Axonal regeneration proceeds through specific axonal fusion in transected C. elegans neurons

A C. elegans mechanosensory neuron expressing GFP undergoes regenerative growth following laserinduced injury. A growth cone, sprouting from the proximal side of the axon, extends towards the separated distal fragment in an attempt to reconnect and fuse. The fluorescent image was digitally duplicated twice and re‐colored (Graphic designer: Dee McGrath). From Neumann et al., Developmental Dynamics 240:1365–1372, 2011     

Behavioral Plasticity in the C. elegans Mechanosensory Circuit

This review outlines research into the cellular and molecular mechanisms underlying a simple behavior in the soil-dwelling nematode, C. elegans. A tap administered to the side of a petri plate acts as a nonlocalized mechanical stimulus to the worms within. Most adult worms respond to this tap stimulus with backward locomotion, an action known as the tap-withdrawal response. This behavior has been thoroughly characterized and the neural circuit mediating it has been determined. The response habituates following repeated stimulation, and current work is aimed at elucidating the mechanism behind this simple form of nonassociative learning. Changes in cell excitability and the strength of glutamatergic synapses play key roles in mediating this plasticity.     

Involvement of cAMP in neuronal survival and axonal regeneration

In vitro, cAMP elevation alters neuronal responsiveness to diffusible growth factors and overcomes myelin-associated inhibitory molecules. Significant advances have been made recently in understanding the role of increases in cAMP in promoting axonal growth. Importantly, it has now been shown that cAMP elevation can promote axonal regeneration and functional recovery after central nervous system injury. Elevation of cAMP can be achieved via either direct application of cAMP analogs or an inhibitor of the enzyme phosphodiesterase that degrades cAMP in vivo. Current information points to a number of protein kinase A-mediated pathways (mitogen-activated protein kinase/extracellular signal-regulated kinase and phosphatidylinositol 3-kinase/akt pathway activation and Rho inactivation) underlying cAMP elevation-induced neuronal survival and axonal regeneration.     

Genetic analysis of KillerRed in C. elegans identifies a shared role of calcium genes in ROS-mediated neurodegeneration

In C. elegans, neurodegeneration induced by excitotoxicity or aggregation of misfolded proteins is dependent on genes involved in calcium release from the endoplasmic reticulum. Reactive oxygen species (ROS) can also induce neurodegeneration, but the relationship between ROS-mediated neurodegeneration and calcium has not been established. We activated KillerRed in the GABA neurons of C. elegans to produce ROS that leads to functional loss and structural degeneration of these neurons and demonstrated that the severity of neurodegeneration was dependent on extent of KillerRed activation. To genetically examine the role of calcium in ROS-mediated neurodegeneration, we measured functional neurodegeneration in itr-1 (inositol trisphosphate receptor), crt-1 (caltreticulin), and unc-68 (ryanodine receptor) mutants. Similar to other neurotoxic conditions, neurodegeneration triggered by KillerRed was reduced in itr-1 and crt-1 mutants. Somewhat unexpectedly, genetic or pharmacological disruption of unc-68 had a minimal effect on neurodegeneration. Our results indicate ROS-mediated neurodegeneration occurs through a conserved calcium regulated mechanism and suggest that components of the degeneration process have different sensitivities to ROS.     

Zygotic loss of ZEN-4/MKLP1 results in disruption of epidermal morphogenesis in the C. elegans embryo.

ZEN-4/MKLP1 is required maternally for cytokinesis in Caenorhabditis elegans, but was originally identified in a screen for zygotic lethal, enclosure abnormal (Zen) mutants. We report that zen-4(w35) homozygotes exhibit stochastic failures in cytokinesis in multiple lineages. Remarkably, multinucleate epidermal cells show directional migration, even when there are as few as half the normal number of cells. Temperature shift experiments and analysis of zen-4::gfp expression confirm that the epidermal requirement for zen-4 function precedes morphogenesis. Driving expression of wild-type zen-4 by means of an epithelial-specific transgene can rescue many epidermal morphogenetic defects in zen-4 mutants. Early expression of unc-119 in epidermal precursors made this promoter unsuitable as a neuronal-specific driver in this context. Our results indicate that zygotic zen-4 function is required for correct division of epidermal precursors and, hence, indirectly for normal morphogenesis and that the epidermal morphogenetic program is surprisingly robust even in the absence of zen-4 function.     

Lack of galectin-1 results in defects in myoblast fusion and muscle regeneration.

