Primitive Vertebrates Research Articles

Siglec7 functions as an inhibitory receptor of non-specific cytotoxic cells and can regulate the innate immune responses in a primitive vertebrate (Oreochromis niloticus)

In mammals, siglec7, an integral component of the siglecs, is principally found on the surface of natural killer (NK) cells, macrophages, and monocytes, where it interacts with various pathogens to perform immunological regulatory activities. Nonetheless, the immune defense and mechanism of siglec7 in early vertebrates remain unknown. In this study, we identified siglec7 from Oreochromis niloticus (OnSiglec7) and revealed its immune functions. Specifically, OnSiglec7 was abundantly expressed in immune-related tissues of healthy tilapia and its transcription level was strongly activated after being challenged with A. hydrophila, S. agalactiae, and Poly: IC. Meanwhile, OnSiglec7 protein was purified and analyzed, which could recognize multiple pathogens through binding and agglutinating activity. Moreover, OnSiglec7-positive cells were mainly distributed in non-specific cytotoxic cells (NCC) of tilapia HKLs and showed cell membrane localization. Furthermore, OnSiglec7 blockage affected multiple innate immune responses (inflammation, apoptosis, and pyroptosis process) by regulating the activation of MAPK, NF-κB, TLR, and JAK-STAT pathways. Finally, OnSiglec7 blockage also greatly enhanced the cytotoxic effect of tilapia NCC. Summarily, this study uncovers immune functions and mechanisms of siglec7 in primitive vertebrates, thereby enhancing our understanding of the systemic evolution and ancient functions of other siglecs within the host's innate immune system (to our knowledge).

Identification of antibacterial activity of liver-expressed antimicrobial peptide 2 (LEAP2) from primitive vertebrate lamprey

Liver-expressed antimicrobial peptide 2 (LEAP2) is a member of the antimicrobial peptides family and plays a key role in the innate immune system of organisms. LEAP2 orthologs have been identified from a variety of fish species, however, its function in primitive vertebrates has not been clarified. In this study, we cloned and identified Lc-LEAP2 from the primitive jawless vertebrate lamprey (Lethenteron camtschaticum) which includes a 25 amino acids signal peptide and a mature peptide of 47 amino acids. Although sequence similarity was low compared to other species, the mature Lc-LEAP2 possesses four conserved cysteine residues, forming a core structure with two disulfide bonds between the cysteine residues in the relative 1–3 (Cys 58 and Cys 69) and 2–4 (Cys 64 and Cys 74) positions. Lc-LEAP2 was most abundantly expressed in the muscle, supraneural body and buccal gland of lamprey, and was significantly upregulated during LPS and Poly I:C stimulations. The mature peptide was synthesized and characterized for its antibacterial activity against different bacteria. Lc-LEAP2 possessed inhibition of a wide range of bacteria with a dose-dependence, disrupting the integrity of bacterial cell membranes and binding to bacterial genomic DNA, although its inhibitory function is weak compared to that of higher vertebrates. These data suggest that Lc-LEAP2 plays an important role in the innate immunity of lamprey and is of great value in improving resistance to pathogens. In addition, the antimicrobial mechanism of LEAP2 has been highly conserved since its emergence in primitive vertebrates.

CL-K1 Promotes Complement Activation and Regulates Opsonophagocytosis of Macrophages with CD93 Interaction in a Primitive Vertebrate.

