Case report:Thirty-year progression of an EMPF1 encephalopathy due to defective mitochondrial and peroxisomal fission caused by a novel de novo heterozygous DNM1L variant

Mutations in DNM1L (DRP1), which encode a key player of mitochondrial and peroxisomal fission, have been reported in patients with the variable phenotypic spectrum, ranging from non-syndromic optic atrophy to lethal infantile encephalopathy. Here, we report a case of an adult female patient presenting with a complex neurological phenotype that associates axonal sensory neuropathy, spasticity, optic atrophy, dysarthria, dysphasia, dystonia, and ataxia, worsening with aging. Whole-exome sequencing revealed a heterozygous de novo variant in the GTPase domain of DNM1L [NM_001278464.1: c.176C>A p.(Thr59Asn)] making her the oldest patient suffering from encephalopathy due to defective mitochondrial and peroxisomal fission-1. In silico analysis suggested a protein destabilization effect of the variant Thr59Asn. Unexpectedly, Western blotting disclosed profound decrease of DNM1L expression, probably related to the degradation of DNM1L complexes. A detailed description of mitochondrial and peroxisomal anomalies in transmission electron and 3D fluorescence microscopy studies confirmed the exceptional phenotype of this patient

Simultaneous MFN2 and GDAP1 mutations cause major mitochondrial defects in a patient with CMT

Mutations in the MFN2 gene are associated with Charcot-Marie-Tooth disease type 2A (CMT2A), a dominant axonal CMT, whereas mutations in GDAP1 are associated with recessive demyelinating CMT (CMT4A), recessive axonal CMT (AR-CMT2), and dominant axonal CMT (CMT2K). Both proteins are involved in energy metabolism and dynamics of the mitochondrial network. We have previously reported that, in fibroblasts from patients with CMT, MFN2 mutations resulted in a mitochondrial energy coupling defect, whereas dominant mutation in GDAP1 resulted in defective complex I activity. In this study, we investigated mitochondrial bioenergetics from a severely affected patient with CMT harboring combined mutations in both GDAP1 and MFN2 genes

Mutations in the m-AAA proteases AFG3L2 and SPG7 are causing isolated dominant optic atrophy.

OBJECTIVE: To improve the genetic diagnosis of dominant optic atrophy (DOA), the most frequently inherited optic nerve disease, and infer genotype-phenotype correlations. METHODS: Exonic sequences of 22 genes were screened by new-generation sequencing in patients with DOA who were investigated for ophthalmology, neurology, and brain MRI. RESULTS: We identified 7 and 8 new heterozygous pathogenic variants in SPG7 and AFG3L2. Both genes encode for mitochondrial matricial AAA (m-AAA) proteases, initially involved in recessive hereditary spastic paraplegia type 7 (HSP7) and dominant spinocerebellar ataxia 28 (SCA28), respectively. Notably, variants in AFG3L2 that result in DOA are located in different domains to those reported in SCA28, which likely explains the lack of clinical overlap between these 2 phenotypic manifestations. In comparison, the SPG7 variants identified in DOA are interspersed among those responsible for HSP7 in which optic neuropathy has previously been reported. CONCLUSIONS: Our results position SPG7 and AFG3L2 as candidate genes to be screened in DOA and indicate that regulation of mitochondrial protein homeostasis and maturation by m-AAA proteases are crucial for the maintenance of optic nerve physiology

Overexpression of the Mitochondrial T3 Receptor p43 Induces a Shift in Skeletal Muscle Fiber Types

