Failure Of Regeneration Research Articles

Synergistic effects of mechanical stimulation and crimped topography to stimulate natural collagen development for tendon engineering

Suitable scaffold structures and mechanical loading are essential for functional tendon engineering. However, the bipolar fibril structure of native tendon collagen is yet to be recaptured in engineered tendons. This study compared the development of Achilles tendons of postnatal rats with and without (via surgical section) mechanical loading to define the mechanism of mechanical stimulation-mediated tendon development. The results demonstrated that the severed tendons weakened mechanically and exhibited disorganization without a bipolar fibril superstructure. Proteomic analysis revealed differentially expressed key regulatory molecules related to the collagen assembly process, including decreased fibromodulin, keratocan, fibroblast growth factor-1, and increased lumican and collagen5a1 in the severed tendons with immunohistochemical verification. Additionally, a complex regulatory network of mechanical stimulation-mediated collagen assembly in a spatiotemporal manner was also revealed using bioinformatics analysis, wherein PI3K–Akt and HDAC4 may be the predominant signaling pathways. A wavy microgrooved surface (Y = 5.47sin(0.015x)) that biomimics tendon topography was observed to enhance the expression of collagen assembly molecules under mechanical loading, and the aforementioned pathways are particularly involved and verified with their respective inhibitors of LY-294002 and LMK-235. Furthermore, an electrospun crimped nanofiber scaffold (approximately 2 μm fiber diameter and 0.12 crimpness) was fabricated to biomimic the tenogenic niche environment; this was observed to be more effective on enhancing collagen production and assembly under mechanical stimulation. In conclusion, the synergistic effect between topographical niche and mechanical stimulation was observed to be essential for collagen assembly and maturation and should be applied to functional tendon engineering in the future. Statement of significanceIn biomaterial-mediated tendon regeneration, mechanical stimulation is essential for tendon collagen assembly. However, the underlying mechanisms remain not fully defined, leading to the failure of the native-like collagen regeneration. In this study, a mechanical stimulation deprivation model of rat tendon was established to reveal the mechanisms in tendon development and define the key regulatory molecules including small leucine-rich proteoglycans, lysyl oxidase and collagen V. After ensuring the importance of biomimetic structure in tendon remodeling, crimped nanofibers were developed to verify these regulatory molecules, and demonstrated that mechanical stimulation significantly enhanced collagen assembly via PIK3 and HDAC4 pathways in biomaterial-regulated tendon regeneration. This study provides more insightful perspectives in the physiologically remodeling progression of tendon collagen and design of tendon scaffolds.

Immunohistochemistry indicates that persistent inflammation determines failure of tail, limb and finger regeneration in the Lizard Podarcis muralis

BackgroundThe presence of white blood inflammatory cells in injured tissues and their effect on the process of organ regeneration in lizards has been assessed on tail, limb and digits. MethodsThe present immunohistochemical survey analyzes the occurrence of CD68-labeled cells in lizard organs uncapable of regenerating tissues that exhibit strong inflammatory activity. ResultsThis marker mainly identifies macrophages and mast cells present in large number within tissues of injured limbs and digits. Also a high inflammation is associated with amputated tails that do not regenerate, derived from cauterization or infection of tissues of the tail stump. In the healing limbs and fingers at 12–20 days post-amputation, numerous CD68-labeled cells, most likely macrophages, are seen among superficial connective tissues and injured muscles and bones. These cells likely stimulate and give rise to scarring tissues and no regeneration of limb and fingers occurs. In the cauterized or in the infected tail stump a strong accumulation of CD68-positive mast cells and macrophages is observed, where they likely evoke epidermal coagulation, formation of scarring connective tissue, and loss of regeneration. ConclusionsThe present observations provide further cytological evidence that support the notion that a strong and lasting inflammatory condition impedes organ regeneration in specifically lizards and, more generally other vertebrates as well.

Nuclear Magnetic Resonance Treatment Accelerates the Regeneration of Dorsal Root Ganglion Neurons in vitro.

