西方蜜蜂产浆量的动态遗传研究

应用条件和非条件遗传效应分析方法对3个西方蜜蜂(Apis mellifera L.)品种的蜂群产浆量、台浆量和台基接受率进行了发育遗传研究。结果表明：蜂群产浆量、台浆量在各个时期均存在显著或极显著的基因型方差,台基接受率在大部分时期存在显著或极显著的基因型方差,说明这3个性状主要由遗传因素决定。条件遗传分析发现,在某些无法检测到非条件方差的时期存在显著水平的条件方差,证明在产浆期的不同阶段,3个性状都有基因的新表达。同一性状不同时期的遗传相关分析表明：蜂群产浆量以及台浆量在各个时期均检测到显著或极显著的基因型相关,台基接受率在大多时期存在显著的基因型相关,表明控制产浆量和台浆量早期表现的遗传效应总是以相同的方式调节后期的表现,而台基接受率则不然。成对性状之间的相关研究表明：蜂群产浆量和台浆量之间在各个时期存在显著或极显著的遗传相关,说明2性状基因效应之间的协同作用是一致的,而蜂群产浆量和台基接受率之间在大多时期存在显著的遗传相关,但在一些时期没有相关,2性状之间的基因效应协同作用较差。     

细胞壁矿质元素含量在细胞生长中的动态变化

在组织水平上已经描述了许多植物通气组织的形成过程,但对其发育过程的调控仍然知道得很少.利用CSEM-EDX微量分析技术,定点测量慈姑叶柄通气组织不唰发育时期的细胞壁矿质元素的组成.这些元素除了C,O以外,还包括Mg,Ca,Cu,Zn,P等必需的矿质元素.结果发现,在叶柄发育的早期,通气组织细胞壁的K和Cl含量很高,分别高达36%和4.3%细胞壁干重.Mg的含量在第二阶段最高,达到细胞壁干重的0.86%.只有在第三和第四阶段监测到Cu和Zn元素,最高含量分别为2.5%和1.5%细胞干重.仅在第四和第五阶段才能检测到Ca,其最高含量为1.3%细胞壁干重.通气组织横膈膜细胞和圆柱体腔壁细胞的元素构成变化有相似的趋势,说明这种变化与组织的发育阶段关系密切.细胞壁的一些元素间呈现较高的相关性,其中K和Cl及Cu和Zn之间成较高的正相关.在不同发育阶段,细胞壁的元素含量呈现动态变化,说明细胞壁(质外体物质)的元素构成有很大的变动范围.早期的通气组织细胞壁大量积累K和Cl,暗示早期的气体空间充满液体(组织液);Mg可能参与细胞伸展的调控;伸展中细胞的细胞壁积累高浓度的Cu和zn,并不影响细胞的正常功能;而Ca的出现使细胞比硬度增加,将终止细胞伸展.Cu和Zn在细胞壁中的积累呈高度的直线关系,回归分析显示,二者呈现定量关系,推测它们可能有共同的或者类似的转运和吸收机制.     

Peculiarities of the embryonic development of Polypodium hydriforme Ussov (Coelenterata), a parasite of acipenserid oocytes

"Unicellular" stages (107 specimens) and multicellular stages (64 specimens) of embryogenesis of Polypodium, found in 14 sterlet (Acipenser ruthenus L.) females, have been studied with light microscopy, cytophotometry, and autoradiography following incubation with 3H-uridine. All stages of the embryonic development occur inside host oocytes. The "unicellular" stage includes a binucleate cell with unequally sized nuclei; separation inside it of a small cell around the smaller nucleus, i.e. transformation of the single cell into a complex of 2 cells, the larger one enveloping the smaller; formation of a cavity inside the nucleus of the large (outer) cell, and migration of the small cell into it, and "cell-in-a-cell" stage, the small (generative) cell being inside the cavity formed by the nucleus of the large (trophic) cell. The latter gives rise to a hypertrophied but still unicellular envelope around the embryo, the trophamnion. The multicellular stages start with segmentation of the generative cell into blastomeres. These form a morula lying inside the cavity of the trophamnion. Gastrulation occurs by morular delamination. The inversion of the germ layers, typical of parasitic Polypodium stages, apparently arises during gastrulation. Both the generative cell ("egg") and the blastomeres are haploid, at least until the morula stage. The eggs of Polypodium are the smallest ones among coelenterates; they lack yolk and develop without fertilization. Diploidy seems to be restored during segmentation. The trophamnion cell grows, its nucleus becomes highly polypoid, and its cytoplasm accumulates mucoprotein inclusions. Both the blastomere nuclei and the trophamnion nucleus have large nucleoli and actively synthesize RNA. The stages of embryogenesis of Polypodium closely correspond to stages of the host oogenesis. The embryonic development of Polypodium lasts several years and is the slowest among coelenterates. However, it has some features typical of the class Hydrozoa.     

