Nitric Oxide Synthase Imunolabeling in the Molluscan CNS and Peripheral Tissues

Hurst, William J.; Moroz, Leonid L.; Gillette, Martha U.; Gillette, Rhanor

doi:10.1006/bbrc.1999.1247

Cited by 19 publications

(7 citation statements)

References 21 publications

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“…Outside of the rhg, Nos was also found in some neurons in the ceg and plg. This is a broader distribution than would be expected by previous studies using IHC or NADPH diaphorase staining in other nudibranchs (Hurst et al, 1999;Moroz and Gillette, 1996;Newcomb and Watson, 2001).…”

Section: Olfaction-associated Genes Show Spatial Organization In the ...supporting

confidence: 43%

Cellular-resolution gene expression mapping reveals organization in the head ganglia of the gastropod,Berghia stephanieae

Ramirez

Bui

Katz

2023

Preprint

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Gastropod molluscs, such as Aplysia, Lymnaea, and Tritonia have been important for determining fundamental rules of motor control and learning and memory because they have individually identifiable neurons. This has allowed neural circuits to be worked out using pair-wise microelectrode recordings. However, neuronal identification that relies on electrophysiology, dye tracing, and immunohistochemistry, limits research to the small number of large neurons, ignoring the much larger number of small neurons. Here we combined high throughput, single-cell RNA sequencing (scRNAseq) with in situ hybridization chain reaction (HCR) to examine gene expression in the head ganglia of the gastropod, Berghia stephanieae. Cluster analysis of the transcriptomic data uncovered multiple neuronal cell clusters, as well as glial, fibroblast, and endothelial cell clusters in the central ring ganglia (CRG: cerebral, pleural, pedal, and buccal ganglia) and the rhinophore ganglia (rhg). Combinatorial molecular signatures were layered on top of size and position using HCR with multiplexed marker genes to help establish specific neuronal class and type phenotypes. Unannotated genes made up significant proportions of the top differentially expressed genes for each cluster, including the most ubiquitously expressed pan-neuronal marker. Neuronal classes expressing major neurotransmitter phenotypes of glutamatergic, cholinergic, and serotonergic neurons were identified. Some neurons co-expressed genes for two neurotransmitters. Serotonergic neurons, not cholinergic neurons, were found to express Unc-4. Among the neurons found to be glutamatergic were a single photoreceptor in the eye and Soluble guanylate cyclase (Sgc)-expressing neurons in the rhg. A sampling of expression patterns from 8 out of the 40 molluscan neuropeptides identified in the single-cell dataset showed the tremendous diversity of neuron types, with few neuropeptides co-expressed in the same neurons. A visually identifiable giant ventral neuron was found to express three peptides and Chat. A subset of neurons in rhg, which contain only small somata (<15 microns), separated into three clusters: 1) glutamatergic + Sgc neurons, 2) nitric oxide synthase (Nos) + pigment dispersing factor (Pdf), and 3) Nos/Pdf-. The three groups were spatially restricted in the rhg. Transcription factors marked a cluster of newly differentiated neurons and were expressed in mature neurons. Six3/6 was expressed only in neurons from the rhinophore and cerebral ganglia, which are anterior-most of the major head ganglia, consistent with Six3/6 anterior segregation in other animals. This study provides the foundation for understanding the fundamental neuronal organization of the gastropod nervous system.

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Section: Olfaction-associated Genes Show Spatial Organization In the ...supporting

confidence: 43%

Cellular-resolution gene expression mapping reveals organization in the head ganglia of the gastropod,Berghia stephanieae

Ramirez

Bui

Katz

2023

Preprint

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“…NO synthase was found to be widely spread in the CNS, and the enzyme’s activity was as high as in mammalian cortical tissue. − Moreover, unlike the NO synthases characterized in other molluscs, in Pleurobranchaea , it was insensitive to Ca 2+ , not resembling the neuronal and epithelial NO synthases of vertebrates, but being more like the Ca 2+ -insensitive NO synthase of macrophages …”

Section: Intracellular Phmentioning

confidence: 99%

cAMP, Ca²⁺, pH_i, and NO Regulate H-like Cation Channels That Underlie Feeding and Locomotion in the Predatory Sea Slug Pleurobranchaea californica

Green

Huang

Sudlow

et al. 2018

ACS Chem. Neurosci.

