Cytological Mechanisms Research Articles

Integrated transmission electron microscopy and proteomic analyses reveal the cytoarchitectural response to cucumber mosaic virus infection in tobacco

Cucumber mosaic virus (CMV) causes huge economic losses to agriculture every year; thus, understanding the mechanism of plant resistance to CMV is imperative. In this study, an integrated analysis of transmission electron microscopy (TEM) observations and proteomic results was used to identify cytoarchitectural differences in Nicotiana tabacum cv. NC82 (susceptible) and cv. Taiyan 8 (T.T.8; resistant) following infection with CMV. The TEM observations showed that the structure of the chloroplasts and mitochondria was severely damaged at the late stage of infection in NC82. Moreover, the chloroplast stroma and mitochondrial cristae were reduced and disaggregated. However, in T.T.8, organelle structure remained largely intact Selective autophagy predominated in T.T.8, whereas non-selective autophagy dominated in NC82, resembling cellular disorder. Proteomic analysis of T.T.8 revealed differentially expressed proteins (DEPs) mostly associated with photosynthesis, respiration, reactive oxygen species (ROS) scavenging, and cellular autophagy. Biochemical analyses revealed that ROS-related catalase, autophagy-related disulfide isomerase, and jasmonic acid and antioxidant secondary metabolite synthesis-related 4-coumarate:CoA ligase (Nt4CL) exhibited different trends and significant differences in expression in the two cultivars after CMV inoculation. Furthermore, mutant phenotyping verified that reduced Nt4CL expression impaired resistance in T.T.8. The identified DEPs are crucial for maintaining intracellular homeostatic balance and likely contribute to the mechanism of CMV resistance in tobacco. These findings increase our understanding of plant cytological mechanisms conferring resistance to CMV infection.

A group of pancreatic islet β cell‐like neurons are vulnerable to energy deficiency

AbstractBackgroundAlzheimer’s disease (AD) bear resemblance to the diabetes mellitus for that both of them exhibit beta‐amyloid (Aβ) deposition, insulin resistance and glycometabolic disorder. Peripheral islet β cells produce insulin and islet amyloid peptides, but whether there is a group of neurons in the brain like islet β cells is not clear.MethodHybridize APP/PS1 transgenic mice and CaMKII‐CreERT2/+ ;Tpkfl/fl strain, with tamoxifen induced Tpk knockout at 6 months of age, to form a model of energy deficiency. Labeling neuron and microglia with NeuN and Iba1 antibody, and immunofluorescence was used to observe Aβ, insulin‐like growth factor expression. TUNEL staining was applied to test cell apoptosis.ResultWe found that layer 5 cortical pyramidal neurons are strongly marked by Aβ antibody, 4G8, in APP/PS1 transgenic mice. Immunofluorescence showed that these 4G8‐positive neurons colocalized with insulin‐like growth factor 1/2 and highly expressed prohormone convertase 2. Further, we conditionally knocked out Tpk gene, a key gene involved in glucose metabolism, in excitatory neurons of APP/PS1 mice. Over time, NeuN and Iba1 labeling showed that the significant loss of neuron and activation of microglia happened in layer 5 cerebral cortex at the earliest. Additionally, the 4G8‐positive neurons in this layer were stained by TUNEL and markedly reduced in the early stage.ConclusionWe found a group of islet β cell‐like neurons that are vulnerable to glucose energy impairment, suggesting a group of vulnerable neurons in sporadic AD, and implicating the cytological mechanism of association between AD and diabetes.

Apomixis for no bacteria-induced thelytoky in Diglyphus wani (Hymenoptera: Eulophidae).

