Visualizing the Live Drosophila Glial-neuromuscular Junction with Fluorescent Dyes

Our project identified GFP labeled glial structures at the developing larval fly neuromuscular synapse. To look at development of live glial-nerve-muscle synapses, we developed a larval tissue preparation that had features of live intact larvae, but also had good optical properties. This new preparation also allowed for access of perfusates to the synapse. We used fly larvae, immersed them in artificial hemolymph, and relaxed their normal rhythmic body contractions by chilling them. Next we dissected off the posterior segments of each animal and with a blunt insect pin pushed the mouth parts backward through the body cavity. This everted the larval body wall, like turning a sock inside-out. We completed the dissection with ultra-fine dissection scissors and thus exposed the visceral side of the body wall muscles. The glial structures at the NMJ expressed membrane targeted GFP under the control of glial specific promoters. The post-synaptic membrane, the SSR (Subsynaptic Reticula) in muscle expressed synaptically targeted dsRed. We needed to acutely label the motor neuron terminals, the third part of the synapse. To do this we applied primary antibodies to HRP, conjugated to a far-red emitting flurophore. To test for dye diffusion properties into the perisynaptic space between the motor neuron terminals and the SSR, we applied a solution of large Dextran molecules conjugated to far-red emitting flurophore and collected images.     

A new computational model to study mass inhomogeneity and hydrophobicity inhomogeneity in proteins

We propose a simple yet reliable computational framework that characterizes the differential mass and hydrophobicity distribution within structural classes of proteins. Radial partitioning of protein interior that could successfully distinguish the mass and hydrophobicity distribution patterns in extremophilic proteins from that in their structurally aligned mesophilic counterparts. Distance-dependent mass and hydrophobicity magnitudes could retrieve vital structural insights; needed to probe the hidden connections between packing, folding and stability within different structural classes of proteins, with causality. New computational markers; one, to represent the total mass content; other, related to hydrophobic centrality of proteins, are proposed as well. Results reveal that mass and hydrophobicity packing within extremophilic proteins is indeed more compact than that in their mesophilic counterparts. Analysis of structural constraints within them vindicate it. Total mass (and hydrophobicity) content is found to be maximum in α/β thermophilic proteins and minimum for the all-α mesophilic proteins. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

How to keep proliferative neural stem/progenitor cells

It has long been argued that cell cycle regulators such as cyclins, cyclin-dependent kinases and their inhibitors affect the fate of neuronal progenitor cells. Recently, we identified that cyclin D2, which localizes at the basal tip of the radial glial cell (i.e., the neural progenitor in the developing neocortex), functions to give differential cell fates to its daughter cells just after cell division. This basally biased localization is due to transportation of cyclin D2 mRNA via its unique cis-regulatory sequence and local translation into cyclin D2 protein at the basal endfoot. During division of the neural progenitor cells, cyclin D2 protein is inherited by the daughter cell that retain the basal process, resulting in asymmetric distribution of cyclin D2 protein between the two daughter cells. Cyclin D2 is similarly localized in the human fetal cortical primordium, suggesting a common mechanism for the maintenance of neural progenitors and a possible scenario in evolution of primate brains. Here we introduce our recent findings and discuss how cyclin D2 functions in mammalian brain development and evolution.     

Glial precursor cell transplantation therapy for neurotrauma and multiple sclerosis

Traumatic injury to the brain or spinal cord and multiple sclerosis (MS) share a common pathophysiology with regard to axonal demyelination. Despite advances in central nervous system (CNS) repair in experimental animal models, adequate functional recovery has yet to be achieved in patients in response to any of the current strategies. Functional recovery is dependent, in large part, upon remyelination of spared or regenerating axons. The mammalian CNS maintains an endogenous reservoir of glial precursor cells (GPCs), capable of generating new oligodendrocytes and astrocytes. These GPCs are upregulated following traumatic or demyelinating lesions, followed by their differentiation into oligodendrocytes. However, this innate response does not adequately promote remyelination. As a result, researchers have been focusing their efforts on harvesting, culturing, characterizing, and transplanting GPCs into injured regions of the adult mammalian CNS in a variety of animal models of CNS trauma or demyelinating disease. The technical and logistic considerations for transplanting GPCs are extensive and crucial for optimizing and maintaining cell survival before and after transplantation, promoting myelination, and tracking the fate of transplanted cells. This is especially true in trials of GPC transplantation in combination with other strategies such as neutralization of inhibitors to axonal regeneration or remyelination. Overall, such studies improve our understanding and approach to developing clinically relevant therapies for axonal remyelination following traumatic brain injury (TBI) or spinal cord injury (SCI) and demyelinating diseases such as MS.     

