Proteoglycan-specific molecular switch for RPTPσ clustering and neuronal extension

Heparan and chondroitin sulfate proteoglycans (HSPGs and CSPGs, respectively) regulate numerous cell surface signaling events, with typically opposite effects on cell function. CSPGs inhibit nerve regeneration through receptor protein tyrosine phosphatase sigma (RPTPσ). Here we report that RPTPσ acts bimodally in sensory neuron extension, mediating CSPG inhibition and HSPG growth promotion. Crystallographic analyses of a shared HSPG-CSPG binding site reveal a conformational plasticity that can accommodate diverse glycosaminoglycans with comparable affinities. Heparan sulfate and analogs induced RPTPσ ectodomain oligomerization in solution, which was inhibited by chondroitin sulfate. RPTPσ and HSPGs colocalize in puncta on sensory neurons in culture, whereas CSPGs occupy the extracellular matrix. These results lead to a model where proteoglycans can exert opposing effects on neuronal extension by competing to control the oligomerization of a common receptor.     

Ordered and dynamic assembly of single spliceosomes

The spliceosome is the complex macromolecular machine responsible for removing introns from precursors to messenger RNAs (pre-mRNAs). We combined yeast genetic engineering, chemical biology, and multiwavelength fluorescence microscopy to follow assembly of single spliceosomes in real time in whole-cell extracts. We find that individual spliceosomal subcomplexes associate with pre-mRNA sequentially via an ordered pathway to yield functional spliceosomes and that association of every subcomplex is reversible. Further, early subcomplex binding events do not fully commit a pre-mRNA to splicing; rather, commitment increases as assembly proceeds. These findings have important implications for the regulation of alternative splicing. This experimental strategy should prove widely useful for mechanistic analysis of other macromolecular machines in environments approaching the complexity of living cells.     

Quantitative electroencephalography in dogs anesthetized with 2.0% end-tidal concentration of isoflurane anesthesia

Quantitative electroencephalography was assessed in dogs under controlled, 2% end-tidal isoflurane anesthetic conditions, and each variable at each electrode site was tested for normal distribution. With the quantitative electroencephalographic system used, 16 values for each of 21 electrode sites were evaluated. Absolute power ratios also were evaluated. The methods for quantitative electroencephalographic recording and analysis appear to be readily adaptable to the dog. Most of the data do not conform to a normal distribution. Therefore, distribution-free nonparametric statistics should be used when looking for differences under experimental or clinical conditions. Quantitative electroencephalography appears to be a sensitive noninvasive method that could be used to evaluate brain function under anesthetic, clinical, and experimental settings.     

Physiologic responses of feral horses to succinylcholine chloride immobilization

Five captive feral horses were immobilized at 72 hour intervals for 30 days by intramuscular administration of succinylcholine chloride (SCh) using a capture gun and dart system. The serum enzyme activities of creatine phosphokinase, glutamate-oxaloacetate transaminase, and lactate dehydrogenase and serum cortisol concentrations were monitored to assess the response to chemical immobilization in feral horses over time.Reference values for these parameters in feral horses were found to be in close agreement with those of normal, rested horses not in training. The results suggest that single, or infrequently repeated, use of succinylcholine-chloride by intramuscular administration to captive feral horses, or to otherwise unapproachable horses, could be efficiently and practically employed in field situations without major physiologic alterations and with minimal stress occurring in these horses.