In Hymenoptera species, the reproductive mode is usually arrhenotoky, where haploid males arise from unfertilized eggs and diploid females from fertilized eggs. In addition, a few species reproduce by thelytoky, where diploid females arise from unfertilized eggs. Diploid females can be derived through various cytological mechanisms in thelytokous Hymenoptera species. Hitherto, these mechanisms were revealed mainly in endosymbiont-induced thelytokous Hymenoptera species. In contrast, thelytokous Hymenoptera species in which a reproductive manipulator has not been verified or several common endosymbionts have been excluded were paid less attention in their cytological mechanisms, for instance, Diglyphus wani (Hymenoptera: Eulophidae). Here, we investigated the cytological mechanism of D. wani using cytological methods and genetic markers. Our observations indicated that the diploid karyotypes of two strains of D. wani consist of four pairs of relatively large metacentric chromosomes and one pair of short submetacentric chromosomes (2n = 10). The arrhenotokous strains could complete normal meiosis, whereas the thelytokous strain lacked meiosis and did not expulse any polar bodies. This reproductive type of lacking meiosis is classified as apomictic thelytoky. Moreover, a total of 636 microsatellite sequences were obtained from thelytokous D. wani, dominated by dinucleotide repeats. Genetic markers results showed all three generations of offspring from thelytokous strain maintained the same genotype as their parents. Our results revealed that D. wani is the first eulophid parasitoid wasp in Hymenoptera whose thelytoky was not induced by bacteria to form an apomictic thelytoky. These findings provide a baseline for future inner molecular genetic studies of ameiotic thelytoky.

Gene transcriptional profiles in gonads of Bacillus taxa (Phasmida) with different cytological mechanisms of automictic parthenogenesis

The evolution of automixis – i.e., meiotic parthenogenesis – requires several features, including ploidy restoration after meiosis and maintenance of fertility. Characterizing the relative contribution of novel versus pre-existing genes and the similarities in their expression and sequence evolution is fundamental to understand the evolution of reproductive novelties. Here we identify gonads-biased genes in two Bacillus automictic stick-insects and compare their expression profile and sequence evolution with a bisexual congeneric species. The two parthenogens restore ploidy through different cytological mechanisms: in Bacillus atticus, nuclei derived from the first meiotic division fuse to restore a diploid egg nucleus, while in Bacillus rossius, diploidization occurs in some cells of the haploid blastula through anaphase restitution. Parthenogens’ gonads transcriptional program is found to be largely assembled from genes that were already present before the establishment of automixis. The three species transcriptional profiles largely reflect their phyletic relationships, yet we identify a shared core of genes with gonad-biased patterns of expression in parthenogens which are either male gonads-biased in the sexual species or are not differentially expressed there. At the sequence level, just a handful of gonads-biased genes were inferred to have undergone instances of positive selection exclusively in the parthenogen species. This work is the first to explore the molecular underpinnings of automixis in a comparative framework: it delineates how reproductive novelties can be sustained by genes whose origin precedes the establishment of the novelty itself and shows that different meiotic mechanisms of reproduction can be associated with a shared molecular ground plan.

Ultra-microhistological study of nonthermal irreversible electroporation on the esophagus

Esophageal ulceration and even fistula are severe complications of pulmonary vein isolation using traditional thermal ablation. Nonthermal irreversible electroporation (NTIRE) is a new technique for pulmonary vein isolation in patients with atrial fibrillation. NTIRE has been shown to be a safe method for pulsed electroporation near the esophagus. NTIRE preserves the structural framework of the esophagus and allows for rapid recovery of the whole layers of the esophagus. The purpose of this study was to elucidate the ultrastructural changes and cytological mechanisms of cell regeneration and tissue repair after esophageal electroporation. The parameter combination of 2000 V/cm multiplied by 90-pulse output was directly applied to the esophagus in 60 New Zealand rabbits, and ultrastructure analysis of the esophagus was implemented subsequently. NTIRE predominantly triggered apoptosis of esophageal cells shortly after electroporation. Since the tissue structural framework was preserved, esophageal cells could regenerate through self-replication within 4 weeks. Complete anatomical repair can eventually be achieved through structural remodeling, and no lumen stenosis, ulcer, or fistula was observed in the ablated segment. Monophasic, bipolar NTIRE pulses delivered using plate electrodes in an esophageal model demonstrates no irreversible ultra-micropathological changes to the esophagus after 4 weeks.

Low Concentration of Aluminum-Stimulated Pollen Tube Growth of Apples (Malus domestica).

