Dysfunction in endoplasmic reticulum-mitochondria crosstalk underlies SIGMAR1 loss of function mediated motor neuron degeneration.

Mutations in Sigma 1 receptor (SIGMAR1) have been previously identified in patients with amyotrophic lateral sclerosis and disruption of Sigmar1 in mouse leads to locomotor deficits. However, cellular mechanisms underlying motor phenotypes in human and mouse with disturbed SIGMAR1 function have not been described so far. Here we used a combination of in vivo and in vitro approaches to investigate the role of SIGMAR1 in motor neuron biology. Characterization of Sigmar1(-/-) mice revealed that affected animals display locomotor deficits associated with muscle weakness, axonal degeneration and motor neuron loss. Using primary motor neuron cultures, we observed that pharmacological or genetic inactivation of SIGMAR1 led to motor neuron axonal degeneration followed by cell death. Disruption of SIGMAR1 function in motor neurons disturbed endoplasmic reticulum-mitochondria contacts, affected intracellular calcium signalling and was accompanied by activation of endoplasmic reticulum stress and defects in mitochondrial dynamics and transport. These defects were not observed in cultured sensory neurons, highlighting the exacerbated sensitivity of motor neurons to SIGMAR1 function. Interestingly, the inhibition of mitochondrial fission was sufficient to induce mitochondria axonal transport defects as well as axonal degeneration similar to the changes observed after SIGMAR1 inactivation or loss. Intracellular calcium scavenging and endoplasmic reticulum stress inhibition were able to restore mitochondrial function and consequently prevent motor neuron degeneration. These results uncover the cellular mechanisms underlying motor neuron degeneration mediated by loss of SIGMAR1 function and provide therapeutically relevant insight into motor neuronal diseases.

Dysfunctional SEMA3E signaling underlies gonadotropin-releasing hormone neuron deficiency in Kallmann syndrome.

Individuals with an inherited deficiency in gonadotropin-releasing hormone (GnRH) have impaired sexual reproduction. Previous genetic linkage studies and sequencing of plausible gene candidates have identified mutations associated with inherited GnRH deficiency, but the small number of affected families and limited success in validating candidates have impeded genetic diagnoses for most patients. Using a combination of exome sequencing and computational modeling, we have identified a shared point mutation in semaphorin 3E (SEMA3E) in 2 brothers with Kallmann syndrome (KS), which causes inherited GnRH deficiency. Recombinant wild-type SEMA3E protected maturing GnRH neurons from cell death by triggering a plexin D1-dependent (PLXND1-dependent) activation of PI3K-mediated survival signaling. In contrast, recombinant SEMA3E carrying the KS-associated mutation did not protect GnRH neurons from death. In murine models, lack of either SEMA3E or PLXND1 increased apoptosis of GnRH neurons in the developing brain, reducing innervation of the adult median eminence by GnRH-positive neurites. GnRH neuron deficiency in male mice was accompanied by impaired testes growth, a characteristic feature of KS. Together, these results identify SEMA3E as an essential gene for GnRH neuron development, uncover a neurotrophic function for SEMA3E in the developing brain, and elucidate SEMA3E/PLXND1/PI3K signaling as a mechanism that prevents GnRH neuron deficiency.

Technical Progress, Sorting, and Early Retirement

[cat] El progrés tecnològic afecta a la jubilació anticipada a través de dues forces oposades. Per una banda, augmenta el salari real i, per tant, crea incentius per posposar la jubilació. Per altra banda, causa l’erosió de les habilitats dels treballadors, el qual augmenta la probabilitat de jubilació anticipada. En aquest article reexaminem l’efecte del progrés tecnològic en la jubilació anticipada tenint en compte que, al principi de la vida laboral, els individus es distribueixen en diferents sectors segons la seva habilitat. Obtenim dos resultats principals: 1) per petits (grans) canvis tecnològics, l’efecte salari (erosió) domina, i 2) els individus més capaços resisteixen millor l’efecte erosió.

Ras-association domain of sorting nexin 27 is critical for regulating expression of GIRK potassium channels

G protein-gated inwardly rectifying potassium (GIRK) channels play an important role in regulating neuronal excitability. Sorting nexin 27b (SNX27b), which reduces surface expression of GIRK channels through a PDZ domain interaction, contains a putative Ras-association (RA) domain with unknown function. Deleting the RA domain in SNX27b (SNX27b-DRA) prevents the down-regulation of GIRK2c/GIRK3 channels. Similarly, a point mutation (K305A) in the RA domain disrupts regulation of GIRK2c/GIRK3 channels and reduces H-Ras binding in vitro. Finally, the dominant-negative H-Ras (S17N) occludes the SNX27b-dependent decrease in surface expression of GIRK2c/GIRK3 channels. Thus, the presence of a functional RA domain and the interaction with Ras-like G proteins comprise a novel mechanism for modulating SNX27b control of GIRK channel surface expression and cellular excitability.

