Dense bodies in silver-stained spermatocytes of the chinese hamster: behavior and cytochemical nature

From the silver staining behavior of various organelles in the nucleus we have divided meiotic prophase (leptotene to the diffuse stage) of the male Chinese hamster into five stages. Components within the nucleus, such as synaptonemal complex (SC), sex bivalent (SB), nucleolus organizer regions (NORs), chromatin and the dense bodies, showed a characteristic feature in each stage of meiotic prophase. The lampbrush chromosome stage was found to be followed by the diffuse stage. The chromatin around SC began to be organized at early pachytene and formed a brush-like structure at late pachytene. During early prophase stages a dramatic change in SB morphology occurred. Three types of morphology of SB were recognized: (1) the XY pair with long synapsis and fusiform or diffuse thickening of the unpaired portions (late zygotene and early pachytene), (2) desynapsed, thread-like axes seen at midpachytene, and (3) multistranded, branched, and anastomosed axes seen at late pachytene.Two types of the dense body were found during meiotic prophase; the double body in early stage (leptotene to early pachytene) and the single body in later stages (mid pachytene to diffuse stage). The small precursors of the double body existed at early leptotene but they increased in size and also changed the silver stainability during zygotene, becoming the characteristic double body consisted of one light body (L-body) and one dark body (D-body). These two bodies can also be recognized after Giemsa or acridine orange (AO) staining. The L-body fluoresced reddish orange after AO staining. The single body, which is probably formed by amalgamation of the D- and the L-bodies, showed a staining reaction similar to that of the D-body.Data from pancreatic lipase and protease treatments suggest that the D-body contained a lipoprotein.     

四川盆地亚热带常绿阔叶林不同物候期凋落物分解与土壤动物群落结构的关系

为了解植物生长不同物候时期凋落物分解过程中土壤动物群落结构动态及其与凋落物分解的关系,以四川盆地亚热带常绿阔叶林典型人工林树种马尾松和柳杉,次生林树种香樟和麻栎凋落物为研究对象,采用凋落物分解袋试验研究,凋落物分解过程中土壤动物的群落特征。4种凋落物分解袋共获得土壤动物8047只,其中,柳杉(2341只)>香樟(2105只)>马尾松(2046只)>麻栎(1555只)。其中,秋末落叶期、萌动期和展叶期,马尾松凋落物袋中主要以捕食性土壤动物为优势类群,而后以菌食性土壤动物为主;香樟凋落物袋中除秋末落叶期和叶衰期以菌食性土壤动物为主要优势类群外,其他各时期均以捕食性土壤动物为主要优势类群;柳杉凋落物分解各时期均以菌食性土壤动物为主要优势类群;麻栎凋落物分解在前3个时期以菌食性为主,而后以植食性土壤动物为主要优势类群。相关分析表明,在秋末落叶期和萌动期土壤动物的个体密度主要和氮、磷含量及其格局密切相关,叶衰期主要和难分解组分木质素显著相关。除在秋末落叶期土壤动物对凋落物分解的贡献率与土壤动物的个体密度显著相关外,其余主要物候关键时期均与土壤动物的类群密度及其食性显著相关。     

Differential photosynthetic characteristics between seedlings and saplings of Abies sachalinensis and Picea glehnii, in the field

Abies sachalinensis and Picea glehnii are co-dominant tree species and major components of the forests of Hokkaido, Japan. Recent work suggests that a reversal in potential competitive superiority at different developmental stages could be important to explain their coexistence. Such shifts in competitive advantage can be mechanistically understood by studying the corresponding physiological differences between distinct life stages. Accordingly, our objective was to investigate in the field the photosynthesis of shade-growing juveniles of these species from two different size-classes, seedlings and saplings. Our results show that seedlings of both species had higher concentrations of photoprotective xanthophylls than saplings, especially in spring, and suggest that seedlings have a lower threshold of stress tolerance than saplings. Photosynthetic capacity per needle area and lateral shoot extension rate decreased from the seedling to the sapling stage in A. sachalinensis, while in P. glehnii, both increased from the seedling to the sapling stage. Abies sachalinensis had higher photosynthetic rates at the seedling stage but lower rates at the sapling stage than P. glehnii. Nevertheless, A. sachalinensis had a higher lateral shoot extension rate than P. glehnii at both stages. Our physiological results support previous ecological observations that A. sachalinensis is a superior competitor to P. glehnii in the understory, and show that its competitive advantage is higher at the seedling stage than at the sapling stage.     