A flow fluorimetric analysis of the cell cycle during growth and differentiation inDictyostelium discoideum

Summary We report a flow fluorimetric analysis of the DNA content of cells and nuclei from vegetative populations and various developmental stages of the cellular slime mouldDictyostelium discoideum using the dyes Hoechst 33258 and mithramycin. Nuclei from all of these populations showed an identical single DNA-content peak, indicating that most vegetative cells and most cells in all developmental stages are in one phase of the cell cycle. Our own data and findings in the literature indicate that this phase is G₂. On the other hand, we also found that various stages, subpopulations of cells at early stages and the different differentiated cell types in the slug stage differ in DNA content per cell. Any particular population typically has one major peak of DNA content, with a modal value that is characteristic for the cell type and for the developmental stage. These differences presumably reflect differences in mitochondrial DNA content per cell.     

Development of the stages of the cycle in mouse seminiferous epithelium after transplantation of green fluorescent protein-labeled spermatogonial stem cells

To study the mechanism of male germ cell differentiation, testicular germ cells carrying green fluorescent protein (GFP) as a transgene marker were transplanted into infertile mouse testis. Fluorescence-positive seminiferous tubule segments colonized with GFP-labeled donor germ cells were isolated and measured, and differentiated germ cells were analyzed in living squashed preparations. Cell associations in normal stages of the seminiferous epithelial cycle were also studied and used as a reference. Two months after transplantation, the average length of the colonies was 1.3 mm. The cell associations of transplanted colonies were consistent with those of normal stages of the cycle. However, stages of the cycle were not necessarily identical in different colonies. Three months after transplantation, the average length of transplanted colonies was 3.4 mm, and the cell association in every portion of a colony was similar to that of the corresponding stage of the cycle. Even in long fused colonies made by transplantation of a higher concentration of male germ cells, the cell association patterns in various regions of a single colony were similar and consistent with those of some of the normal stages of the cycle. Development of different stages inside the colony was observed by 6 mo after transplantation. These results indicate that the commencement of spermatogonial stem cell differentiation occurs randomly to develop different stages of the cycle in different colonies. Then, each colony shows one single stage of the cycle for a long time, even if it becomes a very large colony or fuses with other colonies. These observations indicate the existence of some kind of synchronization mechanism. By 6 mo, however, normal development of the stages of the cycle appeared in seminiferous tubules.     

Ventral cell rearrangements contribute to anterior-posterior axis lengthening between neurula and tailbud stages in Xenopus laevis

Studies of morphogenesis in early Xenopus embryos have focused primarily on gastrulation and neurulation. Immediately following these stages is another period of intense morphogenetic activity, the neurula-to-tailbud transition. During this period the embryo is transformed from the spherical shape of the early stages into the long, thin shape of the tailbud stages. While gastrulation and neurulation depend largely on active cell rearrangement and cell shape changes in dorsal tissues, we find that the neurula-to-tailbud transition depends in part on activities of ventral cells. Ventral explants of neurula lengthen autonomously as much as the ventral sides of intact embryos, while dorsal explants lengthen less than the dorsal sides of intact embryos. Analyses of cell division, cell shapes, and cell rearrangement by transplantation of labeled cells and by time lapse recordings in live intact embryos concur that cell rearrangements in ventral mesoderm and ectoderm contribute to the autonomous anterior-posterior axis lengthening of ventral explants between neurula and tailbud stages.     