Functional recovery from peripheral nerve injuries depends on a multitude of factors. Schwann cells (SCs) are key players in the regenerative process as they develop repair-specific functions to promote axon regrowth. However, chronically denervated SCs lose their repair phenotype, which is considered as a main reason for regeneration failure. Previous studies reported a modulatory effect of low nuclear magnetic resonance therapy (NMRT) on cell proliferation and gene expression. To provide first insight into a possible effect of NMRT on cells involved in peripheral nerve regeneration, this study investigated whether NMRT is able to influence the cellular behavior of primary SC and dorsal root ganglion (DRG) neuron cultures in vitro. The effect of NMRT on rat SCs was evaluated by comparing the morphology, purity, proliferation rate, and expression levels of (repair) SC associated genes between NMRT treated and untreated SC cultures. In addition, the influence of (1) NMRT and (2) medium obtained from NMRT treated SC cultures on rat DRG neuron regeneration was examined by analyzing neurite outgrowth and the neuronal differentiation status. Our results showed that NMRT stimulated the proliferation of SCs without changing their morphology, purity, or expression of (repair) SC associated markers. Furthermore, NMRT promoted DRG neuron regeneration shown by an increased cell survival, enhanced neurite network formation, and progressed neuronal differentiation status. Furthermore, the medium of NMRT treated SC cultures was sufficient to support DRG neuron survival and neurite outgrowth. These findings demonstrate a beneficial impact of NMRT on DRG neuron survival and neurite formation, which is primarily mediated via SC stimulation. Our data suggest that NMRT could be suitable as a non-invasive auxiliary treatment option for peripheral nerve injuries and encourage future studies that investigate the effect of NMRT in a physiological context.

SPIONs mediated magnetic actuation promotes nerve regeneration by inducing and maintaining repair-supportive phenotypes in Schwann cells

BackgroundSchwann cells, the glial cells in the peripheral nervous system, are highly plastic. In response to nerve injury, Schwann cells are reprogrammed to a series of specialized repair-promoting phenotypes, known as repair Schwann cells, which play a pivotal role in nerve regeneration. However, repair Schwann cells represent a transient and unstable cell state, and these cells progressively lose their repair phenotypes and repair‐supportive capacity; the transience of this state is one of the key reasons for regeneration failure in humans. Therefore, the ability to control the phenotypic stability of repair Schwann cells is of great practical importance as well as biological interest.ResultsWe designed and prepared a type of fluorescent–magnetic bifunctional superparamagnetic iron oxide nanoparticles (SPIONs). In the present study, we established rat sciatic nerve injury models, then applied SPIONs to Schwann cells and established an effective SPION-mediated magnetic actuation system targeting the sciatic nerves. Our results demonstrate that magnetic actuation mediated by SPIONs can induce and maintain repair-supportive phenotypes of Schwann cells, thereby promoting regeneration and functional recovery of the sciatic nerve after crush injury.ConclusionsOur research indicate that Schwann cells can sense these external, magnetically driven mechanical forces and transduce them to intracellular biochemical signals that promote nerve regeneration by inducing and maintaining the repair phenotypes of Schwann cells. We hope that this study will provide a new therapeutic strategy to promote the regeneration and repair of injured peripheral nerves.Graphical

Research advances on stem cell therapy for diabetic foot wounds

Diabetic foot wound repair is a challenging issue in clinical practice. Due to the influence of multiple factors including the damage and regeneration failure of local tissue, the impaired pathways of wound repairing through blood vessels and nerve nutrition, and disorders of a variety of cellular factors, traditional treatment methods are often difficult to achieve good therapeutic effects. Stem cells are a type of cells with potentials of multidirectional differentiation, which also possess functions such as regulating immunity and paracrine to facilitate the comprehensive wound repair, so they have promising application prospect at present for the treatment of diabetic foot wounds. Because the relevant parameters of stem cell treatment are in the exploratory phase, there were no standardized data. This paper reviews the application of stem cells in the research of diabetic foot wound treatment over the past 6 years, analyzing and summarizing the contents in focused aspects including the types and sources of stem cells, effects of donor age and gender on stem cells, mode of administration, transplantation survival rate and safety, which may provide a reference for further application of stem cells in the clinical treatment of diabetic foot wound.

