| 研究実績の概要 |
In 2015/05, we recorded spectral and polarisation sensitivities from the angled sunbeam butterfly (Curetis acuta). We found four (UV, blue, green, orange) receptor classes. Diagonal receptors are likely orange-peaking and have a wide bandwidth distribution, possibly due to self-screening. Light microscopy showed no clear ommatidial types. The eight perirhabdomeral pigment clusters have different lengths. This data was presented at the JSCPBC Meeting in Hiroshima in 2015/12.
In 2015/09, we recorded videos of Curetis butterflies in flight. In 2016/01, we have implemented white light spectroscopy measurements to provide optical estimates of Curetis' main rhodopsin and metarhodopsin. In collaboration with Dr. Belusic in Ljubljana, we are developing a LED-based light source for intracellular recordings of spectral sensitivity and for hyperspectral imaging in general.
In 2015/06, Dr. Pirih attended the Animal ERG course at the ISCEV meeting in Ljubljana. He tested multifocal and pattern ERG stimulation on insects for regionalisation and visual acuity measurements. Dr. Pirih visited the lab of Prof. Stavenga in Groningen twice. In 2015/06, he measured the Curetis visual angles, and in 2016/02, he performed imaging spectroscopy measurements of the eyeshine under different rhodopsin/metarhodopsin states. In 2016/02, Dr. Pirih visited the lab of dr. O'Carroll in Lund, for a demonstration of the wide-field stimulation technique.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Although we have managed to keep a larger number of females during the winter 2015/2016, we are unable to get fertile eggs from them. As a consequence, it may be necessary to skip the early summer experimental season on Curetis and postpone the measurements described below to the autumn season. Because it will not be necessary to keep the caught animals for the next winter, all the animals caught will be used for experiments. The analysis of the electrophysiological and optical data from the previous season will be completed successfully because we have already established the recording method for Curetis. LED-based light source will be in operation soon, which can be used for the electrophysiological recording.
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| 今後の研究の推進方策 |
The methods paper describing LED-based source is currently being finalized. We then plan to build a LED-based source for the electrophysiology and the telemicroscope setup. On Curetis, we plan to perform a TEM study and immunohistological labelling of rhodopsins using antibodies produced against Papilio opsins: we have already checked that the antibodies cross-react with Curetis opsins. We plan to perform optical measurements of rhodopsin/metarhocopsin photochemistry and electrophysiological measurements of spectral sensitivity under different rhodopsin/metarhocopsin adaptation states, in order to test the hypothesis that the sensitivity broadening is due to self-screening. We plan to perform electrophysiological acceptance angle measurements in current clamp mode, in order to test whether hyperpolarisation is just a measurement artefact or is partially due to lateral inhibition. In the case that it will be difficult to obtain Curetis in the summer months, the Curetis measurements will be postponed to autumn, and the available time will be used for data analysis and for optical measurements of rhodopsin photochemistry of selected lepidopteran species. We plan to couple the LED-based source to a DLP projector, with which we will attempt acceptance angle measurements with a bright background, and recording from higher order neurones with spatial stimulation in the autumn season.
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