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Moroz, L. L.

Publications and source records attributed to Moroz, L. L..

26 records · Page 2Linked to original sources
bioRxiv2020

Nitric oxide (NO) signaling in Trichoplax and related species: Microchemical characterization and the lineage-specific diversification

Nitric oxide (NO) is a free radical gaseous messenger with a broad distribution across the animal kingdom. However, the early evolution of nitric oxide-mediated signaling in animals is unclear due to limited information about prebilaterian metazoans such as placozoans. Here, we analyzed NO synthases (NOS) in four different species of placozoans (haplotypes H1, H2, H4, H13). In contrast to all other invertebrates studied, Hoilungia and Trichoplax have three distinct NOS genes, including PDZ domain-containing NOS. To characterize NOS activity in Trichoplax adhaerens, we used capillary electrophoresis for microchemical assays of NO-related metabolites. Specifically, we quantified nitrites (products of NO oxidation) and L-citrulline (co-product of NO synthesis from L-arginine), which were affected by NOS inhibitors confirming the presence of functional NOS. Next, using fluorescent single-molecule in situ hybridization, we showed that distinct NOSs are expressed in different subpopulations of cells, with a noticeable distribution close to the edge regions of Trichoplax. These data suggest the compartmentalized release of this messenger and a greater diversity of cell types in placozoans than anticipated. We also revealed a dramatic diversification of NO receptor machinery, including identification of both canonical and novel NIT-domain containing soluble guanylate cyclases as putative NO/nitrite/nitrate sensors. Thus, although Trichoplax is considered to be one of the morphologically simplest free-living animals, the complexity of NO-cGMP-mediated signaling is greater to those in vertebrates. This situation illuminates multiple lineage-specific diversifications of NOSs and NO/nitrite/nitrate sensors from the common ancestor of Metazoa. Short AbstractNitric oxide (NO) is a ubiquitous gaseous messenger, but we know little about its early evolution. Here, we analyzed NO synthases (NOS) in four different species of placozoans - one of the early-branching animal lineages. In contrast to other invertebrates studied, Trichoplax and Hoilungia have three distinct NOS genes, including PDZ domain-containing NOS. Using ultra-sensitive capillary electrophoresis assays, we quantified nitrites (products of NO oxidation) and L-citrulline (co-product of NO synthesis from L-arginine), which were affected by NOS inhibitors confirming the presence of functional enzymes in Trichoplax. Using fluorescent single-molecule in situ hybridization, we showed that distinct NOSs are expressed in different subpopulations of cells, with a noticeable distribution close to the edge regions of Trichoplax. These data suggest both the compartmentalized release of NO and a greater diversity of cell types in placozoans than anticipated. NO receptor machinery includes both canonical and novel NIT-domain containing soluble guanylate cyclases as putative NO/nitrite/nitrate sensors. Thus, although Trichoplax and Hoilungia exemplify the morphologically simplest free-living animals, the complexity of NO-cGMP-mediated signaling in Placozoa is greater to those in vertebrates. This situation illuminates multiple lineage-specific diversifications of NOSs and NO/nitrite/nitrate sensors from the common ancestor of Metazoa.

evolutionary biology↗
bioRxiv2019

Comparative single-cell transcriptomics of complete insect nervous systems

Molecular profiles of neurons influence information processing, but bridging the gap between genes, circuits, and behavior has been very difficult. Furthermore, the behavioral state of an animal continuously changes across development and as a result of sensory experience. How behavioral state influences molecular cell state is poorly understood. Here we present a complete atlas of the Drosophila larval central nervous system composed of over 200,000 single cells across four developmental stages. We develop polyseq, a python package, to perform cell-type analyses. We use single-molecule RNA-FISH to validate our scRNAseq findings. To investigate how internal state affects cell state, we optogentically altered internal state with high-throughput behavior protocols designed to mimic wasp sting and over activation of the memory system. We found nervous system-wide and neuron-specific gene expression changes. This resource is valuable for developmental biology and neuroscience, and it advances our understanding of how genes, neurons, and circuits generate behavior.

neuroscience↗
bioRxiv2019

Atlas of the Neuromuscular System in the Trachymedusa Aglantha digitale (Hydrozoa.

Cnidaria is the sister taxon to bilaterian animals, and therefore, represents a key reference lineage to understand early origins and evolution of the neural systems. The hydromedusa Aglantha digitale is arguably the best electrophysiologically studied jellyfish because of its system of giant axons and unique fast swimming/escape behaviors. Here, using a combination of scanning electron microscopy and immunohistochemistry together with phalloidin labeling, we systematically characterize both neural and muscular systems in Aglantha, summarizing and expanding further the previous knowledge on the microscopic neuroanatomy of this crucial reference species. We found that the majority, if not all ([~]2500) neurons, that are labeled by FMRFamide antibody are different from those revealed by anti--tubulin immunostaining, making these two neuronal markers complementary to each other and, therefore, expanding the diversity of neural elements in Aglantha with two distinct neural subsystems. Our data uncovered the complex organization of neural networks forming a functional annulus-type central nervous system with three subsets of giant axons, dozen subtypes of neurons, muscles and a variety of receptors fully integrated with epithelial conductive pathways supporting swimming, escape and feeding behaviors. The observed unique adaptations within the Aglantha lineage (including giant axons innervating striated muscles) strongly support an extensive and wide-spread parallel evolution of integrative and effector systems across Metazoa.

evolutionary biology↗
bioRxiv2019

Profiling cellular diversity in sponges informs animal cell type and nervous system evolution