A sequential analysis of meiosis in the male mouse using a restricted spermatocyte population obtained by a hydroxyurea/triaziquone treatment

A method is described to restrict the spermatocyte population in mice and other rodents using hydroxyurea (HU) and triaziquone (T). HU affects cells in S-phase, whereas T is an agent especially active on spermatogonia and not on spermatocytes. An application of three i.p. HU injections with 12 h intervals, followed about nine days later by one i.p. T injection creates two large gaps in the spermatogenic line. The two gaps enclose a small, well-defined group of primary spermatocytes in meiotic interphase. — The development of the restricted spermatocyte population is followed day by day. The analysis of meiosis in male mice has revealed the correct sequence of meiotic, and especially prophase I stages. On account of clearly visible differences in chromosome morphology the diplotene stage could be divided into three periods. It is suggested to use the following nomenclature: pre-diffuse diplotene, diffuse diplotene and post-diffuse diplotene. The experiment was also informative about the timing of the stages in spermatocyte development by correlating the days at which the successive stages were observed with the corresponding stage of the epithelial cycle. The calculation of the position and duration of the diffuse diplotene, enables us to put forward a proposal about the significance of the diffuse diplotene. — A combination of the HU/T method with cell separation techniques provides good perspectives for detailed biochemical studies on processes taking place during meiosis.     

The development of oocytes in the ovary of a parthenogenetic tick, Haemaphysalis longicornis

Haemaphysalis longicornis is an important vector of various pathogens in domestic animals and humans. The tick is a unique species with bisexual and parthenogenetic races. Although mating induces oocyte development, it is possible in the parthenogenetic race to complete oogenesis without copulation. Here we examined the developmental process of oocytes from unfed to the oviposition period in parthenogenetic H. longicornis. We classified the developmental stages of oocytes into five stages: stage I, germinal vesicle occupies more than half of the cytoplasm; stage II, germinal vesicle occupies less than half of the cytoplasm; stage III, germinal vesicle migrates from the center in the oocyte to the vicinity of the pedicel cells; stage IV, the cytoplasm is filled with yolk granules of various sizes; stage V, the cytoplasm is occupied by large yolk granules. Oocytes at the unfed period were undeveloped and classified as stage I. Stage I and II oocytes were observed at the rapid feeding period, indicating that oocyte development began after the initiation of blood feeding. All developmental stages of oocytes were observed at the pre-oviposition period. At 10?days after the beginning of the oviposition period, the ratios of stage I and II oocytes were higher than those of the previous period, suggesting that the ovarian development and activity may be continuing. Based on these findings, we propose classification criteria for the oocyte development in the parthenogenetic H. longicornis. The criteria will be useful for understanding the mechanisms of tick reproduction and transovarial transmission of pathogens.     

Pleonal muscle development in the shrimp Penaeus (Litopenaeus) vannamei (Crustacea: Malacostraca: Decapoda: Dendrobranchiata)

Penaeoidean shrimp pleonal muscle is a valuable economic resource worldwide, but little is known of its development during larval stages. The development of pleonal muscle in Penaeus (Litopenaeus) vannamei was studied by rhodamine-phalloidin staining and laser-scanning confocal microscopy. Dorsal pleonal muscle was first evident at the protozoea I stage while ventral pleonal muscle was present by the protozoea II stage. Identifiable ventral pleonal muscles were evident by the protozoea III stage and all ventral muscle types were present in the mysis I. The tail flex response began at the mysis stage and growth of existing pleonal muscles continued. The pleopods formed during the mysis stages, with coxal and basis muscles developed by mysis III. The pleopods became functional beginning with the first post-larval stage. We conclude that the pleonal muscle pattern of P. vannamei larvae is similar to that of adult Penaeus setiferus, and that homologous muscles are present. The major formation of dorsal pleonal muscles occurs during the protozoea II stage, while significant development of ventral pleonal muscles occurs during the protozoea III stage.     