Collectin is a crucial component of the innate immune system and plays a vital role in the initial line of defense against pathogen infection. In mammals, collectin kidney 1 (CL-K1) is a soluble collectin that has recently been identified to have significant functions in host defense. However, the evolutionary origins of immune defense of CL-K1 and its mechanism in clearance of pathogenic microorganisms remain unclear, especially in early vertebrates. In this study, the Oreochromis niloticus CL-K1 (OnCL-K1) protein was purified and identified, which was capable of binding to two important pathogens of tilapia, Streptococcus agalactiae and Aeromonas hydrophila. Interestingly, OnCL-K1 exhibited direct bactericidal activity by binding to lipoteichoic acid or LPS on cell walls, disrupting the permeability and integrity of the bacterial membrane invitro. Upon bacterial challenge, OnCL-K1 significantly inhibited the proliferation of pathogenic bacteria, reduced the inflammatory response, and improved the survival of tilapia. Further research revealed that OnCL-K1 could associate with OnMASPs to initiate and regulate the lectin complement pathway. Additionally, OnCD93 reduced the complement-mediated hemolysis by competing with OnMASPs for binding to OnCL-K1. More importantly, OnCL-K1 could facilitate phagocytosis by collaborating with cell surface CD93 in a lectin pathway-independent manner. Moreover, OnCL-K1 also promoted the formation of phagolysosomes, which degraded and killed ingested bacteria. Therefore, this study reveals the antibacterial response mechanism of CL-K1 in primitive vertebrates, including promoting complement activation, enhancing opsonophagocytosis, and killing of macrophages, as well as its internal links, all of which provide (to our knowledge) new insights into the understanding of the evolutionary origins and regulatory roles of the collectins in innate immunity.

The ontogenesis and heterogeneity of basophils.

Basophils are the rarest leukocytes, but they have essential roles in protection against helminths, allergic disorders, autoimmune diseases, and some cancers. For years, the clinical significance of basophils has been neglected because of the lack of proper experimental tools to study them. The development of basophil-specific antibodies and animal models, along with genomic advances like single-cell transcriptomics, has greatly enhanced our understanding of basophil biology. Recent discoveries regarding basophils prompted us to write this review, emphasizing the basophil developmental pathway. In it, we chronologically examine the steps of basophil development in various species, which reveals the apparent advent of basophils predating IgE and basophil's IgE-independent regulatory role in primitive vertebrates. Then, we cover studies of basophil development in adult bone marrow, and compare those of murine and human basophils, introducing newly identified basophil progenitors and mature basophil subsets, as well as the transcription factors that regulate the transitions between them. Last, we discuss the heterogeneity of tissue-resident basophils, which may develop through extramedullary hematopoiesis. We expect that this review will contribute to a deeper understanding of basophil biology from the intricate aspects of basophil development and differentiation, offering valuable insights for both researchers and clinicians.

MAPK pathway differentially regulated the apoptosis and inflammatory necrosis of respiratory epithelial cells during bacterial infection in a primitive vertebrate

MAPK pathway differentially regulated the apoptosis and inflammatory necrosis of respiratory epithelial cells during bacterial infection in a primitive vertebrate

Toxicity of per- and polyfluoroalkyl substances to aquatic vertebrates

Rapid industrial development and extensive use of chemicals have resulted in elevated concentrations of emerging contaminants worldwide, posing a substantial threat to the ecological environment and human health. Per- and polyfluoroalkyl substances (PFASs) have been recognized as emerging pollutants that are widely distributed and accumulated in the environment and they have drawn the attention of scholars for several decades. The variety, long-term use, and long-distance transmission of PFASs have resulted in the ubiquitous contamination of global ecosystems, especially in aquatic environments. Ever since perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) were added to the Stockholm Convention on Persistent Organic Pollutants (POPs), they have become the most typical, eye-catching, and frequently investigated PFASs. Owing to the high stability and bioaccumulation of PFASs, as well as the adverse impact on the endocrine, immune, and nervous systems, investigating their contamination levels, risk of transfer along the food chain, and ecotoxicity should be prioritized. In addition to the important evolutionary significance as primitive vertebrates and the main consumers of aquatic environment, fishes generally exist in various aquatic food chains from the bottom to the top and occupy a critical position in terms of aquatic ecology protection; while amphibians, as the key link from aquatic to terrestrial organisms, are highly sensitive to different environmental pollutants. This review is a comprehensive summary of the toxic effects and toxicity-related factors of PFASs on aquatic vertebrates, mainly Pisces and Amphilla organisms, the characteristics of different aquatic vertebrates in toxicity investigations, and the evaluation of the feasibility of PFASs substitute applications.