Therapeutic induction of energy metabolism reduces neural tissue damage and increases microglia activation in severe spinal cord injury

Neural tissue has high metabolic requirements. Following spinal cord injury (SCI), the damaged tissue suffers from a severe metabolic impairment, which aggravates axonal degeneration and neuronal loss. Impaired cellular energetic, tricarboxylic acid (TCA) cycle and oxidative phosphorylation metabolism in neuronal cells has been demonstrated to be a major cause of neural tissue death and regeneration failure following SCI. Therefore, rewiring the spinal cord cell metabolism may be an innovative therapeutic strategy for the treatment of SCI. In this study, we evaluated the therapeutic effect of the recovery of oxidative metabolism in a mouse model of severe contusive SCI. Oral administration of TCA cycle intermediates, co-factors, essential amino acids, and branched-chain amino acids was started 3 days post-injury and continued until the end of the experimental procedures. Metabolomic, immunohistological, and biochemical analyses were performed on the injured spinal cord sections. Administration of metabolic precursors enhanced spinal cord oxidative metabolism. In line with this metabolic shift, we observed the activation of the mTORC1 anabolic pathway, the increase in mitochondrial mass, and ROS defense which effectively prevented the injury-induced neural cell apoptosis in treated animals. Consistently, we found more choline acetyltransferase (ChAT)-expressing motor neurons and increased neurofilament-positive corticospinal axons in the spinal cord parenchyma of the treated mice. Interestingly, oral administration of the metabolic precursors increased the number of activated microglia expressing the CD206 marker suggestive of a pro-resolutive, M2-like phenotype. These molecular and histological modifications observed in treated animals ultimately led to a significant, although partial, improvement of the motor functions. Our data demonstrate that rewiring the cellular metabolism can represent an effective strategy to treat SCI.

Projected changes in fire activity and severity feedback in the spruce–feather moss forest of western Quebec, Canada

As a result of extreme weather conditions associated with anthropogenic climate change, fire regimes are expected to continue to change in the boreal forest over the 21st century and beyond. Consequently, changes in ecological attributes like stand composition, tree density and forest carbon stock can be expected. In the present study, we used an adjusted version of the CanFIRE model to project long-term (1971–2100) changes in burn rates, fire severity and fire-induced shifts in vegetation composition in response to anticipated scenarios of climate change, in the black spruce-feather moss subdomain of Western Quebec. The model provides decadal-scale estimates of the immediate physical effects of fire on forest communities by computing expected fire behavior and the resulting ecological effects. Changes in species composition of the forest is also computed based on mechanisms of succession in natural forest communities and fire-mediated vegetation transitions. Projections suggest an increase in potential burn rates across the study area under future weather conditions and also an overall reduction in percent tree mortality and total fuel consumption. This reduction is caused by negative feedback from vegetation composition that shifts to less-fire prone states. Although common forest communities will remain the same in the studied subdomain until 2100 (recurrence dynamics), significant losses of productive area (LPA) are projected, particularly in forest management units rich in forest communities dominated by black spruce or jack pine, as a result of regeneration failure due to very short intervals between successive fires. While remaining similar under moderate (RCP4.5) and high-end (RCP8.5) warming scenarios in all forest management units, LPA will vary from 25 to 36% of the percent cover by 2100 compared to 1970. These results provide insights to policy makers and land managers, and they attract attention to the pressing need to adjust management practices in the context of climate change.

Natural Regeneration of Maritime Pine: A Review of the Influencing Factors and Proposals for Management

The main objective of the present work was to review the current knowledge about the factors that influence the different phases of the natural regeneration of maritime pine (Pinus pinaster Aiton) and propose scientifically based management schedules. The review focused on the natural regeneration of maritime pine within its most representative areas (Portugal, Spain, and France). First, a brief characterisation of the main phases of the natural regeneration of maritime pine is described. Next, information on the factors affecting the natural regeneration of the species is provided, highlighting its positive and negative effects. Finally, management proposals are presented to promote the natural regeneration of maritime pine species. Numerous factors influencing the natural regeneration of maritime pine were identified. Fires and climatic factors can intervene in all three regeneration phases (seeding, germination, and seedling survival and development). The natural regeneration failure of maritime pine species can be aggravated in drier scenarios. Forest management plays an important role as it can promote the natural regeneration of the species.

