Transplantation of canine olfactory ensheathing cells producing chondroitinase ABC promotes chondroitin sulphate proteoglycan digestion and axonal sprouting following spinal cord injury

Olfactory ensheathing cell (OEC) transplantation is a promising strategy for treating spinal cord injury (SCI), as has been demonstrated in experimental SCI models and naturally occurring SCI in dogs. However, the presence of chondroitin sulphate proteoglycans within the extracellular matrix of the glial scar can inhibit efficient axonal repair and limit the therapeutic potential of OECs. Here we have used lentiviral vectors to genetically modify canine OECs to continuously deliver mammalian chondroitinase ABC at the lesion site in order to degrade the inhibitory chondroitin sulphate proteoglycans in a rodent model of spinal cord injury. We demonstrate that these chondroitinase producing canine OECs survived at 4 weeks following transplantation into the spinal cord lesion and effectively digested chondroitin sulphate proteoglycans at the site of injury. There was evidence of sprouting within the corticospinal tract rostral to the lesion and an increase in the number of corticospinal axons caudal to the lesion, suggestive of axonal regeneration. Our results indicate that delivery of the chondroitinase enzyme can be achieved with the genetically modified OECs to increase axon growth following SCI. The combination of these two promising approaches is a potential strategy for promoting neural regeneration following SCI in veterinary practice and human patients

An Efficient and Versatile System for Visualization and Genetic Modification of Dopaminergic Neurons in Transgenic Mice.

BACKGROUND & AIMS: The brain dopaminergic (DA) system is involved in fine tuning many behaviors and several human diseases are associated with pathological alterations of the DA system such as Parkinson's disease (PD) and drug addiction. Because of its complex network integration, detailed analyses of physiological and pathophysiological conditions are only possible in a whole organism with a sophisticated tool box for visualization and functional modification. METHODS & RESULTS: Here, we have generated transgenic mice expressing the tetracycline-regulated transactivator (tTA) or the reverse tetracycline-regulated transactivator (rtTA) under control of the tyrosine hydroxylase (TH) promoter, TH-tTA (tet-OFF) and TH-rtTA (tet-ON) mice, to visualize and genetically modify DA neurons. We show their tight regulation and efficient use to overexpress proteins under the control of tet-responsive elements or to delete genes of interest with tet-responsive Cre. In combination with mice encoding tet-responsive luciferase, we visualized the DA system in living mice progressively over time. CONCLUSION: These experiments establish TH-tTA and TH-rtTA mice as a powerful tool to generate and monitor mouse models for DA system diseases

Limitations of the spike-triggered averaging for estimating motor unit twitch force: a theoretical analysis.

Contractile properties of human motor units provide information on the force capacity and fatigability of muscles. The spike-triggered averaging technique (STA) is a conventional method used to estimate the twitch waveform of single motor units in vivo by averaging the joint force signal. Several limitations of this technique have been previously discussed in an empirical way, using simulated and experimental data. In this study, we provide a theoretical analysis of this technique in the frequency domain and describe its intrinsic limitations. By analyzing the analytical expression of STA, first we show that a certain degree of correlation between the motor unit activities prevents an accurate estimation of the twitch force, even from relatively long recordings. Second, we show that the quality of the twitch estimates by STA is highly related to the relative variability of the inter-spike intervals of motor unit action potentials. Interestingly, if this variability is extremely high, correct estimates could be obtained even for high discharge rates. However, for physiological inter-spike interval variability and discharge rate, the technique performs with relatively low estimation accuracy and high estimation variance. Finally, we show that the selection of the triggers that are most distant from the previous and next, which is often suggested, is not an effective way for improving STA estimates and in some cases can even be detrimental. These results show the intrinsic limitations of the STA technique and provide a theoretical framework for the design of new methods for the measurement of motor unit force twitch.Open-Access-Publikationsfonds 2014peerReviewe

Repetitive Hops Induce Postactivation Potentiation in Triceps Surae as well as an Increase in the Jump Height of Subsequent Maximal Drop Jumps

Postactivation potentiation (PAP) has been defined as the increase in twitch torque after a conditioning contraction. The present study aimed to investigate the effectiveness of hops as conditioning contractions to induce PAP and increase performance in subsequent maximal drop jumps. In addition, we wanted to test if and how PAP can contribute to increases in drop jump rebound height. Twelve participants performed 10 maximal two-legged hops as conditioning contractions. Twitch peak torques of triceps surae muscles were recorded before and after the conditioning hops. Then, subjects performed drop jumps with and without 10 conditioning hops before each drop jump. Recordings included ground reaction forces, ankle and knee angles and electromyographic activity in five leg muscles. In addition, efferent motoneuronal output during ground contact was estimated with V-wave stimulation. The analyses showed that after the conditioning hops, twitch peak torques of triceps surae muscles were 32% higher compared to baseline values (P < 0.01). Drop jumps performed after conditioning hops were significantly higher (12%, P < 0.05), but V-waves and EMG activity remained unchanged. The amount of PAP and the change in drop jump rebound height were positively correlated (r(2) = 0.26, P < 0.05). These results provide evidence for PAP in triceps surae muscles induced by a bout of hops and indicate that PAP can contribute to the observed performance enhancements in subsequent drop jumps. The lack of change in EMG activity and V-wave amplitude suggests that the underlying mechanisms are more likely intramuscular than neural in origin