A new look at an old visual system: structure and development of the compound eyes and optic ganglia of the brine shrimp Artemia salina Linnaeus, 1758 (Branchiopoda,anostraca)

Compared to research carried out on decapod crustaceans, the development of the visual system in representatives of the entomostracan crustaceans is poorly understood. However, the structural evolution of the arthropod visual system is an important topic in the new debate on arthropod relationships, and entomostracan crustaceans play a key role in this discussion. Hence, data on structure and ontogeny of the entomostracan visual system are likely to contribute new aspects to our understanding of arthropod phylogeny. Therefore, we explored the proliferation of neuronal stem cells (in vivo incorporation of bromodeoxyuridine) and the developmental expression of synaptic proteins (immunohistochemistry against synapsins) in the developing optic neuropils of the brine shrimp Artemia salina Linnaeus, 1758 (Crustacea, Entomostraca, Branchiopoda, Anostraca) from hatching to adulthood. The morphology of the adult visual system was examined in serial sections of plastic embedded specimens. Our results indicate that the cellular material that gives rise to the visual system (compound eyes and two optic ganglia) is contributed by the mitotic activity of neuronal stem cells that are arranged in three band-shaped proliferation zones. Synapsin-like immunoreactivity in the lamina ganglionaris and the medulla externa initiated only after the anlagen of the compound eyes had already formed, suggesting that the emergence of the two optic neuropils lags behind the proliferative action of these stem cells. Neurogenesis in A. salina is compared to similar processes in malacostracan crustaceans and possible phylogenetic implications are discussed.     

Neurogenesis in the developing visual system of the branchiopod crustacean Triops longicaudatus (LeConte, 1846): corresponding patterns of compound-eye formation in Crustacea and Insecta?

In the discussion on arthropod phylogeny, the structural evolution of compound eyes and optic ganglia in Crustacea and Insecta is an important topic. On the one hand, many morphological features as well as developmental aspects of the visual system in Insecta and Crustacea correspond in so much detail that eye design in these two groups is likely to have a common euarthropodan ancestor. On the other hand, however, some authors advocate a convergent evolution of the crustacean and insect visual system founding their arguments on differences in the arrangement of the visual neuropils and the fibre connections between Malacostraca and Entomostraca (the "entomostracan enigma"). Therefore, information about cellular aspects of visual system formation in entomostracan Crustacea is likely to enliven this debate, but is not yet available. To fill this gap, we examined the proliferation of neuronal stem cells in the developing visual system of the tadpole shrimp Triops longicaudatus (LeConte, 1846) (Entomostraca, Branchiopoda, Phyllopoda, Calmanostraca, Notostraca) by in vivo incorporation of the proliferation marker bromodeoxyuridine and subsequent immunohistochemical detection. Our results indicate that in the developing visual system of T. longicaudatus, three band-shaped zones containing neuronal stem cells are present corresponding to the proliferation zones found in Malacostraca. We therefore conclude that the ontogenetic mechanisms of visual-system formation are evolutionarily conserved (homologous) in Branchiopoda, Malacostraca, and Insecta.     

The evolution of crustacean and insect optic lobes and the origins of chiasmata

In malacostracan crustaceans and insects three nested optic lobe neuropils are linked by two successive chiasmata that reverse and then reverse again horizontal rows of retinotopic columns. Entomostracan crustaceans possess but two retinotopic neuropils connected by uncrossed axons: a distal lamina and an inner plate-like neuropil, here termed the visual tectum that is contiguous with the protocerebrum. This account proposes an evolutionary trajectory that explains the origin of chiasmata from an ancestral taxon lacking chiasmata. A central argument employed is that the two optic lobe neuropils of entomostracans are homologous to the lamina and lobula plate of insects and malacostracans, all of which contain circuits for motion detection—an archaic attribute of visual systems. An ancestral duplication of a cell lineage originally providing the entomostracan lamina is proposed to have given rise to an outer and inner plexiform layer. It is suggested that a single evolutionary step resulted in the separation of these layers and, as a consequence, their developmental connection by a chiasma with the inner layer, the malacostracan-insect medulla, still retaining its uncrossed connections to the deep plate-like neuropil. It is postulated that duplication of cell lineages of the inner proliferation zone gave rise to a novel neuropil, the lobula. An explanation for the second chiasma is that it derives from uncrossed axons originally supplying the visual tectum that subsequently supply collaterals to the opposing surface of the newly evolved lobula. A cladistic analysis based on optic lobe anatomy of taxa possessing compound eyes supports a common ancestor of the entomostracans, malacostracan crustaceans, and insects.     

