Cholesterol Synthesis and Nerve Regeneration

In this report, we examine the requirement of cholesterol biosynthesis and its axonal transport for goldfish optic nerve regeneration. Cholesterol, labeled by intraocular injection of [ 3 H]mevalonolactone. exhibited a delayed appearance in the optic tectum. Squalene and other minor components were labeled but not transported. Following optic nerve crush, the amount of labeled cholesterol transport was elevated, while retinal labeling was not altered relative to control fish. A requirement for cholesterol biosynthesis is inferred from the inhibition of neurite outgrowth in retinal explants caused by the cholesterol synthesis inhibitor, 20, 25-diazacholes-terol. The inhibition of growth could be overcome by addition of mevalonolactone, but not cholesterol, to the medium. Intraperitoneal administration of 200 nmol of dia-zacholesterol resulted in 92-98% inhibition of retinal cholesterol synthesis and accumulation of labeled des-mosterol and other lipids in fish retina and brain which persisted for 2 weeks. Diazacholesterol-treated fish showed no reduction in the amount of lipid-soluble radioactivity transported following intraocular injection of [ 3 H]mevalonolactone, but there were alterations in the chromatographic pattern of the transported labeled lipids. In contrast to its effects on neurite outgrowth in vitro , diazacholesterol did not inhibit optic nerve regeneration in vivo , as measured both by arrival of labeled rapidly transported protein at the tectum and by time required for the return of visual function.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/65710/1/j.1471-4159.1984.tb12701.x.pd

Decoupling of a Neutron Interferometer from Temperature Gradients

Neutron interferometry enables precision measurements that are typically operated within elaborate, multi-layered facilities which provide substantial shielding from environmental noise. These facilities are necessary to maintain the coherence requirements in a perfect crystal neutron interferometer which is extremely sensitive to local environmental conditions such as temperature gradients across the interferometer, external vibrations, and acoustic waves. The ease of operation and breadth of applications of perfect crystal neutron interferometry would greatly benefit from a mode of operation which relaxes these stringent isolation requirements. Here, the INDEX Collaboration and National Institute of Standards and Technology demonstrates the functionality of a neutron interferometer in vacuum and characterize the use of a compact vacuum chamber enclosure as a means to isolate the interferometer from spatial temperature gradients and time-dependent temperature fluctuations. The vacuum chamber is found to have no depreciable effect on the performance of the interferometer (contrast) while improving system stability, thereby showing that it is feasible to replace large temperature isolation and control systems with a compact vacuum enclosure for perfect crystal neutron interferometry

Lasting Effects of a Graduate Pedagogy Course on the Development of Teacher-Scholars

While graduate pedagogy courses may result in specific teaching behaviors in the short term, little is known whether these scholarly teaching behaviors and attitudes endure over time. Multiple evidenced-based assessments of pedagogy courses which continue well past the completion of the course can demonstrate the ongoing development of students’ perceptions of teaching and learning and students’ progression as teacher-scholars. This poster summarizes an investigation exploring the lasting effects of a graduate pedagogy course entitled MSCI M620: Pedagogical Methods in Health Sciences. This course has been offered for two semesters and a total of sixteen graduate students from a variety of science-related disciplines enrolled in the course. We examined whether the course caused students to: 1) Develop more complex and refined attitudes regarding student learning; 2) Evaluate their current teaching methods and explore alternative teaching strategies; 3) Assess their teaching effectiveness; 4) Plan to be involved in a public teaching resource network; and 5) Develop lasting attitudes about scholarly teaching

Telomere dynamics and fusion of critically shortened telomeres in plants lacking DNA ligase IV

In the absence of the telomerase, telomeres undergo progressive shortening and are ultimately recruited into end-to-end chromosome fusions via the non-homologous end joining (NHEJ) double-strand break repair pathway. Previously, we showed that fusion of critically shortened telomeres in Arabidopsis proceeds with approximately the same efficiency in the presence or absence of KU70, a key component of NHEJ. Here we report that DNA ligase IV (LIG4) is also not essential for telomere joining. We observed only a modest decrease (3-fold) in the frequency of chromosome fusions in triple tert ku70 lig4 mutants versus tert ku70 or tert. Sequence analysis revealed that, relative to tert ku70, chromosome fusion junctions in tert ku70 lig4 mutants contained less microhomology and less telomeric DNA. These findings argue that the KU-LIG4 independent end-joining pathway is less efficient and mechanistically distinct from KU-independent NHEJ. Strikingly, in all the genetic backgrounds we tested, chromosome fusions are initiated when the shortest telomere in the population reaches ∼1 kb, implying that this size represents a critical threshold that heralds a detrimental structural transition. These data reveal the transitory nature of telomere stability, and the robust and flexible nature of DNA repair mechanisms elicited by telomere dysfunction

Measurement of Receptor-Activated Phosphoinositide Turnover in Rat Brain: Nonequivalence of Inositol Phosphate and CDP-Diacylglycerol Formation

Two methods for the measurement of receptor-activated phosphoinositide turnover were evaluated for their degree of correspondence in slices of rat brain; they involved the Li + -dependent accumulations of either [ 3 H]-inositol-labeled inositol phosphates or [ 3 H]cytidine-labeled CDP-diacylglycerol. In contrast to the expectation that the ratio of these two responses would remain approximately constant, varying degrees of correspondence were obtained. The two extremes are exemplified by carbachol, which elicited large increases in both inositol phosphate and CDP-diacylglycerol labeling, and endothelin, which gave a robust inositol phosphate response with little or no accumulation of 3 H-CDP-diacylglycerol. No instance of the presence of the latter response in the absence of 3 H-inositol phosphate accumulation was observed. Measurement of 3 H-CDP-diacylglycerol accumulation thus may add additional insight into the regulation of phosphoinositide turnover and the complex actions of Li + .Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66135/1/j.1471-4159.1993.tb03258.x.pd