Tryptophan metabolism can modulate immunologic tolerance in primitive vertebrate lamprey via IDO-kynurenine-AHR pathway

Tryptophan is mainly degraded through kynurenine pathway (KP) in vertebrates which is closely related to the nerve and depression, while the studies on immunity is still limited. This study aims to explore the functions of tryptophan in the innate immunity of primitive vertebrate lamprey. MTT (3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide) assay showed that tryptophan had no obvious effect on cell viability. Tryptophan was transported into leukocytes and degraded via the KP after tryptophan supplement. Tryptophan treatment (T1x and T2x) failed to alter the total antioxidant capacity regardless of stimulation and exposure time. Real-time quantitative PCR and western blotting results revealed that tryptophan was not only able to reduce the expression of pro-inflammatory factors Lj-TNF-α, Lj-IL1β and Lj–NF–κB, but also to upregulate the expression of anti-inflammatory factor Lj-TGF-β independent of stimulation and time. In addition, tryptophan can exert immune tolerance function by inhibiting TLR-MyD88 and promoting (Indoleamine 2, 3-Dioxygenase) IDO-kynurenine-AHR (aryl hydrocarbon receptor) pathways. This study provides a new understanding for tryptophan-kynurenine metabolism and mechanism of immune tolerance function in primitive vertebrate lamprey.

Review of the unique and dominant lectin pathway of complement activation in agnathans

As the most primitive vertebrates, lampreys are significant in understanding the early origin and evolution of the vertebrate innate and adaptive immune systems. The complement system is a biological response system with complex and precise regulatory mechanisms and plays an important role in innate and adaptive immunity. It consists of more than 30 distinct components, including intrinsic components, regulatory factors, and complement receptors. Complement system is the humoral backbone of the innate immune defense and complement-like factors have also been found in cyclostomes. Our knowledge as such in lamprey has dramatically increased in the recent years. The searching for complement components in the reissner lamprey Lethenteron reissneri genome database, together with published data, has unveiled the existence of all the orthologues of mammalian complement components identified thus far, including the complement regulatory proteins and complement receptors, in lamprey. This review, summarizes the key themes and recent updates on the complement system of agnathans and discusses the individual complement components of lampreys, and critically compare their functions to that of mammalian complement components. Interestingly, the adaptive immune system of agnathans differs from that of gnathostomes. Lamprey complement components also display some distinctive features, such as lampreys are characterized by the variable lymphocyte receptors (VLRs)-based alternative adaptive immunity. This review may serve as important literature for deducing the evolution of the immune system from invertebrates to vertebrates.

Phylogenetic evolutionary aspects of the vertebrate skull - an approach for teaching: vertebrate skull

In this work, the fundamental data will be placed as logically as possible within a simple, but not simplistic philosophy, therefore, the objective was to indicate the basic ontogenetic and phylogenetic data in terms of the evolutionary history of the vertebrate skull starting from the initial, often idealized, structures that formed the framework of the primitive vertebrate skull. Therefore, the methodology used was a literature review, in order to refine the information. From this methodology, the expected result was to reduce the excess or oversimplification of information that could fail in the teaching processes, to avoid the danger of errors of the interpretation. An overview with basic data, but well ordered in terms of substitutions and phylogenetic history is essential to prepare future scientists in the areas of morphology, taxonomy, evolution, comparative anatomy, paleontology, forensic anatomy; therefore, this text was builded objectively as possible to be consulted for the teaching/learning of the cranium comparative anatomy.

