Phosphatidylethanolamine-binding Protein Research Articles

The Roles And Signaling Pathways Of Phosphatidylethanolamine-Binding Protein 4 In Tumors.

Phosphatidylethanolamine-binding protein 4 (PEBP4) has been found to be highly expressed in many tumors and to be closely related to the proliferation, differentiation, and metastasis of tumors. PEBP4 has also been found to be involved in many cancer-activated signaling pathways and to cause therapeutic resistance. In this study, we first reviewed the morphological structure and expression of PEBP4, then discussed the roles of PEBP4 in individualized treatment of some cancers, and finally explored the possibilities of cultivating PEBP4 as a therapeutic target.We also identified the main signaling pathways in which PEBP4 affects different cancers. It is here concluded that over-expression of PEBP4 can enhance the proliferation and metastasis of the cancer cells and the resistance to radiotherapy/chemotherapy in cancers.

Ready for the journey: a comparative proteome profiling of porcine cauda epididymal fluid and spermatozoa

In the present study, we describe the proteome of porcine cauda epididymis fluid and spermatozoa by means of Multidimensional Protein Identification Technology (MudPIT). Ten sexually mature healthy boars were surgically castrated and epididymides were dissected to obtain the cauda epididymal content. Polled protein extracts of cauda epididymal fluid (CEF) and spermatozoa (CESperm) were loaded in an Agilent 1100 quaternary HPLC and peptides eluted from the microcapillary column were electro-sprayed directly into a LTQ Orbitrap XL mass spectrometer. Using bioinformatics, identified proteins were classified by their molecular functions, involvement in biological processes and participation in relevant metabolic pathways associated with spermatozoa physiology, fertility potential and protection. A total of 645 proteins were identified in the CEF, with epididymal-specific lipocalin-5, beta-hexosaminidase subunit beta precursor and phosphatidylethanolamine-binding protein 4 being the most abundant proteins found. A total of 2886 proteins were identified in the CESperm proteome with 81 proteins being considered more abundant (spectral counts > 100). CEF and CESperm data were compared and 345 proteins were present in both proteomes. Phosphatidylethanolamine-binding protein 4 precursor was the only protein found most abundant in both CEF and CESperm proteomes. Based on Gene Ontology analysis, we identified CEF and CESperm proteins associated with sperm protection against ROS and immune mediated response, glycosaminoglycan degradation, ubiquitin-proteasome system, metabolic process and maturation, modulation of acrosome reaction and ZP binding and oocyte penetration. These results provide a better comprehension about the molecular process and biological pathways involved in sperm epididymis maturation and establishment of the cauda epididymis sperm reservoir.

Critical Periods and Changes Involved in Hemerocallis lilioasphodelus Floral Transition

Hemerocallis is a valuable commercial perennial with prominent flowers. In this study, a wild variety of Hemerocallis lilioasphodelus L. cv. Licheng 7 (H. lilioasphodelus, LC7) from China was chosen as the experimental material due to its stable flowering trait in cultivation. Three main conclusions were obtained. Our observations revealed that the transition from vegetative to reproductive growth had a strong relationship with shoot apical meristem (SAM) fasciation, and the process was outlined in four morphological transitional stages. In addition, we detected a temporary increase in soluble sugar in the shoot tip, which was suggested to be a physiological indicator of inflorescence primordium initiation. In addition, the full-length sequences of two phosphatidylethanolamine-binding protein (PEBP) family genes from the wild species LC7 were cloned. From quantitative RT-PCR (qRT-PCR) and correlation analyses, we elucidated that the two PEBP family genes were correlated with some SAM morphological traits and involved in the regulation of the floral transition of the wild species LC7. To the best of our knowledge, this study provided a precious resource investigation of critical periods in the floral transition of H. lilioasphodelus, which will be recommended for regulation of the flowering time in perennials in the future.

Identification and Characterization of the FLOWERING LOCUS T/TERMINAL FLOWER 1 Gene Family in Petunia.

