The mechanism of Drosophila leg development along the proximodistal axis

During development of higher organisms, most patterning events occur in growing tissues. Thus, unraveling the mechanism of how growing tissues are patterned into final morphologies has been an essential subject of developmental biology. Limb or appendage development in both vertebrates and invertebrates has attracted great attention from many researchers for a long time, because they involve almost all developmental processes required for tissue patterning, such as generation of the positional information by morphogen, subdivision of the tissue into distinct parts according to the positional information, localized cell growth and proliferation, and control of adhesivity, movement and shape changes of cells. The Drosophila leg development is a good model system, upon which a substantial amount of knowledge has been accumulated. In this review, the current understanding of the mechanism of Drosophila leg development is described.     

Maintenance of segment and appendage primordia by the Tribolium gene knödel

For homeotic and segment-polarity genes in Drosophila, a switch in gene regulation has been described that distinguishes patterning and maintenance phases. Maintenance of segment and organ primordia involves secondary patterning and differentiation steps, as well as survival factors regulating proliferation and organ size. In a screen for embryonic lethal mutations in the flour beetle Tribolium castaneum, we have recovered two alleles of the kn?del gene, which result in short, bag-like embryos. These embryos have severely reduced appendages and differentiate a cuticle that lacks most overt signs of segmentation. In addition, they lack bristles and display defects in the nervous system. Early patterning in kn?del mutant embryos is normal up to the extended germ band stage, as indicated by the formation of regular even-skipped (Tc'eve) and wingless (Tc'wg) stripes. Afterwards, however, these patterns degenerate. Similarly, proximo-distal growth and patterning of limbs are nearly normal initially, but limb primordia shrink, and proximo-distal patterns degenerate, during subsequent stages. kn?del could be a segment polarity gene required for segment border maintenance in both trunk and appendages. Alternatively, it may have a more general role in tissue or organ maintenance.     

No effect of passive integrated transponder tagging method on survival or body condition in a northern population of Black‐capped Chickadees (Poecile atricapillus)

Passive integrated transponder (PIT) tags allow a range of individual‐level data to be collected passively and have become a commonly used technology in many avian studies. Although the potential adverse effects of PIT tags have been evaluated in several species, explicit investigations of their impacts on small (<12 g) birds are limited. This is important, because it is reasonable to expect that smaller birds could be impacted more strongly by application of PIT tags. In this study, we individually marked Black‐capped Chickadees (Poecile atricapillus), a small (circa 10 g) passerine, at the University of Alberta Botanic Garden to evaluate potential lethal and sublethal effects of two PIT tagging methods: attachment to leg bands or subcutaneous implantation. We used a Cox proportional hazards model to compare the apparent survival of chickadees with leg band (N = 79) and implanted PIT tags (N = 77) compared with control birds that received no PIT tags (N = 76) over the subsequent 2 years based on mist net recaptures. We used radio‐frequency identification (RFID) redetections of leg band PIT tags to evaluate sex‐specific survival and increase the accuracy of our survival estimates. We also used a generalized linear regression model to compare the body condition of birds recaptured after overwintering with leg band PIT tags, implanted PIT tags, or neither. Our analysis found no evidence for adverse effects of either PIT tagging method on survival or body condition. While we recommend carefully monitoring study animals and evaluating the efficacy of different PIT tagging methods, we have shown that both leg band and subcutaneously implanted PIT tags ethical means of obtaining individualized information in a small passerine.     

