Eye regeneration in the mystery snail

Mystery snails (Family Ampullariidae) are aquatic prosobranchs which possess structurally complex eyes at the tip of a cephalic eyestalk. No other sensory organs are found in association with this stalk. These snails possess the ability to regenerate the eye completely after amputation through the mid-eyestalk. Amputation induces gross changes in the cellular character of the entire eyestalk; in particular, an invagination of integumentary epithelium at the apex of the eyestalk stump produces a shallow cleft or "eyecup." Differentiation of all components of the eye apparently occurs by transdetermination of these epithelial cells. Retinal differentiation and the appearance of a new lens is observed as soon as 14 days postamputation. Complete eyes (by external observation), although smaller than the originals, have regenerated by 25 days postamputation. We compare this regeneration to the reconstruction of other animal tissues, in particular the regeneration of amphibian limbs.     

Restoration of morphological and functional integrity in the regenerating eye of the giant African land snail Achatina fulica

To determine whether vision returns to its original state following eye removal in Achatina fulica, light and electron microscope examinations, electrophysiological recordings and behavioural tests were carried out on the regenerating snails. Reparative morphogenesis can result in the restoration of the peripheral sense organ even in the absence of complete regrowth of the tentacle, but it can also lead to the formation of aberrant regenerates. We found that anatomically and ultrastructurally the eyes of the ‘most normal’ regenerates were basically the same as the original eyes. Under normal conditions each eye is composed of a principal and an accessory eye, both sharing a common cornea. The only difference between regenerated and native eyes is the smaller size of the former, as a result of a reduced number of retinal cells. Electroretinographic responses revealed that the molecular mechanism of phototransduction is restored, in principle, but that flicker fusion frequency in the regenerated eye is significantly lower than in the normal eye. The directional movement to a visual stimulus (a black stripe of 45° width) had not completely recovered even 6 months after amputation. This suggests that the central projections of the optic nerve had not become fully re‐established at the time of testing.     

Electroretinographic and ultrastructural study of the regenerated eye of the snail cryptomphallus aspersa

The electroretinographic responses of regenerated eyes of snails (C. aspersa) were studied by means of suction electrodes and single or repetitive flash stimulation. The eyes were fixed and observed under light and electron microscopy. The results indicate that the electroretinographic response of regenerated eyes does not differ from control eyes after dark adaptation. However, the repetitive stimulation of the regenerated eyes induced an earlier fatigue of the response, evident after the fifth stimulus. This fatigue is a function of light intensity. Ultrastructural features of the regenerated eyes are similar to those of the control eyes but regenerated eyes show smaller rhabdomeres, fewer photic vesicles, and fewer paracrystalline bodies. It is suggested that the regenerated eye lability to the repetitive stimulation might be due to the decrease in the amount of photic vesicles and paracrystalline bodies, to the decrease in membrane surface of the rhabdomeres, or to some other metabolic failure not distinguished at electron microscopic level, like the membrane ionic regulation. The appearance of photic vesicles in the axon cones of regenerated and stimulated cells also suggests a possible failure in the mechanism of transport of vesicles.     

Repair process and enzymatic activity associated with induction of shell regeneration in the invasive species Pomacea canaliculata

ABSTRACT

This study investigated the repair process and enzymatic activity responses after induction of shell regeneration in Pomacea canaliculata. The survival rates, height of regenerated shell, and activities of alkaline phosphatase (ALP) and carbonic anhydrase (CA) were investigated and recorded for 15 days. A triangular piece of shell about 8?mm wide was excised from the ventral region of the shell. On the basis of photographs and enzyme assay, it was concluded both yellow-shell apple snails and black-shell apple snails had high survival rates (100%). The regenerated new shell was initially thin, with the wound sealed 5 days after induction of shell regeneration; after that the regenerated shell thickened. Three snails were sampled at 0, 0.5, 1, 2, 3, 5, 10 and 15 days after shell excision for analysis of enzymatic activity of ALP and CA. Trends of the activity of ALP and CA in the mantle were similar. Initially they both showed a decline on the first day, then rose on following days. Activity of ALP declined to a minimum on the first day, and an activity peak in both colour forms appeared on the fifth day (P?<?0.05) and was maintained for the rest of the experiment. The minimum value of CA occurred after 0.5 days, and a significant increase was also observed on the fifth day (P?<?0.05). This study has revealed that apple snails have good shell regeneration abilities, and that ALP and CA probably play a crucial role in shell growth and regeneration.     