The phosphatidylethanolamine-binding protein (PEBP) gene family exists in all eukaryote kingdoms, with three subfamilies: FT (FLOWERING LOCUS T)-like, TFL1 (TERMINAL FLOWER 1)-like, and MFT (MOTHER OF FT AND TFL1)-like. FT genes promote flowering, TFL1 genes act as a repressor of the floral transition, and MFT genes have functions in flowering promotion and regulating seed germination. We identified and characterized orthologs of the Arabidopsis FT/TFL1 gene family in petunia to elucidate their expression patterns and evolution. Thirteen FT/TFL1-like genes were isolated from petunia, with the five FT-like genes mainly expressed in leaves. The circadian rhythms of five FT-like genes and PhCO (petunia CONSTANS ortholog) were figured out. The expression of PhFT1 was contrary to that of PhFT2, PhFT3, PhFT4, and PhFT5. PhCO had a circadian clock different from Arabidopsis CO, but coincided with PhFT1; it decreased in daytime and accumulated at night. Two of the FT-like genes with differential circadian rhythm and higher expression levels, PhFT1 and PhFT4, were used to transform Arabidopsis. Eventually, overexpressing PhFT1 strongly delayed flowering, whereas overexpression of PhFT4 produced extremely early-flowering phenotype. Different from previous reports, PhTFL1a, PhTFL1b, and PhTFL1c were relatively highly expressed in roots. Taken together, this study demonstrates that petunia FT-like genes, like FT, are able to respond to photoperiod. The expression pattern of FT/TFL1 gene family in petunia contributes to a new insight into the functional evolution of this gene family.

Identification of proteomic markers for ram spermatozoa motility using a tandem mass tag (TMT) approach

Sperm motility is crucial for ram fertility; however, differences in the proteome of sperm with high- and low- motility in rams (Ovis aries) has yet to be achieved. Thus, the aim of this study was to identify and characterize ram spermatozoa proteins with different abundances between high- and low- motility, and identify of proteomic markers for ram spermatozoa motility using tandem mass tag (TMT) protein labeling and LC-MS/MS. In this study, the abundance of 150 proteins in high-motility (HM) ram sperm was significantly different compared with low-motility (LM) sperm. Proteins involved in sperm motility, mitochondrial activity, and spermatogenesis showed higher abundance in HM ram spermatozoa; proteins involved in protein processing and spliceosome were more abundant in LM ram spermatozoa. In conclusion, Phosphatidylethanolamine binding protein 4 is a potential proteomic marker for ram sperm motility; CCTs/HSPs are hallmarks of the LM spermatozoa in rams. Our findings highlight the functional differences between HM and LM ejaculated spermatozoa and has identified candidate proteins of interest linked to sperm motility in rams. SignificanceInadequate sperm motility is one of the most important reasons for male subfertility or infertility. In this study, we found that the abundance of 150 proteins in high-motility ram sperm was significantly different compared with low-motility sperm. Proteomic biomarkers were discovered to reflect the motility variation in ram spermatozoa; these biomarkers may assist in illuminating the molecular mechanisms underlying sperm motility. This study expands the potential direction for sperm quality screening and animal breeding.

PhePEBP family genes regulated by plant hormones and drought are associated with the activation of lateral buds and seedling growth in Phyllostachys edulis.

Development of lateral buds on the underground rhizome in moso bamboo is essentially the early stage of the development of aboveground branching, which is regulated by Phosphatidyl-Ethanolamine Binding Protein (PEBP) family genes, but it is unknown whether the PEBP family genes are involved in the activation and development of lateral buds underground. By scanning the whole-genome sequence of moso bamboo, we identified 25 PhePEBP family genes and amplified their full-length open reading frames (ORFs). A sequence analysis revealed that they are composed of four exons and three introns, except for PheFT10, which contains six exons and five introns. PheFT10 underwent alternative splicing, resulting in at least four transcripts (PheFT10α, PheFT10β, PheFT10γ and PheFT10δ). Although PhePEBP genes are generally expressed at low levels and show dramatically organ-specific expressions, the transcription levels of most PhePEBP genes, including the transcripts of PheFT10, change with plant age. Together with the observation that the expression of PhePEBP family genes can be regulated by plant hormones and drought, our data suggest that PhePEBP family genes might be involved in the activation of lateral buds and seedling growth. Particularly, PheFT9, PheTFL2 and PheTFL8 may play vital roles during the activation of dormant buds based on the analysis of amino acid substitution and expression profile. These findings provide insights for in-depth exploration of the biological functions of the PhePEBP family genes in regulating the activation of dormant bud and the development of seedling in moso bamboo.

Identification of cotton MOTHER OF FT AND TFL1 homologs, GhMFT1 and GhMFT2, involved in seed germination.