Aluminum (Al) is an important element in soil constitution. Previous studies have shown that high concentration of Al affects the normal growth of crops, resulting in crop yield reduction and inferior quality. Nevertheless, Al has also been referred to as a beneficial element, especially when used at low concentrations, but the cytological mechanism is not clear. Influences of low concentration AlCl3 on the pollen tube growth of apple (Malus domestica) and its possible cytological mechanism were investigated in this study. The results showed that 20 μM AlCl3 promoted pollen germination and tube elongation; 20 μM AlCl3 enhanced Ca2+ influx but did not affect [Ca2+]c of the pollen tube tip; and 20 μM AlCl3 decreased acid pectins in pollen tubes but increased esterified pectins and arabinan pectins in pollen tubes. According to the information provided in this research, 20 μM AlCl3 stimulated growth of pollen tubes by enhancing Ca2+ influx and changing cell wall components.

Decay of homologous chromosome pairs and discovery of males in the thelytokous fungus-growing ant Mycocepurus smithii

The prevalent mode of reproduction among ants is arrhenotokous parthenogenesis where unfertilized eggs give rise to haploid males and fertilized eggs develop into diploid females. Some ant species are capable of thelytokous parthenogenesis, a type of asexual reproduction where females develop from unfertilized diploid eggs. Thelytoky is well-documented in more than 20 ant species. Cytogenetic data are available for six species demonstrating that some thelytokous ant species are capable of producing males occasionally as well as maintaining their chromosome numbers and proper chromosome pairings. Mycocepurus smithii is a thelytokous fungus-growing ant species that inhabits large parts of Central and South America. Cytogenetic data are unavailable for M. smithii and male individuals were never documented for this species, although the presence of males is expected because genetic recombination was observed in a few sexually reproducing populations in Brazil and haploid sperm was documented from the spermathecae of M. smithii queens. This study aims at comparatively studying asexual and sexual populations of M. smithii using classical and molecular cytogenetic methods to test whether karyotype configuration is modified according to the mode of reproduction in M. smithii. Moreover, we report the discovery of M. smithii males from a sexually reproducing population in the Brazilian state Pará, diagnose the male of M. smithii, and morphologically characterize their spermatozoa. Karyotypic variation was observed within the asexual population (2n = 9, 10, or 11), whereas the chromosome number was fixed in the sexual population (2n = 14, n = 7). Identical karyotypes were maintained within individual M. smithii colonies and karyotype variation was only observed between colonies. In asexual individuals, the karyomorphs showed a decay of homologous chromosome pairs, especially in individuals with the karyomorph 2n = 11, which is potentially caused by relaxed natural selection on proper chromosome pairing. In contrast, females in the sexual population showed proper homologous chromosome pairings. In individuals of both asexual and sexual populations, we find that heterochromatin was localized in centromeric regions and on the short arms of the chromosomes, GC-rich regions were associated with heterochromatic regions, and 18S rDNA genes were located on the largest chromosome pair. This comparative cytogenetic analysis contributes to our understanding about the cytological mechanisms associated with thelytokous parthenogenesis in ants and suggests the decay of chromosome structure in the absence of meiosis and genetic recombination.

Cytological study on haploid male fertility in maize

Doubled haploid (DH) breeding technology, which relies on haploid genome doubling, is widely used in commercial maize breeding. Spontaneous haploid genome doubling (SHGD), a more simplified and straightforward method, is gaining popularity among maize breeders. However, the cytological mechanism of SHGD remains unclear. This study crossed inbred lines RL36 and RL7, which have differing SHGD abilities, with inducer line YHI-1 to obtain haploid kernels. The meiotic processes of pollen mother cells (PMCs) in the haploid plants were compared with diploid controls. The results suggested that three main pathways, the early doubling of haploid PMCs, the first meiotic metaphase chromosomal segregation distortion, and anomaly of the second meiosis, are responsible for SHGD. Furthermore, flow cytometry analysis of ploidy levels in leaves and PMCs from haploids and diploid controls revealed that somatic cell chromosome doubling and germ cell chromosome doubling are independent processes. These findings provide a foundation for further studies on the underlying mechanism of SHGD, aiding the application of DH technology in maize breeding practices.