Repair of the Injured Spinal Cord by Schwann Cell Transplantation

Spinal cord injury (SCI) can result in sensorimotor impairments or disability. Studies of the cellular response to SCI have increased our understanding of nerve regenerative failure following spinal cord trauma. Biological, engineering and rehabilitation strategies for repairing the injured spinal cord have shown impressive results in SCI models of both rodents and non-human primates. Cell transplantation, in particular, is becoming a highly promising approach due to the cells’ capacity to provide multiple benefits at the molecular, cellular, and circuit levels. While various cell types have been investigated, we focus on the use of Schwann cells (SCs) to promote SCI repair in this review. Transplantation of SCs promotes functional recovery in animal models and is safe for use in humans with subacute SCI. The rationales for the therapeutic use of SCs for SCI include enhancement of axon regeneration, remyelination of newborn or sparing axons, regulation of the inflammatory response, and maintenance of the survival of damaged tissue. However, little is known about the molecular mechanisms by which transplanted SCs exert a reparative effect on SCI. Moreover, SC-based therapeutic strategies face considerable challenges in preclinical studies. These issues must be clarified to make SC transplantation a feasible clinical option. In this review, we summarize the recent advances in SC transplantation for SCI, and highlight proposed mechanisms and challenges of SC-mediated therapy. The sparse information available on SC clinical application in patients with SCI is also discussed.

DHATUROOPI SHUKRA AND BEEJAROOPI SHUKRA: A CRITICAL REVIEW

Shukra is studied in Ayurveda both as a dhatu and beeja. As a mammalian human body comprises both somatic and gonadal cells. Somatic cells help for growth and regeneration through mitosis. Meiotic cell division causes equal contribution for the inheritance from maternal and paternal sides. Beejartham (reproduction) is the supreme function attributed to Shukra. Reproduction refers to the formation of new cells for tissue growth, repair/replacement (sukshmavayavantarotpatti), or the production of a new individual (shareerantarotpatti). Regenerative capacity is distributed unequally among species, individuals, and tissues. The affliction of shukrastana by kusthadosha (skin disease) causes a failure in regeneration. The affliction of parents' shukra and artava (gametes) by kusthadosha (skin disease) inherits to the next generation. Vrushan (testis) and medru (penis) are the moola of the shukravahavaha srotus, which is meant to fertilise the ovum (beejarupishukra). Majja (bone marrow) and stana (breasts) are the moola of the shukravaha srotus of the one pervading the entire body (dhaturupishukra).

Macrophage plasticity in Duchenne muscular dystrophy: a nexus of pathological remodelling with therapeutic implications.

Duchenne muscular dystrophy (DMD) is characterized by chronic skeletal muscle necrosis, leading to muscle regeneration failure and fibrosis. Although macrophages (MPs) are normally essential for muscle regeneration, dysregulated MP function promotes pathological muscle remodelling. Infiltrating MPs can be predominantly pro-inflammatory (M1 biased), anti-inflammatory (M2 biased) or of a mixed phenotype and can originate from the adult bone marrow (monocyte dependent) or embryonic precursors (monocyte independent). In mdx mice (genetic model of DMD) lacking either Toll-like receptor (Tlr) 2 or Tlr4, it is found that MP infiltration of dystrophic muscle is significantly reduced and that the MP phenotype is shifted toward a more anti-inflammatory profile. This is accompanied by significant improvements in muscle histology and force production. Lack of the chemokine receptor CCR2, which impedes monocyte release from the bone marrow, leads to similar beneficial effects in mdx mice. Evidence was also found for Tlr4-regulated induction of trained innate immunity in MPs cultured from the bone marrow of mdx mice before their entry into the muscle. These MPs exhibit epigenetic and metabolic alterations, accompanied by non-specific hyper-responsiveness to multiple stimuli, which is manifested by potentiated upregulation of both pro- and anti-inflammatory genes. In summary, exaggerated recruitment of monocyte-derived MPs and signs of trained innate immunity at the level of the bone marrow are features of the immunophenotype associated with dystrophic muscle disease. These phenomena are regulated by Toll-like receptors that bind endogenous damage-associated molecular pattern (DAMP) molecules, suggesting that DAMP release from dystrophic muscles modulates MP plasticity at the bone marrow level through Toll-like receptor-driven mechanisms.