Plant phosphatidylethanolamine-binding protein (PEBP) is comprised of three clades: FLOWERING LOCUS T (FT), TERMINAL FLOWER1 (TFL1) and MOTHER OF FT AND TFL1 (MFT). FT/TFL1-like clades regulate identities of the determinate and indeterminate meristems, and ultimately affect flowering time and plant architecture. MFT is generally considered to be the ancestor of FT/TFL1, but its function is not well understood. Here, two MFT homoeologous gene pairs in Gossypium hirsutum, GhMFT1-A/D and GhMFT2-A/D, were identified by genome-wide identification of MFT-like genes. Detailed expression analysis revealed that GhMFT1 and GhMFT2 homoeologous genes were predominately expressed in ovules, and their expression increased remarkably during ovule development but decreased quickly during seed germination. Expressions of GhMFT1 and GhMFT2 homoeologous genes in germinating seeds were upregulated in response to abscisic acid (ABA), and their expressions also responded to gibberellin (GA). In addition, ectopic overexpression of GhMFT1 and GhMFT2 in Arabidopsis inhibited seed germination at the early stage. Gene transcription analysis showed that ABA metabolism genes ABA-INSENSITIVE3 (ABI3) and ABI5, GA signal transduction pathway genes REPRESSOR OF ga1-3 (RGA) and RGA-LIKE2 (RGL2) were all upregulated in the 35S:GhMFT1 and 35S:GhMFT2 transgenic Arabidopsis seeds. GhMFT1 and GhMFT2 localize in the cytoplasm and nucleus, and both interact with a cotton bZIP transcription factor GhFD, suggesting that both of GhMFT1, 2 have similar intracellular regulation mechanisms. Taken together, the results suggest that GhMFT1 and GhMFT2 may act redundantly and differentially in the regulation of seed germination.

Functional analysis and characterization of antimicrobial phosphatidylethanolamine-binding protein BmPEBP in the silkworm Bombyx mori

Phosphatidylethanolamine-binding proteins (PEBPs) are a class of highly conserved, biologically diverse proteins, which are widely distributed in plants, insects, and mammals. In this study, a Bombyx mori PEBP (BmPEBP) gene was reported, which encodes a protein composed of 209 amino acid residues. BmPEBP includes a predicted signal peptide, indicating that it is an extracellular protein, which differs from the cytoplasmic PEBPs of plants and mammals. Recombinant soluble BmPEBP was successfully synthesized using a prokaryotic expression system and was then purified effectively by Ni2+-NTA affinity chromatography and gel filtration. Far-ultraviolet circular dichroism spectra indicated that BmPEBP had a well-defined β-sheet structure, with the β-sheet content accounting for about 41% of the protein. BmPEBP had a relatively stable structure at temperatures ranging from 15 °C to 57.5 °C. The Tm, ΔH, and ΔS of BmPEBP were 62.27 °C ± 0.14 °C, 570.10 ± 0.17 kJ/mol, and 1.70 ± 0.03 KJ/(mol·K), respectively. Homology modeling analysis suggested that the active sites of BmPEBP were conserved, comprising Pro96, His111, and His143. Quantitative real-time PCR showed that BmPEBP was highly expressed in the silk gland and had very low expression in other tissues. However, BmPEBP expression was significantly upregulated in the larval fat body after infection with two kinds of fungi, Beauveria bassiana and Candida albicans. Moreover, in vitro fungal inhibition tests showed that BmPEBP could significantly inhibit the sporular growth of Saccharomyces cerevisiae, C. albicans, B. bassiana, and Aspergillus fumigatus. To our knowledge, this is the first report to reveal the antifungal role of a PEBP in insects.

A Protein-Linger Strategy Keeps the Plant On-Hold After Rehydration of Drought-Stressed Beta vulgaris.