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A systems approach to regulation of neuronal excitation in the mollusc Pleurobranchaea has described novel interactions of cyclic AMP-gated cation current (I), Ca, pH, and NO. I appears in many neurons of feeding and locomotor neuronal networks. It is likely one of the family of hyperpolarization-activated, cyclic-nucleotide-gated currents (h-current) of vertebrate and invertebrate pacemaker networks. There are two isoforms. Ca regulates both voltage dependence and depolarization-sensitive inactivation in both isoforms. The Type 1 I of the feeding network is enhanced by intracellular acidification. A direct dependence of I on cAMP allows the current to be used as a reporter on cAMP concentrations in the cell, and from there to the intrinsic activities of the synthetic adenyl cyclase and the degradative phosphodiesterase. Type 2 I of the locomotor system is activated by serotonergic inputs, while Type 1 of the feeding network is thought to be regulated peptidergically. NO synthase activity is high in the CNS, where it differs from standard neuronal NO synthase in not being Ca sensitive. NO acidifies pH, potentiating Type 1, and may act to open proton channels. A cGMP pathway does not mediate NO effects as in other systems. Rather, nitrosylation likely mediates its actions. An integrated model of the action of cAMP, Ca, pH, and NO in the feeding network postulates that NO regulates proton conductance to cause neuronal excitation in the cell body on the one hand, and relief of activity-induced hyperacidification in fine dendritic processes on the other.

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“…The molecular weight of this CNS protein is close to that of the ∼65‐kDa protein that we detected in the gastrointestinal tract, so it is predicted that both antibodies detected the same protein in Helix . On the other hand, different NOS isoforms were suggested to be present in Lymnaea (Korneev et al,1998), Aplysia , Pleurobranchaea , and Tritonia (Hurst et al,1999) CNS and PNS, which raises the possibility that the antibody used by us and that applied by Huang et al (1997) labeled different NOS isoforms in the caecum and the CNS of Helix .…”

Section: Discussionmentioning

confidence: 95%

Characterization of nitric oxidergic neurons in the alimentary tract of the snail Helix pomatia L.: Histochemical and physiological study

Serfözö

Szentmiklósi

Elekes

2007

J of Comparative Neurology

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By using NADPH-diaphorase (NADPH-d) histochemistry, nitric oxide synthase (NOS) immunohistochemistry, Western blotting, and NO pharmacology, we investigated the distribution and possible function of NOS-containing neurons in different units of the alimentary tract of the snail, Helix pomatia. Discrete populations of neurons in the buccal ganglia displayed NADPH-d reactivity. NADPH-d-reactive and NOS-immunoreactive (NOS-IR) neurons were present in the caecum, and labeled fibers were found to innervate the circular muscles of the proesophagus and caecum and to form axosomatic connections with neurons of the myenteric and submucosal plexi of the caecum. A 65-kDa protein was found to be nNOS-IR in the caecum protein extract. The majority of the NADPH-d-reactive neurons also displayed FMRFamide immunoreactivity, whereas a mutual innervation by NADPH-diaphorase-reactive and catch-relaxing peptide (CARP)-IR neurons was observed in the caecum. Application of NO-donors [glyceryl trinitrate, S-nitroso-N-acetyl-DL-penicillamine, sodium nitroprusside (SNP)] evoked a dose-dependent increase in tension, frequency, and amplitude of the spontaneous muscle contractions of the proesophagus and caecum. Contractions could be blocked by applying the NO scavenger 2-phenyl-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide. FMRFamide evoked a response of the caecum similar to that with NO, and its simultaneous application was additive. Preincubation with CARP blocked the increase of tension evoked by SNP, whereas Mytilus inhibitory peptide (MIP) decreased the rhythmic contractions induced by the NO donor. Our findings indicate that NO is an important signal molecule in the feeding system of Helix, involved, partially in cooperation with different molluscan neuropeptides, in the regulation of both neuronal and muscular activities.

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Nitric Oxide Synthase Imunolabeling in the Molluscan CNS and Peripheral Tissues

Abstract: NOS immunoreactivity was assayed in CNS and

Cited by 19 publications

References 21 publications

Cellular-resolution gene expression mapping reveals organization in the head ganglia of the gastropod,Berghia stephanieae

Cellular-resolution gene expression mapping reveals organization in the head ganglia of the gastropod,Berghia stephanieae

cAMP, Ca²⁺, pH_i, and NO Regulate H-like Cation Channels That Underlie Feeding and Locomotion in the Predatory Sea Slug Pleurobranchaea californica

Characterization of nitric oxidergic neurons in the alimentary tract of the snail Helix pomatia L.: Histochemical and physiological study

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Nitric Oxide Synthase Imunolabeling in the Molluscan CNS and Peripheral Tissues

Abstract: NOS immunoreactivity was assayed in CNS and

Cited by 19 publications

References 21 publications

Cellular-resolution gene expression mapping reveals organization in the head ganglia of the gastropod,Berghia stephanieae

Cellular-resolution gene expression mapping reveals organization in the head ganglia of the gastropod,Berghia stephanieae

cAMP, Ca2+, pHi, and NO Regulate H-like Cation Channels That Underlie Feeding and Locomotion in the Predatory Sea Slug Pleurobranchaea californica

Characterization of nitric oxidergic neurons in the alimentary tract of the snail Helix pomatia L.: Histochemical and physiological study

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cAMP, Ca²⁺, pH_i, and NO Regulate H-like Cation Channels That Underlie Feeding and Locomotion in the Predatory Sea Slug Pleurobranchaea californica