Intranasal Administration of GRP78 Protein (HSPA5) Confers Neuroprotection in a Lactacystin-Induced Rat Model of Parkinson's Disease.

The accumulation of misfolded and aggregated α-synuclein can trigger endoplasmic reticulum (ER) stress and the unfolded protein response (UPR), leading to apoptotic cell death in patients with Parkinson's disease (PD). As the major ER chaperone, glucose-regulated protein 78 (GRP78/BiP/HSPA5) plays a key role in UPR regulation. GRP78 overexpression can modulate the UPR, block apoptosis, and promote the survival of nigral dopamine neurons in a rat model of α-synuclein pathology. Here, we explore the therapeutic potential of intranasal exogenous GRP78 for preventing or slowing PD-like neurodegeneration in a lactacystin-induced rat model. We show that intranasally-administered GRP78 rapidly enters the substantia nigra pars compacta (SNpc) and other afflicted brain regions. It is then internalized by neurons and microglia, preventing the development of the neurodegenerative process in the nigrostriatal system. Lactacystin-induced disturbances, such as the abnormal accumulation of phosphorylated pS129-α-synuclein and activation of the pro-apoptotic GRP78/PERK/eIF2α/CHOP/caspase-3,9 signaling pathway of the UPR, are substantially reversed upon GRP78 administration. Moreover, exogenous GRP78 inhibits both microglia activation and the production of proinflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), via the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway in model animals. The neuroprotective and anti-inflammatory potential of exogenous GRP78 may inform the development of effective therapeutic agents for PD and other synucleinopathies.

Aquaporin-4 and Parkinson's Disease.

The water-selective channel aquaporin-4 (AQP4) is implicated in water homeostasis and the functioning of the glymphatic system, which eliminates various metabolites from the brain tissue, including amyloidogenic proteins. Misfolding of the α-synuclein protein and its post-translational modifications play a crucial role in the development of Parkinson's disease (PD) and other synucleopathies, leading to the formation of cytotoxic oligomers and aggregates that cause neurodegeneration. Human and animal studies have shown an interconnection between AQP4 dysfunction and α-synuclein accumulation; however, the specific role of AQP4 in these mechanisms remains unclear. This review summarizes the current knowledge on the role of AQP4 dysfunction in the progression of α-synuclein pathology, considering the possible effects of AQP4 dysregulation on brain molecular mechanisms that can impact α-synuclein modification, accumulation and aggregation. It also highlights future directions that can help study the role of AQP4 in the functioning of the protective mechanisms of the brain during the development of PD and other neurodegenerative diseases.

Helminths of three species of White Sea fishes.

Parasitic fauna of the White Sea cod, Gadus morhua marisalbi; the navaga, Eleginus nawaga; and the shorthorn sculpin, Myoxocephalus scorpius, in the White Sea was repeatedly studied, but no large-scale parasitological surveys have been made in the recent three decades. To fill this gap, we conducted a survey of the helminths of these three fish species at the White Sea Biological Station (Karelia, Russia) of the Lomonosov Moscow State University in August 2021. The navaga (50 specimens studied) was found to be infected with 13 species of helminths; the White Sea cod (50 specimens), with 12 species; and the shorthorn sculpin (21 specimens), with 13 species. Plerocercoids of Diphyllobothrium schistochilus and third-stage juveniles of Pseudoterranova bulbosa were recorded in the White Sea for the first time. The helminth infracommunities of the navaga and the White Sea cod were closer in structure to each other than to those of the shorthorn sculpin. In general, the levels of helminth infection of the White Sea cod, the navaga, and shorthorn sculpin have been consistently high over 85 years of observations in the White Sea, but long-term trends in the abundance of some helminth species were multidirectional.

Nematocyst sequestration within the family Fionidae (Gastropoda: Nudibranchia) considering ecological properties and evolution.

