The CD4 T cell-deficient mouse mutation nackt (nkt) involves a deletion in the cathepsin L (Ctsl) gene

We recently reported a novel autosomal recessive mouse mutation designated nackt (nkt). Homozygous mutant mice have diffuse alopecia and a marked reduction in the proportion of CD4+ T cells in the thymus and peripheral lymphoid tissues. Here we show that the CD4 T-cell deficiency is due to a defect in the thymic microenvironment rather than the hematopoietic compartment. Furthermore, we identified the molecular basis of the mutant phenotype by demonstrating that the nkt mutation represents a 118-bp deletion of the cathepsin L (Ctsl) gene which is required for degradation of the invariant chain, a critical chaperone for major histocompatibility complex class II molecules. This finding explains the similarities in skin and immune defects observed in nkt/nkt and Ctsl -/- mice. The data reported here provide further in vivo evidence that the lysosomal cysteine protease cathepsin L plays a critical role in CD4+ T-cell selection in the thymus.     

Phylogenetic relationships and reticulation among Asian Elymus (Poaceae) allotetraploids: Analyses of three nuclear gene trees

This phylogenetic study focuses on a subset of the species in Elymus—specifically, the endemic Asian tetraploids presumed to combine the St genome from Pseudoroegneria with the Y genome from an unknown donor. The primary goals were to (1) determine whether the St and Y genomes are derived from phylogenetically distinct donors; (2) identify the closest relative, and potentially the likely donor, of the Y genome; and (3) interpret variation among StStYY species in terms of multiple origins and/or introgression. The goals were addressed using phylogenetic analyses of sequences from three low-copy nuclear genes: phosphoenolpyruvate carboxylase, β-amylase, and granule-bound starch synthase I. Data sets include 16 StStYY individuals representing nine species, along with a broad sample of representatives from most of the monogenomic (i.e., non-allopolyploid) genera in the tribe. To briefly summarize the results: (1) the data clearly support an allopolyploid origin for the Asian tetraploids, involving two distinct donors; (2) the Y genome was contributed by a single donor, or multiple closely-related donors; (3) the phylogenetic position of the Elymus Y genome varies among the three trees and its position is not strongly supported, so the identity of the donor remains a mystery; and (4) conflicts among the gene trees with regard to the St-genome sequences suggest introgression involving both Elymus and Pseudoroegneria.     

Cadmium-induced ovarian pathophysiology is mediated by change in gene expression pattern of zinc transporters in zebrafish (Danio rerio)

This study explored the potential for expression pattern of genes encoding zinc (Zn) transporters to be involved in the cadmium (Cd)-induced reproductive toxicity in female of zebrafish. For this purpose, oocytes maturity and ovarian histology as well as Cd, Zn and metallothioneins (MTs) accumulation and expression of genes encoding Zrt-,Irt-related protein 10 (ZIP10), Zn transporter 1 (ZnT1) and zebrafish metallothionein (zMT) were examined in ovaries of adult zebrafish exposed to 0.4 mg/L Cd in water and supplemented with Zn (5 mg kg⁻¹) in their diet for 21 days. Cd-exposure decreased the expression of ZnT1 and caused up-regulation of ZIP10 and zMT gene expression. These changes were accompanied by increased Cd and MTs accumulation, decreased Zn contents as well as by histopathological damages in ovarian tissues. The co-exposure of fish to Cd and Zn abolished ZnT1 down-regulation and rendered a persistently increased ZIP10 mRNA level. This treatment also decreased Cd and MTs accumulation, reversed Cd-induced Zn depletion and partially restored Cd-induced histological changes in ovarian tissues. These results imply that the downregulation of ZnT1 as well as the overexpression of ZIP10, in responses to the ovarian Zn depletion induced by Cd, play a major role in Cd accumulation and consequently in its toxicity. The protective effect of dietary Zn supplementation against Cd-induced toxicity is mediated, at least in part, by the increase of Zn availability and subsequently the induction of ZnT1 gene expression.     

Cloning and expression pattern of the lysozyme C gene in zebrafish

Here, we report isolation and developmental expression pattern of the zebrafish lysozyme C gene. Amino acid sequence analysis showed that the zebrafish lysozyme C protein shared approximately 37-80% identities with the mouse, human, chicken, and carp counterparts. Whole-mount in situ hybridization showed that the lysozyme C gene was expressed in macrophages, as its expression was co-localized with the known myeloid lineage markers L-plastin and PU.1. At 20 hours postfertilization (hpf), most of the lysozyme C positive cells were localized in the yolksac and head mesenchyme but not in the intermediate cell mass, supporting the notion that the primitive macrophage originated from the yolksac (Development 126 (1999) 3735). At 36hpf, the lysozyme C positive cells scattered within the head and yolksac, and began to appear in the caudal part of axial vein. By 6 days postfertilization (dpf), the lysozyme C positive cells accumulated in the kidney where hematopoiesis had been indicated to take place after 4dpf (Dev. Dyn. 214 (1999) 323). Taken together, our results demonstrate that the lysozyme C gene is specifically expressed in myeloid lineage, suggesting that it could serve as an excellent marker for genetic screening of both primitive and definitive myeloid lineage development in zebrafish.     

