ABSTRACT
Mutations in GLE1, RNA export mediator (GLE1) gene have previously been shown to cause motor neuron diseases such as lethal congenital contracture syndrome 1 (LCCS1) and lethal arthrogryposis with anterior horn cell disease (LAAHD), including arthrogryposis, fetal akinesis and motor neuron loss as common clinical features. The homozygous FinMajor mutation p.T144_E145insPFQ has been described as one of the causes for LCCS1 whereas LAAHD is caused by a heterocompound FinMajor mutation together with p.R569H, p.V617M or p.I684T missense mutation. None of these heterocompound missense mutations have previously been reported as homozygous states. Here we present the clinical features of 2 siblings with a homozygous p.I684T mutation in GLE1. The patients suffered from similar, but milder symptoms than in LCCS1 and LAAHD, surviving up to 6 months before they died due to a progressive disease course including respiratory failure. Arthrogryposis, lack of spontaneous movements, and epilepsy were notable in both cases and lack of anterior horn cells was identified in autopsy samples. Our studies on patient-derived fibroblasts show that the homozygous p.I684T impairs the nuclear localization of GLE1 further confirming the pathogenic role of this mutation.
Subject(s)
Arthrogryposis/genetics , Genetic Predisposition to Disease/genetics , Motor Neuron Disease/genetics , Mutation, Missense , Nucleocytoplasmic Transport Proteins/genetics , Base Sequence , Consanguinity , Fatal Outcome , Female , Homozygote , Humans , Infant , Male , Pedigree , SiblingsABSTRACT
Blindness due to retinal degeneration affects millions of people worldwide, but many disease-causing mutations remain unknown. PNPLA6 encodes the patatin-like phospholipase domain containing protein 6, also known as neuropathy target esterase (NTE), which is the target of toxic organophosphates that induce human paralysis due to severe axonopathy of large neurons. Mutations in PNPLA6 also cause human spastic paraplegia characterized by motor neuron degeneration. Here we identify PNPLA6 mutations in childhood blindness in seven families with retinal degeneration, including Leber congenital amaurosis and Oliver McFarlane syndrome. PNPLA6 localizes mostly at the inner segment plasma membrane in photoreceptors and mutations in Drosophila PNPLA6 lead to photoreceptor cell death. We also report that lysophosphatidylcholine and lysophosphatidic acid levels are elevated in mutant Drosophila. These findings show a role for PNPLA6 in photoreceptor survival and identify phospholipid metabolism as a potential therapeutic target for some forms of blindness.
Subject(s)
Blindness/genetics , Mutation , Phospholipases/genetics , Phospholipases/physiology , Amino Acid Sequence , Animals , Child , Child, Preschool , Drosophila , Female , Humans , Male , Mice , Mice, Inbred C57BL , Microscopy, Fluorescence , Molecular Sequence Data , Pedigree , Phenotype , Phospholipids/chemistry , Retina/pathology , Retinal Degeneration/genetics , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Spectrometry, Mass, Electrospray IonizationABSTRACT
Whole exome sequencing (WES) has greatly facilitated the identification of causal mutations for diverse human genetic disorders. We applied WES as a molecular diagnostic tool to identify disease-causing genes in consanguineous families in Qatar. Seventeen consanguineous families with diverse disorders were recruited. Initial mutation screening of known genes related to the clinical diagnoses did not reveal the causative mutations. Using WES approach, we identified the definitive disease-causing mutations in four families: (i) a novel nonsense homozygous (c.1034C>G) in PHKG2 causing glycogen storage disease type 9C (GSD9C) in a male with initial diagnosis of GSD3; (ii) a novel homozygous 1-bp deletion (c.915del) in NSUN2 in a male proband with Noonan-like syndrome; (iii) a homozygous SNV (c.1598C>G) in exon 11 of IDUA causing Hurler syndrome in a female proband with unknown clinical diagnosis; (iv) a de novo known splicing mutation (c.1645+1G>A) in PHEX in a female proband with initial diagnosis of autosomal recessive hypophosphatemic rickets. Applying WES as a diagnostic tool led to the unambiguous identification of disease-causing mutations in phenotypically complex disorders or correction of the initial clinical diagnosis in Ë25% of our cases.
Subject(s)
Consanguinity , Disease/genetics , Exome/genetics , Genetic Predisposition to Disease , Sequence Analysis, DNA , Family , Female , Humans , Male , Parents , Pedigree , QatarABSTRACT
Follicular fluid provides the microenvironment within which somatic cells proliferate and differentiate, and the oocyte matures. It contains a number of soluble factors implicated in various stages of follicular development, most of them being functionally unknown. The presence of several high-abundance proteins, mainly originating from the blood circulation, is a major challenge of follicular fluid proteomic analysis, as these proteins can mask or decrease the visualization of follicle-specific proteins. In this study, we evaluated the efficiency of two immunodepletion columns (ProteomeLab™ IgY-HSA and MARS-6) on follicular fluids of human, porcine and canine prior to 2D-PAGE. Our results showed that both columns were suitable to remove some of the high-abundance proteins present in human and canine follicular fluid. In conclusion, we demonstrated that the immunodepletion strategy enables the detection of new protein spots, increases resolution and highly improves the intensity of low-abundance proteins by 2D-PAGE.
Subject(s)
Dogs , Electrophoresis, Gel, Two-Dimensional/methods , Follicular Fluid/chemistry , Swine , Animals , Chromatography, Affinity , Female , Humans , Immunoglobulin G , Immunoglobulins , Proteins/analysis , Proteins/isolation & purification , Proteomics/methodsABSTRACT
The major challenge of follicular fluid proteomic analysis is the presence of high-abundance proteins that originate from plasma. These proteins can prevent the detection of lower abundant ones, produced locally by follicle cells and that may have important roles in follicular activity. In this study, the novel technology called hexapeptide ligand library was evaluated to enrich the low-abundance proteins in follicular fluid of human (HFF), porcine (PFF) and equine (EFF) prior 2D-PAGE. Our results showed that the new strategy enabled detection of many new protein spots, increased resolution and highly improved the intensity of low-abundance proteins by 2D-PAGE.
Subject(s)
Electrophoresis, Gel, Two-Dimensional/veterinary , Follicular Fluid/chemistry , Horses , Peptide Library , Proteins/analysis , Swine , Animals , Female , Humans , Proteomics/methodsABSTRACT
Follicular fluid is a complex extracellular fluid, semi-viscous and yellow in colour, which accumulates in the antrum of ovarian follicles during their growing phase. Follicular fluid provides the microenvironment within which the cumulus-oocyte complex matures and granulosa cells differentiate. Scientists agree that follicular fluid derives mainly from plasma via the vascular compartment in the follicle wall. However, it also contains factors produced locally by the follicle cells, the production of which varies during different reproductive states. The aim of this paper is to review current knowledge on the formation, composition and roles of follicular fluid.