Aeolid nudibranchs are well-known for their ability to incorporate cnidarian nematocysts and use them for defense; this process is tightly linked with the feeding preferences of molluscs. As many nudibranch groups show signs of ecology-based adaptive radiation, studies of prey-based defensive mechanisms can provide valuable insight into details of nudibranch evolutionary history. The main goal of this study is to test the correlation of ecological traits, feeding mechanisms, and prey preferences with cnidosac fine morphology and to pinpoint the phylogenetic value of these traits. We study the cnidosac morphology in thirteen species-representatives of the main lineages within the family Fionidae s.l. The morphological analysis includes histological sections, transmission electron microscopy, confocal laser scanning microscopy, and scanning electron microscopy. For phylogenetic study, available molecular data from public repositories were used, and phylogenetic trees were produced based on Bayesian Inference and Maximum likelihood analysis for a concatenated dataset of three molecular markers (COI, 16S, H3). In general, fionid cnidosacs fit the common aeolid pattern, but among different species we detected a high variation in type of obtained nematocysts, their arrangement within cnidophages, and in number of cell types within cnidosacs. We report on presence of cellules speciale in the haemocoel of all studied species, and for the first time, we report on cells with chitinous spindles in the haemocoel of all fionids except Eubranchus. The function of both these cell types remains unknown. The loss of functional cnidosacs occurred at least three times within Fionidae, and in case of the genera Phestilla, Calma, and Fiona, this loss is linked to their non-cnidarian diet. The diversity of cnidosac fine structure within Fionidae s.l. correlates with that of the radular morphology and feeding preferences of each species. Prey shifts between cnidarian and non-cnidarian prey (both through evolutionary shifts and individual variation) rarely occur within Fionidae s.l.; however, microevolutionary shifts between different hydrozoan species within a single genus are more common. Cnidosac morphology demonstrates considerable resulting changes even when switching between similar hydrozoan species, or changing the feeding site on same prey species. These data indicate that cnidosac morphology likely follows microevolutionary prey shifts-in other words, it is affected by switches in prey species and changes in feeding sites with a single prey species. Thus, the cnidosac morphology may be a useful indicator when studying ecological features of particular species.

Chaperone Hsp70 (HSPA1) Is Involved in the Molecular Mechanisms of Sleep Cycle Integration.

The molecular mechanisms of sleep cycle integration at the beginning and the end of the inactive period are not clear. Sleep cycles with a predominance of deep slow-wave sleep (SWS) seem to be associated with accelerated protein synthesis in the brain. The inducible Hsp70 chaperone corrects protein conformational changes and has protective properties. This research explores (1) whether the Hspa1 gene encoding Hsp70 protein activates during the daily rapid-eye-movement sleep (REMS) maximum, and (2) whether a lower daily deep SWS maximum affects the Hspa1 expression level during the subsequent REMS. Combining polysomnography in male Wistar rats, RT-qPCR, and Western blotting, we reveal a three-fold Hspa1 upregulation in the nucleus reticularis pontis oralis, which regulates REMS. Hspa1 expression increases during the daily REMS maximum, 5-7 h after the natural peak of deep SWS. Using short-term selective REMS deprivation, we demonstrate that REMS rebound after deprivation exceeds the natural daily maximum, but it is not accompanied by Hspa1 upregulation. The results suggest that a high proportion of deep SWS, usually observed after sleep onset, is a necessary condition for Hspa1 upregulation during subsequent REMS. The data obtained can inform the understanding of the molecular mechanisms integrating SWS and REMS and key biological function(s) of sleep.

A comparison of food sources of nudibranch mollusks at different depths off the Kuril Islands using fatty acid trophic markers.

Gastropod molluscs such as nudibranchs are important members of deep-sea benthic ecosystems. However, data on the trophic ecology and feeding specialization of these animals are limited to date. The method of fatty acid trophic markers (FATM) was applied to determine the dietary preferences of nudibranchs off the Kuril Islands. Fatty acid (FA) compositions of Dendronotus sp., Tritonia tetraquetra, and Colga pacifica collected from deep waters were analyzed and compared with those of Aeolidia papillosa and Coryphella verrucosa from the offshore zone. The high level of FATM such as 22:5n-6 and C20 monounsaturated FAs indicated that Dendronotus sp. preys on sea anemones and/or anthoathecates hydroids similarly to that of shallow-water species A. papillosa and C. verrucosa. The high percentage of tetracosapolyenoic acids and the ratio 24:6n-3/24:5n-6 indicated that T. tetraquetra preys on soft corals such as Gersemia and/or Acanella at a depth of 250 m, but soft corals of the family Primnoidae may be the main item in the diet of T. tetraquetra at a depth of 500 m. The high content of Δ 7,13-22:2 and 22:6n-3 shows that C. pacifica can feed on bryozoans. In C. pacifica, 22:5n-6 may be synthesized intrinsically by the mollusks, whereas odd-chain and branched saturated FAs originate from associated bacteria.

General and fine structure of Aeolidia papillosa cnidosacs (Gastropoda: Nudibranchia).