Neurogenesis in the crustacean ventral nerve cord: homology of neuronal stem cells in Malacostraca and Branchiopoda?

SUMMARY In Insecta and malacostracan Crustacea, neurons in the ventral ganglia are generated by the unequal division of neuronal stem cells, the neuroblasts (Nbs), which are arranged in a stereotyped, grid‐like pattern. In malacostracans, however, Nbs originate from ectoteloblasts by an invariant lineage, whereas Nbs in insects differentiate without a defined lineage by cell‐to‐cell interactions within the neuroectoderm. As the ventral ganglia in entomostracan crustaceans were thought to be generated by a general inward proliferation of ectodermal cells, the question arose as to whether neuroblasts in Euarthropoda represent a homologous type of stem cell. In the current project, neurogenesis in metanauplii of the entomostracan crustaceans Triops cancriformis Fabricius, 1780 (Branchiopoda, Phyllopoda) and Artemia salina Linné, 1758 (Branchiopoda, Anostraca) was examined by in vivo incorporation of the mitosis marker bromodeoxyuridine (BrdU) and compared to stem cell proliferation in embryos of the malacostracan Palaemonetes argentinus Nobili, 1901 (Eucarida, Decapoda). The developmental expression of synaptic proteins (synapsins) was studied immunohistochemically. Results indicate that in the ventral neurogenic zone of Branchiopoda, neuronal stem cells with cellular characteristics of malacostracan neuroblasts are present. However, a pattern similar to the lineage‐dependent, grid‐like arrangement of the malacostracan neuroblasts was not found. Therefore, the homology of entomostracan and malacostracan neuronal stem cells remains uncertain. It is now well established that during arthropod development, identical and most likely homologous structures can emerge, although the initiating steps or the mode of generation of these structures are different. Recent evidence suggests that adult Entomostraca and Malacostraca share corresponding sets of neurons so that the present report provides an example that those homologous neurons may be generated via divergent developmental pathways. In this perspective, it remains difficult at this point to discuss the question of common patterns of stem cell proliferation with regard to the phylogeny and evolution of Atelocerata and Crustacea.     

A new look at embryonic development of the visual system in decapod crustaceans: neuropil formation, neurogenesis, and apoptotic cell death

In recent years, comparing the structure and development of the central nervous system in crustaceans has provided new insights into the phylogenetic relationships of arthropods. Furthermore, the structural evolution of the compound eyes and optic ganglia of adult arthropods has been discussed, but it was not possible to compare the ontogeny of arthropod visual systems, owing to the lack of data on species other than insects. In the present report, we studied the development of the crustacean visual system by examining neurogenesis, neuropil formation, and apoptotic cell death in embryos of the American lobster, Homarus americanus, the spider crab, Hyas araneus, and the caridean shrimp, Palaemonetes argentinus, and compare these processes with those found in insects. Our results on the patterns of stem cell proliferation provide evidence that in decapod crustaceans and hemimetabolous insects, there exist considerable similarities in the mechanisms by which accretion of the compound eyes and growth of the optic lobes is achieved, suggesting an evolutionary conservation of these mechanisms.     