The evolutionary origin of metazoan cell types such as neurons, muscles, digestive, and immune cells, remains unsolved. Using whole-body single-cell RNA sequencing in a sponge, an animal without nervous system and musculature, we identify 18 distinct cell types comprising four major families. This includes nitric-oxide sensitive contractile cells, digestive cells active in macropinocytosis, and a family of amoeboid-neuroid cells involved in innate immunity. We uncover presynaptic genes in an amoeboid-neuroid cell type, and postsynaptic genes in digestive choanocytes, suggesting asymmetric and targeted communication. Corroborating this, long neurite-like extensions from neuroid cells directly contact and enwrap choanocyte microvillar collars. Our data indicate a link between neuroid and immune functions in sponges, and suggest that a primordial neuro-immune system cleared intruders and controlled ciliary beating for feeding.

developmental biology↗
bioRxiv2018

Neural System and Receptor Diversity in the ctenophore Beroe abyssicola

Although, neuro-sensory systems might evolve independently in ctenophores, very little is known about their neural organization. Most of the ctenophores are pelagic and deep-water species and cannot be bred in the laboratory. Thus, it is not surprising that neuroanatomical data are available for only one genus within the group - Pleurobrachia. Here, using immunohistochemistry and scanning electron microscopy, we describe the organization of two distinct neural subsystems (subepithelial and mesogleal) and the structure of different receptor types in the comb jelly Beroe abyssicola - the voracious predator from North Pacific. A complex subepithelial neural network of Beroe, with five receptor types, covers the entire body surface and expands deeply into the pharynx. Three types of mesogleal neurons are comparable to the cydippid Pleurobrachia. The predatory lifestyle of Beroe is supported by the extensive development of ciliated and muscular structures including the presence of giant muscles and feeding macrocilia. The obtained neuroanatomy atlas provides unique examples of lineage-specific innovations within these enigmatic marine animals, and remarkable complexity of sensory and effector systems in this clade of basal Metazoa.\n\nGraphical AbstractAlthough, neuro-sensory systems might evolve independently in ctenophores, very little is known about their neuroanatomy. Here, using immunohistochemistry and scanning electron microscopy, we describe the organization of two neural systems and five different receptor types in the comb jelly Beroe abyssicola - the voracious predator from North Pacific. The predatory lifestyle of Beroe is supported by the extensive development of ciliated, muscular, and nervous systems including the presence of giant muscles and exceptional feeding macrocilia. The obtained neuroanatomy atlas provides unique examples of lineage-specific innovations within this enigmatic group of marine animals.\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=150 SRC=\"FIGDIR/small/419218_figG.gif\" ALT=\"Figure 1\">\nView larger version (94K):\norg.highwire.dtl.DTLVardef@1ce7930org.highwire.dtl.DTLVardef@766e6dorg.highwire.dtl.DTLVardef@14bc5baorg.highwire.dtl.DTLVardef@1a27072_HPS_FORMAT_FIGEXP M_FIG C_FIG

evolutionary biology↗
bioRxiv2018

Atlas of Neuromuscular Organization in the Ctenophore, Pleurobrachia bachei (A. Agassiz, 1860)

Enigmatic ctenophores are descendants of one of the earliest branching metazoan lineage. Their nervous systems are equally elusive. The lack of convenient neurogenic molecules and neurotransmitters suggests an extensive parallel evolution and independent origins of neurons and synapses. However, the field is logged behind due to the lack of microanatomical data about the neuro-muscular systems in this group of animals. Here, using immunohistochemistry and scanning electron microscopy, we describe the organization of both muscular and nervous systems in the sea gooseberry, Pleurobrachia bachei, from North Pacific. The diffused neural system of Pleurobrachia consists of two subsystems: the subepithelial neural network and the mesogleal net with about 5000-7000 neurons combined. Our data revealed the unprecedented complexity of neuromuscular organization in this basal metazoan lineage. The anatomical diversity of cell types includes at least nine broad categories of neurons, five families of surface receptors and more than two dozen types of muscle cells as well as regional concentrations of neuronal elements to support ctenophore feeding, complex swimming, escape and prey capture behaviors. In summary, we recognize more than 80 total morphological cell types. Thus, in terms of cell type specification and diversity, ctenophores significantly exceed what we currently know about other prebilaterian groups (placozoan, sponges, and cnidarians), and some basal bilaterians.\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=150 SRC=\"FIGDIR/small/385435_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (114K):\norg.highwire.dtl.DTLVardef@151a0f4org.highwire.dtl.DTLVardef@1e0b71org.highwire.dtl.DTLVardef@813415org.highwire.dtl.DTLVardef@827e25_HPS_FORMAT_FIGEXP M_FIG C_FIG

evolutionary biology↗
bioRxiv2018

Polymorphism within the mitochondrial genome of the ctenophore, Pleurobrachia bachei, and its ongoing rapid evolution

The mitochondrial genomes in ctenophores are among the most compact in the animal kingdom with multiple rearrangements and examples of gene loss. Here, by resequencing of the Pleurobrachia bachei mitochondrial genome, we show that the high level of polymorphism (>10%) in Pleurobrachia might contribute to the ongoing fast evolution of ctenophores including the presence of truncated versions of apparently canonical genes such as cox1. Second, the codon interpretations in ctenophores, without robust proteomic data related to mitochondrial genes, is still a challenging issue, which is open for future experimental analyses.

evolutionary biology↗