Nudibranch mollusks (Gastropoda: Heterobranchia) are widely known for their ability to incorporate some active biochemical compounds of their prey, or even organelles and symbionts of the prey, which assured biological success of this group. At the same time, the process of nematocysts obtaining and incorporation into specific structures called cnidosacs by cladobranch mollusks remain poorly studied. This highlights a necessity of additional ultrastructural studies of cnidosac and adjacent organs in various aeolid mollusks using modern microscopic methods as they may provide new insight into the cnidosac diversity and fine-scale dynamics of nematocysts sequestration process. The present study is focused on the general and fine structure of the cnidosac area in cladobranch Aeolidia papillosa (Aeolidiidae). Specific goals of our study were to provide a detailed description of histological and ultrafine structure of epidermis, upper parts of the digestive glands and the cnidosac, its innervation and proliferation using standard histological techniques, confocal laser scanning microscopy (CLSM) and transmission electron microscopy. Our results clearly demonstrated that A. papillosa cnidosac is a much more complex structure, than it was thought, especially compared with simple cnidosacs found in flabellinids and facelinids. Using CLSM for functional morphological analysis provides a better resolution in visualization of structural elements within a cnidosac compared with traditional histological techniques. We revealed the presence of two cell types in the cnidophage zone: cnidophages and interstitial cells, which differ in ultrastructure and function. Our results also document the presence of a specific cnidopore zone, lined with differentiated cuboid epithelium bearing long microvilli, which likely provides a unidirectional flow of nematocysts during kleptocnides extrusion. For the first time, occurrence of vacuoles containing protective chitinous spindles in the cnidosac epithelium was shown.

Functional morphology and post-larval development of the buccal complex in Eubranchus rupium (Nudibranchia: Aeolidida: Fionidae).

Nudibranch molluscs represent an interesting model group to study the evolution of feeding apparatus and feeding modes, being characterized by specialized buccal complex in combination with extremely diverse dietary preferences and multiply prey shifts in evolutionary history. However, the plasticity of the buccal complex morphology in response to diet and specific feeding modes remains understudied. Here we study the general morphology and ontogenesis of the buccal complex in Eubranchus rupium (Nudibranchia: Fionidae). Specific goals are to provide a detailed description of buccal structures morphology in post-larval stages, suggest the feeding mechanism and discuss the phylogenetic value of the morphological characteristics of buccal armature within the genus Eubranchus. Methods included in vivo observations of the feeding process for E. rupium, light microscopic methods, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and confocal laser scanning microscopy (cLSM). According to our results, E. rupium is a mechanical driller, boring holes in hydrozoan perisarc and sucking internal content. The mechanical drilling is supplied by functionally uniserial radula with plate-like laterals teeth of exclusively supportive function and by massive buccal musculature. Comparative phylogeny-based analysis suggests that the drilling feeding mode is common for the genus Eubranchus and indicates radular characters may have a high phylogenetic signal. The buccal complex morphology and feeding mode were found to be similar in both adults and post-metamorphic specimens, its general structures occur even in settled veligers. Juveniles and adults compete for food source, but the competition is smoothed due to characteristics of prey species growth and life cycle.

Meloscaphander grandis (Heterobranchia: Cephalaspidea), a deep-water species from the North Pacific: Redescription and taxonomic remarks.

Meloscaphander grandis is a little-known species missing from databases and papers on taxonomic revision and phylogenetic analysis of Scaphandridae. This species is redescribed herein, based on the type specimen and specimens from the abyssal plain adjacent to the Kuril-Kamchatka Trench. A phylogenetic analysis of COI, 16S, and 28S markers show M. grandis to nest within the Scaphander clade. Additionally, Scaphander lignarius and S. bathymophilus are suggested to be a complex of cryptic species. Morphological differences between the genera Meloscaphander and Scaphander are of dubious significance and, when coupled with molecular data, give a strong reason for reconsidering Meloscaphander as a junior synonym of Scaphander. Thus, according to an integrative taxonomic analysis, Meloscaphander grandis has been transferred to the genus Scaphander. The diagnosis of the genus Scaphander is expanded. We propose new combinations as follows: Scaphander grandis (Minichev, 1967) comb. n. for Meloscaphander grandis, Scaphander sibogae (Schepman, 1913) comb. n. for Meloscaphander sibogae, and Scaphander imperceptus (Bouchet, 1975) comb. n. for Meloscaphander imperceptus. Due to the homonymy of Scaphander sibogae Schepman, 1913 (with a sunken spire) and Scaphander sibogae (Schepman, 1913) comb. n. (with an elevated spire), the name S. attenuatus Schepman, 1913 becomes valid for the former species (with a sunken spire).

