Intensive resistance by females before copulation induces insemination failure in the West Indian sweet potato weevil <Emphasis Type="Italic">Euscepes postfasciatus</Emphasis>

Persistent mating attempts by males (sexual harassment) are frequently observed among animals. For females, resisting persistent males can be costly because vigorous resistance increases both energy expenditure and the possibility of injury. Although one tactic for coping with male harassment is to cease resistance and mate with the persistent partner, the females of several species are able to prevent the fertilization of their egg(s) despite copulation. In this study, we used three different sex ratios to investigate whether a male’s mating persistence affects his mating success in the West Indian sweet potato weevil Euscepes postfasciatus, in which males mount females both before and after copulation. Consistent with our predictions, we found that female weevils resist and manipulate sperm transfer either before or during copulation according to their preferences. Female weevils were able to reject the sperm of persistent males despite having copulated with them. However, neither copulation and/or post-copulatory mounting affected insemination success. We speculate that the intensive resistance shown by females before copulation may induce mechanical sterility in E. postfasciatus.     

改良CCI大鼠痛行为学检测与损伤区自发放电活动再观察

目的：应用改良CCI模型研究外周神经损伤后痛觉过敏和自发放电各自特征及相互关系。方法：雄性SD大鼠,随机分为CCI组和Sham组,分别于术前1天和术后1、4、7、9、11、14天测定机械刺激缩足反射阈值和热缩腿反射潜伏期,同时选取术侧机械刺激缩腿反射阈值低于4g或者术侧和健侧热缩腿反射潜伏期差异大于2s的CCI模型大鼠观察术后4-14天损伤区自发放电活动。结果：神经纤维损伤后,机械痛敏和热痛敏随时间表现为逐渐增强,同时在损伤区观察到三类放电模式：整数倍放电﹑阵发放电和周期放电。结论：术后大鼠机械痛阈和热痛阈逐渐降低,机械痛敏的产生和损伤区自发放电活动关系密切,不同的放电模式可能代表不同的传入信息。     

Structural changes in the femur with the transition to agriculture on the Georgia coast

Structural characteristics of the femur are compared in preagricultural (2200 B.C.–A.D. 1150) and agricultural (A.D. 1150–1550) subsistence strategy groups from the Georgia coast. Using an automated technique, cross-sectional geometric properties used in structural analyses (areas, second moments of area) were determined at midshaft and distal to the lesser trochanter in 20 adults from each group. A significant decline in magnitude of almost every geometric property occurs in the agricultural group. The differences between groups are reduced but still significant for many properties after standardizing for bone length differences. In addition, a remodeling of the femoral cortex to one of relatively smaller medullary and subperiosteal diameter, as well as a more circular cross-sectional shape, is characteristic of agricultural femora. Thus, while the relative cross-sectional area of bone remains the same, the spatial distribution of bone area is different in the two groups. The results strongly suggest a relative reduction in mechanical loadings of the femur in the agricultural group, implying different levels and possibly types of activity involving the lower limb in the two groups. The data are also compared with similar data available for the Pecos Pueblo (agricultural) sample. The comparison indicates that types of activity may have been more similar in the two agricultural samples, but that general levels of activity were more similar in the Pecos Pueblo and Georgia coast preagricultural samples.     

Autonomous changes in the orientation of cortical microtubules underlying the helicoidal cell wall of the sunflower hypocotyl epidermis: spatial variation translated into temporal changes

Summary. Angles (λ) at which parallel cortical microtubules (cMTs) were oriented with respect to the longitudinal direction were measured in Helianthus annuus hypocotyl epidermal cells. Histograms showing λ frequencies in cell populations at the instant of epidermis fixation were obtained. Analysis of the histograms indicates that, in a particular position within a cell, the angle λ changes periodically with time, i.e., there is a cycle of λ change at that position. This cycle is most likely rotational rather than oscillatory, i.e., the change in λ has a defined chirality (clockwise or counterclockwise). The full diversity of histograms can be consistently explained by rotational cycles with a variable velocity of λ change, and with a cMT rebuilding stage taking place at a different phase of the cycle. The rotational cycles also provide the simplest explanation of cMT arrays in which the angle λ changes along a cell (fixed) and no parallel orientation of cMTs is apparent at a certain position. This explanation assumes a gradient in the phase of the rotational cycle along the cell. The symmetry of the angular characteristics of the rotational cycle, with respect to the morphological directions in cells, leads to the concept that these directions typically represent the principal directions of a certain tensor quantity, which may control the cycling. Possible interactions between the rotational cycle of cMT reorientation and the helicoidal cycle during cell wall formation are discussed. Correspondence and reprints: Department of Biophysics and Cell Biology, Silesian University, ulica Jagiellonska 28, 40-032 Katowice, Poland.     