Aerial and aquatic visual acuity of the grey bichir Polypterus senegalus,as estimated by optokinetic response

The present study assessed the aerial and aquatic visual abilities of juvenile grey bichir Polypterus senegalus, fish capable of terrestrial locomotion, by measuring the optokinetic response to stimuli of varying speed and spatial frequency. In water, fish tracked slow-moving (2° s⁻¹) stimuli moderately well and fast-moving stimuli very poorly. Spatial acuity was very low compared with many other species, with maximum response observed at 0.05–0.075 stimulus cycles per degree of visual arc; however, it should be noted that adult fish, with their larger eyes, are likely to have somewhat improved spatial acuity. Low spatial acuity and limited stimulus tracking ability might be expected in a nocturnal ambush predator such as P. senegalus, where gaze stabilization may be less crucial and other sensory inputs may have greater importance in perception of the environment. In air, spatial and temporal acuity were both poorer by every measure, but some visual ability persisted. As the eye shows no anatomical specialization for aerial vision, poor vision was expected; however, the large decrease in saccade velocity observed in air trials was unexpected. Stimulus parameters typically have little effect on the characteristics of the saccade, so this finding may suggest that the function of the reflex system itself could be compromised in the aerial vision of some fishes capable of terrestrial locomotion.     

The effects of Acanthaster planci predation on populations of two species of massive coral

Species of the freshwater planarian genus Polycelis have a variable number of eyes in the head, typically more than a hundred. To elucidate the mechanisms determining the number of eyes, we investigated the relation between eye number and body length in Polycelis sapporo (Ijima & Kaburaki), a non-fissioning species, and P. auriculata Ijima & Kaburaki, a fissioning species. In P. sapporo reared at 7–8 °C, a positive correlation existed between number of eyes and body length. Eye number decreased with starvation. A similar correlation was true of P. auriculata. In specimens of P. auriculata undergoing regeneration, the rate of eye formation was higher in newly formed heads originating from larger tail-pieces than in those from smaller pieces. As a head regenerated from a tail piece or as the body size increased with feeding, the number of eyes increased. These results suggest that the number of eyes in an individual of Polycelis is determined by body length. The fine structure of the photoreceptor and pigment cells in the eyes of P. auriculata is similar to that of comparable cells in the pair of eyes in Dugesia despite the difference in the number of cells comprising an eye.     

Regeneration of excised shell by the invasive apple snail Pomacea canaliculata

After excision of shell from Pomacea canaliculata, survival rates, repair process, haemocyte response, and calcium content were observed and quantified for three weeks. The following experimental treatments were used: black-colored individuals with large (BL-group) or small (BS-group) shell heights, yellow-colored individuals with large (YL-group) or small (YS-group) shell heights. The survival rates in BL-, YL-, BS-, and YS-groups after making partial excisions of snail shells were, respectively, 93.59, 94.87, 92.31, and 96.15%. Freshly regenerated shell could be seen at 1 day among the four groups after induction of shell regeneration. Regeneration shell filled the wound after 5–10 days. The area of regenerated shell in yellow-colored individuals was larger than that of black-colored individuals, but the thickness of regenerated shell was not significantly different. Multiple (four times) inductions of shell regeneration significantly enhanced the thickness of newly formed shells. Two types of circulating haemocytes (hyalinocytes and granulocytes) were identified under light microscopy based on Wright’s staining. Artificial induction of shell regeneration rendered a significant increase in the number of total circulating haemocytes which was followed by a decrease to pre-wounding levels. This peak value of total circulating haemocytes was 2.86, 2.43, 1.69, and 1.71-fold higher than pre-wounding levels in group BL, YL, BS, and YS, respectively. The number of total circulating haemocytes in yellow-colored individuals was significantly higher than in black-colored. Calcium concentrations in the mantles of snails are favorable for calcium nucleation. It is postulated that calcium content increased in the mantle increase at the start of the experiment and then returned to pre-wounding levels at the end of the experiment. This study shows that apple snails have effective shell regeneration abilities and that haemocytes and calcium transportation appear to play an important role in shell growth and regeneration.     

