Latest papers on Horseshoe Crabs, radiation effects
Opsin co-expression in Limulus photoreceptors: differential regulation by light and a circadian clock.
Whitney Laboratory for Marine Bioscience, University of Florida, 9505 Ocean Shore Boulevard, St Augustine, FL 32080, USA.
A long-standing concept in vision science has held that a single photoreceptor expresses a single type of opsin, the protein component of visual pigment. However, the number of examples in the literature of photoreceptors from vertebrates and invertebrates that break this rule is increasing. Here, we describe a newly discovered Limulus opsin, Limulus opsin5, which is significantly different from previously characterized Limulus opsins, opsins1 and 2. We show that opsin5 is co-expressed with opsins1 and 2 in Limulus lateral and ventral eye photoreceptors and provide the first evidence that the expression of co-expressed opsins can be differentially regulated. We show that the relative levels of opsin5 and opsin1 and 2 in the rhabdom change with a diurnal rhythm and that their relative levels are also influenced by the animal's central circadian clock. An analysis of the sequence of opsin5 suggests it is sensitive to visible light (400-700 nm) but that its spectral properties may be different from that of opsins1 and 2. Changes in the relative levels of these opsins may underlie some of the dramatic day-night changes in Limulus photoreceptor function and may produce a diurnal change in their spectral sensitivity.
Biomed Sci Instrum. 2003 ;39 :105-10 12724877
Do weak adapting backgrounds uncover multiple components in the electroretinogram of the horseshoe crab?
Department of Applied Biology and Biomedical Engineering, Rose-Hulman Institute of Technology, Terre Haute, IN 47803, USA.
The lateral eye of the horseshoe crab, Limulus polyphemus, has been used as a model system for over a century to study visual and circadian processes. One advantage of this system is the relative simplicity of the retina. The input pathway of the retina consists of photoreceptor cells that are electrically coupled to the dendrite of a second-order cell, which sends action potentials to the brain. Electroretinograms (ERGs) recorded from the lateral eye show a biphasic shape, with a leading negative wave and a later positive peak. The purpose of these experiments was to determine whether adapting backgrounds could be used to uncover multiple adaptation mechanisms within the ERG. To test this idea, ERGs were elicited using variable intensity flashes presented under dark-adapted conditions, as well as in the presence of weak adapting backgrounds. Flashes and backgrounds were generated using green LEDs (lambda max = 525 nm) under software control. ERGs were recorded using a corneal wick electrode placed on the lateral eye of the horseshoe crab. Preliminary results suggest that ERGs recorded in the presence of adapting backgrounds are linearly scaled versions of dark-adapted FRGs. This suggests that there is a single adaptation stage in the Limulus retina. This is in contrast with analogous results from mammals, including mouse, cat and monkey, which show multiple stages of adaptation within their more complex retinas.
Categorical and prolonged potentials are evoked when brief, intermediate-intensity flashes stimulate horseshoe crab lateral eye photoreceptors during octopamine neuromodulation.
Sensory Coding Laboratory, Department of Psychological Sciences, Purdue University, West Lafayette, IN 47907-1364, USA. email@example.com
Octopamine, a major efferent neurotransmitter in the lateral eye of the horseshoe crab (Limulus polyphemus), has previously been shown to modulate photoreceptor responses evoked by long flashes. Quantification of these data indicates that this modulation produced a genuine increase in sensitivity to light which cannot be entirely due to an increase in optical efficiency consequent on an anatomical alteration. Other previous studies demonstrated that extrinsic current can modulate Limulus lateral eye photoreceptor cells by inducing a bistable membrane potential with two distinct states. The present study was therefore undertaken to find out if octopamine could modulate visual responses by inducing prolonged and bistable polarization shifts similar to those demonstrated in several other neural systems. Intracellular microelectrodes were used to execute an electrophysiological study of the receptor potentials evoked in the lateral eye of Limulus when brief (20-ms) flashes were delivered while 50 microM octopamine perfused dark-adapted photoreceptors. The combined chemical and optical stimuli prolonged photoreceptor responses to light to the degree that they often exceeded the duration of the brief stimulus by hundreds of milliseconds. Moreover, these prolonged potentials were clearly bistable because they were categorical--either a prolongation was perceptually clear-cut and present or it was not, with no intermediate patterns being observed. During seawater control perfusions, such prolongations were absent. This appears to be the first demonstration of such categorical and prolonged potentials in a photoreceptor neuron. This finding particularly suggests that efferent-driven neuromodulation can enable the development of a persisting short-term representation of a brief stimulus, with this representation being retained at the most distal possible neural site.