Inhibition of central axon regeneration: perspective from chondroitin sulfate proteoglycans in lamprey spinal cord injury.

Inhibition of central axon regeneration: perspective from chondroitin sulfate proteoglycans in lamprey spinal cord injury.

Brain delivering RNA-based therapeutic strategies by targeting mTOR pathway for axon regeneration after central nervous system injury.

Injuries to the central nervous system (CNS) such as stroke, brain, and spinal cord trauma often result in permanent disabilities because adult CNS neurons only exhibit limited axon regeneration. The brain has a surprising intrinsic capability of recovering itself after injury. However, the hostile extrinsic microenvironment significantly hinders axon regeneration. Recent advances have indicated that the inactivation of intrinsic regenerative pathways plays a pivotal role in the failure of most adult CNS neuronal regeneration. Particularly, substantial evidence has convincingly demonstrated that the mechanistic target of rapamycin (mTOR) signaling is one of the most crucial intrinsic regenerative pathways that drive axonal regeneration and sprouting in various CNS injuries. In this review, we will discuss the recent findings and highlight the critical roles of mTOR pathway in axon regeneration in different types of CNS injury. Importantly, we will demonstrate that the reactivation of this regenerative pathway can be achieved by blocking the key mTOR signaling components such as phosphatase and tensin homolog (PTEN). Given that multiple mTOR signaling components are endogenous inhibitory factors of this pathway, we will discuss the promising potential of RNA-based therapeutics which are particularly suitable for this purpose, and the fact that they have attracted substantial attention recently after the success of coronavirus disease 2019 vaccination. To specifically tackle the blood-brain barrier issue, we will review the current technology to deliver these RNA therapeutics into the brain with a focus on nanoparticle technology. We will propose the clinical application of these RNA-mediated therapies in combination with the brain-targeted drug delivery approach against mTOR signaling components as an effective and feasible therapeutic strategy aiming to enhance axonal regeneration for functional recovery after CNS injury.

HDAC Inhibitor Titration of Transcription and Axolotl Tail Regeneration.

New patterns of gene expression are enacted and regulated during tissue regeneration. Histone deacetylases (HDACs) regulate gene expression by removing acetylated lysine residues from histones and proteins that function directly or indirectly in transcriptional regulation. Previously we showed that romidepsin, an FDA-approved HDAC inhibitor, potently blocks axolotl embryo tail regeneration by altering initial transcriptional responses to injury. Here, we report on the concentration-dependent effect of romidepsin on transcription and regeneration outcome, introducing an experimental and conceptual framework for investigating small molecule mechanisms of action. A range of romidepsin concentrations (0–10 μM) were administered from 0 to 6 or 0 to 12 h post amputation (HPA) and distal tail tip tissue was collected for gene expression analysis. Above a threshold concentration, romidepsin potently inhibited regeneration. Sigmoidal and biphasic transcription response curve modeling identified genes with inflection points aligning to the threshold concentration defining regenerative failure verses success. Regeneration inhibitory concentrations of romidepsin increased and decreased the expression of key genes. Genes that associate with oxidative stress, negative regulation of cell signaling, negative regulation of cell cycle progression, and cellular differentiation were increased, while genes that are typically up-regulated during appendage regeneration were decreased, including genes expressed by fibroblast-like progenitor cells. Using single-nuclei RNA-Seq at 6 HPA, we found that key genes were altered by romidepin in the same direction across multiple cell types. Our results implicate HDAC activity as a transcriptional mechanism that operates across cell types to regulate the alternative expression of genes that associate with regenerative success versus failure outcomes.

Mitigating post-fire regeneration failure in boreal landscapes with reforestation and variable retention harvesting: At what cost?

