[Imaging Blood Flow and Pulsation of Retinal Vessels with Full-Field Swept-Source OCT]. / Darstellung von Blutfluss und Pulsation in retinalen Gefäßen mit Full-Field-Swept-Source-OCT.

Optical coherence tomography (OCT) uses interference to image the retina with high axial resolution. In the last 25 years, new technologies have permitted a steady increase in imaging speed, which made it possible to enlarge the imaged field and to avoid motion artefacts. The speed and precision of retinal imaging is now limited by photodamage of the retina caused by the focused OCT beam and by the speed of the scanning mechanics. Full-field swept-source (FF-SS)-OCT decreases irradiance on the retina and dispenses moving parts by using a camera to acquire the full volume of the retina in parallel. Here we show that FF-SS-OCT is rapid and precise enough to image pulsation in the retina induced by the heart beat. Series of OCT volumes 1.8 × 0.7 mm wide and 1.8 mm deep were recorded in young volunteers over a few cycles of the heart beat. Morphology of the retinal vessels, blood flow and tissue motion as caused by vessel pulsation were calculated from the OCT data. FF-SS-OCT was able to visualise the main structures of the neuronal retina, including vessels and small capillaries and without any motion artefacts. Information on three different dynamic processes was obtained from only one recorded series of OCT volumes: pulsation of blood flow and blood pressure in retinal vessels as well as pulsation of the choroid. Delays between arterial and venous pulse and delay between pulsation in retinal and choroidal vessels were calculated. With a time resolution of 0.5 ms, FF-SS-OCT is able to visualise previously unmeasurably fast changes in the retina, including the propagation of pulse waves.

Purification and characterization of RNA polymerase II holoenzyme from Schizosaccharomyces pombe.

We have purified the RNA polymerase II holoenzyme from Schizosaccharomyces pombe to near homogeneity. The Mediator complex is considerably smaller than its counterpart in Saccharomyces cerevisiae, containing only nine polypeptides larger than 19 kDa. Five of these Mediator subunits have been identified as the S. pombe homologs to Rgr1, Srb4, Med7, and Nut2 found in S. cerevisiae and the gene product of a previously uncharacterized open reading frame, PMC2, with no clear homologies to any described protein. The presence of Mediator in a S. pombe RNA polymerase II holoenzyme stimulated phosphorylation of the C-terminal domain by TFIIH purified from S. pombe. This stimulation was species-specific, because S. pombe Mediator could not stimulate TFIIH purified from S. cerevisiae. We suggest that the overall structure and mechanism of the Mediator is evolutionary conserved. The subunit composition, however, has evolved to respond properly to physiological signals.

Identification of new mediator subunits in the RNA polymerase II holoenzyme from Saccharomyces cerevisiae.

Mediator was isolated from yeast on the basis of its requirement for transcriptional activation in a fully defined system. We have now identified three new members of mediator in the low molecular mass range by peptide sequence determination. These are the products of the NUT2, CSE2, and MED11 genes. The product of the NUT1 gene is evidently a component of mediator as well. NUT1 and NUT2 were earlier identified as negative regulators of the HO promoter, whereas mutations in CSE2 affect chromosome segregation. MED11 is a previously uncharacterized gene. The existence of these proteins in the mediator complex was verified by copurification and co-immunoprecipitation with RNA polymerase II holoenzyme.

Analysis of Schizosaccharomyces pombe mediator reveals a set of essential subunits conserved between yeast and metazoan cells.

With the identification of eight new polypeptides, we here complete the subunit characterization of the Schizosaccharomyces pombe RNA polymerase II holoenzyme. The complex contains homologs to all 10 essential gene products present in the Saccharomyces cerevisiae Mediator, but lacks clear homologs to any of the 10 S. cerevisiae components encoded by nonessential genes. S. pombe Mediator instead contains three unique components (Pmc2, -3, and -6), which lack homologs in other cell types. Presently, pmc2(+) and pmc3(+) have been shown to be nonessential genes. The data suggest that S. pombe and S. cerevisiae share an essential protein module, which associates with nonessential speciesspecific subunits. In support of this view, sequence analysis of the conserved yeast Mediator components Med4 and Med8 reveals sequence homology to the metazoan Mediator components Trap36 and Arc32. Therefore, 8 of 10 essential genes conserved between S. pombe and S. cerevisiae also have a metazoan homolog, indicating that an evolutionary conserved Mediator core is present in all eukaryotic cells. Our data suggest a closer functional relationship between yeast and metazoan Mediator than previously anticipated.