Detecting Intestinal Goblet Cells of the Broadgilled Hagfish Eptatretus cirrhatus (Forster, 1801): A Confocal Microscopy Evaluation

Simple SummaryThe intestinal epithelium of fish, similar to mammals, consists mainly of enterocytes and goblet cells. Goblet cells play a key role in the secretion of mucus, which, in addition to promoting the digestion of nutrients, is the first protective barrier against bacteria, viruses, and pathogens. Our study aims to evaluate the presence, localization, and co-localization of 5-HT, TLR2, iNOS, and Piscidin1 in goblet cells of the intestine of Eptatretus cirrhatus. The results obtained by confocal microscopy show, for the first time, the positivity of goblet cells to the antibodies tested, suggesting the involvement of these cells in the intestinal immunity of broadgilled hagfish.The fish intestine operates as a complicated interface between the organism and the environment, providing biological and mechanical protections as a result of a viscous layer of mucus released by goblet cells, which serves as a barrier against bacteria, viruses, and other pathogens, and contributes to the functions of the immune system. Therefore, goblet cells have a role in preserving the health of the body by secreting mucus and acting as sentinels. The ancient jawless fish broadgilled hagfish (Eptatretus cirrhatus, Forster, 1801) has a very basic digestive system because it lacks a stomach. By examining the presence, localization, and co-localization of 5-HT, TLR2, iNOS, and Piscidin1, this study intends to provide insight into the potential immune system contributions arranged by the gut goblet cells of broadgilled hagfish. Our results characterize intestinal goblet cells of broadgilled hagfish, for the first time, with the former antibodies, suggesting the hypothesis of conservation of the roles played by these cells also in primitive vertebrates. Moreover, this study deepens the knowledge about the still little-known immune system of hagfish.

Affinity-Driven Site-Specific High Mannose Modification Determines the Structural Polymerization and Function of Tetrameric IgM in a Primitive Vertebrate.

Teleost tetramer IgM is the predominant Ig in the immune system and plays essential roles in host defense against microbial infection. Due to variable disulfide polymerization of the monomeric subunits, tetrameric IgM possesses considerable structural diversity. Previous work indicated that the teleost IgM H chain was fully occupied with complex-type N-glycans. However, after challenge with trinitrophenyl (TNP) Ag, the complex N-glycans in the Asn-509 site of Oreochromis niloticus IgM H chain transformed into high mannose. This study, therefore, was conducted to examine the functional roles of the affinity-related high-mannose modification in tilapia IgM. The TNP-specific IgM Ab affinity maturation was revealed in tilapia over the response. A positive correlation between TNP-specific IgM affinity and its disulfide polymerization level of isomeric structure was demonstrated. Mass spectrometric analysis indicated that the relationship between IgM affinity and disulfide polymerization was associated with the Asn-509 site-specific high-mannose modification. Furthermore, the increase of high mannose content promoted the combination of IgM and mannose receptor (MR) on the surface of phagocytes. Moreover, the increased interaction of IgM and MR amplified the phagocytic ability of phagocytes to Streptococcus agalactiae. To our knowledge, this study demonstrates that site-specific high-mannose modification associates with IgM Ab affinity and its structural disulfide polymerization and amplifies the phagocytosis of phagocytes by the combination of IgM and MR. The present study provides evidence for understanding the association of IgM structure and function during the evolution of the immune system.

Eph/ephrin signaling controls cell contacts and formation of a structurally asymmetrical tissue boundary in the Xenopus gastrula

In the primitive vertebrate gastrula, the boundary between ectoderm and mesoderm is formed by Brachet's cleft. Here we examine Brachet's cleft and its control by Eph/ephrin signaling in Xenopus at the ultrastructural level and by visualizing cortical F-actin. We infer cortical tension ratios at tissue surfaces and their interface in normal gastrulae and after depletion of receptors EphB4 and EphA4 and ligands ephrinB2 and ephrinB3. We find that cortical tension downregulation at cell contacts, a normal process in adhesion, is asymmetrically blocked in the ectoderm by Eph/ephrin signals from the mesoderm. This generates high interfacial tension that can prevent cell mixing across the boundary. Moreover, it determines an asymmetric boundary structure that is suited for the respective roles of ectoderm and mesoderm, as substratum and as migratory layers. The Eph and ephrin isoforms also control different cell-cell contact types in ectoderm and mesoderm. Respective changes of adhesion upon isoform depletion affect adhesion at the boundary to different degrees but usually do not prohibit cleft formation. In an extreme case, a new type of cleft-like boundary is even generated where cortical tension is symmetrically increased on both sides of the boundary.