Successive disturbances such as fire can affect post-disturbance regeneration density, with documented adverse effects on subsequent stand productivity. We conducted a simulation study to assess the potential of reactive (reforestation) and proactive (variable retention harvesting) post-fire regeneration failure mitigation strategies in a 1.37 Mha fire-prone boreal landscape dominated by black spruce (Picea mariana (Mill.) B.S.P.) and jack pine (Pinus banksiana Lamb.). We quantified their respective capacity to maintain landscape productivity and post-fire resilience, as well as their associated financial returns under current and projected (RCP 8.5) fire regimes. While post-fire reforestation with jack pine revealed to be the most effective strategy to maintain potential production, associated costs quickly became prohibitive when applied over extensive areas. Proactive strategies such as an extensive use of variable retention harvesting, combined with replanting of fire-adapted jack pine only in easily accessible areas, appeared as a more promising approach. Despite this, our results suggest an inevitable erosion of forest productivity due to post-fire regeneration failure events, highlighting the importance of integrating fire a priori in strategic forest management planning as well as its effects on long-term regeneration dynamics.

Stem cell therapy in the treatment of organic and dysfunctional endometrial pathology.

Intrauterine adhesions caused by postpartum curettage, spontaneous abortions, interrupted pregnancies, endometrial ablations, infections and inflammations, can lead to a loss of endometrial function, with consequent hypomenorrhea and infertility in women of reproductive age. In a non-negligible percentage of cases, the available surgical methods and hormone therapy, with sequential administration of estrogen and progesterone, are ineffective. In fact, severe damage to the basal layer of the endometrium causes the loss of endometrial cell precursors and leads to the failure of regeneration of the functional layer to which the endometrium is cyclically exposed. Today, many researchers are evaluating the use of stem cells of different origins as a potential therapy to restore endometrial function. Our interest has been focused on adipose-derived stromal/stem cells (ADSCs) obtained by collecting subcutaneous adipose tissue and subsequently treating it with the MilliGraft® method. This procedure produces a cell suspension, the stromal vascular fraction (SVF), which includes ADSCs and soluble factors such as proteins and extracellular vesicles (exosomes). The SVF thus obtained was characterized in its cellular composition and its functional factors. Our clinical protocol for the future use of adipose tissue in endometrial regeneration in its different phases is presented. The data obtained, even though they still require further support and implementation, show the regenerative properties of SVF obtained from adipose tissue using a mechanical method. These findings can contribute to the development of cell therapies using stem cells of different derivations which are increasingly being utilized in the treatment of endometrial lesions from adherent or dysfunctional pathologies.

EP News: Basic and Translational

Wang et al (J Clin Invest 2021;131:e138637, PMID 34596051) developed a genetic system for ablating cardiomyocytes (CMs) that reside within the atrioventricular conduction system (AVCS). Adult mouse AVCS ablation resulted in regenerative failure characterized by persistent atrioventricular conduction defects and contractile dysfunction. In contrast, AVCS in neonatal mice exhibited evidence of plasticity with recovery from injury in a subset of mice. CM proliferation did not completely account for the observed functional recovery, suggesting that mechanisms regulating recovery are distinct from cardiac regeneration associated with ventricular injury.

Regeneration failure of Scots pine changes the species composition of young forests

ABSTRACT How seedling mortality and browsing affects species composition of regenerating forests has been mostly studied on a small scale. Yet, large-scale analyses based on extensive data are essential for robust prediction of species composition in young forests. In this study, we used a dataset from a national inventory of young forests (1–4 metres in height) to investigate the species composition of young forests across Sweden. We found that most of the regenerated forest area (almost 90%) was planted with Norway spruce (southern Sweden) and Scots pine (northern Sweden). Regeneration of Norway spruce was generally relatively successful but as a consequence of seedling mortality and browsing, almost 40% of the area regenerated with Scots pine will probably not develop into pine-dominated stands. Thus, low survival of Scots pine seedlings and trees can profoundly change the trajectory of species composition of the young forest from what was originally intended, and a large proportion of the young stands may develop into mixtures of conifers and broadleaves. While such mixtures may benefit certain biodiversity and ecosystem services, a loss of Scots pine dominated stands may also have adverse impacts on the economic returns as well as pine-dependent biodiversity and recreational values.

