Amphibian Limb Regeneration and its Relation to Nerves

Much circumstantial evidence points to a neurotrophic influencein amphibian limb regeneration. Although fine-structural observationsof nerves in regenerating limbs have indicated the possibilitythat neurosecretory vesicles accumulate distally in these axons,there is no clear-cut demonstration available that these organellesare neurotropic. Evidence is accumulating that the neural influencein newt limb regeneration is transneuronal. There is also evidencethat trophic substances other than those found in the nerveitself may be involved in the supporting limb regeneration.The characterization of the neurotrophic substance is considereda central task for students of regeneration in the future.     

The global regulatory logic of organ regeneration: circuitry lessons from skin and its appendages

In organ regeneration, the regulatory logic at a systems level remains largely unclear. For example, what defines the quantitative threshold to initiate regeneration, and when does the regeneration process come to an end? What leads to the qualitatively different responses of regeneration, which restore the original structure, or to repair which only heals a wound? Here we discuss three examples in skin regeneration: epidermal recovery after radiation damage, hair follicle fate choice after chemotherapy damage, and wound-induced feather regeneration. We propose that the molecular regulatory circuitry is of paramount significance in organ regeneration. It is conceivable that defects in these controlling pathways may lead to failed regeneration and/or organ renewal, and understanding the underlying logic could help to identify novel therapeutic strategies.     

Spontaneous regeneration of older dystrophic muscle does not reflect its regenerative capacity

Young dystrophic (dy) murine muscle is capable of "spontaneous" regeneration (i.e., regeneration in the absence of external trauma); however, by the time the mice are 8 weeks old, this regeneration ceases. It has been suggested that the cessation of regeneration in dystrophic muscle may be due to exhaustion of the mitotic capability of myosatellite cells during the early stages of the disease. To test this hypothesis, orthotopic transplantation of bupivacaine treated, whole extensor digitorum longus muscles has been performed on 14 to 16-week-old 129 ReJ/++ and 129 ReJ/dydy mice. The grafted dystrophic muscle is able to produce and maintain for 100 days post-transplantation 356 +/- 22 myofibers, a number similar to that found in age-matched dystrophic muscle. The ability of old dystrophic muscle to regenerate subsequent to extreme trauma indicates that the cessation of "spontaneous" regeneration is due to factor(s) other than the exhaustion of mitotic capability of myosatellite cells. Moreover, there is no significant difference in myosatellite cell frequencies between grafted normal and dystrophic muscles (100 days post-transplantation). Myosatellite cell frequencies in grafted muscles are similar to those in age-matched, untraumatized muscles. While grafting of young dystrophic muscle modifies the phenotypic expression of histopathological changes usually associated with murine dystrophy, grafts of older dystrophic muscle show extensive connective-tissue infiltration and significantly fewer myofibers than do grafts of age-matched normal muscle. As early as 14 days post-transplantation, it is possible to distinguish between grafts of old, normal and dystrophic muscles. It is suggested that the connective tissue stroma, present in the dystrophic muscle at the time of transplantation, may survive the grafting procedure.     

Regeneration of the acorn worm pygochord with the implication for its convergent evolution with the notochord

The origin of the notochord is a central issue in chordate evolution. This study examined the development of the acorn worm pygochord, a putative homologue of the notochord. Because the pygochord differentiates only after metamorphosis, the developmental was followed process by inducing regeneration after artificial amputation in Ptychodera flava. It was found that although the regeneration of the posterior part of the body did not proceed via formation of an obvious regeneration bud, pygochord regeneration was observed within a few weeks, possibly via trans-differentiation of endoderm cells. The expression of the fibrillary collagen gene (Fcol) and elav in the pygochord during regeneration was detected. This indicates that pygochord cells are not part of gut epithelial cells, but that they differentiated as a distinct cell type. Our gene expression analyses do not provide supporting evidence for the homology between the pygochord and notochord, but rather favored the convergent evolution between them.     