Sequential feeding with variations in energy and protein levels improves gait score in meat-type chickens

Feeding broilers by alternating different diets for 1 or 2 days is known as sequential feeding, and it possibly reduces leg problems since it slows down early growth and may enhance general activity. The present study compared continuous feeding with a standard diet (C: metabolisable energy = 12.55 MJ/kg, crude protein = 190 g/kg) with alternations of a high-energy/low-protein diet (E+P-:+7% ME; -20% CP) and a low-energy/high-protein diet (E-P+: -7% ME,+20% CP) and investigated its effects on growth, behaviour and gait score in 352 male Ross broiler chickens. Sequential feeding was carried out during ten 48-h sequential-feeding cycles from 8 to 28 days of age. Three treatments were compared: complete diet (C) and two alternations of diets varying in protein and energy contents (S1: E+P- followed by E-P+; and S2: E-P+ followed by E+P-). Chickens received the same feed during the starter and finisher periods (0 to 7 and 29 to 38 days of age). Body weight (BW), feed intake, general activity and gait score, bone quality and carcass conformation were measured to evaluate leg condition and general performance. Sequential feeding significantly reduced BW at 28 days of age (S1: -9.1%; S2: -3.7%/C group; P < 0.05) and S1 were lighter than S2. In both sequential groups, time spent standing increased (C: 28%; S1:33%; S2: 35%; P < 0.05) and leg abnormalities decreased (mean gait score: C: 2.61; S1: 2.45; S2: 2.38; P < 0.02). This improvement was not related to changes in bone quality. BW at slaughter was impaired in Group S1 only, and the feed conversion ratio throughout the rearing period was not significantly impaired by sequential feeding. However, abdominal fat was higher in the S2 group. Sequential feeding using diets varying in energy and crude protein can be a useful method of reducing leg problems in broilers since it improves gait score without impairing growth performance when used as early as 8 days of age and up to not less than 8 days before slaughter in order to compensate for reduced growth. This improvement can be explained by reduced early growth and enhanced motor activity. However, it appears that the low-energy diet should be given first in order to avoid a reduction in BW at slaughter.     

EGFR signaling is required for re-establishing the proximodistal axis during distal leg regeneration in the cricket Gryllus bimaculatus nymph

Nymphs of hemimetabolous insects, such as cockroaches and crickets, possess functional legs with a remarkable capacity for epimorphic regeneration. In this study, we have focused on the role of epidermal growth factor receptor (EGFR) signaling in regeneration of a nymphal leg in the cricket Gryllus bimaculatus. We performed loss-of-function analyses with a Gryllus Egfr homolog (Gb'Egfr) and nymphal RNA interference (RNAi). After injection of double-stranded RNA for Gb'Egfr in the body cavity of the third instar cricket nymph, amputation of the leg at the distal tibia resulted in defects of normal distal regeneration. The regenerated leg lacked the distal tarsus and pretarsus. This result indicates that EGFR signaling is required for distal leg patterning in regeneration during the nymphal stage of the cricket. Furthermore, we demonstrated that EGFR signaling acts downstream of the canonical Wnt/Wg signaling and regulates appendage proximodistal (PD) patterning genes aristaless and dachshund during regeneration. Our results suggest that EGFR signaling influences positional information along the PD axis in distal leg patterning of insects, regardless of the leg formation mode.     

How locomotion sub-functions can control walking at different speeds?

Inspired from template models explaining biological locomotory systems and Raibert׳s pioneering legged robots, locomotion can be realized by basic sub-functions: elastic axial leg function, leg swinging and balancing. Combinations of these three can generate different gaits with diverse properties. In this paper we investigate how locomotion sub-functions contribute to stabilize walking at different speeds. Based on this trilogy, we introduce a conceptual model to quantify human locomotion sub-functions in walking. This model can produce stable walking and also predict human locomotion sub-function control during swing phase of walking. Analyzing experimental data based on this modeling shows different control strategies which are employed to increase speed from slow to moderate and moderate to fast gaits.     