Fixational Eye Movements in the Earliest Stage of Metazoan Evolution

All known photoreceptor cells adapt to constant light stimuli, fading the retinal image when exposed to an immobile visual scene. Counter strategies are therefore necessary to prevent blindness, and in mammals this is accomplished by fixational eye movements. Cubomedusae occupy a key position for understanding the evolution of complex visual systems and their eyes are assumedly subject to the same adaptive problems as the vertebrate eye, but lack motor control of their visual system. The morphology of the visual system of cubomedusae ensures a constant orientation of the eyes and a clear division of the visual field, but thereby also a constant retinal image when exposed to stationary visual scenes. Here we show that bell contractions used for swimming in the medusae refresh the retinal image in the upper lens eye of Tripedalia cystophora. This strongly suggests that strategies comparable to fixational eye movements have evolved at the earliest metazoan stage to compensate for the intrinsic property of the photoreceptors. Since the timing and amplitude of the rhopalial movements concur with the spatial and temporal resolution of the eye it circumvents the need for post processing in the central nervous system to remove image blur.     

Resprouting after disturbance in the short-lived herb Rorippa palustris (Brassicaceae): an experiment with juveniles

The impact of plant age, severity of injury and nutrient levels on the ability to resprout from roots was experimentally assessed in juveniles of the short-lived herb Rorippa palustris (L.) Besser. In a chamber experiment, six cohorts of young plants (1–6 week old) were injured to obtain data on the threshold age for the ability to resprout from roots. We found that plant age was an important factor influencing resprouting ability: injured individuals older than 5 weeks were able to resprout, but not plants younger than 3 weeks. The impact of injury severity (defoliation and removal of axillary buds) and nutrient levels on resprouting ability was assessed on juveniles in a greenhouse experiment. Injury induced growth of new shoots from root buds, while the number of adventitious buds on roots was not influenced by injury. Both injury treatments had a similar effect in this respect, and the amount of regenerated biomass and the extent of regeneration were not different among injury treatments. The number of new shoots produced after injury was higher at the high nutrient level, but the number of formed adventitious buds on roots was not influenced by nutrient level. Nutrient level also influenced the amount of regenerated biomass, but the extent of regeneration (regenerated/removed biomass) was not influenced. The short-lived monocarpic species R. palustris is able to resprout from roots relatively easily. This ability seems to be advantageous in disturbed habitats and this idea is discussed throughout the paper.     

Functional Recovery of Regenerating Motor Axons is Delayed in Mice Heterozygously Deficient for the Myelin Protein P0 Gene

Mice with a heterozygous knock-out of the myelin protein P₀ gene (P₀+/?) develop a neuropathy similar to human Charcot–Marie–Tooth disease. They are indistinguishable from wild-types (WT) at birth and develop a slowly progressing demyelinating neuropathy. The aim of this study was to investigate whether the regeneration capacity of early symptomatic P₀+/? is impaired as compared to age matched WT. Right sciatic nerves were lesioned at the thigh in 7–8 months old mice. Tibial motor axons at ankle were investigated by conventional motor conduction studies and axon excitability studies using threshold tracking. To evaluate regeneration we monitored the recovery of motor function after crush, and then compared the fiber distribution by histology. The overall motor performance was investigated using Rotor-Rod. P₀+/? had reduced compound motor action potential amplitudes and thinner myelinated axons with only a borderline impairment in conduction and Rotor-Rod. Plantar muscle reinnervation occurred within 21 days in all mice. Shortly after reinnervation the conduction of P₀+/? regenerated axons was markedly slower than WT, however, this difference decayed with time. Nevertheless, after 1 month, regenerated P₀+/? axons had longer strength-duration time constant, larger threshold changes during hyperpolarizing electrotonus and longer relative refractory period. Their performance at Rotor-Rod remained also markedly impaired. In contrast, the number and diameter distribution of regenerating myelinated fibers became similar to regenerated WT. Our data suggest that in the presence of heterozygously P₀ deficient Schwann cells, regenerating motor axons retain their ability to reinnervate their targets and remyelinate, though their functional recovery is delayed.     

Changes in visual fields associated with web reduction in the spider family uloboridae

When visual fields of the primitive orb-weaver, Waitkera waitkerensis, are reconstructed using measurements taken from intact lenses and cross and longitudinal sections of the prosoma, they show that this species has complete visual surveillance, but that none of the visual fields of its eight eyes overlap. The more advanced orb-weaver, Uloborus glomosus, also has eight eyes, but each eye has a greater visual angle, giving this species a complex pattern of overlapping visual fields. Uloborids that spin reduced webs are characterized by reduction or loss of the four anterior eyes and other carapace modifications necessary for them to effectively monitor and manipulate their reduced webs. The eyes of these uloborids have greater visual angles than those of orb-weavers, resulting primarily from perimetric expansion of their retinal hemispheres. Additionally, the axes of their visual fields are more ventrally directed due to greater dorsal than ventral retinal expansion and to ventral redirection of the entire eye. Consequently, even though the anterior lateral eyes of the triangle-weaver Hyptiotes cavatus lack retinae, the species' six functional eyes permit complete visual surveillance and exhibit visual overlap. The single-line-weaver, Miagrammopes animotus, has lost its four anterior eyes, and with them much of the anterior vision and all of the visual overlap found in the other species. However, changes similar to those of H. cavatus permit this species to retain most if its dorsal and ventral visual surveillance. Thus, ocular changes act in consort to maintain relatively complete visual surveillance in the face of eye loss and other major carapace modifications necessary for the operation of reduced webs.     

