Mutations in EFHC1 cause juvenile myoclonic epilepsy

Juvenile myoclonic epilepsy (JME) is the most frequent cause of hereditary grand mal seizures. We previously mapped and narrowed a region associated with JME on chromosome 6p12-p11 (EJM1). Here, we describe a new gene in this region, EFHC1, which encodes a protein with an EF-hand motif. Mutation analyses identified five missense mutations in EFHC1 that cosegregated with epilepsy or EEG polyspike wave in affected members of six unrelated families with JME and did not occur in 382 control individuals. Overexpression of EFHC1 in mouse hippocampal primary culture neurons induced apoptosis that was significantly lowered by the mutations. Apoptosis was specifically suppressed by SNX-482, an antagonist of R-type voltage-dependent Ca(2+) channel (Ca(v)2.3). EFHC1 and Ca(v)2.3 immunomaterials overlapped in mouse brain, and EFHC1 coimmunoprecipitated with the Ca(v)2.3 C terminus. In patch-clamp analysis, EFHC1 specifically increased R-type Ca(2+) currents that were reversed by the mutations associated with JME.     

Conserved modes of recruitment of ATM, ATR and DNA-PKcs to sites of DNA damage

Ataxia-telangiectasia mutated (ATM), ataxia-telangiectasia and Rad3-related (ATR) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) are members of the phosphoinositide-3-kinase-related protein kinase (PIKK) family, and are rapidly activated in response to DNA damage. ATM and DNA-PKcs respond mainly to DNA double-strand breaks, whereas ATR is activated by single-stranded DNA and stalled DNA replication forks. In all cases, activation involves their recruitment to the sites of damage. Here we identify related, conserved carboxy-terminal motifs in human Nbs1, ATRIP and Ku80 proteins that are required for their interaction with ATM, ATR and DNA-PKcs, respectively. These motifs are essential not only for efficient recruitment of ATM, ATR and DNA-PKcs to sites of damage, but are also critical for ATM-, ATR- and DNA-PKcs-mediated signalling events that trigger cell cycle checkpoints and DNA repair. Our findings reveal that recruitment of these PIKKs to DNA lesions occurs by common mechanisms through an evolutionarily conserved motif, and provide direct evidence that PIKK recruitment is required for PIKK-dependent DNA-damage signalling.     

Mutant dynactin in motor neuron disease

Impaired axonal transport in motor neurons has been proposed as a mechanism for neuronal degeneration in motor neuron disease. Here we show linkage of a lower motor neuron disease to a region of 4 Mb at chromosome 2p13. Mutation analysis of a gene in this interval that encodes the largest subunit of the axonal transport protein dynactin showed a single base-pair change resulting in an amino-acid substitution that is predicted to distort the folding of dynactin's microtubule-binding domain. Binding assays show decreased binding of the mutant protein to microtubules. Our results show that dysfunction of dynactin-mediated transport can lead to human motor neuron disease.     

Mutations in a novel gene,NPHP3, cause adolescent nephronophthisis,tapeto-retinal degeneration and hepatic fibrosis

Nephronophthisis (NPHP), a group of autosomal recessive cystic kidney disorders, is the most common genetic cause of progressive renal failure in children and young adults. NPHP may be associated with Leber congenital amaurosis, tapeto-retinal degeneration, cerebellar ataxia, cone-shaped epiphyses, congenital oculomotor apraxia and hepatic fibrosis. Loci associated with an infantile type of NPHP on 9q22-q31 (NPHP2), juvenile types of NPHP on chromosomes 2q12-q13 (NPHP1) and 1p36 (NPHP4) and an adolescent type of NPHP on 3q21-q22 (NPHP3) have been mapped. NPHP1 and NPHP4 have been identified, and interaction of the respective encoded proteins nephrocystin and nephrocystin-4 has been shown. Here we report the identification of NPHP3, encoding a novel 1,330-amino acid protein that interacts with nephrocystin. We describe mutations in NPHP3 in families with isolated NPHP and in families with NPHP with associated hepatic fibrosis or tapeto-retinal degeneration. We show that the mouse ortholog Nphp3 is expressed in the node, kidney tubules, retina, respiratory epithelium, liver, biliary tract and neural tissues. In addition, we show that a homozygous missense mutation in Nphp3 is probably responsible for the polycystic kidney disease (pcy) mouse phenotype. Interventional studies in the pcy mouse have shown beneficial effects by modification of protein intake and administration of methylprednisolone, suggesting therapeutic strategies for treating individuals with NPHP3.     