Nanomechanical variability in the early evolution of vertebrate dentition

Conodonts are an extinct group of primitive jawless vertebrates whose elements represent the earliest examples of a mineralized feeding apparatus in vertebrates. Their relative relationship within vertebrates remains unresolved. As teeth, conodont elements are not homologous with the dentition of vertebrates, but they exhibit similarities in mineralization, growth patterns, and function. They clearly represent an early evolutionary experiment in mineralized dentition and offer insight into analogous dentition in other groups. Unfortunately, analysis of functional performance has been limited to a handful of derived morphologies and material properties that may inform ecology and functional analysis are virtually unknown. Here we applied a nanoscale approach to evaluate material properties of conodont bioapatite by utilizing Atomic Force Microscopy (AFM) nanoindentation to determine Young’s modulus (E) along multiple elements representing different ontogenetic stages of development in the coniform-bearing apparatus of Dapsilodus obliquicostatus. We observed extreme and systematic variation in E along the length (oral to aboral) of each element that largely mirrors the spatial and ontogenetic variability in the crystalline structure of these specimens. Extreme spatial variability of E likely contributed to breakage of elements that were regularly repaired/regrown in conodonts but later vertebrate dentition strategies that lacked the ability to repair/regrow likely required the development of different material properties to avoid structural failure.

Resemblance in Physiological Systems during Phylogeny and Ontogeny

Although it is not certain whether some characteristics of the mammalian fetus/embryo may repeat the adult stage of ancestral vertebrates, comparison of these two time‐dependent processes should provide new insight into the molecular and functional evolution and its significance. Here we discuss two important biological models: 1) Renin‐angiotensin system (RAS) and 2) Urine concentration mechanism.Renin is a hormone and substrate‐dependent enzyme secreted mostly by the juxtaglomerular (JG) cells of the afferent arteriole located at the vascular pole of the glomerulus. Renin acts on renin substrates and forms angiotensin (Ang) I which is subsequently converted to Ang II by Ang converting enzyme (ACE). Renin containing cells evolved early both in phylogeny and ontogeny, whereas, the macula densa (tubule component), and extracellular mesangium appeared at later stages.Renin containing cells are distributed more widely intrarenally and/or extrarenally in early stages during phylogeny and ontogeny. Granulated cells and renin expression become progressively restricted to JG areas with phylogenic and ontogenic advancement. Kidney renin activities tend to be higher in primitive fishes than in more advanced, while the relative level of extrarenal renin expression to that of the kidney is higher in embryos than in more mature chicken and mice. Extra renal renin is noted in primitive vertebrates. Also, in both the mammalian fetus and nonmammalian vertebrates, the vasopressor and vascular action of Ang II evolved early. A single application of ACE inhibitors decreases the basal level of blood pressure and increase plasma renin activity in conscious adult teleost fish. Ang II stimulates, whereas ACE inhibitors inhibit, arteriolar branching in zebrafish. Likewise, the RAS is important during early ontogeny of mammals in vascular growth and development.Distal tubules of freshwater trout show secondary active NaCl cotransport mechanism without accompaniment of water and act as the so‐called “diluting segment.” Teleost fish cannot concentrate urine because they lack structural and functional countercurrent mechanisms. Only birds and mammals can form urine clearly hyperosmotic to plasma and conserve body water. The structure and function of adult quail kidneys have similarities to those of developing rodents. Both have a loop of Henle, consisting of descending limb and thick ascending limb, but they lack the thin ascending limb. Countercurrent urine concentration of adult quail and developing rodents heavily depends on NaCl recycling without participation of urea transport. Moreover, in both adult quail and developing rodent kidneys, aquaporin 2 gene expression and protein are present in the apical membrane of the collecting tubules, but the water conservation effect of neurohypophysial hormones is smaller than that of adult rodent kidneys. Hence, comparing these two time‐dependent processes may provide new insights into the interpretation of existing findings and provide perspective for future investigations.