A case of nascent speciation: unique polymorphism of gonophores within hydrozoan Sarsia lovenii.

Revealing the mechanisms of life cycle changes is critical for understanding the processes driving hydrozoan evolution. Our analysis of mitochondrial (COI, 16S) and nuclear (ITS1 and ITS2) gene fragments resulted in the discovery of unique polymorphism in the life cycle of Sarsia lovenii from the White Sea. This polymorphic species exhibits two types of gonophores: hydroids produce both free-swimming medusae and attached medusoids (phenotypic polymorphism). Our phylogenetic analysis revealed the intrinsic genetic structure of S. lovenii (genetic polymorphism). Two haplogroups inhabiting the White Sea differ in their reproductive modes. Haplogroup 1 produces attached medusoids, and haplogroup 2 produces free-swimming medusae. Our experiments indicated the possibility of free interbreeding between haplogroups that likely is a rare event in the sea. We propose that inter-haplogroup crossing of S. lovenii in the White Sea may be limited by discordance in periods of spawning or by spatial differences in habitat of spawning specimens. Our finding can be interpreted as a case of nascent speciation that illustrates the patterns of repeated medusa loss in hydrozoan evolution. Life cycle traits of S. lovenii may be useful for elucidating the molecular mechanisms of medusa reduction in hydrozoans.

Diet-driven ecological radiation and allopatric speciation result in high species diversity in a temperate-cold water marine genus Dendronotus (Gastropoda: Nudibranchia).

While the majority nudibranch clades are more species rich in the tropics, the genus Dendronotus is mainly represented in Arctic and boreal regions. This distribution pattern remains poorly understood. An integrative approach and novel data provided valuable insights into processes driving Dendronotus radiation and speciation. We propose an evolutionary scenario based on molecular phylogenetics and morphological, ecological, ontogenetic data, combined with data on complex geology and paleoclimatology of this region. Estimated phylogenetic relationships based on four molecular markers (COI, 16S, H3 and 28S) shows strong correlation with radular morphology, diet and biogeographical pattern. Ancestral area reconstruction (AAR) provides evidence for a tropical Pacific origin of the genus. Based on AAR and divergence time estimates we conclude that the evolution of Dendronotus has been shaped by different processes: initial migration out of the tropics, diet-driven adaptive radiation in the North Pacific influenced by Miocene climate change, and subsequent allopatric speciation resulting from successive closings of the Bering strait and cooling of the Arctic Ocean during the Pliocene-Pleistocene. At the same time, contemporary amphiboreal species appear to have dispersed into the Atlantic fairly recently.

Drilling in the dorid species Vayssierea cf. elegans (Gastropoda: Nudibranchia): Functional and comparative morphological aspects.

The drilling mode of feeding is known from two clades of Gastropoda: Caenogastropoda and Heterobranchia. However, the level of convergence and parallelism or homology among these two lineages is unclear. The morphology of the buccal complex is well studied for drilling caenogastropods, but poorly known for drilling nudibranchs. It is also unclear whether the drilling feeding mechanism is similar between inside gastropods. Accordingly, a comparison between the feeding mechanisms of drilling nudibranchs and caenogastropods can help to understand the evolutional trends inside gastropods. In this study, we redescribe the morphology of the buccal complex of drilling dorid nudibranch Vayssierea cf. elegans, and compare it to that of previous investigations on this species and closely related dorid species. We describe the feeding mechanism of this species based on the obtained morphological and literature data and compare it to the feeding mechanisms described for drilling caenogastropods. The feeding apparatus of Vayssierea cf. elegans corresponds to the general morphology of the dorid buccal complex; that is, it has a similar arrangement of the buccal musculature and pattern of radular morphology. However, there are also adaptations to the drilling feeding mode similar to those found in Caenogastropoda: that is, specialized dissolving glands and lateral teeth with elongated pointed cusps; and even Sacoglossa: the specialized muscle for sucking. The feeding process of Vayssierea cf. elegans includes the same two stages as those described for drilling caenogastropods: (a) the boring stage, which is provided by mechanical and chemical activity, and (b) the swallowing stage.

New HSF1 inducer as a therapeutic agent in a rodent model of Parkinson's disease.