Technical Progress, Sorting, and Early Retirement

[cat] El progrés tecnològic afecta a la jubilació anticipada a través de dues forces oposades. Per una banda, augmenta el salari real i, per tant, crea incentius per posposar la jubilació. Per altra banda, causa l’erosió de les habilitats dels treballadors, el qual augmenta la probabilitat de jubilació anticipada. En aquest article reexaminem l’efecte del progrés tecnològic en la jubilació anticipada tenint en compte que, al principi de la vida laboral, els individus es distribueixen en diferents sectors segons la seva habilitat. Obtenim dos resultats principals: 1) per petits (grans) canvis tecnològics, l’efecte salari (erosió) domina, i 2) els individus més capaços resisteixen millor l’efecte erosió.

Multimodal stimulus coding by a gustatory sensory neuron in Drosophila larvae.

Accurate perception of taste information is crucial for animal survival. In adult Drosophila, gustatory receptor neurons (GRNs) perceive chemical stimuli of one specific gustatory modality associated with a stereotyped behavioural response, such as aversion or attraction. We show that GRNs of Drosophila larvae employ a surprisingly different mode of gustatory information coding. Using a novel method for calcium imaging in the larval gustatory system, we identify a multimodal GRN that responds to chemicals of different taste modalities with opposing valence, such as sweet sucrose and bitter denatonium, reliant on different sensory receptors. This multimodal neuron is essential for bitter compound avoidance, and its artificial activation is sufficient to mediate aversion. However, the neuron is also essential for the integration of taste blends. Our findings support a model for taste coding in larvae, in which distinct receptor proteins mediate different responses within the same, multimodal GRN.

A monolithic glass chip for active single-cell sorting based on mechanical phenotyping

The mechanical properties of biological cells have long been considered as inherent markers of biological function and disease. However, the screening and active sorting of heterogeneous populations based on serial single-cell mechanical measurements has not been demonstrated. Here we present a novel monolithic glass chip for combined fluorescence detection and mechanical phenotyping using an optical stretcher. A new design and manufacturing process, involving the bonding of two asymmetrically etched glass plates, combines exact optical fiber alignment, low laser damage threshold and high imaging quality with the possibility of several microfluidic inlet and outlet channels. We show the utility of such a custombuilt optical stretcher glass chip by measuring and sorting single cells in a heterogeneous population based on their different mechanical properties and verify sorting accuracy by simultaneous fluorescence detection. This offers new possibilities of exact characterization and sorting of small populations based on rheological properties for biological and biomedical applications.

Three-dimensional supersonic flow over a spike-nosed body of revolution

The unsteady, viscous, supersonic flow over a spike-nosed body of revolution is numerically investigated by solving the Navier-Stokes equations. The time-accurate computations are performed employing an implicit algorithm based on the second-order time-accurate LU-SGS scheme with the incorporation of a subiteration procedure to maintain time accuracy. The characteristics of the flow field for a Mach number of 3.0, Reynolds number of 7.87 x 10(6)/m, and angles of attack of 5 and 10 degrees are described. Self-sustained asymmetric shock wave oscillations were observed in the numerical computations for these angles of attack. The main characteristic of the flow field, as well as its influence on drag coefficient is discussed.

Effects of mineral sorting on concentrator design and economics

Ore sorting after crushing is an effective way to enhance the feed quality of a concentrator. Sorting by hand is the oldest way of concentrating minerals but it has become outdated because of low capacities. Older methods of sorting have also been difficult to use in large scale productions due to low capacities of sorters. Data transfer and processing and the speed of rejection mechanisms have been the bottlenecks for effective use of sorters. A fictive chalcopyrite ore body was created for this thesis. The properties of the ore were typical of chalcopyrite ores and economical limit was set for design. Concentrator capacity was determined by the size of ore body and the planned mine life. Two concentrator scenarios were compared, one with the sorting facility and the other without sorting. Comparison was made for quality and amount of feed, size of equipment and economics. Concentrator with sorting had lower investment and operational cost but also lower incomes due to the ore loss in sorting. Net cash flow, net present value and internal rate of interest were calculated for comparison of the two scenarios.

