Evidence of lophophore diversity in early Cambrian brachiopoda

The lophophore, an essential organ of the Brachiopoda, has been used widely in evolutionary and advanced phylogenetic studies, but is hitherto unknown in the fossil record. Here, the extraordinarily well-preserved lophophores of two inarticulated brachiopods Lingulella chengjiangensis and Heliomedusa orienta, from the Lower Cambrian Chengjiang fauna (Yunnan, China) are described. These primitive lophophores, respectively, trocholophous and schizolophous, have some key characters that may be plesiomorphies inherited by their recent descendants. This discovery provides direct evidence regarding the taxonomy, ecosystems and early evolution of inarticulated brachiopods.     

PKA activation bypasses the requirement for UNC-31 in the docking of dense core vesicles from C. elegans neurons

The nematode C. elegans provides a powerful model system for exploring the molecular basis of synaptogenesis and neurotransmission. However, the lack of direct functional assays of release processes has largely prevented an in depth understanding of the mechanism of vesicular exocytosis and endocytosis in C. elegans. We address this technical limitation by developing direct electrophysiological assays, including membrane capacitance and amperometry measurements, in primary cultured C. elegans neurons. In addition, we have succeeded in monitoring the docking and fusion of single dense core vesicles (DCVs) employing total internal reflection fluorescence microscopy. With these approaches and mutant perturbation analysis, we provide direct evidence that UNC-31 is required for the docking of DCVs at the plasma membrane. Interestingly, the defect in DCV docking caused by UNC-31 mutation can be fully rescued by PKA activation. We also demonstrate that UNC-31 is required for UNC-13-mediated augmentation of DCV exocytosis.     

UNC-31/CAPS docks and primes dense core vesicles in C. elegans neurons

UNC-31 or its mammalian homologue, Ca²⁺-dependent activator protein for secretion (CAPS), is indispensable for exocytosis of dense core vesicle (DCV) and synaptic vesicle (SV). From N- to the C-terminus, UNC-31 contains putative functional domains, including dynactin 1 binding domain (DBD), C2, PH, (M)UNC-13 homology domain (MHD) and DCV binding domain (DCVBD), the last four we examined in this study. We employed UNC-31 null mutant C. elegans worms to examine whether UNC-31 functions could be rescued by ectopic expression of full length UNC-31 vs each of these four domain-deleted mutants. Full length UNC-31 cDNA rescued the phenotypes of C. elegans null mutants in response to Ca²⁺-elevation in ALA neurons. Surprisingly, MHD deletion also rescued UNC-31 exocytotic function in part because the relatively high Ca²⁺ level (pre-flash Ca²⁺ was 450 nM) used in the capacitance study could bypass the MHD defect. Nonetheless, the three other domain-truncation cDNAs had almost no rescue on Ca²⁺ evoked secretion. Importantly, this genetic null mutant rescue strategy enabled physiological studies at levels of whole organism to single cells, such as locomotion assay, pharmacological study of neurotransmission at neuromuscular junction, in vivo neuropeptide release measurement and analysis of vesicular docking. Our results suggest that each of these UNC-31 domains support distinct sequential molecular actions of UNC-31 in vesicular exocytosis, including steps in vesicle tethering and docking that bridge vesicle with plasma membrane, and subsequently priming vesicle by initiating the formation of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) core complex.     

The Establishment of an In Vivo HIV-1 Infection Model in Humanized B-NSG Mice

Suitable animal models for human immunodeficiency virus type 1(HIV-1) infection are important for elucidating viral pathogenesis and evaluating antiviral strategies in vivo. The B-NSG(NOD-PrkdcscidIl2rgtm1/Bcge) mice that have severe immune defect phenotype are examined for the suitability of such a model in this study. Human peripheral blood mononuclear cells(PBMCs) were engrafted into B-NSG mice via mouse tail vein injection, and the repopulated human T-lymphocytes were observed at as early as 3-weeks post-transplantation in mouse peripheral blood and several tissues.The humanized mice could be infected by HIV-1, and the infection recapitulated features of T-lymphocyte dynamic observed in HIV-1 infected humans, meanwhile the administration of combination antiretroviral therapy(cART) suppressed viral replication and restored T lymphocyte abnormalities. The establishment of HIV-1 infected humanized B-NSG mice not only provides a model to study virus and T cell interplays, but also can be a useful tool to evaluate antiviral strategies.     