Molecular chaperone HSP70 (HSPA1A) has therapeutic potential in conformational neurological diseases. Here we evaluate the neuroprotective function of the chaperone in a rat model of Parkinson's disease (PD). We show that the knock-down of HSP70 (HSPA1A) in dopaminergic neurons of the Substantia nigra causes an almost 2-fold increase in neuronal death and multiple motor disturbances in animals. Conversely, pharmacological activation of HSF1 transcription factor and enhanced expression of inducible HSP70 with the echinochrome derivative, U-133, reverses the process of neurodegeneration, as evidenced by а increase in the number of tyrosine hydroxylase-containing neurons, and prevents the motor disturbances that are typical of the clinical stage of the disease. The neuroprotective effect caused by the elevation of HSP70 in nigral neurons is due to the ability of the chaperone to prevent α-synuclein aggregation and microglia activation. Our findings support the therapeutic relevance of HSP70 induction for the prevention and/or deceleration of PD-like neurodegeneration.

A Radical Solution: The Phylogeny of the Nudibranch Family Fionidae.

Tergipedidae represents a diverse and successful group of aeolid nudibranchs, with approximately 200 species distributed throughout most marine ecosystems and spanning all biogeographical regions of the oceans. However, the systematics of this family remains poorly understood since no modern phylogenetic study has been undertaken to support any of the proposed classifications. The present study is the first molecular phylogeny of Tergipedidae based on partial sequences of two mitochondrial (COI and 16S) genes and one nuclear gene (H3). Maximum likelihood, maximum parsimony and Bayesian analysis were conducted in order to elucidate the systematics of this family. Our results do not recover the traditional Tergipedidae as monophyletic, since it belongs to a larger clade that includes the families Eubranchidae, Fionidae and Calmidae. This newly recovered clade is here referred to as Fionidae, the oldest name for this taxon. In addition, the present molecular phylogeny does not recover the traditional systematic relationships at a generic level, and therefore, systematic changes are required. We recognize the following clades within Fionidae: Calma, Cuthona, Cuthonella, Eubranchus, Fiona, Murmania, Tenellia, Tergipes, Tergiposacca gen. nov., Rubramoena gen. nov. and Abronica gen. nov. The type species of Tergiposacca, T. longicerata nov. sp. is described. The other two new genera have a previously described species as their type species. Most of these taxa, with the exceptions of Eubranchus, Tergipes and Fiona are composed of radically different constituent species from their traditional membership, but appear to be supported by morphological synapomorphies as well as molecular data. Aenigmastyletus, Catriona, Phestilla, Tenellia and Trinchesia are nested within other clades and, thus are here considered as synonyms of the larger clades. The phylogenetic position and validity of Myja, Guyvalvoria, Leostyletus and Subcuthona still need to be tested in future studies when material becomes available.

Exogenous protein Hsp70/Hsc70 can penetrate into brain structures and attenuate the severity of chemically-induced seizures.

Heat shock protein 70 kDa (Hsp70) possesses a remarkable neuroprotective activity and the results of recent studies demonstrated its efficacy in the attenuation of epileptic seizures. The aim of this study was to explore the effects of a pure Hsp70/Hsc70 preparation delivered to the brain regions involved in generalized seizures induced in rats by intracerebroventricular microinjections of NMDA or systemic injections of pentylenetetrazole. Purified Hsp70/Hsc70 was administered (intracerebroventricular) 2 h before the induction of seizures. Compared to the vehicle-treated control animals, Hsp70/Hsc70-pretreated rats demonstrated reduced severity of NMDA- and pentylenetetrazole-induced seizures. To identify the brain structures potentially implicated in the Hsp70/Hsc70-mediated anticonvulsant effect, we analysed the localization of a fluorescently-labelled chaperone in the brain. Labelled Hsp70/Hsc70 was found in neurons and terminals of the limbic seizure complex of the brain and was co-localized in these regions with NMDA receptors, synaptophysin and the GABA-synthesizing enzyme, L-glutamic acid decarboxylase 67. An immunoprecipitation assay confirmed interactions between Hsp70 and both synaptophysin and L-glutamic acid decarboxylase 67 in brain tissue. We suggest that the anticonvulsant effect of exogenous Hsp70/Hsc70 is not only based on its protective capacity but is also related to its ability to modulate GABA neurotransmission, which in turn contributes to the maintenance of the excitatory-inhibitory balance of the CNS.