Two visual systems in one eyestalk: The unusual optic lobe metamorphosis in the stomatopod Alima pacifica

The compound eyes of adult stomatopod crustaceans have two to six ommatidial rows at the equator, called the midband, that are often specialized for color and polarization vision. Beneath the retina, this midband specialization is represented as enlarged optic lobe lamina cartridges and a hernia‐like expansion in the medulla. We studied how the optic lobe transforms from the larvae, which possess typical crustacean larval compound eyes without a specialized midband, through metamorphosis into the adults with the midband in a two midband‐row species Alima pacifica. Using histological staining, immunolabeling, and 3D reconstruction, we show that the last‐stage stomatopod larvae possess double‐retina eyes, in which the developing adult visual system forms adjacent to, but separate from, the larval visual system. Beneath the two retinas, the optic lobe also contains two sets of optic neuropils, comprising of a larval lamina, medulla, and lobula, as well as an adult lamina, medulla, and lobula. The larval eye and all larval optic neuropils degenerate and disappear approximately a week after metamorphosis. In stomatopods, the unique adult visual system and all optic neuropils develop alongside the larval system in the eyestalk of last‐stage larvae, where two visual systems and two independent visual processing pathways coexist. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 78: 3–14, 2018     

Brain organization and the origin of insects: an assessment

Within the Arthropoda, morphologies of neurons, the organization of neurons within neuropils and the occurrence of neuropils can be highly conserved and provide robust characters for phylogenetic analyses. The present paper reviews some features of insect and crustacean brains that speak against an entomostracan origin of the insects, contrary to received opinion. Neural organization in brain centres, comprising olfactory pathways, optic lobes and a central neuropil that is thought to play a cardinal role in multi-joint movement, support affinities between insects and malacostracan crustaceans.     

Wiring a periscope--ocelli, retinula axons, visual neuropils and the ancestrality of sea spiders

The Pycnogonida or sea spiders are cryptic, eight-legged arthropods with four median ocelli in a ‘periscope’ or eye tubercle. In older attempts at reconstructing phylogeny they were Arthropoda incertae sedis, but recent molecular trees placed them as the sister group either to all other euchelicerates or even to all euarthropods. Thus, pycnogonids are among the oldest extant arthropods and hold a key position for the understanding of arthropod evolution. This has stimulated studies of new sets of characters conductive to cladistic analyses, e.g. of the chelifores and of the hox gene expression pattern. In contrast knowledge of the architecture of the visual system is cursory. A few studies have analysed the ocelli and the uncommon “pseudoinverted” retinula cells. Moreover, analyses of visual neuropils are still at the stage of Hanström''s early comprehensive works. We have therefore used various techniques to analyse the visual fibre pathways and the structure of their interrelated neuropils in several species. We found that pycnogonid ocelli are innervated to first and second visual neuropils in close vicinity to an unpaired midline neuropil, i.e. possibly the arcuate body, in a way very similar to ancestral euarthropods like Euperipatoides rowelli (Onychophora) and Limulus polyphemus (Xiphosura). This supports the ancestrality of pycnogonids and sheds light on what eyes in the pycnogonid ground plan might have ‘looked’ like. Recently it was suggested that arthropod eyes originated from simple ocelli similar to larval eyes. Hence, pycnogonid eyes would be one of the early offshoots among the wealth of more sophisticated arthropod eyes.     

Optic lobes of the larval and imaginal scorpionfly Panorpa vulgaris (Mecoptera,Panorpidae): A neuroanatomical study of neuropil organization,retinula axons,and lamina monopolar cells

Panorpa larvae possess stemmata (lateral ocelli), which have the structure of compound eyes, and stemma lamina and stemma medulla neuropils. A distinct lobula neuropil is lacking. The stemma neuropils have a columnar organization. They contain lamina monopolar cells, and both short and long visual fibers. All the identified larval monopolar neurons have radially arranged dendrites along the entire depth of the lamina neuropil and a single terminal arborization within the medulla (L1/L2-type). The terminals of visual fibers have short spiny lateral projections. Long fibers possess en passant synapses within the lamina. The same principles of organization of first and second order visual neuropils are found in Panorpa imagines. In contrast to the larvae, a lobula neuropil is present. Adults have monopolar cells of the L1-type that are similar to the L1-neurons found in Diptera. The columnar organization, the presence of short and long visual fibers, and lamina monopolar neurons are thus features common to both visual systems, viz., the larval (stemmata) and the imaginal (compound eyes).     