Cytological mechanism of nanowire reinforced biological bone cement in the treatment of vertebral osteoporotic fractures

Cytological mechanism of nanowire reinforced biological bone cement in the treatment of vertebral osteoporotic fractures

Cytological mechanism of nanowire reinforced biological bone cement in the treatment of vertebral osteoporotic fractures

Cytological mechanism of nanowire reinforced biological bone cement in the treatment of vertebral osteoporotic fractures

Zerumbone’s Effects on Mouse Spleen T Cell Migration and Proliferation

Zerumbone is the main ingredient extracted from wild ginger Zingiber zermbet Smith. It is known that the root of ginger contains the most zerumbone. It has been reported to function in several biological environments, including tumors and inflammation. In addition, it is known to affect the functions of several types of cells, including immune cells. In this study, it was confirmed that the migration phenomenon induced by SDF-1α was inhibited by zerumbone using T cells isolated from mouse spleen. On the other hand, the proliferation of mouse T cells through antiCD3 and anti-CD28 antibodies was not affected by zerumbone. Zerumbone inhibited phosphorylation of Erk1/2 signaling protein induced by SDF1α. This result has the meaning of confir ming that the effect of zerumbone confirmed in T cell lines also occurs in physiological T cells, and more detailed biochemical and cytological mechanisms are needed.

Flavors of Non-Random Meiotic Segregation of Autosomes and Sex Chromosomes

Segregation of chromosomes is a multistep process occurring both at mitosis and meiosis to ensure that daughter cells receive a complete set of genetic information. Critical components in the chromosome segregation include centromeres, kinetochores, components of sister chromatid and homologous chromosomes cohesion, microtubule organizing centres, and spindles. Based on the cytological work in the grasshopper Brachystola, it has been accepted for decades that segregation of homologs at meiosis is fundamentally random. This ensures that alleles on chromosomes have equal chance to be transmitted to progeny. At the same time mechanisms of meiotic drive and an increasing number of other examples of non-random segregation of autosomes and sex chromosomes provide insights into the underlying mechanisms of chromosome segregation but also question the textbook dogma of random chromosome segregation. Recent advances provide a better understanding of meiotic drive as a prominent force where cellular and chromosomal changes allow autosomes to bias their segregation. Less understood are mechanisms explaining observations that autosomal heteromorphism may cause biased segregation and regulate alternating segregation of multiple sex chromosome systems or translocation heterozygotes as an extreme case of non-random segregation. We speculate that molecular and cytological mechanisms of non-random segregation might be common in these cases and that there might be a continuous transition between random and non-random segregation which may play a role in the evolution of sexually antagonistic genes and sex chromosome evolution.

Effects of Qingre Huatan Huoxue prescription on biological characteristics and cell cycle proteins of esophageal cancer stem cells.

