Use of axonal projection patterns for the homologisation of cerebral nerves in Opisthobranchia, Mollusca and Gastropoda

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Abstract

Introduction: Gastropoda are guided by several sensory organs in the head region, referred to as cephalic sensory organs (CSOs). These CSOs are innervated by distinct nerves. This study proposes a unified terminology for the cerebral nerves and the categories of CSOs and then investigates the neuroanatomy and cellular innervation patterns of these cerebral nerves, in order to homologise them. The homologisation of the cerebral nerves in conjunction with other data, e.g. ontogenetic development or functional morphology, may then provide insights into the homology of the CSOs themselves.Results: Nickel-lysine axonal tracing (" backfilling" ) was used to stain the somata projecting into specific nerves in representatives of opisthobranch Gastropoda. Tracing patterns revealed the occurrence, size and relative position of somata and their axons and enabled these somata to be mapped to specific cell clusters. Assignment of cells to clusters followed a conservative approach based primarily on relative location of the cells. Each of the four investigated cerebral nerves could be uniquely identified due to a characteristic set of soma clusters projecting into the respective nerves via their axonal pathways.Conclusions: As the described tracing patterns are highly conserved morphological characters, they can be used to homologise nerves within the investigated group of gastropods. The combination of adequate number of replicates and a comparative approach allows us to provide preliminary hypotheses on homologies for the cerebral nerves. Based on the hypotheses regarding cerebral nerve homology together with further data on ultrastructure and immunohistochemistry of CSOs published elsewhere, we can propose preliminary hypotheses regarding homology for the CSOs of the Opisthobranchia themselves. © 2013 Klussmann-Kolb et al.; licensee BioMed Central Ltd.

Figures

  • Table 1 Terminology of cerebral nerves in Opisthobranchia
  • Table 2 Innervation of CSOs by cerebral nerves; categories of
  • Figure 1 Cephalic sensory organs (CSOs) within the Opisthobranchia. A: Aplysia punctata, picture showing the rhinophores and the labial tentacles. B: Pleurobranchaea meckeli, picture showing the rhinophores, the labial tentacles and the oral veil. C: Archidoris pseudoargus, picture showing the rhinophores, the labial tentacles and the oral veil, usually the labial tentacles and the oral veil are completely covered by the massive mantle. In the picture shown, part of the mantle is upturned and the labial tentacles and the oral veil are highlighted. Abbreviations (referring to all subfigures, colors indicate the nervous innervation of the CSOs). N2 (N2a= inner branch, N2b= outer branch) in blue and the N3 in red, LT – labial tentacle, OV – oral veil, RH – rhinophore.
  • Figure 2 Neuroanatomy. A: Neuroanatomical scheme of the four cerebra Aplysia show no significant differences (only right hemisphere shown). B: N Pleurobranchomorpha Pleurobranchaea meckeli (only right hemisphere sho Archidoris pseudoargus (only right hemisphere shown). Abbreviations (referr yellow, N2 (N2a= inner branch, N2b= outer branch) in blue, N3 in red and
  • Figure 3 Scheme for the development of the axonal tracing patterns as shown in Figures 4, 5, 6. The first image (A) shows a photograph of part of the right hemisphere of the cerebral ganglion for an axonal tracing of the Nclc of Aplysia punctata. The stained nerve, axons and somata are labeled. B shows a close-up of A and several clusters are indicated. The white asterisks mark somata which are not found in all replicates and the cluster Cnclc2 (see C and D) is not visible here. The schematic in Figure C shows the first step in developing the schematics shown in Figures 4, 5, 6. First the location of the clusters is determined by axonal tracings in several replicates viewed from different angles. After the common position is defined, the clusters are color-coded, and finally in D mirrored to the other hemisphere and the common axonal pathways and somata are added in the original hemisphere. It is important to understand that D is a schematic based on multiple replicates (including B) and that somata which have not been found in all replicates are not shown in D.
  • Figure 4 Cellular innervation patterns of the cerebral nerves in Aplysia spp. Schematic outline of somata and their axons projecting into the N1 (A), N2 (B), N3 (C) and Nclc (D). The size and position of the somata are digitalized from camera lucida drawings, the distribution of the axons are averaged over all replicates. CG - cerebral ganglia, N1 - Nervus oralis, N2 - Nervus labialis, N3 - Nervus rhinophoralis, Nclc - Nervus clypei capitis, N. Opt. - Nervus opticus, , PdG - pedal ganglia, PlG - pleural ganglia.
  • Figure 5 Cellular innervation patterns of the cerebral nerves in Pleurobranchaea meckeli. Schematic outline of somata and their axons projecting into the N1 (A), N2 (B), N3 (C) and Nclc (D). The size and position of the somata are digitalized from camera lucida drawings, the distribution of the axons are averaged over all replicates. CG - cerebral ganglia, N1 - Nervus oralis, N2 - Nervus labialis, N3 - Nervus rhinophoralis, Nclc - Nervus clypei capitis, N. Opt. - Nervus opticus, PdG - pedal ganglia, PlG - pleural ganglia.
  • Figure 6 Cellular innervation patterns of the cerebral nerves in Archidoris pseudoargus. Schematic outline of somata and their axons projecting into the N1 (A), N2 (B), N3 (C) and Nclc (D). The size and position of the somata are digitalized from camera lucida drawings, the distribution of the axons are averaged over all replicates. CG - cerebral ganglia, N1 - Nervus oralis, N2 - Nervus labialis, N3 - Nervus rhinophoralis, Nclc - Nervus clypei capitis, PdG - pedal ganglia, PlG - pleural ganglia.

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Klussmann-Kolb, A., Croll, R. P., & Staubach, S. (2013). Use of axonal projection patterns for the homologisation of cerebral nerves in Opisthobranchia, Mollusca and Gastropoda. Frontiers in Zoology, 10(1). https://doi.org/10.1186/1742-9994-10-20

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