Mechanistic models for muscle diseases and disorders originating in the sarcoplasmic reticulum

AbstractThis review focuses on muscle disorders and diseases caused by defects in the Ca2+ release channels of the sarcoplasmic reticulum, the ryanodine receptors, and in the luminal, low affinity, high capacity Ca2+-binding proteins, calsequestrins. It provides a time line over the past half century of the highlights of research on malignant hyperthermia (MH), central core disease (CCD) and catecholaminergic polymorphic ventricular tachycardia (CPVT), that resulted in the identification of the ryanodine receptor (RYR), calsequestrin (CASQ) and dihydropyridine receptor (CACNA1S) genes as sites of disease-causing mutations. This is followed by a description of approaches to functional analysis of the effects of disease-causing mutations on protein function, focusing on studies of how mutations affect spontaneous (store overload-induced) Ca2+-release from the sarcoplasmic reticulum, the underlying cause of MH and CPVT. Subsequent sections describe results obtained by analysis of knockin mouse lines carrying MH- and CCD-causing mutations, including a Casq1 knockout. The review concludes with the presentation of two mechanistic models. The first shows how dysregulation of Ca2+ homeostasis can lead to muscle diseases involving both RyR and Casq proteins. The second describes a theory of central core formation wherein non-uniformity of Ca2+ release, resulting in non-uniformity of muscle contraction, is presented as an intrinsic property of the specific tertiary structure of mutant heterotetrameric ryanodine receptors and as the underlying cause of core formation in skeletal muscle. This article is part of a Special Issue entitled: 11th European Symposium on Calcium

Rabbit cardiac and slow-twitch muscle express the same phospholamban gene

AbstractThe nucleotide sequences of cDNAs encoding phospholamban were found to be virtually identical when the cDNA clones were isolated from rabbit slow-twitch (soleus) and rabbit cardiac muscle libraries. These findings demonstrate that both types of muscle express the same phospholamban gene. The deduced amino acid sequences of rabbit and dog phospholamban were identical except for a change from Asp (dog) to Glu (rabbit) at position 2. The nucleotide sequences of the 5′- and the very long 3′-untranslated regions of rabbit and dog phospholamban cDNAs also exhibited a high percentage of identity

Deletion of NH2− and COOH-terminal sequences destroys function of the Ca2+ ATPase of rabbit fast-twitch skeletal muscle sarcoplasmic reticulum

AbstractDeletion mutants of the Ca2+ ATPase of rabbit fast-twitch skeletal muscle sarcoplasmic reticulum (SERCA1a) were constructed and expressed in COS-1 cells. The mutants were expressed at levels 7- to 15-fold lower than the wild-type and were inactive. In vitro transcription-translation-insertion experiments showed that deletion of transmembrane sequences M1 and M2, but not of M8, M9, M10 or the NH2−terminal 30 amino acids inhibited the stable insertion of the enzyme into the membrane. Thus there was no correlation between loss of function and membrane insertion. A signal sequence for membrane insertion may exist in M1 and M2

Interleukin (IL)–12 and IL-23 Are Key Cytokines for Immunity against Salmonella in Humans

Patients with inherited deficiency of the interleukin (IL)–12/IL-23–interferon (IFN)–g axis show increased susceptibility to invasive disease caused by the intramacrophage pathogens salmonellae and mycobacteria. We analyzed data on 154 patients with such deficiency. Significantly more patients with IL-12/IL-23–component deficiency had a history of salmonella disease than did those with IFN-g–component deficiency. Salmonella disease was typically severe, extraintestinal, and caused by nontyphoidal serovars. These findings strongly suggest that IL-12/IL-23 is a key cytokine for immunity against salmonella in humans and that IL-12/IL-23 mediates this protective effect partly through IFN-g–independent pathways. Investigation of the IL-12/IL-23–IFN-g axis should be considered in patients with invasive salmonella disease

A Gravitational Theory of the Quantum

The synthesis of quantum and gravitational physics is sought through a finite, realistic, locally causal theory where gravity plays a vital role not only during decoherent measurement but also during non-decoherent unitary evolution. Invariant set theory is built on geometric properties of a compact fractal-like subset $I_U$ of cosmological state space on which the universe is assumed to evolve and from which the laws of physics are assumed to derive. Consistent with the primacy of $I_U$, a non-Euclidean (and hence non-classical) state-space metric $g_p$ is defined, related to the $p$-adic metric of number theory where $p$ is a large but finite Pythagorean prime. Uncertain states on $I_U$ are described using complex Hilbert states, but only if their squared amplitudes are rational and corresponding complex phase angles are rational multiples of $2 \pi$. Such Hilbert states are necessarily $g_p$-distant from states with either irrational squared amplitudes or irrational phase angles. The gappy fractal nature of $I_U$ accounts for quantum complementarity and is characterised numerically by a generic number-theoretic incommensurateness between rational angles and rational cosines of angles. The Bell inequality, whose violation would be inconsistent with local realism, is shown to be $g_p$-distant from all forms of the inequality that are violated in any finite-precision experiment. The delayed-choice paradox is resolved through the computational irreducibility of $I_U$. The Schr\"odinger and Dirac equations describe evolution on $I_U$ in the singular limit at $p=\infty$. By contrast, an extension of the Einstein field equations on $I_U$ is proposed which reduces smoothly to general relativity as $p \rightarrow \infty$. Novel proposals for the dark universe and the elimination of classical space-time singularities are given and experimental implications outlined

Ca2+ signaling in HEK-293 and skeletal muscle cells expressing recombinant ryanodine receptors harboring malignant hyperthermia and central core disease mutations.

Malignant hyperthermia (MH) and central core disease (CCD) are caused by mutations in the RYR1 gene encoding the skeletal muscle isoform of the ryanodine receptor (RyR1), a homotetrameric Ca(2+) release channel. Rabbit RyR1 mutant cDNAs carrying mutations corresponding to those in human RyR1 that cause MH and CCD were expressed in HEK-293 cells, which do not have endogenous RyR, and in primary cultures of rat skeletal muscle, which express rat RyR1. Analysis of intracellular Ca(2+) pools was performed using aequorin probes targeted to the lumen of the endo/sarcoplasmic reticulum (ER/SR), to the mitochondrial matrix, or to the cytosol. Mutations associated with MH caused alterations in intracellular Ca(2+) homeostasis different from those associated with CCD. Measurements of luminal ER/SR Ca(2+) revealed that the mutations generated leaky channels in all cases, but the leak was particularly pronounced in CCD mutants. Cytosolic and mitochondrial Ca(2+) transients induced by caffeine stimulation were drastically augmented in the MH mutant, slightly reduced in one CCD mutant (Y523S) and completely abolished in another (I4898T). The results suggest that local Ca(2+) derangements of different degrees account for the specific cellular phenotypes of the two disorders