Price spike forecasting in a competitive day-ahead energy market

Electricity price forecasting has become an important area of research in the aftermath of the worldwide deregulation of the power industry that launched competitive electricity markets now embracing all market participants including generation and retail companies, transmission network providers, and market managers. Based on the needs of the market, a variety of approaches forecasting day-ahead electricity prices have been proposed over the last decades. However, most of the existing approaches are reasonably effective for normal range prices but disregard price spike events, which are caused by a number of complex factors and occur during periods of market stress. In the early research, price spikes were truncated before application of the forecasting model to reduce the influence of such observations on the estimation of the model parameters; otherwise, a very large forecast error would be generated on price spike occasions. Electricity price spikes, however, are significant for energy market participants to stay competitive in a market. Accurate price spike forecasting is important for generation companies to strategically bid into the market and to optimally manage their assets; for retailer companies, since they cannot pass the spikes onto final customers, and finally, for market managers to provide better management and planning for the energy market. This doctoral thesis aims at deriving a methodology able to accurately predict not only the day-ahead electricity prices within the normal range but also the price spikes. The Finnish day-ahead energy market of Nord Pool Spot is selected as the case market, and its structure is studied in detail. It is almost universally agreed in the forecasting literature that no single method is best in every situation. Since the real-world problems are often complex in nature, no single model is able to capture different patterns equally well. Therefore, a hybrid methodology that enhances the modeling capabilities appears to be a possibly productive strategy for practical use when electricity prices are predicted. The price forecasting methodology is proposed through a hybrid model applied to the price forecasting in the Finnish day-ahead energy market. The iterative search procedure employed within the methodology is developed to tune the model parameters and select the optimal input set of the explanatory variables. The numerical studies show that the proposed methodology has more accurate behavior than all other examined methods most recently applied to case studies of energy markets in different countries. The obtained results can be considered as providing extensive and useful information for participants of the day-ahead energy market, who have limited and uncertain information for price prediction to set up an optimal short-term operation portfolio. Although the focus of this work is primarily on the Finnish price area of Nord Pool Spot, given the result of this work, it is very likely that the same methodology will give good results when forecasting the prices on energy markets of other countries.

Facilitation of the main generator source of earthworm muscle contraction by a peripheral neuron

A constant facilitation of responses evoked in the earthworm muscle contraction generator neurons by responses evoked in the neurons of its peripheral nervous system was demonstrated. It is based on the proposal that these two responses are bifurcations of an afferent response evoked by the same peripheral mechanical stimulus but converging again on this central neuron. A single-peaked generator response without facilitation was demonstrated by sectioning the afferent route of the peripheral facilitatory modulatory response, or conditioning response (CR). The multipeaked response could be restored by restimulating the sectioned modulatory neuron with an intracellular substitutive conditioning stimulus (SCS). These multi-peaked responses were proposed to be the result of reverberating the original single peaked unconditioned response (UR) through a parallel (P) neuronal circuit which receives the facilitation of the peripheral modulatory neuron. This peripheral modulatory neuron was named "Peri-Kästchen" (PK) neuron because it has about 20 peripheral processes distributed on the surface of a Kästchen of longitudinal muscle cells on the body wall of this preparation as revealed by the Lucifer Yellow-CH-filling method.

Persistent attenuation and enhancement of the earthworm main muscle contraction generator response induced by repeated stimulation of a peripheral neuron

Responses evoked in the earthworm, Amynthas hawayanus, main muscle contraction generator M-2 (postsynaptic mechanical-stimulus-sensitive) neuron by threshold mechanical stimuli in 2-s intertrial intervals (ITI) were used as the control or unconditioned responses (UR). Their attenuation induced by decreasing these intervals in non-associative conditioning and their enhancement induced by associating the unconditioned stimuli (US) to a train of short (0.1 s) hyperpolarizing electrical substitutive conditioning stimuli (SCS) in the Peri-Kästchen (PK) neuron were measured in four parameters, i.e., peak numbers (N) and amplitude ()averaged from 120 responses, sum of these amplitudes (SAMP) and the highest peak amplitude (V) over a period of 4 min. Persistent attenuation similar to habituation was induced by decreasing the control ITI to 0.5 s and 2.0 s in non-associative conditioning within less than 4 min. Dishabituation was induced by randomly pairing one of these habituated US to an electrical stimulus in the PK neuron. All four parameters of the UR were enhanced by forward (SCS-US), but not backward (US-SCS), association of the US with 25, 100 and 250-Hz trains of SCS with 40-ms interstimulus intervals (ISI) for 4 min and persisted for another 4 min after turning off the SCS. The enhancement of these parameters was proportional to the SCS frequencies in the train. No UR was evoked by the SCS when the US was turned off after 4 min of classical conditioning.