In previous studies, we have characterized a new hormonal pathway involving a mitochondrial T3 receptor (p43) acting as a mitochondrial transcription factor and consequently stimulating mitochondrial activity and mitochondrial biogenesis. We have established the involvement of this T3 pathway in the regulation of in vitro myoblast differentiation.We have generated mice overexpressing p43 under control of the human α-skeletal actin promoter. In agreement with the previous characterization of this promoter, northern-blot and western-blot experiments confirmed that after birth p43 was specifically overexpressed in skeletal muscle. As expected from in vitro studies, in 2-month old mice, p43 overexpression increased mitochondrial genes expression and mitochondrial biogenesis as attested by the increase of mitochondrial mass and mt-DNA copy number. In addition, transgenic mice had a body temperature 0.8°C higher than control ones and displayed lower plasma triiodothyronine levels. Skeletal muscles of transgenic mice were redder than wild-type animals suggesting an increased oxidative metabolism. In line with this observation, in gastrocnemius, we recorded a strong increase in cytochrome oxidase activity and in mitochondrial respiration. Moreover, we observed that p43 drives the formation of oxidative fibers: in soleus muscle, where MyHC IIa fibers were partly replaced by type I fibers; in gastrocnemius muscle, we found an increase in MyHC IIa and IIx expression associated with a reduction in the number of glycolytic fibers type IIb. In addition, we found that PGC-1α and PPARδ, two major regulators of muscle phenotype were up regulated in p43 transgenic mice suggesting that these proteins could be downstream targets of mitochondrial activity. These data indicate that the direct mitochondrial T3 pathway is deeply involved in the acquisition of contractile and metabolic features of muscle fibers in particular by regulating PGC-1α and PPARδ

Dominant ACO2 mutations are a frequent cause of isolated optic atrophy.

Biallelic mutations in ACO2, encoding the mitochondrial aconitase 2, have been identified in individuals with neurodegenerative syndromes, including infantile cerebellar retinal degeneration and recessive optic neuropathies (locus OPA9). By screening European cohorts of individuals with genetically unsolved inherited optic neuropathies, we identified 61 cases harbouring variants in ACO2, among whom 50 carried dominant mutations, emphasizing for the first time the important contribution of ACO2 monoallelic pathogenic variants to dominant optic atrophy. Analysis of the ophthalmological and clinical data revealed that recessive cases are affected more severely than dominant cases, while not significantly earlier. In addition, 27% of the recessive cases and 11% of the dominant cases manifested with extraocular features in addition to optic atrophy. In silico analyses of ACO2 variants predicted their deleterious impacts on ACO2 biophysical properties. Skin derived fibroblasts from patients harbouring dominant and recessive ACO2 mutations revealed a reduction of ACO2 abundance and enzymatic activity, and the impairment of the mitochondrial respiration using citrate and pyruvate as substrates, while the addition of other Krebs cycle intermediates restored a normal respiration, suggesting a possible short-cut adaptation of the tricarboxylic citric acid cycle. Analysis of the mitochondrial genome abundance disclosed a significant reduction of the mitochondrial DNA amount in all ACO2 fibroblasts. Overall, our data position ACO2 as the third most frequently mutated gene in autosomal inherited optic neuropathies, after OPA1 and WFS1, and emphasize the crucial involvement of the first steps of the Krebs cycle in the maintenance and survival of retinal ganglion cells

Etude des relations entre fonctionnement mitochondrial et types contractiles des fibres musculaires (approche sur mitochondries isolées et fibres perméabilisées)

RENNES1-BU Sciences Philo (352382102) / SudocSudocFranceF

Iron Deficiency without Anemia Decreases Physical Endurance and Mitochondrial Complex I Activity of Oxidative Skeletal Muscle in the Mouse

Iron deficiency (ID), with or without anemia, is responsible for physical fatigue. This effect may be linked to an alteration of mitochondrial metabolism. Our aim was to assess the impact of ID on skeletal striated muscle mitochondrial metabolism. Iron-deficient non-anemic mice, obtained using a bloodletting followed by a low-iron diet for three weeks, were compared to control mice. Endurance was assessed using a one-hour submaximal exercise on a Rotarod device and activities of mitochondrial complexes I and IV were measured by spectrophotometry on two types of skeletal striated muscles, the soleus and the quadriceps. As expected, ID mice displayed hematologic markers of ID and reduced iron stores, although none of them were anemic. In ID mice, endurance was significantly reduced and activity of the respiratory chain complex I, normalized to citrate synthase activity, was significantly reduced in the soleus muscle but not in the quadriceps. Complex IV activities were not significantly different, neither in the soleus nor in the quadriceps. We conclude that ID without anemia is responsible for impaired mitochondrial complex I activity in skeletal muscles with predominant oxidative metabolism. These results bring pathophysiological support to explain the improved physical activity observed when correcting ID in human. Further studies are needed to explore the mechanisms underlying this decrease in complex I activity and to assess the role of iron therapy on muscle mitochondrial metabolism