Most crop plants are exposed to intermittent drought periods. To cope with these continuous changes, plants need strategies to prevent themselves from exhaustive adjustment maneuvers. Drought stress recovery has been shown to be an active process, possibly involved in a drought memory effect allowing plants to better cope with recurrent aridity. An integrated understanding of the molecular processes of enhanced drought tolerance is required to tailor key networks for improved crop protection. During summer, prolonged periods of drought are the major reason for economic yield losses of sugar beet (Beta vulgaris) in Europe. A drought stress and recovery time course experiment was carried out under controlled environmental conditions. In order to find regulatory key mechanisms enabling plants to rapidly react to periodic stress events, beets were either subjected to 11 days of progressive drought, or were drought stressed for 9 days followed by gradual rewatering for 14 days. Based on physiological measurements of leaf water relations and changes in different stress indicators, plants experienced a switch from moderate to severe water stress between day 9 and 11 of drought. The leaf proteome was analyzed, revealing induced protein pre-adjustment (prior to severe stress) and putative stress endurance processes. Three key protein targets, regulatory relevant during drought stress and with lingering levels of abundance upon rewatering were further exploited through their transcript performance. These three targets consist of a jasmonate induced, a salt-stress enhanced and a phosphatidylethanolamine-binding protein. The data demonstrate delayed protein responses to stress compared to their transcripts and indicate that the lingering mechanism is post-transcriptionally regulated. A set of lingering proteins is discussed with respect to a possible involvement in drought stress acclimation and memory effects.

Effect of RKIP on proliferation and migration of malignant melanoma cells and potential mechanism

Objective: To explore the effect and mechanism of Rafkinase inhibitor protein (RKIP) on proliferation and migration of malignant melanoma cells in vitro. Methods: The RKIP overexpression and down-regulated stable transfected strains of mouse malignant melanoma cell line B16 were constructed by recombinant lentiviral transfection technique and established as RKIP overexpression group and RKIP down-regulation group, the mouse malignant melanoma B16 cells without any treatment were used as a blank control group, and the proliferation activity and migration ability of each group were detected by cell counting kit-8 (CCK-8) and cell scratch test. The relative expression levels of CyclinD1, Calcium-dependent cell adhesin, Ki-67, Matrix metalloproteinase (MMP)-9, MMP-13, MMP-2 and Phosphatidylethanolamine binding protein (PEBP-1) were detected by quantitative real-time polymerase chain reaction (qPCR). Western blot was used to detect the difference of RKIP expression and the protein expression level of nuclear factor-kappa B (NF-κB) signaling pathway in each group. Results: Comparison of RKIP overexpression group and blank control group shown cell proliferation and migration were significantly inhibited in RKIP overexpression group (0.794±0.038 vs 1.200±0.081) (P<0.001). However, there was no significant difference in cell proliferation between RKIP down-regulation group and blank control group (1.077±0.084 vs 1.200±0.081) (P>0.05), and the cell migration ability of RKIP down-regulation group was significantly higher than that of the blank control group (P<0.001). In addition, there was no significant difference between the RKIP down-regulation group and the blank control group in PEBP-1 expression (P>0.05), while the expression levels of the remaining genes in the RKIP overexpression group were significantly lower than those in the blank control group, and the expression levels in the RKIP down-regulation group were significantly higher than those in the blank control group (P<0.001). Furthermore, the protein level of phosphorylated P65 (p-P65) in RKIP overexpression group was significantly lower than that in blank control group (0.080±0.000 vs 0.236±0.000), and RKIP down-regulation group was significantly higher than that in blank control group (1.139±0.001 vs 0.236±0.000) (both P<0.001). Conclusion: RKIP overexpression can inhibit the proliferation and migration of malignant melanoma cells, which may be related to the regulation of NF-κB signaling pathway-related protein p-P65.

Retraction: Human Phosphatidylethanolamine-Binding Protein 4 Promoted the Radioresistance of Human Rectal Cancer by Activating Akt in an ROS-Dependent Way.

Retraction: Human Phosphatidylethanolamine-Binding Protein 4 Promoted the Radioresistance of Human Rectal Cancer by Activating Akt in an ROS-Dependent Way.

Cloning and Characterization of Two FLOWERING LOCUS T-like Genes from Rubber Tree (Hevea brasiliensis)

Two Flower locus T-like genes were isolated and characterized in rubber tree. Both of them were found to have conserved genomic characteristics of the phosphatidylethanolamine-binding protein family and their encoding proteins possess conserved key amino acid residues of FT. RT-qPCR results showed that HbFT1 was restricted in mature leaves, male and female flowers whereas HbFT2 showed the main expression in reproductive organs. Both of them can be induced by short-day conditions but with different expression patterns and regulated by temperature. Transgenic wild-type Arabidopsis can reduce the flowering time and change plant morphogenesis, with fewer rosette leaves and terminal inflorescence to some degrees, as well as the phenotype of ft-10 can also be successfully rescued by ectopic expressing each of them, respectively, suggesting that both of them can exert the function of AtFT. However, they displayed different seasonal-change expression patterns, with HbFT1 mainly accumulating in September for adult trees and in October for juvenile trees and HbFT2 transcript accumulating in March, which was the main floral initiation period, even though both of them showed higher transcript levels in adult trees than 2-year-old trees, implying their different biological functions during growth and development as well as flowering transition in rubber tree.

