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Glycogen storage disease type V (GSDV, McArdle disease) is a rare genetic myopathy caused by deficiency of the muscle isoform of glycogen phosphorylase (PYGM). This results in a block in the use of muscle glycogen as an energetic substrate, with subsequent exercise intolerance. The pathobiology of GSDV is still not fully understood, especially with regard to some features such as persistent muscle damage (i.e., even without prior exercise). We aimed at identifying potential muscle protein biomarkers of GSDV by analyzing the muscle proteome and the molecular networks associated with muscle dysfunction in these patients. Muscle biopsies from eight patients and eight healthy controls showing none of the features of McArdle disease, such as frequent contractures and persistent muscle damage, were studied by quantitative protein expression using isobaric tags for relative and absolute quantitation (iTRAQ) followed by artificial neuronal networks (ANNs) and topology analysis. Protein candidate validation was performed by Western blot. Several proteins predominantly involved in the process of muscle contraction and/or calcium homeostasis, such as myosin, sarcoplasmic/endoplasmic reticulum calcium ATPase 1, tropomyosin alpha-1 chain, troponin isoforms, and alpha-actinin-3, showed significantly lower expression levels in the muscle of GSDV patients. These proteins could be potential biomarkers of the persistent muscle damage in the absence of prior exertion reported in GSDV patients. Further studies are needed to elucidate the molecular mechanisms by which PYGM controls the expression of these proteins.

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[Background]: McArdle disease is caused by myophosphorylase deficiency and results in complete inability for muscle glycogen breakdown. A hallmark of this condition is muscle oxidation impairment (e.g., low peak oxygen uptake (VO2peak)), a phenomenon traditionally attributed to reduced glycolytic flux and Krebs cycle anaplerosis. Here we hypothesized an additional role for muscle mitochondrial network alterations associated with massive intracellular glycogen accumulation. [Methods]: We analyzed in depth mitochondrial characteristics-content, biogenesis, ultrastructure-and network integrity in skeletal-muscle from McArdle/control mice and two patients. We also determined VO2peak in patients (both sexes, N = 145) and healthy controls (N = 133). [Results]: Besides corroborating very poor VO2peak values in patients and impairment in muscle glycolytic flux, we found that, in McArdle muscle: (a) damaged fibers are likely those with a higher mitochondrial and glycogen content, which show major disruption of the three main cytoskeleton components-actin microfilaments, microtubules and intermediate filaments-thereby contributing to mitochondrial network disruption in skeletal muscle fibers; (b) there was an altered subcellular localization of mitochondrial fission/fusion proteins and of the sarcoplasmic reticulum protein calsequestrin-with subsequent alteration in mitochondrial dynamics/function; impairment in mitochondrial content/biogenesis; and (c) several OXPHOS-related complex proteins/activities were also affected. [Conclusions]: In McArdle disease, severe muscle oxidative capacity impairment could also be explained by a disruption of the mitochondrial network, at least in those fibers with a higher capacity for glycogen accumulation. Our findings might pave the way for future research addressing the potential involvement of mitochondrial network alterations in the pathophysiology of other glycogenoses.

Background: McArdle disease is caused by myophosphorylase deficiency and results in complete inability for muscle glycogen breakdown. A hallmark of this condition is muscle oxidation impairment (e.g., low peak oxygen uptake (VO2peak)), a phenomenon traditionally attributed to reduced glycolytic flux and Krebs cycle anaplerosis. Here we hypothesized an additional role for muscle mitochondrial network alterations associated with massive intracellular glycogen accumulation. Methods: We analyzed in depth mitochondrial characteristics-content, biogenesis, ultrastructure-and network integrity in skeletal-muscle from McArdle/control mice and two patients. We also determined VO2peak in patients (both sexes, N = 145) and healthy controls (N = 133). Results: Besides corroborating very poor VO2peak values in patients and impairment in muscle glycolytic flux, we found that, in McArdle muscle: (a) damaged fibers are likely those with a higher mitochondrial and glycogen content, which show major disruption of the three main cytoskeleton components-actin microfilaments, microtubules and intermediate filaments-thereby contributing to mitochondrial network disruption in skeletal muscle fibers; (b) there was an altered subcellular localization of mitochondrial fission/fusion proteins and of the sarcoplasmic reticulum protein calsequestrin-with subsequent alteration in mitochondrial dynamics/function; impairment in mitochondrial content/biogenesis; and (c) several OXPHOS-related complex proteins/activities were also affected. Conclusions: In McArdle disease, severe muscle oxidative capacity impairment could also be explained by a disruption of the mitochondrial network, at least in those fibers with a higher capacity for glycogen accumulation. Our findings might pave the way for future research addressing the potential involvement of mitochondrial network alterations in the pathophysiology of other glycogenoses.