Carbodiimide fixation for electron microscopy and immunoelectron cytochemistry

Zusammenfassung In mit wasserlöslichen Carbodiimiden unter bestimmten Bedingungen fixiertem Gewebe können Polypeptidhormone immunfluoreszenzoptischnachgewiesen werden. Unter optimalen Bedingungen fixiertes Material ist auch zur Herstellung elektronenmikroskopischer Präparate vorzüglich geeignet. Carbodiimide werden deshalb als Fixierungsmittel in elektronenoptischen, immunzytochemischen Studien vorgeschlagen.

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We wish to thank Dr.S. Bloom for preparing the anti-percine secretin used in this study. This work was supported by the Wellcome Trust and the Cancer Research Campain.     

Primary LAMP-2 deficiency causes X-linked vacuolar cardiomyopathy and myopathy (Danon disease)

"Lysosomal glycogen storage disease with normal acid maltase" which was originally described by Danon et al., is characterized clinically by cardiomyopathy, myopathy and variable mental retardation. The pathological hallmark of the disease is intracytoplasmic vacuoles containing autophagic material and glycogen in skeletal and cardiac muscle cells. Sarcolemmal proteins and basal lamina are associated with the vacuolar membranes. Here we report ten unrelated patients, including one of the patients from the original case report, who have primary deficiencies of LAMP-2, a principal lysosomal membrane protein. From these results and the finding that LAMP-2-deficient mice manifest a similar vacuolar cardioskeletal myopathy, we conclude that primary LAMP-2 deficiency is the cause of Danon disease. To our knowledge this is the first example of human cardiopathy-myopathy that is caused by mutations in a lysosomal structural protein rather than an enzymatic protein.     

Growing knowledge: Epistemic objects in agricultural extension work

We introduce a novel form of experimental knowledge that is the result of institutionally structured communication practices between farmers and university- and local community-based agronomists (agricultural extension specialists). This form of knowledge is exemplified in these communities’ uses of the concept of grower standard. Grower standard is a widely used but seldom discussed benchmark concept underpinning protocols used within agricultural experiments. It is not a one-size-fits-all standard but the product of local and active interactions between farmers and agricultural extension specialists. Grower standard is in some ways similar to more familiar epistemic objects discussed in philosophy of experiment, such as controls or background conditions. However, we argue that grower standard is epistemically novel, due to how knowledge arising from it is coproduced by farmers and agricultural extension specialists. Further, in the United States, this knowledge coproduction is institutionally structured by federal legislature dating back to the 19th century. We use our analysis of grower standard to focus a discussion of the positionality of the coproducers as well as the epistemic products of this form of knowledge coproduction, and we explore the role extension work plays in shaping agricultural science more broadly.     

Colligation in modelling practices: From Whewell’s tides to the San Francisco Bay Model

“Colligation”, a term first introduced in philosophy of science by William Whewell (1840), today sparks a renewed interest beyond Whewell scholarship. In this paper, we argue that adopting the notion of colligation in current debates in philosophy of science can contribute to our understanding of scientific models. Specifically, studying colligation allows us to have a better grasp of how integrating diverse model components (empirical data, theory, useful idealization, visual and other representational resources) in a creative way may produce novel generalizations about the phenomenon investigated. Our argument is built both on the theoretical appraisal of Whewell’s philosophy of science and the historical rehabilitation of his scientific work on tides. Adopting a philosophy of science in practice perspective, we show how colligation emerged from Whewell’s empirical work on tides. The production of idealized maps (“cotidal maps”) illustrates the unifying and creative power of the activity of colligating in scientific practice. We show the importance of colligation in modelling practices more generally by looking at its epistemic role in the construction of the San Francisco Bay Model.