Different divergence events for three pairs of PEBPs in Gossypium as implied by evolutionary analysis.

The phosphatidylethanolamine-binding protein (PEBP) gene family plays a crucial role in seed germination, reproductive transformation, and other important developmental processes in plants, but its distribution in Gossypium genomes or species, evolutionary properties, and the fates of multiple duplicated genes remain unclear. The primary objectives of this study were to elucidate the distribution and characteristics of PEBP genes in Gossypium, as well as the evolutionary pattern of duplication and deletion, and functional differentiation of PEBPs in plants. Using the PEBP protein sequences in Arabidopsis thaliana as queries, blast alignment was carried out for the identification of PEBP genes in four sequenced cotton species. Using the primers designed according to the PEBP genome sequences, PEBP genes were cloned from 15 representative genomes of Gossypium genus, and the gene structure, CDS sequence, protein sequence and properties were predicted and phylogenetic analysis was performed. Taking PEBP proteins of grape as reference, grouping of orthologous gene, analysis of phylogeny and divergence of PEBPs in nine species were conducted to reconstruct the evolutionary pattern of PEBP genes in plants. We identified and cloned 160 PEBPs from 15 cotton species, and the phylogenetic analysis showed that the genes could be classified into the following three subfamilies: MFT-like, FT-like and TFL1-like. There were eight single orthologous group (OG) members in each diploid and 16 double OG members in each tetraploid. An analysis of the expression and selective pressure indicated that expression divergence and strong purification selection within the same OG presented in the PEBP gene family. An evolutionary pattern of duplication and deletion of the PEBP family in the evolutionary history of Gossypium was suggested, and three pairs of genes resulted from different whole-genome duplication events.

Identification of FT family genes that respond to photoperiod, temperature and genotype in relation to flowering in cassava (Manihot esculenta, Crantz)

Cassava is a starch-storing root crop that is an important source of dietary energy in tropical regions of the world. Genetic improvement of cassava by breeding is hindered by late flowering and sparse flower production in lines that are needed as parents. To advance understanding of regulatory mechanisms in cassava, this work sought to identify and characterize homologs of the FLOWERING LOCUS T (FT) gene. Ten members of the phosphatidylethanolamine-binding protein gene family, to which FT belongs, were obtained from the cassava genome database. Phylogenetic and sequence analysis of these proteins was used to identify two putative FT homologs which had amino acid sequences at key positions in accordance with those predicted for functional FTs. Expression of these ten genes was determined in mature leaves, immature leaves, flower buds, fibrous roots, storage roots and stem. The FT transcripts were expressed in mature leaves, as expected for their possible role in leaf-to-apical meristem signaling. In growth chamber studies, plants flowered earlier in long-day photoperiod than in short-day photoperiod. Expression studies indicated that while MeFT1 was expressed in leaves without a clear-cut photoperiod response, MeFT2 was expressed in a photoperiod-dependent manner, consistent with its involvement in photoperiodic control of flowering. In growth chambers that subjected plants to a range of temperatures from 22 to 34 °C, flowering was delayed by warmer temperatures although MeFT1 and MeFT2 expression declined in only one genotype, indicating other factors regulate this response. The earliest flowering genotype, IBA980002, had high levels of MeFT1 and MeFT2 expression, suggesting that both homologs contribute to earliness of this genotype.

Phosphatidylethanolamine-binding protein 4 promotes the epithelial-to-mesenchymal transition in non-small cell lung cancer cells by activating the sonic hedgehog signaling pathway.