不同密度华北落叶松林天然更新及其影响因子

以关帝山区8种林分密度(128、240、320、400、480、560、640和720株·hm^-2)华北落叶松纯林为研究对象,分析了不同密度下华北落叶松天然更新优劣及其影响因子。结果表明: 更新指数随林分密度增大先升高后降低,中密度(400～560株·hm^-2)林分更新效果明显优于过疏和过密林分;林下光环境、草本植物及凋落物对落叶松更新有双重作用,光照过强、草本植物盖度过大或凋落物过厚不利于林下更新,土壤有机质可使土壤质地松软、水分含量充足并形成低磷高氮环境促进林分更新;各因子对更新指数的影响依次为:土壤含水量(0.798)>全氮(0.621)>凋落物厚度(-0.597)>土壤孔隙度(0.504)>土壤有机质(0.493)>全磷(-0.404)>林下总光照(-0.274)>草本植物盖度(-0.021)。在后续森林经营中,可通过抚育间伐或补植将林分密度控制在480株·hm^-2左右,并适当清理林下凋落物,改善土壤条件,促进华北落叶松天然更新。     

When one tail isn't enough: abnormal caudal regeneration in lepidosaurs and its potential ecological impacts

Abnormal caudal regeneration, the production of additional tails through regeneration events, occurs in lepidosaurs as a result of incomplete autotomy or sufficient caudal wound. Despite being widely known to occur, documented events generally are limited to opportunistic single observations – hindering the understanding of the ecological importance of caudal regeneration. Here we compiled and reviewed a robust global database of both peer‐reviewed and non‐peer reviewed records of abnormal regeneration events in lepidosaurs published over the last 400 years. Using this database, we qualitatively and quantitatively assessed the occurrence and characteristics of abnormal tail regeneration among individuals, among species, and among populations. We identified 425 observations from 366 records pertaining to 175 species of lepidosaurs across 22 families from 63 different countries. At an individual level, regenerations ranged from bifurcations to hexafurcations; from normal regeneration from the original tail to multiple regenerations arising from a single point; and from growth from the distal third to the proximal third of the tail. Species showing abnormal regenerations included those with intra‐vertebral, inter‐vertebral or no autotomy planes, indicating that abnormal regenerations evidently occur across lepidosaurs regardless of whether the species demonstrates caudal autotomy or not. Within populations, abnormal regenerations were estimated at a mean ± SD of 2.75 ± 3.41% (range 0.1–16.7%). There is a significant lack of experimental studies to understand the potential ecological impacts of regeneration on the fitness and life history of individuals and populations. We hypothesised that abnormal regeneration may affect lepidosaurs via influencing kinematics of locomotion, restrictions in escape mechanisms, anti‐predation tactics, and intra‐ and inter‐specific signalling. Behaviourally testing these hypotheses would be an important future research direction.     

昆明西山半湿润常绿阔叶林及其〖HJ*3〗〖HJ*2〗次生演替群落的更新特征

对昆明西山国家森林公园处于不同演替阶段的云南油杉林、云南油杉-滇青冈混交林和滇青冈林内木本植物幼苗的种类组成、数量及更新方式等进行了调查分析。结果表明:在所调查的共144个25m2的样方中,共记录到木本植物幼苗32科45属共49种。随着演替的进行,幼苗总密度表现出先增加后降低的趋势,处于演替中期的云南油杉-滇青冈混交林内幼苗最丰富,平均密度达80株.25m-2。在不同演替阶段各植被类型中,实生和萌生两种更新方式同时存在,单一的更新方式可能使群落的更新面临较大的风险。幼苗发生了顶死或梢枯后,从其根颈处生出多个萌枝形成多干基株的更新方式结合了实生和萌生更新各自的优点,是幼苗在与环境长期作用过程中进化出的一种有效对策。     

Regeneration of neuron processes of the spinal ganglion after its experimental section in rats

Sensory fibers regeneration after dorsal root ganglion (DRG) section was studied in the rat. After middle cross-section of DRG-13 (left side) there were neurons in the proximal part with damage to peripheral processes and in the distal part with damage to central processes. Axonal ionophoresis of cobalt salts was used for the study of sensory fibers regeneration through the scar in DRG during 3, 7, 15, 30, 120 and 180 days after the damage. Peculiarity regeneration of the sensory fibers was shown in spite of damage localization near ganglion cells body, the regeneration of peripheral and central processes of ganglion cells started already on the 3rd day, and sprouting sensory fibers through the scar of DRG--on the 7th day.     