e16052 Background: Esophageal cancer (EC) stem cells characterized with immature differentiation, high invasion, high tumorigenesis and other biological characteristics, which are the root of the occurrence, development, recurrence, metastasis of EC. In this study, its purpose is to select and identify EC stem cells from serum-free cultured EC9706 cells, explore the effect of Chinese herbal prescription and its separated prescriptions on the biological characteristics of EC9706 stem cells, and to reveal the cytological and molecular mechanisms of Chinese herbal prescription anti-cancer. Methods: Firstly, MTT method was applied to explore the serum-free culture conditions of EC9706 stem cells in diameter of 60 mm low adhesion culture dish, at the same time, applying flow cytometry to identify the p75NTR mark ratio of each passage of cell to screenEC9706 stem cells. Next, Plotting the growth curve of EC9706 stem cells, detecting the proliferation inhibition rate EC9706 stem cells by cisplatin, soft agar clone formation, cell cycle to identify the characteristics of EC9706 stem cells by MTT method, soft agar assay, flow cytometry. Then applying MTT method and orthogonal test to screen and optimize Qingre Huatan Huoxue prescription (QHHP), using flow cytometry and western blot to test the effects of QHHP and its separated prescriptions on EC9706 stem cells appraisal indicators and Cell Cycle Proteins. Results: EC9706 cells with concentration of 8×104 cells/dish inoculateed on diameter of 60mm low adhesion culture dish, with adding 5 ml DMEM/F12 medium, a daily supplement of 20ng EGF, 10ng bFGF, per 10 to 14 days to passage one generation. After 5 generations of cell passage, the expression rate of p75NTR was 4-5%, was selected as the experimental object.EC9706 stem cells with the biological characteristics of slower adhesive growth, stronger cisplatin resistance, higher soft agar clone formation and p75NTR expression rate than that of EC9706 cells, and The cell cycle distribution was concentrated in the G0/G1 phase. 6 Chinese herbs were screened out ( Coptis coptidis, Rhizoma officinalis, Cucurbita pedicle, Toona bark, Cantharidopsis cantharidis, Augustia japonica) and compose QHHP. QHHP and its separated prescriptions could change the morphology, colony formation ability and cycle distribution of EC9706 stem cells in serum-free culture condition, which were related to the BMI1/CyclinB1 signaling pathway. Conclusions: Serum-free cell culture is a method to enrich stem cell-like cells in EC9706 cells. Qingre Huatan Huoxue prescription and its separated prescriptions inhibited the proliferation of EC9706 stem cells and change esophageal cancer stem cells characters. The mechanism of QHHP and its separated prescriptions on stem cells in EC9706 cells was related to BMI1 /CyclinB1 signaling pathway.

A Review of Unreduced Gametes and Neopolyploids in Alfalfa: How to Fill the Gap between Well-Established Meiotic Mutants and Next-Generation Genomic Resources.

The gene flow mediated by unreduced gametes between diploid and tetraploid plants of the Medicago sativa-coerulea-falcata complex is pivotal for alfalfa breeding. Sexually tetraploidized hybrids could represent the best way to exploit progressive heterosis simultaneously derived from gene diversity, heterozygosity, and polyploidy. Moreover, unreduced gametes combined with parthenogenesis (i.e., apomixis) would enable the cloning of plants through seeds, providing a unique opportunity for the selection of superior genotypes with permanently fixed heterosis. This reproductive strategy has never been detected in the genus Medicago, but features of apomixis, such as restitutional apomeiosis and haploid parthenogenesis, have been reported. By means of an original case study, we demonstrated that sexually tetraploidized plants maintain apomeiosis, but this trait is developmentally independent from parthenogenesis. Alfalfa meiotic mutants producing unreduced egg cells revealed a null or very low capacity for parthenogenesis. The overall achievements reached so far are reviewed and discussed along with the efforts and strategies made for exploiting reproductive mutants that express apomictic elements in alfalfa breeding programs. Although several studies have investigated the cytological mechanisms responsible for 2n gamete formation and the inheritance of this trait, only a very small number of molecular markers and candidate genes putatively linked to unreduced gamete formation have been identified. Furthermore, this scenario has remained almost unchanged over the last two decades. Here, we propose a reverse genetics approach, by exploiting the genomic and transcriptomic resources available in alfalfa. Through a comparison with 9 proteins belonging to Arabidopsis thaliana known for their involvement in 2n gamete production, we identified 47 orthologous genes and evaluated their expression in several tissues, paving the way for novel candidate gene characterization studies. An overall view on strategies suitable to fill the gap between well-established meiotic mutants and next-generation genomic resources is presented and discussed.

Reproductive Isolation Between Taxonomically Controversial Forms of the Gray Voles (Microtus, Rodentia; Arvicolinae): Cytological Mechanisms and Taxonomical Implications.