Galectin-1 has been implicated in the development of skeletal muscle, being maximally expressed at the time of myofiber formation. Furthermore, in the presence of exogenous galectin-1, mononuclear myoblasts show increased fusion in vitro. In the current study, we have used the galectin-1 null mouse to elucidate the role of galectin-1 in skeletal muscle development and regeneration. Myoblasts derived from the galectin-1 mutant showed a reduced ability to fuse in vitro. In galectin-1 null mutants, there was evidence of a delay in muscle fiber development at the neonatal stage and muscle fiber diameter was reduced when compared with wild-type at the adult stage. Muscle regeneration was also compromised in the galectin-1 mutant with the process being delayed and a reduced fiber size being maintained. These results, therefore, show a definitive role for galectin-1 in fusion of myoblasts both in vitro, in vivo, and in regeneration after recovery from induced injury.     

Axonal transport of synapsin I- and cholinergic synaptic vesicle-like material; further immunohistochemical evidence for transport of axonal cholinergic transmitter vesicles in motor neurons

The axonal transport of organelles in motor axons in the sympathectomized rat sciatic has been studied using two antisera which recognize specific components of synaptic vesicles. Anti-synapsin I recognizes synapsin I (SYN I) which is affiliated with the external membrane of synaptic vesicles, while rabbit-anti-synaptic vesicle antiserum (RASVA) recognizes integral membrane glycoproteins in cholinergic synaptic vesicles. Immunofluorescence studies, including cytofluorimetric scanning, show that immuno-reactive (IR) material recognized by both antisera: (1) rapidly accumulate proximal to a crush; (2) the material has a granular appearance in the microscope; (3) is redistributed in an isolated segment, and (4) that the transport of the material is sensitive to vinblastine. Thus the proximodistal transport has the characteristics of fast axonal transport. Furthermore, recycling organelles, accumulating on the distal side of a crush are recognized by RASVA, but carry only very little SYN I-IR. The results give further support to the hypothesis that motor cholinergic axons transport axonal cholinergic vesicles towards the motor endplates.     

Homeodomain interacting protein kinase (HPK‐1) is required in the soma for robust germline proliferation in C. elegans

Background: Tightly regulated pathways maintain the balance between proliferation and differentiation within stem cell populations. In Caenorhabditis elegans, the germline is the only tissue that is maintained by stem‐like cells into adulthood. In the current study, we investigated the role played by a member of the Homeodomain interacting protein kinase (HIPK) family of serine/threonine kinases, HPK‐1, in the development and maintenance of the C. elegans germline. Results: We report that HPK‐1 is required for promotion of germline proliferation during development and into adulthood. Additionally, we show that HPK‐1 is required in the soma for regulation of germline proliferation. We also show that HPK‐1 is a predominantly nuclear protein expressed in several somatic tissues including germline‐interacting somatic cells. Conclusions: Our observations are consistent with a conserved role for HIPKs in the control of cellular proliferation and identify a new context for such control in germ cell proliferation. Developmental Dynamics 242:1250–1261, 2013. © 2013 Wiley Periodicals, Inc.     

Effect of Schwann cell transplantation combined with electroacupuncture on axonal regeneration and remyelination in rats with spinal cord injury

Axonal impairment and demyelination after compressed spinal cord injury lead to serious neurological dysfunction. Increasing studies have suggested that Schwann cells (SCs) transplantation is a reliable, effective, and promising method for treating spinal cord injury. However, single SCs transplantation is insufficient to promote the full recovery of neurological function. Additional approaches are required to support SCs transplantation as a treatment for spinal cord injury. In the study, we investigated whether the combination of electroacupuncture (EA) and SCs transplantation was a reliable intervention for spinal cord injury. We found that rats in the combination group had significantly higher functional locomotor scores than those received single treatment. By immunostaining, we found EA can not only improve survival and proliferation of transplanted SCs but also inhibit SC apoptosis and block the formation of an astrocytic scar. Additionally, EA promoted regenerated axons extending “bullet-shaped” growth cones into the lesion. Remarkably, EA can modify astrogliosis to promote axonal regeneration following SCs transplantation through inducing extension of astrocytic processes in the SCs graft interface. More importantly, the combination of SCs engraftment and EA can enhance corticospinal-tract axonal regeneration and remyelination after spinal cord injury through up-regulating neuregulin 1 type III in SCs and its downstream signaling mediators. Thus, it is concluded that SCs effectively promote axonal recovery after spinal cord injury when combined with EA stimulation. The experimental results have reinforced the theoretical basis of EA for its clinical efficacy in patients with spinal cord injury and merited further investigation for potential clinical application.     