家蚕组织蛋白酶基因家族的鉴定及表达特征分析

家蚕是鳞翅目完全变态昆虫,在其变态过程中伴随着巨大的形态变化,包括旧组织的解离和新组织的形成,在这过程中有多种组织蛋白酶参与。组织蛋白酶是一类细胞内蛋白酶,广泛存在于各个物种中,包括组织蛋白酶B、H、L等几个亚家族。对家蚕组织蛋白酶的研究将有利于阐明家蚕变态发育的详细过程。通过对家蚕基因组数据库进行筛选,共在家蚕中鉴定到13种组织蛋白酶,并对这13种组织蛋白酶的基本信息和表达模式进行了分析。另外,利用家蚕基因芯片数据和荧光定量PCR分析,鉴定编号为BGIBMGA004622的基因为卵巢特异表达的组织蛋白酶L亚家族基因。该基因全长1 209 bp,编码402个氨基酸。经过序列分析,该酶与其他物种的组织蛋白酶L具有较高的同源性,其活性位点高度保守,且与鳞翅目的组织蛋白酶L在进化上聚为一支。同时,对该基因进行克隆并原核表达,结果显示重组蛋白以包涵体的形式表达。定量PCR结果显示,该酶在蛹发育初期表达量逐渐升高,至蛹3 d达到最高值,推测其可能参与卵巢与卵母细胞的发育过程。     

Cloning and expression pattern of vat-1 homolog gene in zebrafish

The VAT-1 protein is present in the electric organ of marine rays where it is suggested to play a central role in nerve signal transmission. VAT-1 homolog protein was also identified in mouse and human but its function remains to be determined. We have investigated VAT-1 homolog in zebrafish Danio rerio since it is an excellent model amenable to the combination of genetic, molecular and embryological studies. Amino acid sequence analysis shows that the zebrafish VAT-1 homolog shares approximately 51-61% identity with the electric ray, mouse, and human counterparts. By in situ hybridization, vat-1 homolog mRNA is first observed in the trigeminal nuclei at the 8-somite stage. At 20-somite stage, vat-1 homolog is detected in the brain, namely in primary clusters of neurons, in the epiphysis and in the hindbrain. vat-1 homolog is also present in the neural tube but this expression disappears after 72 h post-fertilization. At 24 h post-fertilization, vat-1 homolog starts to be expressed in the developing gut. At later stages, vat-1 homolog is present throughout the brain, appears in the maturing retina and the pharyngeal cavity.     

Phylogenetic relationships of allotetraploid Hordelymus europaeus (L.) Harz (Poaceae: Triticeae)

Phylogenetic analyses of sequence data from two plastid genes (rbcL and ndhF) and two single-copy nuclear genes (DMC1 and EF-G) are used to elucidate the origin of the tetraploid, monotypic Hordelymus europaeus. Previous data have mostly shown an allopolyploid origin of Hordelymus, but very recently it was suggested that Hordelymus is autoploid. The present analysis, including representatives from all basic genome types accepted in the Triticeae, lends support to an alloploid origin. There is substantial support for the progenitor of Psathyrostachys as female genome donor of Hordelymus. Individual nuclear gene trees disagree about the male genome donor, but combined analysis of all data weakly supports the common progenitor of Pseudoroegneria and Henrardia as the male genome donor.     

Phylogenetic relationships of the Temnocephaloidea (Platyhelminthes)

Temnocephala novaezealandiae (family Temnocephalidae) and Troglocaridicola mrazeki and Scutariella georgica (family Scutariellidae) were studied by electron microscopy in an attempt to reveal characters that would indicate their phylogenetic relationship to other members of the Platyhelminthes. Ultrastructural features of the epidermis in these temnocephaloideans are like those of the neoophoran turbellarians. The epidermis is syncytial, is honeycombed by a multitude of gland necks whose secretions produce an epidermal surface film, and is underlaid by a thick basement membrane. Some cells in the parenchyma are compartmentalized by intrusive cell processes from neighboring parenchymal cells in a fashion similar to parenchymal structure in the Monogenea and Digenea. The spermatozoa have a pair of free 9+1 flagella and contain aligned dense bodies. The Temnocephaloidea is evidently derived from an early rhabdocoel-turbellarian-like ancestor.     

