Deletion of Ripk3 Prevents Motor Neuron Death In Vitro but not In Vivo.

Increasing evidence suggests that necroptosis, a form of programmed cell death (PCD), contributes to neurodegeneration in several disorders, including ALS. Supporting this view, investigations in both in vitro and in vivo models of ALS have implicated key molecular determinants of necroptosis in the death of spinal motor neurons (MNs). Consistent with a pathogenic role of necroptosis in ALS, we showed increased mRNA levels for the three main necroptosis effectors Ripk1, Ripk3, and Mlkl in the spinal cord of mutant superoxide dismutase-1 (SOD1G93A) transgenic mice (Tg), an established model of ALS. In addition, protein levels of receptor-interacting protein kinase 1 (RIPK1; but not of RIPK3, MLKL or activated MLKL) were elevated in spinal cord extracts from these Tg SOD1G93A mice. In postmortem motor cortex samples from sporadic and familial ALS patients, no change in protein levels of RIPK1 were detected. Silencing of Ripk3 in cultured MNs protected them from toxicity associated with SOD1G93A astrocytes. However, constitutive deletion of Ripk3 in Tg SOD1G93A mice failed to provide behavioral or neuropathological improvement, demonstrating no similar benefit of Ripk3 silencing in vivo. Lastly, we detected no genotype-specific myelin decompaction, proposed to be a proxy of necroptosis in ALS, in either Tg SOD1G93A or Optineurin knock-out mice, another ALS mouse model. These findings argue against a role for RIPK3 in Tg SOD1G93A-induced neurodegeneration and call for further preclinical investigations to determine if necroptosis plays a critical role in the pathogenesis of ALS.

Hatha-yoga as a psychological adjuvant for women undergoing IVF: a pilot study.

OBJECTIVES: To evaluate the influence of Hatha-yoga (HY) practice on distress of women before starting their first in vitro fertilization (IVF) cycle. STUDY DESIGN: We offered 143 consecutive women with couple infertility the opportunity to attend a free HY course lasting 3 months as a psychological support before starting their first IVF cycle. All women were asked to complete the State-Trait Anxiety Inventory-Y1 (STAY-Y1), Edinburgh Depression Scale (EDS) and General Health Questionnaire-12 (GHQ-12) at baseline (T1) and after 3 months (T2), to evaluate symptoms of anxiety, depression and distress, respectively. RESULTS: Of the 143 women, 120 completed all three questionnaires. Of these, 45 attended the HY course and 75 did not. At T1, EDS and GHQ-12 scores were significantly higher in the HY group than in the non-HY group. There were no group differences in STAI-Y1 scores. At T2 there were no group differences. When, in each group, the score of each questionnaire at T1 was compared to the score at T2, a significant T1 to T2 reduction was observed in the HY group (p<0.0001 for STAY-Y1 and GHQ-12, p<0.001 for EDS). CONCLUSIONS: Our data suggest that women who are more distressed are more likely to accept psychological support before starting an IVF cycle and that in these women HY practice is associated with distress reduction.

A cell system for phenotypic screening of modifiers of SMN2 gene expression and function.

Spinal muscular atrophy (SMA) is an inherited neurodegenerative disease caused by homozygous inactivation of the SMN1 gene and reduced levels of the survival motor neuron (SMN) protein. Since higher copy numbers of the nearly identical SMN2 gene reduce disease severity, to date most efforts to develop a therapy for SMA have focused on enhancing SMN expression. Identification of alternative therapeutic approaches has partly been hindered by limited knowledge of potential targets and the lack of cell-based screening assays that serve as readouts of SMN function. Here, we established a cell system in which proliferation of cultured mouse fibroblasts is dependent on functional SMN produced from the SMN2 gene. To do so, we introduced the entire human SMN2 gene into NIH3T3 cell lines in which regulated knockdown of endogenous mouse Smn severely decreases cell proliferation. We found that low SMN2 copy number has modest effects on the cell proliferation phenotype induced by Smn depletion, while high SMN2 copy number is strongly protective. Additionally, cell proliferation correlates with the level of SMN activity in small nuclear ribonucleoprotein assembly. Following miniaturization into a high-throughput format, our cell-based phenotypic assay accurately measures the beneficial effects of both pharmacological and genetic treatments leading to SMN upregulation. This cell model provides a novel platform for phenotypic screening of modifiers of SMN2 gene expression and function that act through multiple mechanisms, and a powerful new tool for studies of SMN biology and SMA therapeutic development.