Genome-wide analysis of metallothionein gene family in maize to reveal its role in development and stress resistance to heavy metal

BACKGROUND: Maize (Zea mays L.) is a widely cultivated cereal and has been used as an optimum heavy metal phytoremediation crop. Metallothionein (MT) proteins are small, cysteine-rich, proteins that play important roles in plant growth and development, and the regulation of stress response to heavy metals. However, the MT genes for maize have not been fully analyzed so far. METHODS: The putative ZmMT genes were identified by HMMER. The heat map of ZmMT genes spatial expression analysis was generated by using R with the log2 (FPKM + 1). The expression profiles of ZmMT genes under three kinds of heavy metal stresses were quantified by using qRT-PCR. The metallothionein proteins was aligned using MAFFT and phylogenetic analysis were constructed by ClustalX 2.1. The protein theoretical molecular weight and pI, subcellular localization, TFs binding sites, were predicted using ProtParam, PSORT, PlantTFDB, respectively. RESULTS: A total of 9 ZmMT genes were identified in the whole genome of maize. The results showed that eight of the nine ZmMT proteins contained one highly conserved metallothio_2 domain, while ZmMT4 contained a Metallothio_PEC domain. All the ZmMT proteins could be classified into three major groups and located on five chromosomes. The ZmMT promoters contain a large number of hormone regulatory elements and hormone-related transcription factor binding sites. The ZmMT genes exhibited spatiotemporal specific expression patterns in 23 tissues of maize development stages and showed the different expression patterns in response to Cu, Cd, and Pb heavy metal stresses. CONCLUSIONS: We identified the 9 ZmMT genes, and explored their conserved motif, tissue expression patterns, evolutionary relationship. The expression profiles of ZmMT genes under three kinds of heavy metal stresses (Cu, Cd, Pb) were analyzed. In summary, the expression of ZmMTs have poteintial to be regulated by hormones. The specific expression of ZmMTs in different tissues of maize and the response to different heavy metal stresses are revealed that the role of MT in plant growth and development, and stress resistance to heavy metals.

Expression patterns of mesenchymal stem cell-specific proteins in adipose tissue-derived cells: possible immunosuppressing agent in partial allograft for restoring the urinary bladder in rabbits / Padrão de expressão de proteínas específicas das células-tronco mesenquimais derivadas de tecido adiposo: possível agente imunossupressor em aloenxerto parcial para restauração de vesícula urinária de coelhos

Adipose tissue-derived stem cells (ADSCs) are an attractive source of mesenchymal stem cells (MSCs) for use in tissue engineering and clinical applications. This paper focuses on the characterization of ADSCs used as immunosuppressive agent in rabbits undergoing partial allograft for urine bladder restorage. For this study highlighted the characterization of the ADSCs used as immunosuppressive agents in rabbits submitted to partial allograft for restoration of the urinary vesicle, using 25 animals, six months old, New Zealand. ADSCs at the third peal were characterized by the MSC-specific CD105, CD73 and CD90 expression and by the absence of the hematopoietic marker CD45, as revealed by flow cytometry analysis. Moreover, ADSCs were efficient in preventing allograft rejection from the urinary bladder, as judged by biochemical, clinical and ultrasonography analysis. Together, these results compose characterization of protein expression profiles and immunosuppressive functionality of ADSCs in rabbits, which had undergone partial allografts of the urinary bladder, foreseeing future applications in clinical practice.(AU)

As células mesenquimais derivadas de tecido adiposo (ADSCs) são uma fonte atraente de células-tronco mesenquimais (MSCs) para uso na engenharia de tecidos e suas aplicações clínicas. Este trabalho destacou a caracterização das ADSCs utilizadas como agentes imunossupressores em coelhos submetidos a aloenxerto parcial para restauração da vesícula urinária, sendo utilizados 25 animais, de seis meses de idade, Nova Zelândia. As ADSCs, após o terceiro repique, foram caracterizadas pela expressão específica de MSC CD105, CD73 e CD90 e pela ausência do marcador hematopoiético CD45, tal como revelado por análise de citometria de fluxo. Além disso, os ADSCs foram eficientes na prevenção da rejeição de aloenxertos da vesícula urinária, conforme avaliado por análises clínica, bioquímica e ultrassonográfica. Juntos, esses resultados compõem a caracterização dos perfis de expressão proteica e a funcionalidade imunossupressora de ADSCs em coelhos, que sofreram aloenxertos parciais da bexiga, prevendo futuras aplicações na prática clínica.(AU)

Different expression patterns of β-catenin and its correlation with clinicopathological facters in colorectal cancer / 临床与实验病理学杂志