Functional groups based on leaf physiology: are they spatially and temporally robust?

The functional grouping concept, which suggests that complexity in ecosystem function can be simplified by grouping species with similar responses, was tested in the Florida scrub habitat. Functional groups were identified based on how species regulate exchange of carbon and water with the atmosphere as indicated by both instantaneous gas exchange measurements and integrated measures of function (%N, δ¹³C, δ¹⁵N, C:N ratio) in fire-maintained Florida scrub, which was considered the natural state for scrub habitat. Using cluster analysis, five distinct physiologically based functional groups were identified in the fire-maintained scrub and were determined to be distinct clusters and not just arbitrary divisions in a continuous distribution by the non-parametric multivariate analysis of similarities (ANOSIM; R=0.649, P=0.005). These functional groups were tested for robustness spatially, temporally, and with management regime using ANOSIM. The physiological functional groups remained distinct clusters in this broader array of sites (R=0.794, P=0.001) and were not altered by plot differences, primarily, water table depth (R=−0.115, P=0.893) or by the three different management regimes: prescribed burn, mechanically treated and burned, and fire-suppressed (R=0.018, P=0.349). The physiological groupings also remained robust between the two climatically different years, with 1999 being a much wetter year than 2000 (R=−0.027, P=0.725). Easy-to-measure morphological characteristics, if they indicate the same functional groups, would be more practical for scaling and modeling ecosystem processes than detailed gas exchange measurements; therefore, we tested a variety of morphological characteristics as functional indicators. A combination of non-parametric multivariate techniques were used to compare the ability of life form, leaf thickness (LT), and specific leaf area (SLA) classifications to identify the physiologically based functional groups. Life form classifications (ANOSIM; R=0.629, P=0.001) were able to depict the physiological groupings more adequately than either SLA (ANOSIM; R=0.426, P=0.001) or LT (ANOSIM; R=0.344, P=0.001). The ability of life forms to depict the physiological groupings was improved by separating the parasitic Ximenia americana from the shrub category (ANOSIM; R=0.794, P=0.001). Therefore, a life form classification including parasites was determined to be a good indicator of the physiological processes of scrub species and would be a useful method of grouping species for scaling physiological processes to the ecosystem level.     

Mechanical regulation of HB-GAM expression in bone cells

Bone adaption upon mechanical stimulation is accompanied by changes in gene expression. In this context we investigated the influence of mechanical loading on heparin binding growth associated molecule (HB-GAM) expression, an extracellular matrix molecule which in cell culture has been shown to stimulate the differentiation of osteoblasts. We obtained information on the participating signal transduction pathways using a mitogenic loading regimen. Specific inhibitors of various signal transduction pathways were added to loaded cells and to unloaded controls. By semi-quantitative PCR studies we demonstrated a rapid decrease of HB-GAM expression in primary osteoblasts and SaOs-2 cells by 20-30% upon mechanical loading within 30min. We showed that the RGD-integrin interaction is involved in the regulation of HB-GAM expression. Furthermore, integrity of the cytoskeleton, stretch-activated, and voltage-sensitive Ca(2+) channels as well as gap junctional communication are necessary for the downregulation of HB-GAM expression by mechanical loading.     

The effect of hormones on bone growth is mediated through mechanical stress

Mechanical stresses play a key role in regulating cell growth and cell differentiation. Using mechanical and physiological data available in the literature, we are able to construct a growth curve of a child, which we compare to the standard curve. It appears likely that the impact of hormones on pubertal growth rate sprout followed by growth arrest can be solely explained by increased mechanical stresses. The uptake of hormones by the muscles results in increased mechanical stress on the chondrocyte before and at the puberty, resulting in a peak in growth followed by growth cessation.     