Tandem Duplication and Random Loss for mitogenome rearrangement in Symphurus (Teleost: Pleuronectiformes)

Background

The mitochondrial genomes (mitogenomes) of flatfishes (Pleuronectiformes) exhibit highly diversified types of large-scale gene rearrangements. We have reported that the mitogenomes of Crossorhombus azureus (Bothidae), Samariscus latus (Samaridae) and Cynoglossus fishes (Cynoglossidae) show different types of gene rearrangements.

Results

In the present study, the complete mitogenomes of two Symphurus species (Cynoglossidae), Symphurus plagiusa and Symphurus orientalis, were determined. The gene order in the S. plagiusa mitogenome is the same as that of a typical vertebrate (without any gene rearrangements). Surprisingly, large-scale gene rearrangements have occurred in S. orientalis. In the rearranged fragment from the control region (CR) to the WANCY tRNA cluster (tRNA cluster of tRNA-W, tRNA-A, tRNA-N, tRNA-C and tRNA-Y) in the S. orientalis mitogenome, tRNA-V and tRNA-M have been translocated to the 3’ end of the 16S rRNA gene, with six large intergenic spacers over 20 bp in length. In addition, an origin for light-strand replication (O_L) structure that is typically located in the WANCY region was absent in both the S. plagiusa and S. orientalis mitogenomes. It is generally recognized that a sequence in the WANCY region that encodes tRNAs forms a hairpin structure (O_L-like structure) and can act as the O_L when the typical locus is lost. Moreover, an additional O_L-like structure was identified near the control region in the S. plagiusa mitogenome.

Conclusions

The positions of the intergenic spacers and the rearranged genes of the S. orientalis mitogenome strongly indicate that the mechanism underlying the rearrangement of this mitogenome was Tandem Duplication and Random Loss. Additionally, two O_L-like regions substituting for the typical locus were found in the S. plagiusa mitogenome. We speculate that the ancestral mitogenomes of S. plagiusa and S. orientalis also had this characteristic, such that if both O_L-like structures functioned during mitochondrial replication, they could initiate duplicate replications of the light strand (L-strand), leading to duplication of the region between the two structures. We consider that this mechanism may account for the gene duplication that occurred during the gene rearrangement process in the evolution of the ancestral mitogenome to the S. orientalis mitogenome.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1581-6) contains supplementary material, which is available to authorized users.     

A novel model of double replications and random loss accounts for rearrangements in the Mitogenome of Samariscus latus (Teleostei: Pleuronectiformes)

Background

Although more than one thousand complete mitochondrial DNA (mtDNA) sequences have been determined in teleostean fishes, only a few gene rearrangements have been observed, and genome-scale rearrangements are even rarer. However, flatfishes (Pleuronectiformes) have been identified as having diverse types of mitochondrial gene rearrangements. It has been reported that tongue soles and the blue flounder mitogenomes exhibit different types of large-scale gene rearrangements.

Results

In the present study, the complete mitochondrial genome of another flatfish, Samariscus latus, was sequenced, and genome-scale rearrangements were observed. The genomic features of this flounder are different from those of any other studied vertebrates, including flatfish species too. The mitogenome of S. latus is characterized by the duplication and translocation of the control region (CR). The genes located between the two CRs are divided into two clusters in which their relative orders are maintained.

Conclusions

We propose a “Double Replications and Random Loss” model to explain the rearrangement events in S. latus mitogenome. This model consists of the following steps. First, the CR was duplicated and translocated. Subsequently, double replications of the mitogenome were successively initiated from the two CRs, leading to the duplication of the genes between the two CRs. Finally, one of each pair of duplicated genes was lost in a random event.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-352) contains supplementary material, which is available to authorized users.     