Optimization of conditions and cell feeding procedures for alcohol fermentation

Alcohol fermentation was studied with an emphasis on the separation of cell growth and alcohol production stages. Experiments were conducted to establish the optimal conditions for alcohol production in batch fermentations and to simulate continuous fermentations with cell feeding at various stages. It was found that the glucose concentration should be kept under 10% (w/v), and the temperature should be between 40 and 42.5 degrees C for maximum specific alcohol productivity. If the cell concentration is increased, a decrease in specific alcohol productivity is observed. Higher cell concentrations are needed for higher final alcohol concentrations. Among the cell feeding procedures into alcohol production stages, a cocurrent design was found to be better than recycle and countercurrent designs.     

Distinct mechanisms mediate the initial and sustained phases of cell migration in epidermal growth factor receptor-overexpressing cells

Elevated levels of epidermal growth factor receptor (EGFR) are predictive of increased invasion and metastasis in many human cancers. In the present study, we have shown that two distinct pathways regulate cell migration in EGFR-overexpressing invasive cells such as MDA 468 breast cancer cells: mitogen-activated protein kinase (MAPK or ERK 1 and 2) pathways play a major role in early stages to cell migration; and protein kinase C delta isoforms (PKC-delta) play a significant role in later stages of sustained cell migration. Inhibition of MAPK activity with MAP kinase kinase (MEK) inhibitor PD98059 blocks early stages of cell migration (up to 4 h); however, cells revert back to enhanced cell migration after 4 h. While inhibition of PKC-delta activity with rottlerin or dominant-negative PKC-delta expression blocks sustained cell migration after 4 h and up to 12 h, the combination of MAPK and PKC inhibitors completely blocked transforming growth factor alpha (TGF-alpha)-induced cell migration in EGFR-overexpressing breast cancer cells. However, inhibition of MAPK activity completely blocked cell migration in low EGFR-expressing non-invasive breast cancer cells such as MCF-7 cells. Forced overexpression of EGFR in MCF-7 cells (EGFR/MCF-7 cells) resulted in cell migration patterns seen in MDA 468 cells, that is, MAPK pathways play a major role in early stages to cell migration, and PKC-delta plays a major role in later stages of sustained cell migration. The above data demonstrate that EGFR-overexpressing invasive cells have the ability to compensate the loss of MAPK-mediated signaling through activation of PKC-delta signaling for cell migration, which plays a major role in invasion and metastasis. In addition, data suggest that inhibition of MAPK and PKC-delta signaling pathways should abrogate cell migration and invasion in EGFR-overexpressing human breast cancer cells.     

The shape and distribution of lysosomes and endocytosis in the ciliary epithelial cells of rats

Stages of the spermatogenic cycle in human seminiferous tubules were evaluated in men with varied efficiencies of spermatogenesis to determine if the architectural arrangement of stages or the atypical cell types contributed to variation in sperm production rates. Testes were selected from men with low, intermediate, and high daily sperm production per g parenchyma (DSP/g). Round tubular cross sections were photographed by bright-field microscopy. Stages were identified for each cross section by two observers and the number of stages represented in each cross section was recorded. Number of stages per cross section in men with low efficiency of spermatogenesis were significantly (P<0.05) fewer than men with intermediate and high efficiency of spermatogenesis. Further, the percentage of stages with atypical cell types in men with high DSP/g was significantly (P<0.05) higher than men with low DSP/g. There was a significant relationship (P<0.01) between the percentages of stages with atypical cell types per stage and number of stages per cross section. The atypical cell types appear to result from high density of stages per cross section in men with high DSP/g. There was no significant difference observed between groups for tubular volume, diameter, length, volume density, and volume density of seminiferous epithelium. However, a significant (P<0.05) positive correlation between percent seminiferous epithelium per testis with DSP/g or with the number of stages per cross section was found. These findings reveal that the architectural makeup of stages within seminiferous tubules and atypical cell types within stages varies with the level of efficiency of spermatogenesis, and this variation may reflect differences in yield of early spermatogonial divisions that are responsible for generating the different stages.     