The formation of hybrid sterility is an important stage of speciation. The voles of the genus Microtus, which is the most speciose genus of rodents, provide a good model for studying the cytological mechanisms of hybrid sterility. The voles of the “mystacinus” group of the subgenus Microtus (2n = 54) comprising several recently diverged forms with unclear taxonomic status are especially interesting. To resolve the taxonomic status of Microtus mystacinus and Microtus kermanensis, we crossed both with Microtus rossiaemeridionalis, and M. kermanensis alone with Microtus arvalis “obscurus” and M. transcaspicus and examined the reproductive performance of their F1 hybrids. All interspecies male hybrids were sterile. Female M. kermanensis × M. arvalis and M. kermanensis × M. transcaspicus hybrids were sterile as well. Therefore, M. mystacinus, M. kermanensis, and M. rossiaemeridionalis could be considered valid species. To gain an insight into the cytological mechanisms of male hybrid sterility, we carried out a histological analysis of spermatogenesis and a cytological analysis of chromosome synapsis, recombination, and epigenetic chromatin modifications in the germ cells of the hybrids using immunolocalization of key meiotic proteins. The hybrids showed wide variation in the onset of spermatogenesis arrest stage, from mature (although abnormal) spermatozoa to spermatogonia only. Chromosome asynapsis was apparently the main cause of meiotic arrest. The degree of asynapsis varied widely across cells, individuals, and the crosses—from partial asynapsis of several small bivalents to complete asynapsis of all chromosomes. The asynapsis was accompanied by a delayed repair of DNA double-strand breaks marked by RAD51 antibodies and silencing of unpaired chromatin marked by γH2A.X antibodies. Overall, the severity of disturbances in spermatogenesis in general and in chromosome synapsis in particular increased in the hybrids with an increase in the phylogenetic distance between their parental species.

Distinct cytological mechanisms for food availability in three Inga ingoides (Fabaceae)-Cecidomyiidae gall systems.

Gall cytological, metabolic, and structural traits are established due to the feeding habits of the associated galling herbivores, and sometimes are influenced by other organisms involved in the interaction. We tested this assumption on three gall morphotypes, the globoid, the lenticular, and the fusiform, induced by Cecidomyiidae on leaflets of Inga ingoides (Rich.) Willd. (Fabaceae: Caesalpinioideae). Taking for granted that the three Cecidomyiidae galls are induced on the same host plant and organ, we assume that the cytological and histochemical traits of their nutritive cells may be similar, but under the fungi influence, the ambrosia gall cytological profile may be peculiar and reflect on the accumulation of primary metabolites. The ambrosia globoid galls involve three organisms (host plant, gall inducer, and fungi), while the fusiform and the lenticular galls involve two organisms (host plant and gall inducer). The accumulation of primary metabolites is similar among the three gall morphotypes, except for the non-detection of reducing sugars in the fusiform galls. The fungi presence can impact the system but does not define exclusive features for the ambrosia globoid galls when compared to the lenticular and fusiform morphotypes. In fact, the cytological traits have revealed three different cytological mechanisms for food resources availability to the three galling Cecidomyiidae: (a) cell wall destructuring and cell death by fungi intermediation in the ambrosia globoid galls, (b) necrosis-type cell death in the fusiform galls, and (c) maintenance of continuous metabolic activity in the lenticular galls.

Study on Paraffin Section Technology of Fruit Tumor Tissues in Cucumber (<i>Cucumis sativus</i> L.)

Fruit tumor is one of the importantly external quality traits affected to the economic value of cucumber fruit. In order to obtain the paraffin section technology and study the cytological mechanism of morphological formation for fruit tumor in cucumber, we used the samples of warty fruit line (S52) and non-warty fruit lines (S42 with big fruit spine and S06 with small fruit spine) as materials, and improved some key links such as dehydration, dyeing and dewaxing etc., based on traditional paraffin section technology. Results demonstrated that increasing ethanol concentration gradient during dehydration can shorten the treatment time of each stage to 30 minutes. Samples were stained with Ehrlich's hematoxylin for three days, which was convenient for subsequent operation. Optimal thickness of the section was 8 μm. After twice pure xylene dewaxing, sections could be examined and selected by microscope, which shortened the time for obtaining high-quality section. High quality sections with complete tissue and clear structure can be obtained by the optimized paraffin section technology, which is an effective method to observe the cytological structure of fruit tumor. This study provides a set of complete and efficient operation steps of paraffin section technology for cucumber fruit tumor, which will provide technical reference for anatomical researches on other organs in cucumber and structural similar characteristics of other plants.