Retinoic acid and its receptors in limb regeneration

The effects of retinoids on the regenerating amphibian limb are described: the mesenchymal cells of the blastema can be proximalized, posteriorized and ventralized. Ectopic limbs are also induced after retinoid treatment of regenerating tails, but not during limb development unless the limb bud is damaged. The cellular and molecular alterations induced by retinoids are reported as well as experiments which have revealed the importance of endogenous retinoids for normal limb regeneration. Various retinoic acid receptors are expressed in the regeneration blastema and the experiments which have revealed functions for individual isoforms are described. These experiments reveal that retinoids are a crucial component of the normal, regenerating limb and demonstrate the value of the regenerating limb as an experimental system for providing functional data on individual retinoic acid receptors.     

甲酸脱氢酶及其在手性生物制造中的应用

氧化还原生物合成体系在绿色生物制造手性化合物中具有重要应用价值.甲酸脱氢酶(formate dehydrogenase,FDH)能氧化甲酸盐生成二氧化碳,同时将NAD(P)+还原为NAD(P)H,是氧化还原生物合成中辅酶再生体系的关键酶.但天然的FDH催化效率低、稳定性差、辅酶利用率不高等缺点制约了其在工业生产中的应用...     

A new immunoblotting assay for thymidylate synthetase and its application to the regulation of enzyme activity in regenerating rat liver

A highly sensitive and specific immunoblot assay has been developed to quantitate the content of rat liver thymidylate synthetase (EC 2.1.1.45). Applying the method, it is demonstrated that the increase of the activity of thymidylate synthetase in liver regeneration after partial hepatectomy is due to the de novo synthesis of the enzyme protein. Administration of cycloheximide, phenoxybenzamine, phorbol 12-myristate 13-acetate, nifedipine, dexamethasone or indomethacin to partially hepatectomized rats prevented the synthesis of thymidylate synthetase in regenerating liver. Thyroparathyroidectomy also inhibited the increase of the enzyme in liver regeneration. These observations are discussed in relation to the signal transduction concerning the alpha 1-receptor, which was shown to regulate liver regeneration in our previous papers.     

黄土高原三种耐干藓营养繁殖的季节差异及机理

苔藓植物在园林绿化、水土保持等方面的应用日益增加,目前藓类植物营养繁殖的影响因素鲜见报道,限制了其应用研究。本研究以黄土高原常见的耐干藓(扭口藓、土生对齿藓、短叶对齿藓)为对象,研究了3种藓春、夏、秋、冬季营养繁殖能力(以活力指数表征)差异及其叶绿素、可溶性糖、可溶性蛋白质、丙二醛(MDA)含量等生理指标变化,探索了耐干藓营养繁殖的季节差异与其生理特征之间的关系。结果表明: 1)藓类繁殖具有明显的季节差异,3种藓夏季活力指数均低于其他3个季节,平均较秋、冬、春季分别下降56.1%、48.4%、10.1%;相同季节的活力指数具有明显的年际变化。2)3种藓繁殖能力具有明显的种间差异,以短叶对齿藓最高,扭口藓最低,且后者的繁殖能力季节和年际变化最大。3)3种藓不同季节生理特征差异显著,其中夏季MDA含量高于其他季节,可溶性糖和可溶性蛋白质含量低于其他季节。4)耐干藓的营养繁殖季节变化主要与可溶性糖含量有关。本研究明确了藓类营养繁殖能力随季节变化的规律,指出可溶性糖含量是影响藓类营养繁殖的关键因素,可为藓类植物保护和人工培养提供科学依据。     

Melatonin and its ubiquitous anticancer effects

Molecular and Cellular Biochemistry - Mesenchymal stem cell transplantation has been proposed as a promising therapy for regeneration of damaged tissues—especially, bone marrow mesenchymal...     