The synganglion of the jumping spider Marpissa muscosa (Arachnida: Salticidae): Insights from histology,immunohistochemistry and microCT analysis

Jumping spiders are known for their extraordinary cognitive abilities. The underlying nervous system structures, however, are largely unknown. Here, we explore and describe the anatomy of the brain in the jumping spider Marpissa muscosa (Clerck, 1757) by means of paraffin histology, X-ray microCT analysis and immunohistochemistry as well as three-dimensional reconstruction. In the prosoma, the CNS is a clearly demarcated mass that surrounds the esophagus. The anteriormost neuromere, the protocerebrum, comprises nine bilaterally paired neuropils, including the mushroom bodies and one unpaired midline neuropil, the arcuate body. Further ventrally, the synganglion comprises the cheliceral (deutocerebrum) and pedipalpal neuropils (tritocerebrum). Synapsin-immunoreactivity in all neuropils is generally strong, while allatostatin-immunoreactivity is mostly present in association with the arcuate body and the stomodeal bridge. The most prominent neuropils in the spider brain, the mushroom bodies and the arcuate body, were suggested to be higher integrating centers of the arthropod brain. The mushroom body in M. muscosa is connected to first and second order visual neuropils of the lateral eyes, and the arcuate body to the second order neuropils of the anterior median eyes (primary eyes) through a visual tract. The connection of both, visual neuropils and eyes and arcuate body, as well as their large size corroborates the hypothesis that these neuropils play an important role in cognition and locomotion control of jumping spiders. In addition, we show that the architecture of the brain of M. muscosa and some previously investigated salticids differs significantly from that of the wandering spider Cupiennius salei, especially with regard to structure and arrangement of visual neuropils and mushroom body. Thus, we need to explore the anatomical conformities and specificities of the brains of different spider taxa in order to understand evolutionary transformations of the arthropod brain.     

A developmental study of serotonin-immunoreactive neurons in the embryonic brain of the Marbled Crayfish and the Migratory Locust: Evidence for a homologous protocerebral group of neurons

It is well established that the brains of adult malacostracan crustaceans and winged insects display distinct homologies down to the level of single neuropils such as the central complex and the optic neuropils. We wanted to know if developing insect and crustacean brains also share similarities and therefore have explored how neurotransmitter systems arise during arthropod embryogenesis. Previously, Sintoni et al. (2007) had already reported a homology of an individually identified cluster of neurons in the embryonic crayfish and insect brain, the secondary head spot cells that express the Engrailed protein. In the present study, we have documented the ontogeny of the serotonergic system in embryonic brains of the Marbled Crayfish in comparison to Migratory Locust embryos using immunohistochemical methods combined with confocal laser-scan microscopy. In both species, we found a cluster of early emerging serotonin-immunoreactive neurons in the protocerebrum with neurites that cross to the contralateral brain hemisphere in a characteristic commissure suggesting a homology of this cell cluster. Our study is a first step towards a phylogenetic analysis of neurotransmitter system development and shows that, as for the ventral nerve cord, traits related to neurogenesis in the brain can provide valuable hints for resolving the much debated question of arthropod phylogeny.     

Evolution of the central complex in the arthropod brain with respect to the visual system

Modular midline neuropils, termed arcuate body (Chelicerata, Onychophora) or central body (Myriapoda, Crustacea, Insecta), are a prominent feature of the arthropod brain. In insects and crayfish, the central body is connected to a second midline-spanning neuropil, the protocerebral bridge. Both structures are collectively termed central complex. While some investigators have assumed that central and arcuate bodies are homologous, others have questioned this view. Stimulated by recent evidence for a role of the central complex in polarization vision and object recognition, the architectures of midline neuropils and their associations with the visual system were compared across panarthropods. In chelicerates and onychophorans, second-order neuropils subserving the median eyes are associated with the arcuate body. The central complex of decapods and insects, instead, receives indirect input from the lateral (compound) eye visual system, and connections with median eye (ocellar) projections are present. Together with other characters these data are consistent with a common origin of arcuate bodies and central complexes from an ancestral modular midline neuropil but, depending on the choice of characters, the protocerebral bridge or the central body shows closer affinity with the arcuate body. A possible common role of midline neuropils in azimuth-dependent sensory and motor tasks is discussed.     