Altered lower extremity joint mechanics occur during the star excursion balance test and single leg hop after ACL-reconstruction in a collegiate athlete

The effects of ACL-reconstruction on lower extremity joint mechanics during performance of the Star Excursion Balance Test (SEBT) and Single Leg Hop (SLH) are limited. The purpose of this study was to determine if altered lower extremity mechanics occur during the SEBT and SLH after ACL-reconstruction. One female Division I collegiate athlete performed the SEBT and SLH tasks, bilaterally, both before ACL injury and 27 months after ACL-reconstruction. Maximal reach, hop distances, lower extremity joint kinematics and moments were compared between both time points. Musculoskeletal simulations were used to assess muscle force production during the SEBT and SLH at both time points. Compared to the pre-injury time point, SEBT reach distances were similar in both limbs after ACL-reconstruction except for the max anterior reach distance in the ipsilateral limb. The athlete demonstrated similar hop distances, bilaterally, after ACL-reconstruction compared to the pre-injury time point. Despite normal functional performance during the SEBT and SLH, the athlete exhibited altered lower extremity joint mechanics during both of these tasks. These results suggest that measuring the maximal reach and hop distances for these tasks, in combination with an analysis of the lower extremity joint mechanics that occur after ACL-reconstruction, may help clinicians and researchers to better understand the effects of ACL-reconstruction on the neuromuscular system during the SEBT and SLH.     

31P-NMR saturation transfer study of the in vivo kinetics of arginine kinase in Carcinus crab leg muscle

The kinetics of the reaction catalyzed by arginine kinase have been determined at 9.5 and 23°C for in vivo leg muscle of Carcinus maenas (the common shore crab) using the noninvasive technique of ³¹P-NMR spectroscopy. Concentrations of mobile phosphorus metabolites were the same at both temperatures: 78.7 mM for arginine phosphate, 9.0 mM for adenosine triphosphate (ATP), and 2.6 mM for inorganic phosphate (P_i), as estimated from NMR resonance intensities and literature values for ATP concentration as assayed by traditional biochemical methods. Apparent unidirectional rate constants for formation of ATP from arginine phosphate and ADP were 0.09 s^?1 at 9.5°C and 0.27 s^?1 at 23°C. Pseudo-first-order rate constants for arginine phosphate generation from Arg and ATP were 0.38 and 1.10 s^?1 at 9.5 and 23°C, respectively. In vivo Q₁₀ for the arginine kinase reaction between 9.5 and 23°C was thus 2.2 for both directions. When the kinetic data are analyzed using the Arrhenius equation, activation energies of 126 kJ/mol for ATP formation and 105 kJ/mol for arginine phosphate formation are found. The measured chemical fluxes through arginine kinase in the forward reaction (arginine phosphate hydrolysis) were twice those in the reverse reaction, consistent with either compartmentation of substrates or participation of substrates in alternative metabolic pathways.     

中华绒螯蟹蜕壳过程中肌肉的组织学、超微结构及主要蛋白质含量的变化

为探讨中华绒螯蟹(Eriocheir sinensis)蜕壳前后肌肉组织的形态特征变化, 采用石蜡切片、电镜及生物化学方法, 研究了中华绒螯蟹蜕皮过程中步行足和腹部肌肉的组织学、超微结构及主要蛋白质含量的变化。结果显示: 相对于蜕皮间期, 步行足在蜕皮前后组织学形态特征无明显变化; 超微结构在蜕皮前无明显变化, 蜕皮后可见肌原纤维纵裂及肌小节横裂现象, 表明蜕皮后外骨骼硬化的过程伴随着肌肉的生长。相对于蜕皮间期, 腹部肌肉在蜕皮前后组织学特征变化明显: 蜕皮前肌束间隙增大, 蜕皮后肌束内肌纤维间隙增大。电子显微镜观察显示, 蜕皮前肌原纤维在内部降解, 出现空洞, 肌原纤维边缘降解, 导致肌原纤维间隙增大; 蜕皮后肌原纤维重新组装、重建, 恢复到间期正常形态。生物化学研究发现, 蜕皮前后步行足和腹部肌肉中肌原纤维蛋白和可溶性蛋白含量的变化同其结构特征的变化相一致。以上研究结果表明, 中华绒螯蟹肌肉组织的结构特征同蜕皮周期密切相关。