Identification of Pip4k2β as a mechanical stimulus responsive gene and its expression during musculoskeletal tissue healing

To investigate the mechano-transduction system of cells, we identified genes responsive to a cyclic mechanical stimulus. MC3T3.E1 cells were cultured on a computer-controlled vacuum-pump-operated device designed to provide a cyclic mechanical stimulus. A maximum elongation of 15% of membrane at 10 cycles/min (3 s extension followed by 3 s relax per cycle) was repeated for 48 h. By means of a differential display, the gene expression pattern of cells exposed to the stimulus was compared with that of unexposed cells. As a result, a gene fragment that was exclusively expressed in mechanically stressed cells was identified. By using expressed sequence tag walking together with the oligo-capping method, this gene was identified as phosphatidylinositol 4-phosphate 5-kinase type II β (initially known as Pip5k2β but now reclassified as Pip4k2β). The specific up-regulation of Pip4k2β upon mechanical stimulus was also confirmed by using another apparatus, viz. a computer-controlled linearized-stepping motor system. To examine the involvement of the cyclic mechanical stimulus in the regulation of Pip4k2β expression in musculoskeletal tissue, we created an Achilles tendon transection model in rabbits. The temporal expression of Pip4k2β was assessed by means of a quantitative reverse-transcribed polymerase chain reaction. In the gastrocnemius muscle, expression of Pip4k2β rapidly decreased 1 week after transection but was restored to normal levels at 4 weeks. In the Achilles tendon, however, expression remained decreased until 4 weeks after transection. We suggest that the expression of Pip4k2β can be used as a marker for cells receiving a suitable mechanical stimulus. This study was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.     

A rat model of bone cancer pain induced by intra-tibia inoculation of Walker 256 mammary gland carcinoma cells

This study described a modified rat model of bone cancer pain. Syngeneic Walker 256 mammary gland carcinoma cells were injected into the tibia medullary cavity via intercondylar eminence. Series of tests were carried out including bone radiology, bone histology, ambulatory pain, thermal hyperalgesia, mechanical allodynia, weight bearing ability, and electrophysiological recording from primary afferent fibers. The rats inoculated with carcinoma cells showed significant ambulatory pain, mechanical allodynia, and reduction in weight bearing, as well as increased incidence of spontaneous activity in Abeta fibers in affected limb, whereas PBS (vehicle) or heat-killed cells (sham) injected rats showed no significant difference in comparison to normal rats. The pain hypersensitive behaviors were aggravated with time and destruction of bone. Interestingly, mechanical allodynia was also observed in the contralateral limb, indicating the involvement of 'mirror image' pain in bone cancer pain. In summary, the present study provided a useful and easily established rat model of bone cancer pain which will contribute to further study of the mechanisms underlying cancer pain.     

Role of RhoA/ROCK-dependent actin contractility in the induction of tenascin-C by cyclic tensile strain

In chick embryo fibroblasts, the mRNA for extracellular matrix protein tenascin-C is induced 2-fold by cyclic strain (10%, 0.3 Hz, 6 h). This response is attenuated by inhibiting Rho-dependent kinase (ROCK). The RhoA/ROCK signaling pathway is primarily involved in actin dynamics. Here, we demonstrate its crucial importance in regulating tenascin-C expression. Cyclic strain stimulated RhoA activation and induced fibroblast contraction. Chemical activators of RhoA synergistically enhanced the effects of cyclic strain on cell contractility. Interestingly, tenascin-C mRNA levels perfectly matched the extent of RhoA/ROCK-mediated actin contraction. First, RhoA activation by thrombin, lysophosphatidic acid, or colchicine induced tenascin-C mRNA to a similar extent as strain. Second, RhoA activating drugs in combination with cyclic strain caused a super-induction (4- to 5-fold) of tenascin-C mRNA, which was again suppressed by ROCK inhibition. Third, disruption of the actin cytoskeleton with latrunculin A abolished induction of tenascin-C mRNA by chemical RhoA activators in combination with cyclic strain. Lastly, we found that myosin II activity is required for tenascin-C induction by cyclic strain. We conclude that RhoA/ROCK-controlled actin contractility has a mechanosensory function in fibroblasts that correlates directly with tenascin-C gene expression. Previous RhoA/ROCK activation, either by chemical or mechanical signals, might render fibroblasts more sensitive to external tensile stress, e.g., during wound healing.