Identification and functional characterization of an immune adapter molecular TRIF in Northeast Chinese lamprey (Lethenteron morii)

The TIR domain-containing adaptor inducing IFN-β (TRIF) is an adaptor molecule that plays a critical role in the Toll-like receptors (TLRs)-mediated innate immune signaling pathway. Lamprey, as the most primitive jawless vertebrate, rely mainly on innate immunity to defend against various pathogens infection. The function of TRIF in lamprey remains unknown. In this study, a homologous adaptor molecule TRIF, named LmTRIF, was identified in Northeast Chinese lamprey (Lethenteron morii). The LmTRIF coding sequence (cds) is 1242 bp in length and encodes 413 amino acids (aa). Domain analysis showed that LmTRIF is characterized with the classical TIR domain and a lack of TRAF6 binding motif. The results of evolutionary tree indicated that the relationship between LmTRIF and other homologous proteins was consistent with the position of lamprey in the species evolutionary history. The relative expression of LmTRIF was highest in the liver of larvae and in the gill of adults, respectively. Cellular immunofluorescence assays showed that LmTRIF was expressed in the cytoplasma in both mammalian cell line HEK 293T and the fish cell line EPC. The double luciferase reporter gene assay showed that the overexpression of LmTRIF promoted the activity of NF-κB, an immune transcription factor downstream of the classical TLR signaling pathway. In this study, we identified the TLR adaptor molecule TRIF from L. morii, a vertebrate more primitive than fish. Our results suggested an important role of LmTRIF in the innate immune signal transduction process of L. morii and is the basis for the origin and evolution of the TLR signaling pathway in the innate immune system in vertebrates.

The evolutionary development of the jaws and dentition

The development of the jaws and dentition during half a billion years of evolution is described in broad lines and two theories regarding the origin of the dentition are discussed in detail. It is highly likely our dentition developed from dermal ossification 'denticles' originally protecting, in particular, the head of primitive extinct vertebrates. Simple, at first tubercular, later conical teeth developed into complex tooth and molar forms, especially in mammals. Special attention is given to the development of molar cusps. The process of continuous replacement of the dentition is also discussed in more detail.

Discovery of an ancient MHC category with both class I and class II features

Two classes of major histocompatibility complex (MHC) molecules, MHC class I and class II, play important roles in our immune system, presenting antigens to functionally distinct T lymphocyte populations. However, the origin of this essential MHC class divergence is poorly understood. Here, we discovered a category of MHC molecules (W-category) in the most primitive jawed vertebrates, cartilaginous fish, and also in bony fish and tetrapods. W-category, surprisingly, possesses class II-type α- and β-chain organization together with class I-specific sequence motifs for interdomain binding, and the W-category α2 domain shows unprecedented, phylogenetic similarity with β2-microglobulin of class I. Based on the results, we propose a model in which the ancestral MHC class I molecule evolved from class II-type W-category. The discovery of the ancient MHC group, W-category, sheds a light on the long-standing critical question of the MHC class divergence and suggests that class II type came first.

X-ray nanotomography and electron backscatter diffraction demonstrate the crystalline, heterogeneous and impermeable nature of conodont white matter.

Conodont elements, microfossil remains of extinct primitive vertebrates, are commonly exploited as mineral archives of ocean chemistry, yielding fundamental insights into the palaeotemperature and chemical composition of past oceans. Geochemical assays have been traditionally focused on the so-called lamellar and white matter crown tissues; however, the porosity and crystallographic nature of the white matter and its inferred permeability are disputed, raising concerns over its suitability as a geochemical archive. Here, we constrain the characteristics of this tissue and address conflicting interpretations using ptychographic X-ray-computed tomography (PXCT), pore network analysis, synchrotron radiation X-ray tomographic microscopy (srXTM) and electron back-scatter diffraction (EBSD). PXCT and pore network analyses based on these data reveal that while white matter is extremely porous, the pores are unconnected, rendering this tissue closed to postmortem fluid percolation. EBSD analyses demonstrate that white matter is crystalline and comprised of a single crystal typically tens of micrometres in dimensions. Combined with evidence that conodont elements grow episodically, these data suggest that white matter, which comprises the denticles of conodont elements, grows syntactically, indicating that individual crystals are time heterogeneous. Together these data provide support for the interpretation of conodont white matter as a closed geochemical system and, therefore, its utility of the conodont fossil record as a historical archive of Palaeozoic and Early Mesozoic ocean chemistry.