Induction of phosphorylated c‐Jun in neonatal spinal motoneurons after axonal injury is coincident with both motoneuron death and regeneration

c‐Jun activation has been implicated not only in neuronal degeneration, but also in survival and regeneration. Here, we investigated c‐Jun activation in injured motoneurons by using a nerve crush model in neonatal rats. We identified two distinct subpopulations of motoneurons: about 60% underwent degeneration following injury whereas the remaining 40% survived and induced a regeneration response at 3 weeks post injury. However, all motoneurons examined expressed phosphorylated‐c‐Jun‐immunoreactivity (p‐c‐Jun‐IR) at the early stage of 3 days following injury. These results suggest that active c‐Jun was induced in all neonatal motoneurons following nerve crush injury, regardless of whether they were destined to degenerate or undergo successful regeneration at a later stage. Our findings therefore support the hypothesis that active c‐Jun is involved in both neuronal degeneration and regeneration.     

New developmental insights from high-throughput biological analysis in Caenorhabditis elegans

The use of Caenorhabditis elegans as a model system for understanding animal development and human disease has long been recognized as an efficient tool of discovery. Recent developments, particularly in our understanding of RNA-mediated interference and its ability to modify gene activity, have facilitated the use of C. elegans in determining gene function via high-throughput analysis. These new strategies have provided a framework that allows investigators to analyse gene function globally at the genomic level and will likely become a prototypic model for biological analysis in the post-genome era.     

Caenorhabditis elegans germ line: a model for stem cell biology.

Like many stem cell systems, the Caenorhabditis elegans germ line contains a self-renewing germ cell population that is maintained by a niche. Although the exact cellular mechanism for self-renewal is not yet known, three recent studies shed considerable light on the cell cycle behavior of germ cells, including a support for significant and plastic renewal potential. This review brings together the results of the three recent cell-based studies, places them in the context of previous work, and discusses future perspectives for the field.     

Axonal transport in the central axon of sensory neurons during regeneration of their peripheral axon

Following sciatic nerve injury (which provokes prompt regeneration of peripheral sensory axons) there was no change in the ratio between amounts of labeled protein conveyed by fast axonal transport into the central and peripheral axons. A change in composition of the transported protein, characteristic of regenerating peripheral axons, also occurred in the central axons. When the peripheral axon is injured, changes in axonal transport thus affect both processes, and there is no routing of fast-transported protein into or away from the regenerating process.     

Muscle-epidermis interactions affect exoskeleton patterning in Caenorhabditis elegans.

The C. elegans hypodermis is a single epithelial cell layer separated from the musculature by a thin basement membrane on its basal surface. The hypodermis secretes the extracellular material of the cuticle from its apical surface. The regulation of cuticle synthesis and apical secretion is not well understood. UNC-95 is a component of the muscle dense bodies and M-lines, which are integrin-based adhesion complexes required for force transduction to the cuticle. Using gene expression profiling and in vivo assays, we show that, in unc-95 mutant worms, there is an increase in expression levels of a group of hypodermal and pharyngeal genes related to cuticle structure and molting. Moreover, the cuticle structure of unc-95 mutant adult is impaired. Our findings suggest that aberrant force transduction from the structurally impaired muscle attachments across the basement membrane to the underlying hypodermis elicits intercellular signaling that plays a role in regulating cuticle synthesis and patterning.     

Generation and modulation of chemosensory behaviors in C. elegans

C. elegans recognizes and discriminates among hundreds of chemical cues using a relatively compact chemosensory nervous system. Chemosensory behaviors are also modulated by prior experience and contextual cues. Because of the facile genetics and genomics possible in this organism, C. elegans provides an excellent system in which to explore the generation of chemosensory behaviors from the level of a single gene to the motor output. This review summarizes the current knowledge on the molecular and neuronal substrates of chemosensory behaviors and chemosensory behavioral plasticity in C. elegans.     

EGL-4/PKG regulates the role of an interneuron in a chemotaxis circuit of C. elegans through mediating integration of sensory signals

Interneurons, innervated by multiple sensory neurons, need to integrate information from these sensory neurons and respond to sensory stimuli adequately. Mechanisms how sensory information is integrated to form responses of interneurons are not fully understood. In Caenorhabditis elegans, loss-of-function mutations of egl-4, which encodes a cGMP-dependent protein kinase (PKG), cause a defect in chemotaxis to odorants. Our genetic and imaging analyses revealed that the response property of AIY interneuron to an odorant is reversed in the egl-4 mutant, while the responses of two upstream olfactory neurons, AWA and AWC, are largely unchanged. Cell- ablation experiments show that AIY in the egl-4 mutant functions to suppress chemotaxis. Furthermore, the reversal of AIY response occurs only in the presence of sensory signals from both AWA and AWC. These results suggest that sensory signals are inadequately integrated in the egl-4 mutant. We also show that egl-4 expression in AWA and another sensory neuron prevents the reversed AIY response and restores chemotaxis in the egl-4 mutants. We propose that EGL-4/PKG, by suppressing aberrant integration of signals from olfactory neurons, converts the response property of an interneuron to olfactory stimuli and maintains the role of the interneuron in the circuit to execute chemotactic behavior.