoneal) was greater in MT(-/-) trans vs. MT(-/-) lami mice, but protein oxidation (protein carbonyls) was not altered by MT deficiency or SCT. Expression of key antioxidant proteins (catalase, manganese, and copper-zinc superoxide dismutase) was similar between the groups. In summary, MT deficiency did not impact soleus MM loss, but resulted in contractile dysfunction and increased lipid peroxidation following acute SCT. These findings suggest a role of MT in mediating protective adaptations in skeletal muscle following disuse mediated by spinal cord injury.
Asunto(s)
Metalotioneína/deficiencia , Contracción Muscular , Músculo Esquelético/metabolismo , Traumatismos de la Médula Espinal/metabolismo , Enfermedad Aguda , Animales , Antioxidantes/metabolismo , Catalasa/metabolismo , Modelos Animales de Enfermedad , Laminectomía , Peroxidación de Lípido , Masculino , Metalotioneína/genética , Ratones , Ratones Noqueados , Fibras Musculares de Contracción Rápida/metabolismo , Fibras Musculares de Contracción Lenta/metabolismo , Fuerza Muscular , Músculo Esquelético/patología , Músculo Esquelético/fisiopatología , Atrofia Muscular/metabolismo , Atrofia Muscular/fisiopatología , Estrés Oxidativo , Carbonilación Proteica , Traumatismos de la Médula Espinal/genética , Traumatismos de la Médula Espinal/patología , Traumatismos de la Médula Espinal/fisiopatología , Superóxido Dismutasa/metabolismo , Vértebras Torácicas/cirugíaRESUMEN
Pompe disease is a severe form of muscular dystrophy due to glycogen accumulation in all tissues, especially striated muscle. Disease severity is directly related to the deficiency of acid alpha-glucosidase (GAA), which degrades glycogen in the lysosome. Respiratory dysfunction is a hallmark of the disease, muscle weakness has been viewed as the underlying cause, and the possibility of an associated neural contribution has not been evaluated previously. Therefore, we examined behavioral and neurophysiological aspects of breathing in 2 animal models of Pompe disease--the Gaa(-/-) mouse and a transgenic line (MTP) expressing GAA only in skeletal muscle, as well as a detailed analysis of the CNS in a Pompe disease patient. Glycogen content was elevated in the Gaa(-/-) mouse cervical spinal cord. Retrograde labeling of phrenic motoneurons showed significantly greater soma size in Gaa(-/-) mice vs. isogenic controls, and glycogen was observed in Gaa(-/-) phrenic motoneurons. Ventilation, assessed via plethysmography, was attenuated during quiet breathing and hypercapnic challenge in Gaa(-/-) mice (6 to >21 months of age) vs. controls. We confirmed that MTP mice had normal diaphragmatic contractile properties; however, MTP mice had ventilation similar to the Gaa(-/-) mice during quiet breathing. Neurophysiological recordings indicated that efferent phrenic nerve inspiratory burst amplitudes were substantially lower in Gaa(-/-) and MTP mice vs. controls. In human samples, we demonstrated similar pathology in the cervical spinal cord and greater accumulation of glycogen in spinal cord compared with brain. We conclude that neural output to the diaphragm is deficient in Gaa(-/-) mice, and therapies targeting muscle alone may be ineffective in Pompe disease.