A medium for commercial production of the halophilic Micrococcus nuclease.

A simple synthetic medium (glutamate-sucrose medium) was devised for production, during growth in shaken flasks, of extracellular halophilic nuclease (nuclease H) by a moderate halophile, Micrococcus varians subsp. halophilus. A simple medium consisting of 0.7% ammonium sulfate, 1.0% glucose, minerals, three vitamins, and 2 M NaCl gave good growth and excellent production of nuclease H in a jar fermentor when the pH was adjusted to 7.5 to 8.0 during cultivation.     

Effects of long-term physical exercise on skeletal muscles in senescence-accelerated mice (SAMP8)

We examined the effect of long-term exercise on the prevention of sarcopenia using a senescence-accelerated-prone mice (SAMP8) model. Mice were housed in a wheel cage for 25 weeks to increase voluntary exercise. At week 23, endurance running capacity was examined using a treadmill. In a treadmill running test, the wheel cage group had increased endurance running capacity, which suggests that aging-related loss of muscle function was recovered by long-term exercise. Mice were sacrificed and microarray analysis revealed that genes involved in protein synthesis and degradation were upregulated in the skeletal muscles of the wheel cage group, suggesting accelerated protein turnover. Total body and adipose tissue weights decreased following the use of the wheel cage. Thus, long-term, spontaneous physical exercise may assist in recovering from aging-related sarcopenia (loss of muscle function) and obesity.     

Developmental and regional alteration of methionine enkephalin-like immunoreactivity in seizure-susceptible E1 mouse brain

Methionine enkephalin-like immunoreactivity (ME-LI) in the brain of El mice (seizure-susceptible strain) was measured by radioimmunoassay (RIA) to elucidate the relation between seizures and the opioid system. The lyophilized supernatants of tissue extracts were subjected to ME RIA. The concentration of ME-LI in 25-day-old El mice that had no seizures was significantly decreased in the hippocampus. At the age of 50 days when El mice displayed abortive seizures, the levels of ME-LI in both El(+) and nonstimulated El(o) mice were also significantly reduced in the hippocampus and septal area. It was further shown that the ME-LI concentrations in both 150-day-old adult El(+) during interictal periods and El(o) mice were markedly decreased in the cerebral cortex, septal area, and striatum, as compared with the corresponding regions in ddY mice (seizurenonsusceptible strain; the mother strain of El). The decrease of ME-LI in the El mouse brain was generally compatible with our previous findings concerning the up-regulation of opioid delta receptors in this species. These results suggest that the reduction of ME-LI in the El mouse brain is not due to convulsions, but could be associated with the pathogenesis of seizure diathesis and seizure manifestations in the El mouse.     

Evidence That an Unconventional Actin Can Provide Essential F-Actin Function and That a Surveillance System Monitors F-Actin Integrity in Chlamydomonas

Actin is one of the most conserved eukaryotic proteins. It is thought to have multiple essential cellular roles and to function primarily or exclusively as filaments (“F-actin”). Chlamydomonas has been an enigma, because a null mutation (ida5-1) in its single gene for conventional actin does not affect growth. A highly divergent actin gene, NAP1, is upregulated in ida5-1 cells, but it has been unclear whether NAP1 can form filaments or provide actin function. Here, we used the actin-depolymerizing drug latrunculin B (LatB), the F-actin-specific probe Lifeact-Venus, and genetic and molecular methods to resolve these issues. LatB-treated wild-type cells continue to proliferate; they initially lose Lifeact-stained structures but recover them concomitant with upregulation of NAP1. Thirty-nine LatB-sensitive mutants fell into four genes (NAP1 and LAT1–LAT3) in which we identified the causative mutations using a novel combinatorial pool-sequencing strategy. LAT1–LAT3 are required for NAP1 upregulation upon LatB treatment, and ectopic expression of NAP1 largely rescues the LatB sensitivity of the lat1–lat3 mutants, suggesting that the LAT gene products comprise a regulatory hierarchy with NAP1 expression as the major functional output. Selection of LatB-resistant revertants of a nap1 mutant yielded dominant IDA5 mutations that presumably render F-IDA5 resistant to LatB, and nap1 and lat mutations are synthetically lethal with ida5-1 in the absence of LatB. We conclude that both IDA5 and the divergent NAP1 can form filaments and redundantly provide essential F-actin functions and that a novel surveillance system, probably responding to a loss of F-actin, triggers NAP1 expression and perhaps other compensatory responses.