Failure of interocular transfer in two types of learning in the crab Chasmagnathus

Crabs (Chasmagnathus granulatus) were trained monocularly and then tested after 24 h, in order to study interocular transfer (IOT) in two habituation paradigms. 1) Habituation of the escape response to an iterated visual danger stimuli. Monocular crabs exhibited retention after 24 h when trained and tested with the same uncovered eye, but not with different uncovered eyes. 2) Habituation of the exploratory activity to a novel environment. This long term habituation of the exploratory activity is exhibited after 24 h in monocular animals trained and tested with the same uncovered eye but not with different uncovered eyes. An explanation of this failure of IOT will be discussed in terms of retrieval impairment due to a) a different perception of training and testing stimuli, b) lateralization of memory storage, or c) the existence of two symmetrical, independent and redundant lateral storage sites.     

Formation of the function of the photosensing system in early development

The function of simple eyes in two planarian species, two-eyed Girardia tigrina and multi-eyed Polycelis tenuis, has been studied. When exposed to light, planarians display a light avoidance reaction known as negative phototaxis. This reaction has been investigated in intact animals and in head and tail fragments in the course of eye regeneration after their section. Specific features of the phototaxis reaction have been described in all groups of animals. The differences in light response recovery were shown between two planarian species and two regenerating fragments. No correlation has been found between phototactic reactions and restoration of eye structure, the number of eyes, maturation of the ganglion, growth of regenerative blastema, and motor system. The phototactic response occurred two days after the recovery of the morphology of eyes and their connection with the brain. The participation of conserved and novel genes in early development of the eye function is discussed.     

Visual pigment spectra of the comma butterfly,<Emphasis Type="Italic"> Polygonia c-album</Emphasis>, derived from in vivo epi-illumination microspectrophotometry

The visual pigments in the compound eye of the comma butterfly, Polygonia c-album, were investigated in a specially designed epi-illumination microspectrophotometer. Absorption changes due to photochemical conversions of the visual pigments, or due to light-independent visual pigment decay and regeneration, were studied by measuring the eye shine, i.e., the light reflected from the tapetum located in each ommatidium proximal to the visual pigment-bearing rhabdom. The obtained absorbance difference spectra demonstrated the dominant presence of a green visual pigment. The rhodopsin and its metarhodopsin have absorption peak wavelengths at 532 nm and 492 nm, respectively. The metarhodopsin is removed from the rhabdom with a time constant of 15 min and the rhodopsin is regenerated with a time constant of 59 min (room temperature). A UV rhodopsin with metarhodopsin absorbing maximally at 467 nm was revealed, and evidence for a blue rhodopsin was obtained indirectly.     

Visual selection and maintenance of the cell lines with high plant regeneration ability and low ploidy level in Dianthus acicularis by monitoring with flow cytometry analysis

Efficient plant regeneration system from cell suspension cultures was established in D. acicularis (2n=90) by monitoring ploidy level and visual selection of the cultures. The ploidy level of the cell cultures closely related to the shoot regeneration ability. The cell lines comprising original ploidy levels (2C+4C cells corresponding to DNA contents of G1 and G2 cells of diploid plant, respectively) showed high regeneration ability, whereas those containing the cells with 8C or higher DNA C-values showed low or no regeneration ability. The highly regenerable cell lines thus selected consisted of compact cell clumps with yellowish color and relatively moderate growth, suggesting that it is possible to select visually the highly regenerable cell lines with the original ploidy level. All the regenerated plantlets from the highly regenerable cell cultures exhibited normal phenotypes and no variations in ploidy level were observed by flow cytometry (FCM) analysis.     