Spatial Cognition in Teleost Fish: Strategies and Mechanisms.

Simple SummaryThe study of the neurobiological basis of spatial cognition has been demonstrated to be one of the most exciting, successful, and productive research fields in neuroscience. An enormous number of experimental results on the brain mechanisms of navigation have been obtained over several decades and a number of theories and detailed mechanistic computational models have been developed to account for these data obtained mainly in mammals and birds. Recently, the use of teleost fish species as animal models in neurobiology has exponentially increased, nicely complementing the use of traditional mammalian models in basic and translational neuroscience research. Comparative neurobiological research has shown that teleost fish can use a variety of navigational strategies that closely resemble those described in mammals and birds. Although some of these similarities could indicate evolutionary convergence shaped by common environmental constraints and survival requirements, at least some of these strategies seem to be based on conserved neural substrata likely shared with land vertebrates, suggesting that these strategies and their neurobiological basis could have appeared very early on during vertebrate evolution.Teleost fish have been traditionally considered primitive vertebrates compared to mammals and birds in regard to brain complexity and behavioral functions. However, an increasing amount of evidence suggests that teleosts show advanced cognitive capabilities including spatial navigation skills that parallel those of land vertebrates. Teleost fish rely on a multiplicity of sensory cues and can use a variety of spatial strategies for navigation, ranging from relatively simple body-centered orientation responses to allocentric or “external world-centered” navigation, likely based on map-like relational memory representations of the environment. These distinct spatial strategies are based on separate brain mechanisms. For example, a crucial brain center for egocentric orientation in teleost fish is the optic tectum, which can be considered an essential hub in a wider brain network responsible for the generation of egocentrically referenced actions in space. In contrast, other brain centers, such as the dorsolateral telencephalic pallium of teleost fish, considered homologue to the hippocampal pallium of land vertebrates, seem to be crucial for allocentric navigation based on map-like spatial memory. Such hypothetical relational memory representations endow fish’s spatial behavior with considerable navigational flexibility, allowing them, for example, to perform shortcuts and detours.

Identification, molecular evolution, and expression analysis of the transcription factor Smad gene family in lamprey

Transcription factor small mothers against decapentaplegic (Smad) family SMAD proteins are the essential intracellular signal mediators and transcription factors for transforming growth factor β (TGF-β) signal transduction pathway, which usually exert pleiotropic actions on cell physiology, including immune response, cell migration and differentiation. In this study, the Smad family was identified in the most primitive vertebrates through the investigation of the transcriptome data of lampreys. The topology of phylogenetic tree showed that the four Smads (Smad1, Smad3, Smad4 and Smad6) in lampreys were subdivided into four different groups. Meanwhile, homology analysis indicated that most Smads were conserved with typical Mad Homology (MH) 1 and MH2 domains. In addition, Lethenteron reissneri Smads (Lr-Smads) adopted general Smads folding structure and had high tertiary structural similarity with human Smads (H-Smads). Genomic synteny analysis revealed that the large-scale duplication blocks were not found in lamprey genome and neighbor genes of lamprey Smads presented dramatic differences compared with jawed vertebrates. Importantly, quantitative real-time PCR analysis demonstrated that Smads were widely expressed in lamprey, and the expression level of Lr-Smads mRNA was up-regulated with different pathogenic stimulations. Moreover, depending on the weighted gene co-expression network analysis (WGCNA), four Lr-Smads were identified as two meaningful modules (green and gray). The functional analysis of these two modules showed that they might have a correlation with ployI:C. And these genes presented strong positive correlation during the immune response from the results of Pearson's correlation analysis. In conclusion, our results would not only enrich the information of Smad family in jawless vertebrates, but also lay the foundation for immunity in further study.