Ca2+/calmodulin-binding peptides block phototransduction in Limulus ventral photoreceptors: evidence for direct inhibition of phospholipase C.
Biology, Brandeis University, Waltham, MA 02254-9110, USA.
Phototransduction in Limulus photoreceptors involves a G protein-mediated activation of phospholipase C (PLC) and subsequent steps involving InsP3-mediated release of intracellular Ca2+. While exploring the role of calmodulin in this cascade, we found that intracellular injection of Ca2+/calmodulin-binding peptides (CCBPs) strongly inhibited the light response. By chemically exciting the cascade at various stages, we found the primary target of this effect was not in late stages of the cascade but rather at the level of G protein and PLC. That PLCdelta1 contains a calmodulin-like structure raised the possibility that PLC might be directly affected by CCBPs. To test this possibility, in vitro experiments were conducted on purified PLC. The activity of this enzyme was strongly inhibited by CCBPs and also inhibited by calmodulin itself. Our results suggest that the calmodulin-like region of PLC has an important role in regulating this enzyme.
Department of Neurobiology, Alexander Silberman Institute of Life Sciences, Hebrew University of Jerusalem, Israel.
In Limulus ventral photoreceptor cells the time-course of the desensitization of InsP3 response was measured by an injection-pair paradigm. Pressure pulses of InsP3 were delivered into the cell with various interpulse intervals. The desensitization of the response to the second injection of each pair approached totality at 200 ms, which is the duration of the response to a single pressure pulse of InsP3. Lowering extracellular calcium did not affect the time-course of the desensitization. Lowering the temperature slowed down both the time-course of the response to InsP3 and the time-course of the desensitization to the same extent. These findings suggest that the desensitization is powerful enough and its onset fast enough to contribute to the transience of the InsP3 response. The time-course of the desensitization suggests it may influence light adaptation.
Laboratory of Neurobiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892.
The role of light in turnover of photosensitive membranes was studied in isolated photoreceptors maintained in vitro. Ventral photoreceptors of the horseshoe crab, Limulus polyphemus, were used since they have been the subjects of many in vitro physiological studies. This study shows that the two classes of ventral photoreceptors, the large and small photoreceptors (Herman: companion paper), differ in their morphological response to light. The rhabdom of small photoreceptors is remarkable for its regularity, independent of lighting condition. The photosensitive microvilli of the rhabdom of small photoreceptors are narrow and almost always tightly packed in a hexagonal arrangement. In contrast, the morphology of the rhabdom of the large ventral photoreceptors is different in the dark and in the light, and the rhabdom undergoes turnover during lighting transitions. When fully dark-adapted, the photosensitive microvilli of large photoreceptors are narrow and well organized, sometimes in a crystalline array. However, in the light-adapted state, the microvilli are much thicker and very irregular. The transitions between the dark and light-adapted states, examined at midday, are rapid. After 5 minutes light exposure, the microvilli are dilated at their bases and shed membranes are present in the cytoplasm. By 30 minutes after light onset, the appearance of the rhabdom of large photoreceptors is indistinguishable from fully light-adapted cells. The transition to the dark-adapted state is equally rapid. Even at 5 or 12 minutes after light offset, most microvilli are narrow and quite regular, and by 30 minutes, the rhabdom usually appears to be fully dark-adapted. These experiments show that both the synthetic and degradative phases of rhabdom renewal take place in isolated photoreceptors. No efferent neural activity is required to initiate turnover; rather, changes in illumination alone are sufficient to generate rhabdom turnover in large ventral photoreceptors in vitro.