The diffuse diplotene stage of meiotic prophase in Neurospora

The prophase stages of meiosis in Neurospora crassa are re-examined following McClintock (1945) and Singleton (1953). A diffuse chromosome stage occurring between pachynema and diakinesis is described. It is proposed that the diffuse stage does not necessarily represent a condition of intense gene activity in the sense of directing the metabolic activity of the ascus.Supported by U. S. Public Health Service grants AI-01462 and GM-14263.     

Autoinhibition of Ethylene Formation in Nonripening Stages of the Fruit of Sycomore Fig (Ficus sycomorus L.)

Differences in the mechanism of ethylene emanation of Ficus sycomorus L. during various stages of the fruit development were investigated by enclosing the figs in jars. Two distinct patterns of ethylene emanation were found. Pattern a. in stages not capable of ripening, neither spontaneously nor as a result of physiological treatment (nonripening stages A and C), ethylene concentration in the jar increased linearly for a short time and then remained constant. Pattern b. in stages capable of ripening (ripening stages B, D, and E), the linear increase in ethylene concentration continued for the entire period of measurement. In nonripening stages, ethylene emanation stopped when ethylene concentration in the jar reached a constant value (0.6 μl/l at stage C). Aeration of the figs and the jar renewed ethylene emanation. CO₂ concentration in the jar never exceeded 0.5%. Treatment of stage C figs with 0.6 to 10 μl/l exogenous ethylene caused immediate and complete cessation of ethylene emanation whereas the same treatment did not cause any change in rate of ethylene emanation from figs at the ripening stages B and D. Gashing (wounding) of stage C figs temporarily changed the pattern of ethylene emanation from pattern a to pattern b.     

Role of Plasmodium berghei cGMP-dependent Protein Kinase in Late Liver Stage Development

The liver is the first organ infected by Plasmodium sporozoites during malaria infection. In the infected hepatocytes, sporozoites undergo a complex developmental program to eventually generate hepatic merozoites that are released into the bloodstream in membrane-bound vesicles termed merosomes. Parasites blocked at an early developmental stage inside hepatocytes elicit a protective host immune response, making them attractive targets in the effort to develop a pre-erythrocytic stage vaccine. Here, we generated parasites blocked at a late developmental stage inside hepatocytes by conditionally disrupting the Plasmodium berghei cGMP-dependent protein kinase in sporozoites. Mutant sporozoites are able to invade hepatocytes and undergo intracellular development. However, they remain blocked as late liver stages that do not release merosomes into the medium. These late arrested liver stages induce protection in immunized animals. This suggests that, similar to the well studied early liver stages, late stage liver stages too can confer protection from sporozoite challenge.     

Experimental manipulation leading to induction of dorsal ectodermal ridges on normal limb buds results in a phenocopy of the eudiplopodia chick mutant

Elongation of chick limb buds depends on the presence of the apical ectodermal ridge which is induced by subjacent limb bud mesoderm. Recombination experiments have shown that the limb bud mesoderm loses the capacity to induce ridges by late stage 17. Moreover, in normal limb development only one ridge forms. However, in the eudiplopodia chick mutant accessory ectodermal ridges form on the dorsal surface of limb buds as late as stage 22. Tissue recombinant experiments show that the mutation affects the ectoderm, extending the time it responds to ridge induction (Fraser and Abbott, 1971a, Fraser and Abbott, 1971b while the mesoderm is normal. The result is polydactyly, with extra digits dorsal to the normal digits. Because eudiplopodia limb bud dorsal mesoderm can induce ridges at stage 22 but is unaffected by the gene, genetically normal dorsal limb bud mesoderm may also be able to induce ridges after stage 17. To test this possibility we grafted stages 14–18 flank ectoderm to normal limb bud dorsal mesoderm and found that mesoderm from stages 17 through 20 was able to induce a ridge and subsequently dorsal digits developed. Limbs with duplicate digits were similar to eudiplopodia limbs. In other experiments, stage 18, 19, and 20 leg bud dorsal ectoderm did not form ridges when grafted to leg bud dorsal mesoderm of the same stage, indicating a lack of response to the mesoderm. Finally, the inductive capacity of limb bud mesoderm appeared to be reduced compared to mesoderm at pre-limb bud stages. These experiments demonstrate a spatially generalized potential in limb bud dorsal mesoderm to induce ridges during the stages when the apical ridge is induced. The determination of where the ridge will form and the acquired inability of limb bud dorsal ectoderm to respond to induction by underlying mesoderm are necessary early pattern forming events which assure that a single proximodistal limb axis will form.     