Organization of deutocerebral neuropils and olfactory behavior in the centipede Scutigera coleoptrata (Linnaeus, 1758) (Myriapoda: Chilopoda)

Myriapods represent an arthropod lineage, that originating from a marine arthropod ancestor most likely conquered land independently from hexapods and crustaceans. Establishing aerial olfaction during a transition from the ocean to land requires molecules to be detected in gas phase instead of in water solution. Considering that the olfactory sense of myriapods has evolved independently from that in hexapods and crustaceans, the question arises if and how myriapods have solved the tasks of odor detection and odor information processing in air. Comparative studies between arthropod taxa that independently have established a terrestrial life style provide a powerful means of investigating the evolution of chemosensory adaptations in this environment and to understand how the arthropod nervous system evolved in response to new environmental and ecological challenges. In general, the neuroethology of myriapods and the architecture of their central nervous systems are insufficiently understood. In a set of experiments with the centipede Scutigera coleoptrata, we analyzed the central olfactory pathway with serial semi-thin sectioning combined with 3-dimensional reconstruction, antennal backfilling with neuronal tracers, and immunofluorescence combined with confocal laser-scanning microscopy. Furthermore, we conducted behavioral experiments to find out if these animals react to airborne stimuli. Our results show that the primary olfactory and mechanosensory centers are well developed in these organisms but that the shape of the olfactory neuropils in S. coleoptrata is strikingly different when compared with those of hexapods and malacostracan crustaceans. Nevertheless, the presence of distinct neuropils for chemosensory and mechanosensory qualities in S. coleoptrata, malacostracan Crustacea, and Hexapoda could indicate a common architectural principle within the Mandibulata. Furthermore, behavioral experiments indicate that S. coleoptrata is able to perceive airborne stimuli, both from live prey and from a chemical extract of the prey. These results are in line with the morphological findings concerning the well-developed olfactory centers in the deutocerebrum of this species.     

Eye and optic lobe metamorphosis in the sunburst diving beetle, Thermonectus marmoratus (Coleoptera: Dytiscidae)

Nearly nothing is known about the transition that visual brain regions undergo during metamorphosis, except for Drosophila in which larval eyes and the underlying neural structure are strongly reduced. We have studied the larvae of the sunburst diving beetle, Thermonectus marmoratus (Coleoptera: Dytiscidae), which are sophisticated visually oriented predators characterized by six elaborate stemmata on each side of the head and an associated large optic lobe. We used general neurohistological staining and 3D reconstruction to determine how the eyes and optic lobe of T. marmoratus change morphologically during metamorphosis. We find that in third (last) instar larvae, the adult neuropils are already forming de novo dorsally and slightly anteriorly to the larval neuropils, while the latter rapidly degenerate. Larval eyes are eventually reduced to distinct areas with dark pigmentation. This complete reorganization, which may be an evolutionarily conserved trait in holometabolous insects, occurs despite the considerable costs that must apply to such a visually complex animal. Our findings are consistent with the concept that stemmata are homologous to the most posterior ommatidia of hemimetabolous insects, an idea also recently supported by molecular data.     

蜜蜂复眼的视觉通路研究进展

视觉通路的研究在神经科学、 仿生应用和医学治疗上都具有十分重要的意义。西方蜜蜂Apis mellifera作为神经生物学研究的重要模式生物已被广泛地应用于视觉通路的研究。蜜蜂的视觉器官包括1对复眼和3只单眼, 复眼是形成视觉的主要感觉器官。视叶是蜜蜂传递和处理视觉信息的主要神经构造, 它包括视神经节层、 视髓质层、 视小叶和前视结节4个等级的神经纤维网。复杂的视觉信息在经过大脑的各级神经时被分离, 以许多空间隔离的并行连续的视觉通路传递和加工, 然后汇集到高级脑中枢, 部分甚至与其他感觉模态的信息相整合, 最终输出有效信息来调控蜜蜂的各种行为。本文按照信息在视叶中逐级传递的顺序对蜜蜂复眼的视觉通路研究进展进行综述。     