中国东南部不同海拔亚热带森林中马尾松径向生长对气候的响应

为了解我国东南部亚热带森林不同海拔树木生长对气候响应的差异,建立了福建省武夷山脉东麓2个样点的4个马尾松(Pinus massoniana)轮宽年表,对树木径向生长与气候因子进行了bootstrapped相关分析和线性混合模型(LME)拟合。结果表明,在高海拔地区马尾松径向生长对气候因子年际波动敏感性较强,主要表现为与生长季前冬季光温条件以及生长季内7月降水的正相关,生长-气候关系在不同样点间表现出较强的一致性。线性混合模型可以较好地拟合高海拔树木生长变化,当使用前1年12月平均日最高温、当年1月日照时长和当年7月降雨量3个气候变量进行拟合时,模型解释量达到0.5,其中前1年12月最高温和当年1月日照时数在模型中起到主导作用,累积相对贡献率约占80%,说明生长季前冬季的光热条件是限制高海拔马尾松径向生长最主要的气候因子。因此,我国亚热带地区高海拔的树木径向生长可能对未来气候变化有更强的敏感性,相关森林管理政策的制定需要将此纳入考虑;同时我国亚热带地区高海拔森林中的树木有被用于树轮气候重建的潜力。     

A cyp19a1b‐gfp (aromatase B) transgenic zebrafish line that expresses GFP in radial glial cells

Aromatase is an enzyme that catalyzes the synthesis of estrogen in gonads and brain. Teleost fish express aromatase (AroB) strongly in the brain facilitating its detailed examination. To understand the function of AroB in the brain, we generated transgenic zebrafish that expresses green fluorescent protein (GFP) driven by the brain aromatase cyp19a1b promoter. GFP was found in the radial glial cells of transgenic larvae and adult fish that overlap with AroB immunoreactivity in the correct temporal and spatial pattern. GFP was also coexpressed with radial cell marker BLBP, but was not in neurons. In addition, GFP expression in the radial glial cells was stimulated by estrogen, same as endogenous AroB expression. Thus, this transgenic line faithfully mimics the regulation of AroB expression in radial glial cells. It provides a powerful tool to further characterize progenitor radial cells in adult and developing fish and to evaluate estrogenic activities of xenoestrogens and phytoestrogens. genesis 47:67–73, 2009. © 2008 Wiley‐Liss, Inc.     

Radial growth of the Sahara’s oldest trees : Cupressus dupreziana A. Camus

The radial growth of Duprez’ cypress (Cupressus dupreziana A. Camus), a tree endemic to the Tassili n’Ajjer in the Central Sahara, was measured from cores taken in 1967 and 1997 from the same trees. A comparison of these cores, spanning 30 years, reveals that even in this hyper-arid climate, this species has a juvenile vigour comparable to other Mediterranean Cupressaceae. Wide individual variability was found, however, which was induced by the strong influence of microhabitat factors. Traditional dendrochronological methods of cross-dating are impossible to apply in the case of these trees. Ages of eight individuals were estimated with the help of radiocarbon dating, yielding an age range from 600 to 2400 years for trees of the most usual girths.     

Penetration of soil aggregates of finite size

Summary Maximum penetrometer pressure was measured on artificial soil aggregates of finite size (2–29 mm) using blunt probes (total cone angle 60°) driven at 3 mm min⁻¹. Maximum penetrometer pressure increased asymptotically with increase in dimensionless aggregate radius,b/a, wherea andb are the probe and aggregate radii, respectively. A theory was developed for penetration of blunt probes into soil aggregates of finite size. The theory assumed that plastic failure occurs out to a radius,R, and that beyond this only elastic straining occurs. This theory can be applied to estimate the radial and tangential stresses adjacent to a blunt probe. The estimated radial and tangential stresses increased with increase in dimensionless aggregate radius,b/a. The radius of the plastic front,R, around the probe is predicted to increase with increased aggregate size. The results also demonstrate the effect of soil shear cohesion and internal friction angle onR. The results are discussed with reference to root penetration.