Intra- and interannual variation in the diet of the Magellanic penguin (Spheniscus magellanicus) at Martillo Island,Beagle Channel

Knowledge of seabirds’ diet at each breeding site and its temporal variation is key to understanding and evaluating how changes in marine resources affect each seabird population. In this study, we determined the diet of Magellanic penguins (MP, Spheniscus magellanicus) at Martillo Island, accounting for sex, breeding stage and year. We analyzed a total of 144 stomach contents during three consecutive breeding seasons (2006–2007, 2007–2008 and 2008–2009) and stages (incubation, early and late chick-rearing). MP fed mainly on fuegian sprat (Sprattus fuegensis), which represented 75 % of the biomass consumed by birds during the entire study. The next important prey was squat lobster (Munida gregaria), followed by Patagonian squid (Loligo gahi). Both sexes consumed similar prey items. We observed variation in diet relative composition among breeding years and stages. Fuegian sprat consumption decreased throughout the years whereas squat lobster increased. Penguins consumed a higher proportion of squat lobster and Patagonian squid during the incubation stage than in the chick-rearing stages, whereas fuegian sprat was almost the only prey item consumed during the late chick-rearing stage. MPs show certain flexibility in the use of resources probably as a response to changes in prey populations. Variability in the diet among different reproductive stages could be related to changes in the distribution and abundance of their main prey near the colony during the breeding season together with changes in the energy requirements of seabirds.     

The influence of hypotonic treatment on the morphology of meiotic stages I. Prophase of the first meiotic division of male mice,man and Locusta migratoria

Male mice were treated with hydroxyurea (HU) to obtain a restricted germ cell population and a sequential analysis of the first meiotic prophase was performed. The results from HU-treated mice were compared with those from untreated mice, human and grasshopper material with and without hypotonic treatment.In hypotonically treated material the three subclasses of diffuse stages were obvious in all species investigated. In material not treated hypotonically the different classes of prophase stages showed a more gradual course, there was a tendency towards less spreading of the bivalents and no pronounced diffuse substages could be found. The first desynaptic stage in this material could be recognized in most cases as wellspread classical pachytene like bivalents with a pronounced double-threaded structure. The more advanced desynaptic stage in untreated material resembled the diffuse stage as found in plant meiosis.     

Use of a cloned library for the study of abundant poly(A)+RNA during Xenopus laevis development

Over 200 cloned sequences from recombinant DNA libraries prepared from Xenopus laevis embryonic poly(A)⁺RNA have been analyzed by colony hybridization with [³²P]cDNA prepared from poly(A)⁺RNA from several stages of development. The period of early embryogenesis extending through the beginning of gastrulation (stage 10) is marked by the relative constancy of the abundant poly(A)⁺RNA population. Between the gastrula and tailbud stages (stage 24) there is a dramatic change in the pattern of abundant poly(A)⁺RNA species; the new pattern remains fairly constant for at least 2 days of development to the late prefeeding tadpole stages (stage 41). We have also compared nonpolysomal and polysomal poly(A)⁺RNA populations at two different stages. In stage 10 (early gastrula) postribosomal (free ribonucleoprotein) and polysomal poly(A)⁺RNA populations partly overlap; however, many cloned sequences occur in quite different concentrations in one fraction or the other. Among the sequences that are predominantly nonpolysomal at gastrula few become predominantly polysomal at tailbud stages. Thus, we have no evidence for a major recruitment of abundant nonpolysomal RNAs into polysomes with progressing development. We rather observe a general pattern in which a cloned sequence that is nonpolysomal in one stage of development tends to be nonpolysomal (if detectable at all) in other stages as well.     