Phylogenetic relationships in the skin-inhabiting Sarcoptoidea (Acari: Astigmata)

A phylogenetic analysis of relationships is carried out among the skin-inhabiting mites presently included in the families Rhyncoptidae, Audycoptidae and Sarcoptidae. The analysis included eight taxa (five genera of follicle associates and three subfamilies of skin-burrowing sarcoptids) and 41 characters, and generated a single most parsimonious tree of length 60 with a consistency index of 0.745. The genus Caenolestocoptes (Sarcoptidae, Caenolestocoptinae) is the sister group of the assemblage of Ursicoptes, Saimirioptes, Audycoptes (all Audycoptidae) and Rhyncoptes (Rhyncoptidae). Within the latter grouping Audycoptes is the sister group of Rhyncoptes. Based on these results the Caenolestocoptinae Fain & Lukoschus, 1976 and Audycoptidae Lavoipierre, 1964 are synonymised with the Rhyncoptidae Lawrence, 1956. The Rhyncoptidae (sensu nov.) appears to be the sister group of the Sarcoptidae (sensu stricto).The distribution pattern of follicle inhabitation and skin-burrowing in the Sarcoptoidea can be explained by one adaptation to follicle inhabitation (Rhyncoptidae), another to skin-burrowing (Sarcoptidae) and possibly a third in their common ancestor from living on the hairs or on the skin to living in the skin (follicles, burrows or both).New host and locality data for various taxa in the Rhyncoptidae (sensu nov.) are provided. The larva of Audycoptes lawrencei Lavoipierre, 1964, is described and the female redescribed.     

Phylogenetic analysis of the myostatin gene sub-family and the differential expression of a novel member in zebrafish

The myostatin (MSTN)-null phenotype in mammals is characterized by extreme gains in skeletal muscle mass or "double muscling" as the cytokine negatively regulates skeletal muscle growth. Recent attempts, however, to reproduce a comparable phenotype in zebrafish have failed. Several aspects of MSTN biology in the fishes differ significantly from those in mammals and at least two distinct paralogs have been identified in some species, which possibly suggests functional divergence between the different vertebrate classes or between fish paralogs. We therefore conducted a phylogenetic analysis of the entire MSTN gene sub-family. Maximum likelihood, Bayesian inference, and bootstrap analyses indicated a monophyletic distribution of all MSTN genes with two distinct fish clades: MSTN-1 and -2. These analyses further indicated that all Salmonid genes described are actually MSTN-1 orthologs and that additional MSTN-2 paralogs may be present in most, if not all, teleosts. An additional zebrafish homolog was identified by BLAST searches of the zebrafish Hierarchical Tets Generation System database and was subsequently cloned. Comparative sequence analysis of both genes (zebrafish MSTN (zfMSTN)-1 and -2) revealed many differences, primarily within the latency-associated peptide regions, but also within the bioactive domains. The 2-kb promoter region of zfMSTN-2 contained many putative cis regulatory elements that are active during myogenesis, but are lacking in the zfMSTN-1 promoter. In fact, zfMSTN-2 expression was limited to the early stages of somitogenesis, whereas zfMSTN-1 was expressed throughout embryogenesis. These data suggest that zfMSTN-2 may be more closely associated with skeletal muscle growth and development. They also resolve the previous ambiguity in classification of fish MSTN genes.     

Phylogenetic relationships of fantails (Aves: Rhipiduridae)

We explore the phylogenetic relationships of fantails (Aves: Rhipiduridae) using molecular characters derived from two nuclear introns and two mitochondrial genes. Our results indicate that Rhipidura hypoxantha is not a true fantail, but rather a member of the Stenostiridae clade that is morphologically and behaviourally convergent with fantails. Within the true Rhipiduridae, we identified six distinct clades; however, phylogenetic relationships among these groups were unresolved. The only well-supported sister relationship was between members of the grey and the rufous fantail complexes. Clades recovered through our model-based phylogenetic analyses generally correspond to previously proposed fantail complexes based on morphological characters. The phylogenetic position of R. atra and R. diluta remain unclear, as sister relationships varied between analyses for the prior whereas the latter was placed as sister to the New Guinea thicket fantails, R. leucothorax and R. threnothorax ; yet significant node support was not recovered for either taxa. Biogeographically, fantails appear to have radiated rapidly and the six clades are not geographically restricted, but instead span South-east Asia, New Guinea, Australia and Pacific Islands.     

Phylogenetic relationships of Pieridae (Lepidoptera: Papilionoidea) in China based on seven gene fragments

The butterfly family Pieridae includes approximately 1000 described extant species distributed worldwide. Among these, 154 species belonging to 24 genera are recorded in China. There has been no previous comprehensive phylogenetic study of the molecular phylogeny of Chinese pierids based on molecular data. In this study, 52 species representing 21 genera distributed in China were sampled. We reconstructed their evolutionary history based on four mitochondrial (COII, ND1, Cytb and 16S rDNA) and three nuclear (28S rRNA (D2–D3), 28S rRNA (D8) and EF‐1α) gene fragments using maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI). Our results are congruent with recent studies and support the monophyly of three subfamilies, with Dismorphiinae sister to Coliadinae + Pierinae. Relationships among five genera of Coliadinae are: Eurema + (Dercas + (Goneperyx + (Catopsilia + Colias))). Relationships among the largest subfamily Pierinae are consistently recovered as follows: Leptosiaini + ((Nepheroniini + Teracolini) + (Anthocharidini + Pierini)). The division of three subgenera of Aporia (Aporia, Metaporia and Mesapia) is not supported because both the Aporia (Aporia) and Aporia (Metaporia) are found to be strongly paraphyletic, with Mesapia nested within Aporia sensu lato.