Persisting stemma neuropils in Chaoborus crystallinus (Diptera: Chaoboridae): Development and evolution of a bipartite visual system

Stemmata or “larval” eyes are of crucial importance for the understanding of the evolution and ontogeny of the hexapod's main visual organs, the compound eyes. Using classical neuroanatomical techniques, I showed that the persisting stemmata of Chaoborus imagos are connected to persisting stemma neuropils neighboring the first and second order neuropils of the compound eyes, and therefore also the imago possesses a stemma lamina and medulla closely associated with the architecture and the developmental pattern of those of the compound eyes. The findings are compared with other arthropods, e.g. accessory lateral eyes in Amandibulata and Myriapoda, suggesting some ancestral rather than derived character states. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

Proliferation pattern and early differentiation of the optic lobes in Drosophila melanogaster

Summary The larval and early pupal development of the optic lobes in Drosophila is described qualitatively and quantitatively using [³H]thymidine autoradiography on 2-m plastic sections. The optic lobes develop from 30–40 precursor cells present in each hemisphere of the freshly hatched larva. During the first and second larval instars, these cells develop to neuroblasts arranged in two epithelial optic anlagen. In the third larval instar and in the early pupa these neuroblasts generate the cells of the imaginal optic lobes at discrete proliferation zones, which can be correlated with individual visual neuropils.The different neuropils as well as the repetitive elements of each neuropil are generated in a defined temporal sequence. Cells of the medulla are the first to become postmitotic with the onset of the third larval instar, followed by cells of the lobula complex and finally of the lamina at about the middle of the third instar. The elements of each neuropil connected to the most posterior part of the retina are generated first, elements corresponding to the most anterior retina are generated last.The proliferation pattern of neuroblasts into ganglion mother cells and ganglion cells is likely to include equal as well as unequal divisions of neuroblasts, followed by one or two generations of ganglion mother cells. For the lamina the proliferation pattern and its temporal coordination with the differentiation of the retina are shown.     

SIFamide in the brain of the sphinx moth, Manduca sexta

SIFamides form a group of highly conserved neuropeptides in insects, crustaceans, and chelicerates. Beyond their biochemical commonalities, the neuroanatomical distribution of SIFamide in the insect nervous system also shows a remarkable degree of conservation. Thus, expression of SIFamide has been found to be restricted to four neurons of the pars intercerebralis in different holometabolous species. By means of immunohistological stainings, we here show that in Manduca sexta, those four cells are complemented by additional immunoreactive cells located in the vicinity of the mushroom body calyx. Immunopositive processes form arborizations throughout the brain, innervating major neuropils like the antennal lobes, the central complex, and the optic neuropils.     

Nonvisual photoreceptors in arthropods with emphasis on their putative role as receptors of natural Zeitgeber stimuli

In various insect and arachnid species, three different types of photoreceptors that do not serve image processing have been discovered and analyzed by means of neurobiological methods: They can be found for example: (1) as lamina and lobula organs (LaOs and LoOs) next to the optic neuropils in the optic lobes of holo- and hemimetabolous insects; (2) inside the last ganglia of the cord of the scorpion and a marine midge; and (3) as modified visual photoreceptors in metamorphosized larval stemmata and the lateral eyes of scorpions, which have been compound eyes in fossil scorpion relatives. Imnunocytology with various antibodies against proteins of the phototransduction cascade, the rhabdom turnover cycle and neurotransmitters of afferent and efferent pathways, was combined with light- and ultrastructural investigations in well-defined adaptational states, in order to study their photoreceptive function and neuronal wiring. Pilot chronobiological experiments with a newly developed twilight simulating lamp, behavioral studies, and model calculations provide evidence that these photoreceptors may well serve a role in the complex task of detecting time cues out of natural dawn and dusk.