Patterns of Cell Division,Cell Differentiation and Cell Elongation in Epidermis and Cortex of Arabidopsis pedicels in the Wild Type and in erecta

Plant organ shape and size are established during growth by a predictable, controlled sequence of cell proliferation, differentiation, and elongation. To understand the regulation and coordination of these processes, we studied the temporal behavior of epidermal and cortex cells in Arabidopsis pedicels and used computational modeling to analyze cell behavior in tissues. Pedicels offer multiple advantages for such a study, as their growth is determinate, mostly one dimensional, and epidermis differentiation is uniform along the proximodistal axis. Three developmental stages were distinguished during pedicel growth: a proliferative stage, a stomata differentiation stage, and a cell elongation stage. Throughout the first two stages pedicel growth is exponential, while during the final stage growth becomes linear and depends on flower fertilization. During the first stage, the average cell cycle duration in the cortex and during symmetric divisions of epidermal cells was constant and cells divided at a fairly specific size. We also examined the mutant of ERECTA, a gene with strong influence on pedicel growth. We demonstrate that during the first two stages of pedicel development ERECTA is important for the rate of cell growth along the proximodistal axis and for cell cycle duration in epidermis and cortex. The second function of ERECTA is to prolong the proliferative phase and inhibit premature cell differentiation in the epidermis. Comparison of epidermis development in the wild type and erecta suggests that differentiation is a synchronized event in which the stomata differentiation and the transition of pavement cells from proliferation to expansion are intimately connected.     

Origin of the prechordal plate and patterning of the anteroposterior regional specificity of the involuting and extending archenteron roof of a urodele, Cynops pyrrhogaster

We analyzed the notochord formation, formation of the prechordal plate, and patterning of anteroposterior regional specificity of the involuting and extending archenteron roof of a urodele, Cynops pyrrhogaster. The lower (LDMZ) and upper (UDMZ) domains of the dorsal marginal zone (DMZ) of the early gastrula involuted and formed two distinct domains: the anterior fore-notochordal endodermal roof and the posterior domain containing the prospective notochord. Cygsc is expressed in the LDMZ from the onset of gastrulation, and the Cygsc-expressing LDMZ planarly induces the notochord in the UDMZ at the early to mid gastrula stages. At the mid to late gastrula stages, part of the Cygsc-expressing LDMZ is confined to the prechordal plate. On the other hand, Cybra expression only begins at mid gastrula stage, coincident with notochord induction at this stage. Anteroposterior regional specificity of the neural plate was patterned by the posterior domain of the involuting archenteron roof containing the prospective notochord at the mid to late gastrula stages. Cynops gastrulation thus differs significantly from Xenopus gastrulation in that the regions of the DMZ are specified from the onset of gastrulation, while the equivalent state of specification does not occur in Cynops until the middle of gastrulation. Thus we propose that Cynops gastrulation is divided into two phases: a notochord induction phase in the early to mid gastrula, and a neural induction phase in the mid to late gastrula.     

The effect of optimal space allowance on growth performance and physiological responses of pigs at different stages of growth