Cell cycle regulation in mouse heart during embryonic and postnatal stages

The regulation of cardiomyocyte proliferation is important for heart development and function. Proliferation levels of mouse cardiomyocytes are high during early embryogenesis and start to decrease at midgestation. Many cardiomyocytes undergo mitosis without cytokinesis, resulting in binucleated cardiomyocytes during early postnatal stages, following which the cell cycle arrests irreversibly. It remains unknown how the proliferation pattern is regulated, and how the irreversible cell cycle arrest occurs. To clarify the mechanisms, fundamental information about cell cycle regulators in cardiomyocytes and cell cycle patterns during embryonic and postnatal stages is necessary. Here, we show that the expression, complex formation, and activity of main cyclins and cyclin‐dependent kinases (CDKs) changed in a synchronous manner during embryonic and postnatal stages. These levels decreased from midgestation to birth, and then showed one wave in which the peak was around postnatal day 5. Detailed analysis of the complexes suggested that CDK activities were inhibited before the protein levels decreased. Analysis of DNA content distribution patterns in mono‐ and binucleated cardiomyocytes after birth revealed changes in cell cycle distribution patterns and the transition from mono‐ to binucleated cells. These analyses indicated that the wave of cell cycle regulator expression or activities during postnatal stages mainly produced binucleated cells from mononucleated cells. The data obtained should provide a basis for the analysis of cell cycle regulation in cardiomyocytes during embryonic and postnatal stages.     

Genetic damage by bifunctional agents in repair-active pre-meiotic stages of Drosophila males

Most of our understanding of germline mutagenesis in Drosophila is based on the DNA repair-inactive, haploid post-meiotic stages. The diploid, repair-active pre-meiotic stages are more relevant to the situation encountered in somatic cells. DNA mono-adducts induced by agents like methyl methanesulphonate (MMS) and ethylene oxide (EO) are well repaired in the pre-meiotic cell stages, and these agents show therefore, no or considerable lower mutagenic activity in these stages. In contrast, in this study the two bifunctional nitrogen mustards chlorambucil (CAB) and mechlorethamine (MEC) show significantly elevated mutant frequencies of both post- and pre-meiotic germ cells. Results were similar for the X-chromosomal and the autosomal (2nd) recessive lethal (RL) test. CAB and MEC were also active in stem cells, but in comparison with post-stem cell stages they seem to be better protected. The germ cell specific response in post- and pre-meiotic cell stages was for both nitrogen mustards comparable to mutagenic activity patterns observed in the specific locus test in the mouse.It was reported that for diepoxybutane (DEB), another cross-linking agent, the ratio of the RL frequency for the 2nd- and the X-chromosome was increased from 2.1 for post-meiotic stages to 9.5 for pre-meiotic stages. In own experiments aiming to confirm this observation, a high ratio was indeed found. The induction of large deletions by DEB could be the reason for this difference, since such lesions might include both a sex-linked lethal and a vital gene required for the development of spermatocytes into mature sperm. Similar differences were expected for CAB and MEC since they are also inducers of large deletions. But unexpectedly, no differences in 2nd/X RL ratio between post- and pre-meiotic cell stages were found for the nitrogen mustards. Possible causes such as distinct proportions of multi-locus deletions (MLDs), mitotic recombination and the formation of persistent lesions, are discussed.     

Effects of phosphorus and nitrogen on nodulation are seen already at the stage of early cortical cell divisions in Alnus incana

BACKGROUND AND AIMS: The present work aimed to study early stages of nodulation in a chronological sequence and to study phosphorus and nitrogen effects on early stages of nodulation in Alnus incana infected by Frankia. A method was developed to quantify early nodulation stages in intact root systems in the root hair-infected actinorhizal plant A. incana. Plant tissue responses were followed every 2 d until 14 d after inoculation. Cortical cell divisions were already seen 2 d after inoculation with Frankia. Cortical cell division areas, prenodules, nodule primordia and emerging nodules were quantified as host responses to infection. METHODS: Seedlings were grown in pouches and received different levels of phosphorus and nitrogen. Four levels of phosphorus (from 0.03 to 1 mM P) and two levels of nitrogen (0.71 and 6.45 mM N) were used to study P and N effects on these early stages of nodule development. KEY RESULTS: P at a medium concentration (0.1 mM) stimulated cell divisions in the cortex and a number of prenodules, nodule primordia and emerging nodules as compared with higher or lower P levels. A high N level inhibited early cell divisions in the cortex, and this was particularly evident when the length of cell division areas and presence of the nodulation stages were related to root length. CONCLUSIONS: Extended cortical cell division areas were found that have not been previously shown in A. incana. The results show that effects of P and N are already expressed at the stage when the first cortical cell divisions are induced by Frankia.     

