No abstract

Insects are favorable subjects for neuroethological studies. Their nervous systems are relatively small and contain many individually identifiable cells. The CNS is highly compartmentalized with clear separations between multisensory higher order neuropiles in the brain and neuropiles serving sensory-motor routines in the ventral cord (Huber, 1974). The rich behavior of insects includes orientation in space and time, visual, chemical, and mechanical communication, and complex motor routines for flying, walking, swimming, nest building, defense, and attack. Learning and memory, though, are not usually considered to be a strong point of insects. Rather, insect behavior is often regarded as highly stereotyped and under tight control of genetically programmed neural circuits. This view, however, does not do justice to the insect order of Hymenoptera (bees, wasps, ants). Most Hymenopteran species care for their brood either as individual females or as a social group of females. Consequently, they regularly return to their nest site to feed, protect, and nurse the larvae, store food, and hide from adverse environmental conditions. Since they search for food (prey; nectar and pollen on flowers) at unpredictable sites, they have to learn the celestial and terrestrial cues that guide their foraging trips over long distances and allow them to find their nest sites (central place foraging; von Frisch, 1967; Seeley, 1985). They learn to relate the sun's position and sky pattern of polarized light to the time of the day (Lindauer, 1959), and landmarks are learned in relationship to the nest site within the framework of the time- compensated sun compass. The honeybee communicates direction and distance of a feeding place to hive mates by performing a ritualized body movement, the waggle dance (von Frisch, 1967). Associative learning is an essential component of the bee's central place foraging behavior and dance communication. Hive mates attending a dance performance learn the odor emanating from the dancing bee and seek it at the indicated food site. The odor, color, and shape of flowers are learned when the bee experiences these stimuli shortly before it finds food (nectar, pollen). This appetitive learning in bees has many characteristics of associative learning well known from mammalian learning studies (Menzel, 1985, 1990; Bitterman, 1988). It follows the rules of classical and operant conditioning, respectively, so that stimuli or behavioral acts are associated with evaluating stimuli. Since associative learning, especially of the classical type, is well described at the phenomenological and operational level (Rescorla, 1988), it provides a favorable approach in the search for the neural substrate underlying learning and memory.(ABSTRACT TRUNCATED AT 400 WORDS)

The purpose of this study was to measure the hemoglobin oxygenation in retinal vessels and to evaluate the sensitivity and reproducibility of the measurement. Using a fundus camera equipped with a special dual wavelength transmission filter and a color charge-coupled device camera, two monochromatic fundus images at 548 and 610 nm were recorded simultaneously. The optical densities of retinal vessels for both wavelengths and their ratio, which is known to be proportional to the oxygen saturation, were calculated. From 50-deg images, the used semiautomatic vessel recognition and tracking algorithm recognized and measured vessels of 100 microm or more in diameter. On average, arterial and venous oxygen saturations were measured at 98+/-10.1% and 65+/-11.7%, respectively. For measurements in the same vessel segments from the five images per subject, standard deviations of 2.52% and 3.25% oxygen saturation were found in arteries and veins, respectively. Respiration of 100% oxygen increased the mean arterial and venous oxygen saturation by 2% and 7% respectively. A simple system for noninvasive optical oximetry, consisting of a special filter in a fundus camera and software, was introduced. It is able to measure the oxygen saturation in retinal branch vessels with reproducibility and sensitivity suitable for clinical investigations.

Functional alterations are first signs of a starting pathological process. A device that measures parameter for the characterization of the metabolism at the human eye-ground would be a helpful tool for early diagnostics in stages when alterations are yet reversible. Measurements of blood flow and of oxygen saturation are necessary but not sufficient. The new technique of auto-fluorescence lifetime measurement (FLIM) opens in combination with selected excitation and emission ranges the possibility for metabolic mapping. FLIM not only adds an additional discrimination parameter to distinguish different fluorophores but also resolves different quenching states of the same fluorophore. Because of its high sensitivity and high temporal resolution, its capability to resolve multi-exponential decay functions, and its easy combination with laser scanner ophthalmoscopy, multi-dimensional time-correlated single photon counting was used for fundus imaging. An optimized set up for in vivo lifetime measurements at the human eye-ground will be explained. In this, the fundus fluorescence is excited at 446 or 468 nm and the time-resolved autofluorescence is detected in two spectral ranges between 510 and 560 nm as well as between 560 and 700 nm simultaneously. Exciting the fundus at 446 nm, several fluorescence maxima of lifetime t1 were detected between 100 and 220 ps in lifetime histograms of 40 degrees fundus images. In contrast, excitation at 468 nm results in a single maximum of lifetime t1 = 190 +/- 16 ps. Several fundus layers contribute to the fluorescence intensity in the short-wave emission range 510-560 nm. In contrast, the fluorescence intensity in the long-wave emission range between 560 and 700 nm is dominated by the fluorescence of lipofuscin in the retinal pigment epithelium. Comparing the lateral distribution of parameters of a tri-exponential model function in lifetime images of the fundus with the layered anatomical fundus structure, the shortest component (t1 = 190 ps) originates from the retinal pigment epithelium and the second lifetime (t2 = 1,000 ps) from the neural retina. The lifetime t3 approximately 5.5 ns might be influenced by the long decay of the fluorescence in the crystalline lens. In vitro analysis of the spectral properties of expected fluorophores under the condition of the living eye lightens the interpretation of in vivo measurements. Taking into account the transmission of the ocular media, the excitation of NADH is unlikely at the fundus.

The increase of retinal vessel oxygen saturation in diabetic retinopathy points to a diabetic microvascular alteration. This may be due to occlusions and obliterations in the capillary bead and the formation of arterio-venous shunt vessels. On the other hand, hyperglycaemia-induced endothelial dysfunction, with subsequent suppression of the endothelial NO-synthase and disturbance of the vascular auto-regulation, may contribute to retinal tissue hypoxia.