Cytological abnormalities during pollen development in interspecific hybrids of Nicotiana

Nicotiana alata is resistant to Tomato spotted wilt virus (TSWV) and of great value in breeding. However, hybrid sterility constrains the application of interspecific genetic resources. Previously, we obtained interspecific hybrids between a cytoplasmic male sterility (CMS) line of Nicotiana tabacum and N. alata, some of which were pollen sterile. In the present research, we studied the cytological abnormalities during pollen development in sterile hybrids (F1-D) by comparing pollen development with that in fertile hybrids (F1-S) from the same cross. Transmission electron microscopy and DiI staining showed that the membrane structures of microspores and pollen in F1-D sterile hybrids were impaired. Carbol fuchsin staining revealed that cytomixis, chromosome loss and asymmetric callose wall formation occurred with high frequency in the microsporocytes and microspores of the sterile hybrids. The cytoplasm and nucleus were lost in the microspores and pollen of sterile hybrids, leading to mature pollen grains that were vacuous and collapsed in the aperture region. In addition, delayed tapetum degradation was detected in the anther of sterile hybrids, and sporopollenin was deposited in the aperture region. Impaired membrane structures of microspores and pollen in F1-D sterile hybrids affected the integrity of the cells, and might be associated with chromosome, nuclear and cytoplasm loss, vacuous pollen, and sterility in F1-D hybrids. Abnormal tapetum degradation in the anther and irregular sporopollenin deposition in the pollen wall of the F1-D sterile hybrids might also be related to the pollen sterility. This study deepens our understanding of the cytological mechanisms of hybrid sterility, and may facilitate the application of TSWV-resistant resources in cultivated Nicotiana species through hybrid fertility restoration and backcross breeding.

Nerve growth factor against PTSD symptoms: Preventing the impaired hippocampal cytoarchitectures

Although exogenous nerve growth factor (NGF) demonstrated great potential for post-traumatic stress disorder (PTSD) treatment, its therapeutic effect and underlying cytological mechanism were not fully elucidated so far. We employed a controlled, prospectively designed modified single prolonged stress mice model to investigate the role of exogenous NGF on the modified single prolonged stress induced PTSD-like symptoms and hippocampal cytoarchitecture impairment, as well as the potential neuronal signaling modulation. We discovered that the modified single prolonged stress-exposure induced significant PTSD-like symptoms as well as mildly impaired hippocampal Cornu Ammonis 1 (CA1) subregion cytoarchitecture, but not dentate gyrus neurogenesis, together with a gradual inhibition of TrkA-CREB-ERK signalings in hippocampal CA1 subregion. NGF treatment dose-dependently ameliorated the modified single prolonged stress induced PTSD-like symptoms. NGF increased the cytoplasm/nucleus ratio and improved the neuronal plasticity, mainly via the TrkA-ERK-CREB pathway. Our study offered the translational evidence for the potential application of exogenous NGF for treating or early preventing PTSD after stress exposure.

Aluminum toxicity-induced pollen tube growth inhibition in apple (Malus domestica) is mediated by interrupting calcium dynamics and modification of cell wall components

Aluminum (Al) is known the most abundant metal in the Earth’s crust which is a major problem limiting crop production and yield throughout the world. Al could inhibit pollen tube elongation, however the mechanism is largely unknown. The aim of this study was to investigate the influence of Al toxicity on Malus domestica pollen tube growth and possible cytological mechanism. The results showed that Al inhibited pollen germination and tube growth in a concentration dependent manner. An non-invasive ion-selective electrode technique together with fluorescent labeling indicated that Al toxicity decreased pollen tube apex calcium influx and disrupted [Ca2+]c gradient. Al toxicity also altered actin filament orientation. Results from immuno-localization and fluorescence labeling indicated that Al toxicity affected the accumulation and distribution of callose, acid pectins, esterified pectins and arabinogalactan proteins in pollen tubes, which were supported by Fourier-transform infrared analysis. In a word, Al induced a series of the cellular toxic changes concerning calcium influx, [Ca2+]c gradient, actin and cell wall components. Based on the information provided in this article, it is concluded that Al toxicity inhibits apical growth of pollen tubes by changing calcium influx and [Ca2+]c and cell wall components.