Cambrian Onychophora or Xenusians

The Cambrian xenusians have certain similarities to modern onychophorans and tardigrades, and it is tempting to suggest a relationship to one or the other of them. However, similarities are often on a fairly general level. For instance, the three groups all shed their cuticle, but they leave it in three different ways: through a ventral split, through a dorsal split, and through the anterior opening, respectively. Considering also all other characters, we are left with a complicated mosaic character pattern that is indeed difficult to interpret. In addition, the fossils show a great diversity, and there is disagreement on how to interpret some features of them. It seems premature to interpret xenusians as close relatives of any extant group. They may or may not be related: we should also remember the plain fact that an overwhelming majority of characters in organisms have evolved more than once.     

An early Cambrian polyp reveals a potential anemone-like ancestor for medusozoan cnidarians

Cnidarians form a disparate phylum of animals and their diploblastic body plan represents a key step in animal evolution. Cnidarians are split into two main classes; anthozoans (sea anemones, corals) are benthic polyps, while medusozoans (hydroids, jellyfishes) generally have alternating life cycle stages of polyps and medusae. A sessile polyp is present in both groups and is widely regarded as the ancestral form of their last common ancestor. However, the nature and anatomy of the ancestral polyp, particularly of medusozoans, is controversial, owing to the divergent body plans of the extant lineages and the scarcity of medusozoan soft tissues in the fossil record. Here, we redescribe Conicula striata Luo & Hu from the early Cambrian Chengjiang biota, south China, which has previously been interpreted as a polyp, lophophorate or deuterostome. Through re-examination of the holotype and 51 exceptionally preserved specimens, we show that C. striata possessed features of both anthozoans and medusozoan polyps. A conical, annulated organic skeleton (periderm) fully encasing a polyp is found in fossil and living medusozoans, while a tubular pharynx extending from the mouth into a gut partitioned by c. 28 mesenteries, resembling the actinopharynx of anthozoans. Our phylogenetic analyses recover C. striata as a stem-group medusozoan, implying that the wealth of medusozoan diversity derived, ultimately, from an anemone-like ancestor.     

利用PMCA技术实验不同剂量MCT联合DTT对PrP~(sc)转化的抑制效应

蛋白质错误折叠循环扩增(protein misfolding cyclic amplification,PMCA)技术,是一种具有感染性的朊蛋白(PrPsc)体外扩增的技术,可用于抑制细胞型朊蛋白(PrPc)向PrPsc转化药物的筛选.本研究在Saborio等提供的方法基础上,成功优化了最佳扩增时间,利用优化的PMCA技术实验了不同剂量氮芥(mechlorethamine,MCT)联合二硫苏糖醇(dithiothreitol,DTT)对PrPsc转化的抑制效应.结果发现,MCT联合DTT能体外抑制PrPc向PrPsc的转化,抑制作用具有明显的量效关系.利用针对人PrPc不同结构域的抗体发现,MCT联用DTT可使抗体6H4的抗原表位隐蔽,该表位位于PrPc第1个α螺旋区内,是构象转化的主要部位.对其机制的深入探讨,将有助于新一类可传播性海绵状脑病(transmissible spongiform encephalopathy,TSE)治疗药物的研发.     

爬行动物MHC基因研究进展

主要组织相容性复合体(MHC)是有颌脊椎动物中发现的编码免疫球蛋白受体的高度多态的基因群,因其在免疫系统中的重要作用而备受关注。脊椎动物不同支系间MHC的结构和演化差异较大。尽管MHC基因特征在哺乳类、鸟类、两栖类和鱼类中已被较好地描述,但对爬行动物MHC的了解仍较少。鉴于爬行动物对于理解MHC基因的演化占据很重要的系统发育位置,研究其MHC具有重要意义。本文就近年来爬行动物MHC的分子结构、多态性维持机制、功能和主要应用的研究现状进行了系统地回顾和总结,并展望了其研究前景。