Phosphatidylethanolamine-binding protein 4 (PEBP4), a member of the PEBP family, has been reported to play a pivotal role in tumor progression. However, its role in epithelial-to-mesenchymal transition (EMT) in non-small cell lung cancer (NSCLC) cells remains unclear. Here, we investigated the effects and underlying mechanism of PEBP4 in NSCLC EMT. Three human NSCLC cell lines (A549, H1299, and H460) were transfected with pcDNA3.1-PEBP4 or PEBP4-targeting small interfering RNA. Then, cell proliferation was analyzed by the MTT assay, and cell migration and invasion were analyzed by the transwell chamber assay. Protein and messenger RNA expression of the related genes and proteins were assessed by Western blot analysis and quantitative real-time polymerase chain reaction, respectively. Results showed that PEBP4 was highly expressed in the human lung cancer tissues and three human NSCLC cell lines. Pretreatment with pcDNA3.1-PEBP4 promoted the proliferation, invasion, and migration of NSCLC cells and increased EMT in vitro and lung tumor metastasis in vivo. Whereas knockdown of PEBP4 suppressed NSCLC cell migration, PEBP4, and invasion with prevented EMT. Furthermore, PEBP4 overexpression significantly promoted the transcriptional activity of sonic hedgehog (Shh) signaling in NSCLC cells. Further analysis showed that using cyclopamine to inhibit Shh signaling significantly ameliorated the effect on cell proliferation, invasion, and migration, as well as EMT triggered by PEBP4 overexpression. Together, these results suggest that PEBP4 may promote tumorigenesis in NSCLC by regulating cell proliferation and EMT via activation of the Shh signaling pathway.

Hippocampal Cholinergic Neurostimulating Peptide Suppresses Acetylcholine Synthesis in T Lymphocytes

Lymphocytic cholinergic system has important roles in T cell functions, including immune responses and proliferation and differentiation of immune cells. T lymphocytes exclusively produces acetylcholine (ACh) via choline acetyltransferase (ChAT), activating their muscarinic and nicotinic ACh receptors (mAChRs and nAChRs, respectively) in an autocrine and paracrine manners. Hippocampal cholinergic neurostimulating peptide (HCNP) is an undecapeptide cleaved from N-terminal of phosphatidylethanolamine-binding protein 1 (PEBP1). HCNP enhances ACh synthesis through upreglation of ChAT expression in septo-hippocampal cholinergic neurons and participates in neuronal development and differentiation. Although PEBP1 and HCNP appears to be distributed ubiquitously in tissues and cells including spleen, its functions in immune cells have not been understood. In the present study, we observed that PEBP1 is also expressed in human and murine T cells. Long-term exposure to HCNP suppressed ChAT expression in MOLT3 human leukemic T cells, resulting in decreased release of ACh. HCNP also decreased the expression of extracellular signal-regulated kinase (ERK). Thus, HCNP appears to suppress lymphocytic cholinergic signaling, which might act as an immune modulator.

Gynura procumbens Improved Fertility of Diabetic Rats: Preliminary Study of Sperm Proteomic.

Gynura procumbens (GP) is a medicinal herb that has long been known as anti-inflammatory and antihyperglycaemic. Recently, this herbal extract has been associated with a profertility effect, suggesting its applicability in treating both diabetes and male infertility. In this study, the effects of GP aqueous extract (GPAE) on diabetic rats were investigated through evaluating testes histology and androgen hormone levels as well as the implantation sites of female rats on copulation with the treated male rats. Three dosages of GPAE were used (150, 300, and 450 mg/kg), and there were three control groups [normal, diabetic, and metformin-treated diabetic]. Testes histology, androgen hormone levels, and number of implantation sites of the GPAE-treated groups matched those of the normal group in contrast to the diabetic and metformin-treated diabetic controls. Sperm proteomics analysis identified 666 proteins, but only 88 were consistently found in all the control and 450-mg/kg GPAE-treated groups. Four proteins, including cysteine-rich secretory protein 1, carboxylesterase 5A, zona pellucida binding protein, and phosphatidylethanolamine-binding protein 1, were significantly upregulated with GPAE treatment compared with the diabetic control, matching the protein levels of the normal group. These proteins were mainly involved in sperm maturation, sperm capacitation, and sperm-egg interaction, suggesting that GP treatment was able to restore the fertility of male diabetic rats at molecular protein level. In conclusion, GP treatment effectively treats infertility of male diabetic rats, possibly through the upregulation of proteins related to sperm maturation and sperm-egg interaction.

Cloning and functional analysis of five TERMINAL FLOWER 1/CENTRORADIALIS-like genes from Hevea brasiliensis.