Recent advances in stem cell therapy for neurodegenerative disease: Three dimensional tracing and its emerging use

Neurodegenerative disease is a brain disorder caused by the loss of structure and function of neurons that lowers the quality of human life. Apart from the limited potential for endogenous regeneration, stem cell-based therapies hold considerable promise for maintaining homeostatic tissue regeneration and enhancing plasticity. Despite many studies, there remains insufficient evidence for stem cell tracing and its correlation with endogenous neural cells in brain tissue with three-dimensional structures. Recent advancements in tissue optical clearing techniques have been developed to overcome the existing shortcomings of cross-sectional tissue analysis in thick and complex tissues. This review focuses on recent progress of stem cell treatments to improve neurodegenerative disease, and introduces tissue optical clearing techniques that can implement a three-dimensional image as a proof of concept. This review provides a more comprehensive understanding of stem cell tracing that will play an important role in evaluating therapeutic efficacy and cellular interrelationship for regeneration in neurodegenerative diseases.     

Taurine and its Trophic Effects in the Retina Lucimey Lima

The sulphur amino acid taurine possesses variable functions during development and regeneration of the central nervous system. The retina synthesize and uptake taurine, which is the amino acid present in the highest concentration in this tissue. Deficiency of taurine alters the structure and the function of the cerebral and cerebelar cortex, as well as the retina. Taurine increases outgrowth of postcrush goldfish retina in culture, partially by elevating calcium influx, and also by the modulation of protein phosphorylation. Its concentration increases in the retina after the lesion of the optic nerve, and the intraocular injection of it, between the crush and the explantation, stimulates the outgrowth of neurites. Taken together, although there are a great number of unresolved questions on the mechanisms of action of this amino acid as atrophic substance, the results support the role of taurine during regeneration of the optic nerve.     

The regeneration of a protected Sonoran Desert cactus since 1800 A.D. over 50,000 km2 of its range

The tall, branched saguaro cactus (Carnegiea gigantea) is perhaps the best-recognized symbol of the desert. However, little is known about the regeneration and future of the species outside of a few well-studied populations. In 2000, data for 537 saguaros were collected in 30 populations throughout the northern Sonoran Desert. A recently developed technique now provides a mechanism by which to reconstruct individual age at multiple populations. This new technique is based on a general growth curve with a site-specific adjustment factor, which I calculate based on local growth data and a recognized relationship with summer rain. Thus, the year of establishment was estimated for all saguaros in each of the populations individually, followed by the merging of all individuals, to create a single regeneration and survivorship curve for the combined regional dataset. Unlike other studies that only look at regeneration at one site, this is the first study to look at the long-term regeneration of the species over an area of more than 50,000 km², nearly the US portion of its range. The results suggest that over the long term, the population is quite stable, with a favorable period of regeneration in the late 1800s and early 1900s. It is also encouraging that the frequency of individuals that established in the most recent time period is relatively high. However, whether these youngest individuals will persist over the long-term in the face of future extreme freezing events (which can substantially thin populations) is not clear.     

Regeneration of amphioxus oral cirri and its skeletal rods: implications for the origin of the vertebrate skeleton

The oral cirri of amphioxus function as the first filter during feeding by eliminating unwanted large or noxious particulates. In this study, we were able to regenerate cirri following artificial amputation. This is the first firm observation of regeneration in amphioxus. Using this regeneration system, we studied skeletogenesis of the cellular skeleton of amphioxus oral cirri. During regeneration, the skeletal cells showed expression of fibrillar collagen and SoxE genes. These observations suggest that an evolutionarily conserved genetic regulatory system is involved in amphioxus cirrus and vertebrate cartilage skeletogenesis. In addition, Runx and SPARC/osteonectin expression were observed in regenerating cirral skeletal cells, indicating that cirral skeletogenesis is similar to vertebrate osteogenesis. We propose that the common ancestors of chordates possessed a genetic regulatory system that was the prototype of chondrogenesis and osteogenesis in vertebrates. Genome duplications caused divergence of this genetic regulatory system resulting in the emergence of cartilage and mineralized bone. The development of the vertebrate skeleton is an example of the functional segregation and subsequent recruitment of unique genetic materials that may account for the evolutionary diversification of novel cell types.     