GNSS Timing Receiver Performance in Urban Canyons

Time synchronization is critical for the operation of radio base stations (RBS) in telecommunication companies. Global navigation satellite system (GNSS) is an existing technology to provide precise timing information to distributed RBSs. GNSS timing receiver is used for providing higher timing accuracy than normal GNSS receiver in this synchronization domain.In this thesis, an experiment method for GNSS timing receiver performance in urban canyon has been designed and implemented to evaluate information and the quality of the one pulse per second (1PPS) signal generated by two different GNSS timing receivers. Multi-path signals and the gathered satellite geometry caused by poor sky visibility is identified as the main influential factors to the performance of the GNSS timing receivers. A mathematical model has been built for estimating the multi-path effect. GNSS planning tools are used to simulate the number of line-of-sight (LOS) satellites and Dilution of Precision (DOP) value.Sentinel is a 1PPS signal analyzing equipment from Calnex. Sentinel has an embedded rubidium clock, GNSS antenna, and receiver, and it can produce 1PPS signals to be used as a reference. In this report, we installed our GNSS antenna of Sentinel on the roof and test GNSS antenna in two specified positions representing urban canyon and rooftop. Recorded NMEA messages from GNSS receiver can help us to study the number of visible satellites, PDOP value and multi-path signals in realistic situations.The results show how the noise and time phase of 1PPS signals will be influenced in urban canyons. Since, the geometry of used satellites is similar to the rooftop situation, the multi-path effect of signals is identified as the main reason of this difference.This information is useful when telecommunication companies want to install their radio base station in urban canyons. It will help Ericsson to understand how their GNSS timing receiver is working and how the urban canyon will influence its performance.Tidssynkronisering är kritisk för driften av radiobasstationer (RBS) i telekommunikationsföretag. Global Navigation Satellite System (GNSS) är en befintlig teknik för att ge exakt tidsinformation till distribuerade basstationer. GNSS-baserade tidsmottagare används för att ge högre timing-noggrannhet än vanlig GNSS mottagare i denna synkroniseringsdomän. I denna avhandling har en experimentmetod för GNSS-timingmottagarnas prestanda i urban canyon utformats och implementerats för att utvärdera den genererade informationen och kvaliteten på en puls per sekund-signal (1PPS). Flervägssignaler och den samlade satellitgeometrin som orsakas av dålig himmelsynlighet identifieras som de mest inflytelserika faktorerna för GNSS-tidsmottagarnas prestanda. En matematisk modell har donstruerats för att estimera multi-path-effekten. GNSS-planeringsverktyg används för att simulera antalet LOS-satelliter och DOP-värde (Dilution of Precision). Sentinel är en 1PPS signalanalysutrustning från Calnex. Sentinel har en inbyggd rubidiumklocka, GNSS-antenn och mottagare, och den kan producera 1PPS-signaler som ska användas som referens. I den här rapporten installerade vi vår GNSS-antenn på Sentinel på taket och GNSS-testantennen i två angivna positioner som representerar urban canyon och tak. Inspelade NMEA-meddelanden från GNSS-mottagare kan hjälpa oss att studera antalet synliga satelliter, PDOP-värde och flervägssignaler i realistiska scenarier. Resultatet visar att ljud- och tidsfasen för 1PPS-signaler påverkas i urban canyons. Eftersom satellitgeometrin liknar den för antenner placerade på taket, så är identifieras flervägsutbredningen som huvudorsak för denna skillnad. Denna information är användbar när telekommunikationsföretag vill installera sina radiobasstationer i urban canyons. Det kommer att hjälpa Ericsson att förstå hur deras GNSS-timingmottagare arbetar och hur urban canyon påverkar dess prestanda