Five TERMINAL FLOWER 1 (TFL1)/CENTRORADIALIS (CEN)-like genes were isolated and characterized from rubber tree (Hevea brasiliensis). All genes, except HbCEN1, were found to have conserved genomic organization, characteristic of the phosphatidyl ethanolamine-binding protein (PEBP) family. Overexpression of all of them delayed flowering and altered flower architecture compared with the wild-type (wt) counterpart. In addition, as premature-flowering of the terminal bud was successfully overcome in the tfl1-1 mutant of Arabidopsis, all these genes have a potential function similar to TFL1. Quantitative reverse transcriptase-polymerase chain reaction analysis showed higher expressions of HbCEN1 and HbCEN2 in the shoot apices and stems of both immature and mature rubber trees than in reproductive organs. HbTFL1-1 and HbTFL1-2 expression was confined to roots of 3-month-old seedlings and HbTFL1-3 was significantly higher in the shoot apices of these seedlings. These results suggested that HbCEN1 and HbCEN2 could be associated with the development of vegetative growth, whereas HbTFL1-1, HbTFL1-2 and HbTFL1-3 seem to be mainly related with maintenance of juvenility. In addition, four of the five genes displayed variable diurnal expression, HbTFL1-1 and HbTFL1-3 being mainly expressed during the night whereas HbCEN1 and HbCEN2 showed irregular diurnal rhythms.

Raf kinase inhibitor protein protects microglial cells against 1-methyl-4-phenylpyridinium-induced neuroinflammation in vitro

The Raf kinase inhibitor protein (RKIP), belonging to a member of the phosphatidylethanolamine-binding protein (PEBP) family, is involved in regulating neural development. However, the role of RKIP in microglial cells stimulated with 1-methyl-4-phenylpyridinium (MPP+) has not been determined. Thus, in the present study, we investigated the role of RKIP and its underlying mechanism in Parkinson's disease (PD). Our results showed that the expression of RKIP was significantly reduced in BV-2 cells treated with MPP+. Overexpression of RKIP markedly rescued cell viability and inhibited cell apoptosis in BV-2 cells exposed to MPP+. In addition, overexpression of RKIP inhibited MPP+-induced the production of pro-inflammatory molecules in BV-2 cells. Similar results were observed in primary microglial cells isolated from neonatal mice. Exploration of the underlying mechanisms of its action indicated that overexpression of RKIP prevented the activation of NF-κB and MEK/ERK pathways in MPP+-stimulated BV-2 cells. Taken together, these findings indicated that RKIP suppresses apoptosis and inflammation in MPP+-treated microglial cells through the inactivation of NF-κB and MEK/ERK signaling pathways. Thus, RKIP may be a promising target molecular involving in the pathogenesis of PD.

Differentially Expressed Proteins in Rat Hippocampus after Chronic Immobilization Stress and Intervention Using Xiao Yao San Decoction

ABSTRACT Objective To identify differentially expressed proteins in the hippocampus of rats after chronic immobilization stress (CIS) using a proteomics approach, and to study the effect of the Xiao Yao San (XYS) decoction on differentially expressed proteins. Methods Twenty-four Sprague Dawley rats were randomly assigned to one of four groups of equal body weight: control (non-stress), 7-day stress, 21-day stress and 21-day stress + XYS treatment groups. Two-dimensional gel electrophoresis (2-DE) was used to detect differences in protein expression in rat hippocampus. One differentially expressed protein was measured and verified by western blotting. Results Seventeen proteins showed differential expression. Among these, eight could be identified: glial fibrillary acidic protein-2 (GFAP-2), tubulin alpha-1c, cytoplasmic muscle actin 2, 14-3-3 protein, β-2a tubulin, phosphatidylethanolamine binding protein, synuclein α syn3, and a low molecular weight (18 kD) protein. Six of these proteins exhibited increased expression, one showed decreased expression, and the other protein, which comprised five subtypes, were either increased or decreased. These proteins are known to be involved in immunity, signal transduction, cell cycle control, apoptosis, regulation of enzyme activity, cytoskeleton structure, and synaptic plasticity. GFAP-2 was further analyzed, and its differential expression confirmed by western blotting. Conclusion Some proteins are differentially expressed in the hippocampus of rats under chronic stress. The biological functions of these differentially expressed proteins are varied. Finally, the XYS decoction can significantly up- or down-regulate these protein expression levels.