Cell wall extensibility during branch formation in the xanthophycean alga Vaucheria terrestris

In the tip-growing filamentous cell of the xanthophycean alga Vaucheria terrestris sensu Götz, a new growing tip develops in the non-growing, cylindrical region of the cell that was exposed by local illumination. The present study examined changes in the strength and extensibility of the cell wall of the new growing tip and in the matrix components of the inner surface of the cell wall. The internal pressure required to rupture the cell walls decreased remarkably during the early to middle stages of growing tip development, but the cell wall hardly extended before rupture. In contrast, during the middle and late stages of development, cell walls were extended by internal pressure. Atomic force microscopy revealed that protease-resistant, fine granular matrix components were present only at the apical portion of a normal growing tip, and were absent in the non-growing cylindrical region. In the early and middle stages of new growing tip development, these matrix components appeared in the cell walls in patches. These results suggest that first cell wall strength decreases and then cell wall extensibility increases in the development of new growing tips, and that protease-resistant, fine granular matrix components may be involved in rendering a cell wall extensible.     

Comparative characterization of mesenchymal bone marrow stromal cells at early and late stages of culturing

The mesenchymal stromal cell is a multipotent precursor of osteoblasts, adipocytes, and some other cell types. In this study, a comparative analysis of cultured mesenchymal stromal cells from the rat bone marrow at the early and late stages of subculturing has been performed using molecular genetic and cytological methods. The culture has undergone 11 passages during 140 days. Upon long-term culturing, the mesenchymal stromal cells have proved to lose their potential for adipogenic differentiation but preserve the potential for osteogenesis. Morphological characters typical of osteogenic differentiation can be observed at the earlier stages of culturing (passages 1–4) but disappear at later stages (passages 9–11), despite mineralization of the extracellular matrix and the expression of osteogenic differentiation markers. A comparative analysis of the proliferation potential of stromal cells has shown that differences in the period of cell population doubling at the early and later stages of culturing are insignificant. An almost complete arrest of cell growth has been observed in the middle of the culture period (passages 5 and 6).     

Chitin structures of the cell walls of synchronously grown virgin cells of Saccharomyces cerevisiae.

The ability of a lytic beta-glucanase of Arthrobacter GJM-1 to dissolve cell walls of Saccharomyces cerevisiae with exception of the chitin-containing fraction was employed for the isolation of chitin-rich residues of the cell walls of synchronously growing populations of virgin cells. Electron microscopical examination of such wall residues isolated from cells at various stages of the budding cycle showed that the first phase of chitin deposition in the wall corresponds to the formation of an annular structure found as a part of the bud scar after cell division. The annular chitin-rich structure could not be isolated at cell cycle stages preceding the bud emergence and at earliest stages of bud development. The observations confirmed that the annular structure (chitin ring) formed during bud growth represents a major part of total chitin present in the bud scar after septum closure.     

Calpain 2 expression pattern and sub-cellular localization during mouse embryogenesis

Regulation of migration and proliferation by calpain has been shown in various cell types; however, no data are available concerning calpain 2 (capn2) localization in embryonic tissues. Here, we report the expression pattern of capn2 during mouse embryonic development. Expression of the capn2 gene is observed throughout embryonic development. From ES cells and the 8-cell stage to late neurulation stages, CAPN2 is expressed in the cytoplasm and nuclear compartments, with a clear co-localisation with chromatin. Whole-mount in situ hybridization analysis from E8.5 to 14.5 stages indicates high levels of capn2 expression in the nervous system, heart and mesodermal tissues. Up-regulation is maintained during later developmental stages in proliferating cells and in precursor cells involved in muscle (myoblasts) or bone formation (chondrocytes). At later developmental stages, elevated mRNA levels coincided with CAPN2 nuclear localization in these cell types, while differentiated cells maintained cytoplasmic expression. This detailed analysis reveals dynamic expression: nuclear localization was associated either with active cell mitosis in embryonic stem cells and early developmental stages or with precursor cells later during organogenesis. Thus, these data indicate that CAPN2 may represent a key factor in development from the first cell division.     