Promoting axon regeneration in the central nervous system by increasing PI3-kinase signaling.

Much research has focused on the PI3-kinase and PTEN signaling pathway with the aim to stimulate repair of the injured central nervous system. Axons in the central nervous system fail to regenerate, meaning that injuries or diseases that cause loss of axonal connectivity have life-changing consequences. In 2008, genetic deletion of PTEN was identified as a means of stimulating robust regeneration in the optic nerve. PTEN is a phosphatase that opposes the actions of PI3-kinase, a family of enzymes that function to generate the membrane phospholipid PIP3 from PIP2 (phosphatidylinositol (3,4,5)-trisphosphate from phosphatidylinositol (4,5)-bisphosphate). Deletion of PTEN therefore allows elevated signaling downstream of PI3-kinase, and was initially demonstrated to promote axon regeneration by signaling through mTOR. More recently, additional mechanisms have been identified that contribute to the neuron-intrinsic control of regenerative ability. This review describes neuronal signaling pathways downstream of PI3-kinase and PIP3, and considers them in relation to both developmental and regenerative axon growth. We briefly discuss the key neuron-intrinsic mechanisms that govern regenerative ability, and describe how these are affected by signaling through PI3-kinase. We highlight the recent finding of a developmental decline in the generation of PIP3 as a key reason for regenerative failure, and summarize the studies that target an increase in signaling downstream of PI3-kinase to facilitate regeneration in the adult central nervous system. Finally, we discuss obstacles that remain to be overcome in order to generate a robust strategy for repairing the injured central nervous system through manipulation of PI3-kinase signaling.

A short-interval reburn catalyzes departures from historical structure and composition in a mesic mixed-conifer forest

Recent increases in fire frequency and severity across the western US are triggering abrupt changes in ecosystem structure and composition, especially in lower montane forests, but consequences of fire-regime change for mesic, mixed-conifer forests remain uncertain. Glacier National Park (Montana, USA) is characterized by a complex mosaic of species typical of Rocky Mountain, Pacific Northwest and boreal floras, creating opportunities for species composition to shift as species respond individualistically to environmental change. We quantified prefire and postfire stem density and composition in a recent anomalously short-interval fire (15 yrs) and historically typical long-interval fire (88–282 + yrs) to answer two questions: (1) How do the structure and composition of tree regeneration differ after long- and short-interval fires that burned in close proximity? (2) What drivers explain differences in postfire regeneration densities among species? We additionally compared understory cover in each fire to anticipate consequences of contemporary fire for the whole plant community. Mean postfire tree seedling density was 45 times higher after the long- than short-interval fire, and many stands in the short-interval fire lacked any tree regeneration. Postfire tree seedling densities were above prefire stem densities in the long-interval fire but well below in the short-interval fire. After both fires, fire-avoiding conifers (e.g., Picea engelmannii, Abies lasiocarpa) were generally replaced by fire-tolerating or fire-embracing species (e.g., Larix occidentalis, Pinus contorta var. latifolia). Some stands remained dominated by fire-sensitive conifers after long-interval fire but never after short-interval fire. Seedlings of Thuja plicata and Tsuga heterophylla were present after the long-interval fire but absent before and after the short-interval fire, despite historically occupying both burned areas. For all species, seedling density increased with indicators of proximate seed supply. Seedling density of dispersal-dependent species declined with distance to live trees, whereas P. contorta responded to serotiny. Seedlings of drought-intolerant species were more abundant on northerly aspects. Total understory cover was 50% lower after short-interval than long-interval fire, but shrub cover was twice as high. Our study suggests that persistent seed sources will allow L. occidentalis to expand following sequential high-severity fires, while dispersal-limited, drought-intolerant conifers may decline due to loss of seed sources and elevated postfire aridity. Increased fire activity will likely produce sparser forests that are dominated by fire-tolerating and fire-embracing conifers at the expense of fire-avoiding and fire-refuge species. Complete regeneration failure may be delayed by high fire-trait syndrome diversity, but the structure and composition of extant forests will shift.