This study was conducted to determine the optimal space allowance for maximizing the growth performance of pigs at each of the following five growth stages (based on BW ranges): stage 1, 11 to 25 kg BW; stage 2, 25 to 45 kg BW; stage 3, 45 to 65 kg BW; stage 4, 65 to 85 kg BW; and stage 5, 85 to 110 kg BW. A total of 1590 crossbred (Landrace×Yorkshire×Duroc) pigs were assigned to one of four treatments at each growth stage, with three replicates each. Pen areas at each growth stage were 6, 11, 16, 19.5 and 20 m² for stages 1 to 5, respectively. Space allowances for the four treatments at each growth stage were modified by varying the number of pigs per pen (22, 25, 28 and 31 pigs in T1, T2, T3 and T4, respectively). Blood samples were collected on the final day of each growth stage. The average daily gain (ADG) decreased significantly with decreased space allowances at all growth stages, except at stage 2. Average daily feed intake (ADFI) was not significantly affected by space allowances at stages 1 to 4; however, at stage 5, there was a linear effect of space allowance on ADFI. Thus, the feed conversion ratio showed results similar to those for ADG. Serum cortisol concentrations, indicating the level of stress response, increased as space allowances decreased. The highest serum cortisol concentrations were observed in T3 at stages 2 to 5. Serum tumor necrosis factor-α levels were significantly higher in association with a small space allowance than with at large space allowance at stages 2, 4 and 5. Serum interleukin-1β levels also increased in a significant linear manner at every growth stage in pigs reared at a low space allowance, except at stage 4 (P=0.068). This study found that limited space allowance decreases the growth performance of pigs and induces stress and inflammatory responses. We confirmed that no significant effect of space allowance on growth performance and serum cortisol concentrations are observed between T1 and T2 across all growth stages. We suggest that the optimal space allowances for pigs according to their BW are as follows: 0.24, 0.44, 0.64, 0.78 and 0.80 m²/pig for BWs of 11 to 25, 25 to 45, 45 to 65, 65 to 85 and 85 to 115 kg, respectively.     

Variability of local spatial structure in a wave-regenerated Abies forest

Although some studies have demonstrated temporal patterns of changes in spatial structure during forest development, few studies have examined the variability of spatial structure between stands at the same developmental stage. In the present study, we investigated variations of spatial structure between sites at the same developmental stage for three developmental stages (sapling, intermediate, and mature) in a wave-regenerated Abies veitchii and Abies mariesii forest. The spatial structure of tree heights in each plot was analyzed by using the mark correlation and mark variogram methods, and the pattern of tree locations in each plot was analyzed by using the pair-correlation function. Analysis of the spatial height structure indicated that a size hierarchy between neighboring trees (a local size hierarchy) generally did not develop at the sapling stage. A local size hierarchy developed in most plots during the two later stages. There was no obvious difference among developmental stages in the spatial pattern of tree locations because of the large variation within each stage. Our results demonstrate that large variation in spatial structure existed between sites in the wave-regenerated Abies forest, even at the same developmental stage. The variability in spatial structure confirmed the importance of stochastic factors in forest dynamics.     

石羊河下游白刺灌丛演替过程中群落结构及数量特征

白刺灌丛(Nitraria tangutorum Bobr.)是在石羊河下游存活面积最大的天然植被类型,研究不同演替阶段白刺灌丛群落的结构和数量特征可为该区白刺群落保护与恢复提供科学依据.根据白刺灌丛生长及其生境状况,在石羊河下游选择了初期发育、稳定、衰退、严重衰退的群落,研究了各演替过程中各阶段的群落物种组成、生活型谱,群落盖度、群落物种多样性及各演替阶段群落的相似性特征.结果表明,从初期发育阶段到稳定阶段的群落物种数量明显减少,而从稳定阶段至严重衰退阶段的演替过程中群落物种是逐渐增加的.在整个演替过程中,1年生植物所占比例逐渐增加,至严重衰退阶段时1年生植物所占的比重高达80％.不同演替阶段群落盖度最高、最低值出现的月份也不相同,5-10月群落盖度的平均值为稳定阶段＞初期发育阶段＞衰败阶段＞严重衰败阶段.各演替阶段多样性Simpson、Shannon-Wiener指数以及Pielou均匀度指数5-10月均呈现波动变化特征,Simpson、Shannon-Wiener指数大小顺序均表现为初期发育阶段＞严重衰退阶段＞衰退阶段＞稳定阶段,而均匀度Pielou指数为衰退阶段＞严重衰退阶段＞稳定阶段＞初期发育阶段.各演替阶段群落之间的相似性均较低,演替序列越相近,相似性系数越高.