McArdle disease is an autosomal recessive disorder of muscle glycogen metabolism caused by pathogenic mutations in the PYGM gene, which encodes the skeletal muscle-specific isoform of glycogen phosphorylase. Clinical symptoms are mainly characterized by transient acute “crises” of early fatigue, myalgia and contractures, which can be accompanied by rhabdomyolysis. Owing to the difficulty of performing mechanistic studies in patients that often rely on invasive techniques, preclinical models have been used for decades, thereby contributing to gain insight into the patho-physiology and pathobiology of human diseases. In the present work, we describe the existing in vitro and in vivo preclinical models for McArdle disease and review the insights these models have provided. In addition, despite presenting some differences with the typical patient’s phenotype, these models allow for a deep study of the different features of the disease while representing a necessary preclinical step to assess the efficacy and safety of possible treatments before they are tested in patients.

Regulated deficit irrigation (RDI) involves water stress management in different phenological periods throughout the season. Research in olive trees (oil production) suggested RDI during pit hardening based in pre-dawn and midday stem water potential (SWP) thresholds. However, the previous thresholds may not be extrapolated to table olive because fruit size, a very important feature in the table olive yield quality, is very sensitive to water stress. RDI in table olive deserve further research to determine the optimal water potential thresholds and the duration of the RDI periods for the specificity of the crop (low crop load to promote high fruit size). The aim of this work was to study different RDI schedules during pit hardening, considering different levels and durations of water stress. The experiment was performed in the 2015, 2016 and 2017 seasons, in a commercial mature table olive orchard (cv. Manzanilla) in Dos Hermanas (Seville, Spain). Control treatments were based on midday SWP measurement in order to optimize the water status with values around −1.4 MPa. Two RDI treatments were applied during pit hardening, dated (according to the changes in longitudinal fruit growth) from mid-June to the last week of August) to maintain water potential values around −2 MPa (RDI-1) and −3.5 MPa (RDI-3). Another RDI treatment (RDI-2) received irrigation to maintain values around −3.5 MPa but the recovery was performed at early July in order to obtain different durations of water stress. Irrigation strategies were evaluated with water relations measurements (soil moisture, gas exchange), fruit and shoot growth and quality and quantity yield indicators. Yield was not significantly affected in any of the RDI treatments with an ANOVA analysis. However, fruit drop estimated as the percentage of fruit lost only in the period of water deficit was related with water stress parameters (SWP and stress integral, IS). In addition, the relationship between fruits size and these latt

El olivar es un cultivo milenario en la cuenca Mediterránea, asociado habitualmente a condiciones de secano. Esto es debido a su gran resistencia al estrés hídrico, que le permite soportar condiciones extremas de falta de agua, siendo capaz de continuar produciendo. No obstante, el riego e este cultivo supone una mejora en la producción, mayor cuanto más densos son los olivares. Esta mejora productiva ha dado lugar a un incremento paulatino de la superficie en estas condiciones

Deficit irrigation scheduling is becoming increasingly important under commercial conditions. Water status measurement is a useful tool in these conditions. However, the information about water stress levels for olive trees is scarce. The aim of this experiment was to evaluate the effect on yield of a moderate controlled water stress level at the end of the irrigation season. The experiment was conducted in the experimental farm of La Hampa (Coria del Río, Seville, Spain) during three years. A completely randomized block design was performed using three different irrigation treatments. Deficit irrigation was applied several (4 or 2) weeks before harvest. Irrigation was controlled using the midday stem water potential, with a threshold value of −2 MPa and compared with a full irrigated treatment. This water stress did not reduced gas exchange during the deficit period. The effect on yield was not significant in any of the three seasons. In the high-fruit load season, fruit volume was slightly affected (around 10%), but this was not significant at harvest. Results suggest an early affection of fruit growth with water stress, but with a slow rate of decrease. Moderate water stress could be useful for the management of deficit irrigation in table olive trees.