Distribution of the microtubule-dependent motors cytoplasmic dynein and kinesin in rat testis.

To examine the possible role of microtubule-based transport in testicular function, we used immunofluorescent techniques to study the presence and localization of the microtubule mechanoenzymes cytoplasmic dynein (a slow-growing end-directed motor) and kinesin (a fast-growing end-directed motor) within rat testis. Cytoplasmic dynein immunofluorescence was observed in Sertoli cells during all stages of spermatogenesis, with a peak in apical cytoplasm during stages IX-XIV. Cytoplasmic dynein immunofluorescence was also localized within Sertoli cells to steps 9-14 (stages IX-XIV) germ cell-associated ectoplasmic specializations. In germ cells, cytoplasmic dynein immunofluorescence was observed in manchettes of steps 15-17 (stages I-IV) spermatids, and small, hollow circular structures were seen in the cytoplasm of step 17 and step 18 spermatids during stages V and VI. Kinesin immunofluorescence was observed in manchettes of steps 10-18 spermatids (stages X-VI). The stage-dependent apical Sertoli cell cytoplasmic dynein immunofluorescence, in conjunction with the previously reported orientation of Sertoli cell microtubules (slow-growing ends toward the lumen) and peak secretion of androgen-binding protein and transferrin, is consistent with the hypothesis that cytoplasmic dynein is involved in Sertoli cell protein transport and secretion. Further, the localization of cytoplasmic dynein and kinesin to manchettes is consistent with current hypotheses concerning manchette function.     

A Role for phosphoinositide 3-kinase in the control of cell division and survival during retinal development

Neurogenesis in the retina requires the concerted action of three different cellular processes: proliferation, differentiation, and apoptosis. Class IA phosphoinositide 3-kinase (PI3K) is a heterodimer composed of a p85 regulatory and a p110 catalytic subunit. p110alpha has been shown to regulate cell division and survival. Little is known of its function in development, however, as p110alpha knockout mice exhibit CNS defects, but death at early embryonic stages impairs further study. Here, we examine the role of PI3K in mouse retina development by expressing an activating form of PI3K regulatory subunit, p65(PI3K), as a transgene in the retina. Mice expressing p65(PI3K) showed severely disrupted retina morphogenesis, with ectopic cell masses in the neuroepithelium that evolved into infoldings of adult retinal cell layers. These changes correlated with an altered cell proliferation/cell death balance at early developmental stages. Nonetheless, the most affected cell layer in adult retina was that of photoreceptors, which correlated with selectively increased survival of these cells at developmental stages at which cell division has ceased. These results demonstrate the relevance of accurate PI3K regulation for normal retinal development, supporting class IA PI3K involvement in induction of cell division at early stages of neurogenesis. These data also show that, even after cell division decline, PI3K activation mediates survival of differentiated neurons in vivo.     

Modes of programmed cell death during Ceratitis capitata oogenesis

In the present study, we demonstrate the existence of two distinct apoptotic patterns in nurse cells during Ceratitis capitata oogenesis. One is developmentally regulated and normally occurs during stages 12 and 13, and the other is stage specific and is sporadically observed during stages 7 and 8. The pre-apoptotic manifestation of the first pattern begins at stage 11 and is characterized by the formation of actin bundles. Subsequently, at stages 12 and 13, the nurse cell nuclei exhibit condensed chromatin and contain fragmented DNA, as revealed by TUNEL assay. The apoptotic nurse cell remnants are phagocytosed by the neighboring follicle cells at the end of oogenesis during stages 13 and 14. In the second apoptotic pattern, which occurs sporadically during stages 7 and 8, the nurse cells degenerate and are phagocytosed by the follicular epithelium that contains apoptotic cell bodies. The data presented herein, compared to previous reported results in Drosophila melanogaster and Dacus oleae (Nezis et al., 2000, 2001), strongly suggest that nurse cell apoptosis is a developmentally regulated and phylogenetically conserved mechanism in higher Dipteran. They also suggest that, the sporadic apoptotic pattern consists of a possible protective mechanism throughout oogenesis when damaged or abnormal egg chambers, are eliminated before they reach maturity.