To observe different expression patterns of β-catenin and its clinical significance in colorectal cancer (CRC).Methods A total of 181 cases of CRC tissues and 30 cases of normal colorectal tissue were investigated by immunohistochemistry for the expression of β-catenin.Results The expression rate of β-catenin was 56.9％ (103/181) in CRC,and higher than that in normal colorectal tissue (P ＜ 0.05).The overexpression of nuclear β-catenin was significantly correlated with histological differentiation,lymph node metastasis and Dukes' stage in CRC (P ＜ 0.05),and no relationship with other pathological parameters,such as age,gender and the depth of infiltration.The incomplete membranous expression of β-catenin was significantly correlated with histological differentiation,the depth of infiltration,lymph node metastasis and Dukes' stage in CRC (P ＜ 0.05).The high expression of nuclear β-catenin related to histological differentiation and Dukes' stage in CRC (P ＜ 0.05).In the follow-up data of 82 cases of CRC,the expression of nuclear β-catenin was associated with poor prognosis,and the 5-year survival rate was significantly lower than that of self-control groups (P ＜ 0.05).Conclusion β-catenin plays important roles in colorectal carcinogenesis.Abnormal expression of β-catenin was related to the aggressive progression of CRC and may be helpful for evaluating the prognosis of patients with CRC.β-catenin is expected to become a new target for diagnosis and treatment of CRC in future.

Expression analysis of vasa in Asian paddle crab (Charybdis japónica) exposed to Bisphenol A

Background: Bisphenol A (BPA) is an endocrine-disrupting chemical (EDC) with a weak estrogen-like activity in fish that is found ubiquitously in aquatic environments. However, there has been little study about BPA on the endocrine disrupting effects of crab. In the present study, cDNA of vasa was cloned and characterized in the Charybdis japonica. Histological structures of testis and expression patterns of vasa gene in the testis of C. japonica after treatment with BPA were investigated. Results: The cDNA of vasa is composed of 3051 bp with a 2166 bp open reading frame encoding 721 AA. The deduced amino acid sequence contained eight conserved domains of the DEAD-box protein family. The tissue distribution showed that vasa mRNA was specifically expressed in ovary and testis. Histologically, the sperm cells were decreased in number and an acellular zone was seen in the testis. The transcript level of vasa gradually increased with a significant difference between the experimental and control groups. After BPA exposure with 0.50 and 1.00 mg/L for 1,3, 6 and 9 d, the expression levels of vasa increased. Conclusion: These findings suggest that BPA can increase the expression level of vasa mRNA and influence the development of the testis in C. japonica.

Identification and expression analysis of two Wnt4 genes in the spotted scat (Scatophagus argus)

Background: WNT4 is a protein that plays a crucial role in ovarian differentiation and development in mammals, with a relatively well understood function in mammalian gonadal differentiation. The role of WNT4 in teleost fish; however, remains unclear. In the present study, cDNAs of Wnt4a and Wnt4b were cloned and characterized in the spotted scat. The expression patterns of two Wnt4 genes in the gonads at different stages of development and in fish after treatment with 17a-methyltestosterone (MT) were investigated. Results: The tissue distribution showed that Wnt4a was expressed in various tissues, including the gonads, gills, spleen, brain, and fin. Interestingly, Wnt4b not only was expressed in the gills, brain, and spleen, but also was obviously expressed in the ovary. During gonad development, Wnt4a was highly expressed in the testis at stage I and Wnt4b was mainly expressed in the ovary at stages II-III. After MT treatment, the mRNA expression of Wnt4a increased significantly up to 40 d, and the transcript level of Wnt4b decreased at 20 d. Conclusions: These results suggest that Wnt4a may be involved in gonad development and plays a role in the process of spermatogonial proliferation. Our results also demonstrate that Wnt4b is not only expressed in the nervous system, but also in the ovary and it may be involved in ovary development of the spotted scat.

Temporal protein expression pattern in intracellular signalling cascade during T-cell activation: A computational study.

Various T-cell co-receptor molecules and calcium channel CRAC play a pivotal role in the maintenance of cell’s functional responses by regulating the production of effector molecules (mostly cytokines) that aids in immune clearance and also maintaining the cell in a functionally active state. Any defect in these co-receptor signalling pathways may lead to an altered expression pattern of the effector molecules. To study the propagation of such defects with time and their effect on the intracellular protein expression patterns, a comprehensive and largest pathway map of T-cell activation network is reconstructed manually. The entire pathway reactions are then translated using logical equations and simulated using the published time series microarray expression data as inputs. After validating the model, the effect of in silico knock down of co-receptor molecules on the expression patterns of their downstream proteins is studied and simultaneously the changes in the phenotypic behaviours of the T-cell population are predicted, which shows significant variations among the proteins expression and the signalling routes through which the response is propagated in the cytoplasm. This integrative computational approach serves as a valuable technique to study the changes in protein expression patterns and helps to predict variations in the cellular behaviour.