The planarian neoblast: the rambling history of its origin and some current black boxes

First described by Randolph in 1897, the nature and main features of planarian neoblasts have a long rambling history. While their morphologically undifferentiated features have long been recognized, their origin and actual role during regeneration have been highly debated. Here I summarize the main stages of this rambling history: 1) undifferentiated, wandering cells of uncertain origin with a main, albeit undefined, role in regeneration (1890-1940s); 2) quiescent, undifferentiated cells whose main function is to build the blastema during regeneration, an idea which culminated in the 'neoblast theory' of the French School (1940-1960); 3) neoblasts as temporal, undifferentiated cells arising by dedifferentiation from differentiated cells (the 'cell dedifferentiation theory'; 1960-1980s); 4) a new paradigm, starting in the late 1970s-early 1980s, that brought together the role of neoblasts as the main cell for regeneration, with its more important role as somatic stem cells for the daily wear and tear of tissues and as the source of germ cells; and 5) more recent developments that culminate in the report of rescuing lethally irradiated planarians by injection of single neoblasts, which makes of neoblasts an unrivaled toti-, pluripotent somatic stem cell system in the Animal Kingdom. I finally discuss some "black boxes" regarding neoblasts which still baffle us, namely their phylogenetic and ontogenetic origins, their role in body size control, how their pool is regulated during growth and degrowth, the logic of their proliferative control, and some 'old' long-sought missing tools.     

Prevention of the death of the rat axotomized hypoglossal nerve and promotion of its regeneration by bovine brain gangliosides.

We have examined the time course of the neuronal death and regeneration of rat axotomized hypoglossal nerve with various conditions of the nerve resection, and established a useful system to measure neurotrophic activities of bioactive substances. In this system, neuronal death can be evaluated by counting surviving neurons in the nucleus of hypoglossal neuron at the brain stem, and the degree of the regeneration can be measured by counting horseradish peroxidase-positive cells at the same region after injection of horseradish peroxidase into tongue. Using this system, the effects of brain gangliosides on rat hypoglossal nerve regeneration following 5 mm transection were examined. The addition of a ganglioside mixture from bovine brain as well as the autograft strongly prevented the death of neurons and promoted the regeneration of the lesioned nerve at 10 weeks after the operation. Further analyses on the dose effects and injection sites of gangliosides were performed. Although the mechanisms of the neurotrophic effects of the gangliosides are unknown, the therapeutic application of gangliosides for neuronal degeneration is a promising approach.     

The presence of cyclic AMP, and its effects on oral regeneration and in situ ciliary regeneration in Stentor coeruleus.

We have found cyclic AMP in the large, heterotrichous ciliate Stentor coeruleus in amounts per milligram protein similar to those found in another ciliate, Tetrahymena pyriformis. The possible function of cyclic AMP in Stentor was first examined by determining its effects on oral regeneration, the process by which Stentor can replace a missing oral apparatus in eight to ten hours. Once begun (by brief exposure to a 15% sucrose solution, causing shedding of the oral apparatus) regeneration follows eight specific morphological stages visible with the dissecting microscope. Continuous exposure of regenerating cells to either N⁶, 2′-0-dibutyryl adenosine cyclic 3′:5′-monophosphate (DBC) or theophylline begun at the onset of oral regeneration (stage 0) caused delays in the completion of regeneration. The delays induced by DBC occurred in the early stages prior to stage 5. Regenerating cells exposed to DBC or theophylline at various stages of development were delayed, even at stages 5 and 6. Both DBC and theophylline reversibly bleached the cortical pigment of the cells. Guanosine 3′:5′-cyclic monophosphate (cyclic GMP), AMP, GMP, and sodium butyrate neither delayed oral regeneration nor bleached the cortical pigment. Excess extracellular calcium ions alone had no effect on oral regeneration, but 10 mM calcium and DBC caused more delay than DBC alone. Thus, the delay of oral regeneration in Stentor caused by cyclic AMP may involve calcium ions. To determine if cyclic AMP can retard in situ ciliary regeneration by Stentor, as it does in Tetrahymena, a new technique, more accurate than past methods, was developed to monitor ciliary regrowth. Using this procedure we found that both DBC and theophylline significantly delayed the in situ ciliary regeneration by Stentor.