Initial findings on visual acuity thresholds in an African elephant (Loxodonta africana)

There are only a few published examinations of elephant visual acuity. All involved Asian elephants (Elephas maximus) and found visual acuity to be between 8′ and 11′ of arc for a stimulus near the tip of the trunk, equivalent to a 0.50 cm gap, at a distance of about 2 m from the eyes. We predicted that African elephants (Loxodonta africana) would have similarly high visual acuity, necessary to facilitate eye‐trunk coordination for feeding, drinking and social interactions. When tested on a discrimination task using Landolt‐C stimuli, one African elephant cow demonstrated a visual acuity of 48′ of arc. This represents the ability to discriminate a gap as small as 2.75 cm in a stimulus 196 cm from the eye. This single‐subject study provides a preliminary estimate of the visual acuity of African elephants. Zoo Biol 29:30–35, 2010. © 2009 Wiley‐Liss, Inc.     

Lateral asymmetry of eye use in Octopus vulgaris

The lateralization of sensory and motor functions has been recently demonstrated in various groups of vertebrates. We examined lateral asymmetry of eye use in Octopus vulgaris by behavioural methods. Octopus vulgaris uses monocular vision almost exclusively and can move its eyes independently. The amount of binocular vision is small because the eyes are on the sides of the head. We tested eight octopuses in two conditions (one with and one without moving stimuli) where the use of the eye for frontal vision could be determined unequivocally. Data were recorded on videotape. All animals showed a preference for one eye (five left, three right). There was no correlation between eye use and the animal's direction of movement. Pigmentation of the ventral side of the arms tended to be most intense on the side of the preferred eye and the body was most pigmented on the side of the eye currently in use. We found no sex differences for visual lateralization. Pigmentation of the ventral side of the arms was lighter in females than in males. Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.     

Ocular changes accompanying eye loss in the spider family uloboridae

In uloborid spiders, eye loss is accompanied by increased visual angles, optical material investment, and potential visual acuity of the retained eyes. Relative to carapace volume, the six-eyed Hyptiotes cavatus and two four-eyed Miagrammopes species have greater retinal hemisphere areas and lens volumes than do the eight-eyed uloborids Waitkera waitkerensis, Uloborus glomosus, and Octonoba sinensis. In Waitkera, in which the eyes have little visual overlap, and in Miagrammopes, in which eye loss simplifies the spiders' patterns of visual overlap, increased retinal cell density enhances potential visual acuity. However, this occurs at the expense of potential retinal cell sensitivity.     

Low-Vision Rehabilitation by Means of MP-1 Biofeedback Examination in Patients with Different Macular Diseases: A Pilot Study

Macular disease is one of the main causes of visual impairment. We studied the efficacy of low-vision rehabilitation by means of MP-1 biofeedback examination in patients with different macular disease. Five patients were enrolled (3 female and 2 male, mean age 53.8 years) and a total of 9 eyes was examined: 2 eyes with vitelliform dystrophy, 1 with a post-traumatic macular scar, 2 with Stargardt disease, 2 with myopic macular degeneration, 2 with cone dystrophy. All the patients underwent the following tests: visual acuity, reading speed, fixation test, MP-1 microperimetry. Low-vision rehabilitation, which lasted 10 weeks, consisted of 10 training sessions of 10 min for each eye, performed once a week using the MP-1 biofeedback examination. Statistical analysis was performed using Student’s t-test. p values less than 0.05 were considered statistically significant. After training all patients displayed an improvement in visual acuity, fixation behaviour, retinal sensitivity and reading speed. Fixation behaviour within the 2° diameter circle improved and was statistically significant for reading speed (p = 0.01). Reading speed improved from a mean value of 64.3 to 92 words/min. Our results show that audio feedback can, by increasing attentional modulation, help the brain to fix the final preferred retinal locus. Audio feedback facilitates stimuli transmission between intraretinal neurons as well as between the retina and brain, which is where the highest level of stimuli processing occurs, thereby probably supporting a “remapping phenomenon”.     

The knowledge base of the oculomotor system.

In everyday life, eye movements enable the eyes to gather the information required for motor actions. They are thus proactive, anticipating actions rather than just responding to stimuli. This means that the oculomotor system needs to know where to look and what to look for. Using examples from table tennis, driving and music reading we show that the information the eye movement system requires is very varied in origin and highly task specific, and it is suggested that the control program or schema for a particular action must include directions for the oculomotor and visual processing systems. In many activities (reading text and music, typing, steering) processed information is held in a memory buffer for a period of about a second. This permits a match between the discontinuous input from the eyes and continuous motor output, and in particular allows the eyes to be involved in more than one task.