Molecular cloning and expression analysis of the MaASR1 gene in banana and functional characterization under salt stress

Background Abscisic acid (ABA)-, stress- and ripening-induced protein (ASR) is plant-specific hydrophilic transcriptional regulators involved in sucrose stress and wounding in banana. However, it is not known whether banana ASR genes confer salt stress tolerance. The contexts of the study was to analysis the sequence characterization of banana ASR1, and identify its expression patterns and function under salt stress using quantitative real-time PCR (qPCR) and overexpression in Arabidopsis. The purpose was to evaluate the role of banana ASR1 to salt stress tolerance employed by plants. Results A full-length cDNA isolated from banana fruit was named MaASR1, and it had a 432 bp open reading frame (ORF) encoding 143 amino acids. MaASR1 was preferential expression in roots and leaves compared to low expression in fruits, rhizomes and flowers. Under salt stress, the expression of MaASR1 quickly increased and highest expression level was detected in roots and leaves at 4 h, and then gradually decreased. These results suggested that MaASR1 expression was induced under salt stress. MaASR1 protein was localized in the nucleus and plasma membrane. MaASR1 was transformed to Arabidopsis and verified by southern and northern analysis, transgenic lines L14 and L38 integrated one and two copies of MaASR1, respectively, while overexpression in transgenic lines provided evidence for the role of MaASR1 to salt stress tolerance. Conclusions This study demonstrated that overexpression of MaASR1 in Arabidopsis confers salt stress tolerance by reducing the expression of ABA/stress-responsive genes, but does not affect the expression of the ABA-independent pathway and biosynthesis pathway genes.

Molecular cloning, sequence characterization and mRNA tissue expression analysis of TDRP1 gene from the Banna minipig inbred line (BMI) / 中国实验动物学报

Objective To get TDRP1 gene of sterile and fertile boar of the Banna minipig inbred line (BMI), predict its function by bioinformatics analysis, and detect its expression patterns in the fertile boar.Methods Based on the NM_001198925 sequence, we designed specific primers and amplified BMI TDRP1 using RT-PCR method for sequen-cing and bioinformatics analysis.Meanwhile, the expression of TDRP1 in 17 organ tissues ( heart, liver, spleen, lung, kidney, thymus, lymph nodes, skin, duodenum, stomach, cerebrum, cerebellum, testis, epididymis, seminal vesicle, prostate, and bulbourethral gland) of fertile BMI boar and in the testis of sterile and fertile BMI boars was analyzed by semi-quantitative RT-PCR.Results The experiment obtained 680 bp cDNA sequence ( GenBank accession number:KJ186786) of BMI TDRP1, which encodes a protein of 186 amino acids with a predicted molecular weight (Mw) of 20.49 kDa and isoelectric point (pI) 5.86, and no signal peptide.It was a nuclear protein with a probability of 94.1%and had a leucine-rich nuclear export signals.Homology analysis of protein sequences revealed that BMI TDRP1 showed high identi-ty with that of humans, macaca mulatta, mouse and rat.The RT-PCR analysis showed that TDRP1 had a similar expression in the testes of sterile and fertile BMI boars.It was highly abundant in the seminal vesicle and prostate, moderately ex-pressed in cerebellum and testis and weakly expressed in cerebrum and kidney, while undetected in other 11 organ tissues. Conclusions We have cloned TDRP1 complete coding sequence, and found 2 SNPs,showing no difference in sequences and the testis mRNA expression levels between the fertile and sterile BMI boars.The multi-tissue transcription profile shows different expression levels in different organ tissues, being high in the seminal vesicle and prostate.The results of this study provide a foundation for further insight into the role of this gene in spermatogenesis.

Expression Patterns of Tbx1 during Development of the Mouse Mandibular First Molar / 대한해부학회지

A T-box transcription factor gene, Tbx1 is a principal candidate of the most frequent chromosomal deletion syndrome found in human, DiGeorge/velocardiofacial syndrome which is a complex developmental disorder associated with cardiac outflow tract abnormalities, mid facial dysmorphology, velopharyngeal insufficiency and submucosal cleft palate. We performed in situ hybridization against mouse embryo from E13.5 (bud stage) to E18.5 (late bell stage) in order to analyze the expression patterns of Tbx1 in the developing mouse first molar, a derivative of the first pharyngeal arch. Tbx1 transcripts were found in the dental lamina and its surrounding mesenchyme at E13.5 and in the dental organ except enamel knot at E14.5 (cap stage). Tbx1 was strongly expressed in the cervical loop and stratum intermedium but was weak in the dental papilla and dental follicle at E15.5 (early bell stage). At E18.5, Tbx1 was strongly expressed not only in the dental organ (bell stage) except stellate reticulum but also dental papilla and dental follicle adjacent to the inner dental epithelium. In conclusion, Tbx1 transcripts were specifically expressed both in the dental epithelium and surrounding mesenchyme of developing tooth from initiation to bell stage, which were the most similar with those of Sox9 but little different from those of Pitx2 and ectodin. These results strongly suggested that Tbx1 may play a role as a transcription factor regulating proliferation and differentiation of both dental epithelium and mesenchyme through the tooth development.