Functional evaluation of peripheral nerve regeneration and target reinnervation in animal models: a critical overview

Peripheral nerve injuries usually lead to severe loss of motor, sensory and autonomic functions in the patients. Due to the complex requirements for adequate axonal regeneration, functional recovery is often poorly achieved. Experimental models are useful to investigate the mechanisms related to axonal regeneration and tissue reinnervation, and to test new therapeutic strategies to improve functional recovery. Therefore, objective and reliable evaluation methods should be applied for the assessment of regeneration and function restitution after nerve injury in animal models. This review gives an overview of the most useful methods to assess nerve regeneration, target reinnervation and recovery of complex sensory and motor functions, their values and limitations. The selection of methods has to be adequate to the main objective of the research study, either enhancement of axonal regeneration, improving regeneration and reinnervation of target organs by different types of nerve fibres, or increasing recovery of complex sensory and motor functions. It is generally recommended to use more than one functional method for each purpose, and also to perform morphological studies of the injured nerve and the reinnervated targets.

[1]  Xavier Navarro,et al.  Bimodal dose‐dependence of FK506 on the rate of axonal regeneration in mouse peripheral nerve , 2002, Muscle & nerve.

[2]  J. Ilha,et al.  The IBB Forelimb Scale as a tool to assess functional recovery after peripheral nerve injury in mice , 2014, Journal of Neuroscience Methods.

[3]  E. Costanzo,et al.  Co‐regulation of synaptic efficacy at stable polyneuronally innervated neuromuscular junctions in reinnervated rat muscle , 1999, The Journal of physiology.

[4]  C. Krarup,et al.  Functional Recovery of Regenerating Motor Axons is Delayed in Mice Heterozygously Deficient for the Myelin Protein P0 Gene , 2013, Neurochemical Research.

[5]  A. Windebank,et al.  Measurement of forelimb function by digital video motion analysis in rat nerve transection models , 2008, Journal of the peripheral nervous system : JPNS.

[6]  P. Thanos,et al.  Insulin-like growth factor-I promotes nerve regeneration through a nerve graft in an experimental model of facial paralysis. , 1999, Restorative neurology and neuroscience.

[7]  C. Vleggeert-Lankamp,et al.  The role of evaluation methods in the assessment of peripheral nerve regeneration through synthetic conduits: a systematic review. Laboratory investigation. , 2007, Journal of neurosurgery.

[8]  S. Barbay,et al.  An index of topographic normality in rat somatosensory cortex: application to a sciatic nerve crush model. , 2002, Journal of neurophysiology.

[9]  Theofano Eftaxiopoulou,et al.  Gait compensations in rats after a temporary nerve palsy quantified using temporo-spatial and kinematic parameters , 2014, Journal of Neuroscience Methods.

[10]  S. Dunnett,et al.  The “staircase test”: a measure of independent forelimb reaching and grasping abilities in rats , 1991, Journal of Neuroscience Methods.

[11]  X. Navarro,et al.  Comparison of Immunohistochemical and Functional Reinnervation of Skin and Muscle after Peripheral Nerve Injury , 1997, Experimental Neurology.

[12]  J. Terzis,et al.  An experimental model to study the blink reflex. , 1994, Journal of reconstructive microsurgery.

[13]  J. Kerns,et al.  A mathematical model for regeneration rate and initial delay following surgical repair of peripheral nerves , 1993, Journal of Neuroscience Methods.

[14]  X. Navarro,et al.  Neurophysiological, histological and immunohistochemical characterization of bortezomib-induced neuropathy in mice , 2010, Experimental Neurology.

[15]  Paul L. Evans,et al.  Regeneration across preserved peripheral nerve grafts , 1995, Muscle & nerve.

[16]  A. Valero-Cabré,et al.  Superior muscle reinnervation after autologous nerve graft or poly‐L‐lactide‐ϵ‐caprolactone (PLC) tube implantation in comparison to silicone tube repair , 2001, Journal of neuroscience research.

[17]  Pedro Melo-Pinto,et al.  Methods for the experimental functional assessment of rat sciatic nerve regeneration , 2004, Neurological research.

[18]  W. Gispen,et al.  A new approach for the evaluation of recovery after peripheral nerve damage , 1988, Journal of Neuroscience Methods.

[19]  J. Diamond,et al.  Impulse activity evokes precocious sprouting of nociceptive nerves into denervated skin. , 1984, Somatosensory research.

[20]  X. Navarro,et al.  Immediate electrical stimulation enhances regeneration and reinnervation and modulates spinal plastic changes after sciatic nerve injury and repair , 2008, Experimental Neurology.

[21]  C. Krarup,et al.  Comparative electrophysiological, functional, and histological studies of nerve lesions in rats , 2005, Microsurgery.

[22]  F. Schügner,et al.  Evaluation of functional nerve recovery with Visual-SSI—A novel computerized approach for the assessment of the static sciatic index (SSI) , 2008, Journal of Neuroscience Methods.

[23]  J. Fawcett,et al.  Characterization of tests of functional recovery after median and ulnar nerve injury and repair in the rat forelimb , 2007, Journal of the peripheral nervous system : JPNS.

[24]  Electrophysiologic findings and grip strength after nerve injuries in the rat forelimb , 2008, Muscle & nerve.

[25]  J. Vibert,et al.  Maturation of regenerating nerve assessed by evoked potential in the rat: Hyperbolic time course , 1988, Experimental Neurology.

[26]  X. Navarro,et al.  Chapter 6: Methods and protocols in peripheral nerve regeneration experimental research: part III-electrophysiological evaluation. , 2009, International review of neurobiology.

[27]  W. Freed,et al.  An index of the functional condition of rat sciatic nerve based on measurements made from walking tracks , 1982, Experimental Neurology.

[28]  X. Navarro,et al.  On the use of fast blue, fluoro-gold and diamidino yellow for retrograde tracing after peripheral nerve injury: uptake, fading, dye interactions, and toxicity , 2002, Journal of Neuroscience Methods.

[29]  S. Dunnett,et al.  The staircase test of skilled reaching in mice , 2001, Brain Research Bulletin.

[30]  X. Navarro,et al.  Nociceptive responses and spinal plastic changes of afferent C-fibers in three neuropathic pain models induced by sciatic nerve injury in the rat , 2009, Experimental Neurology.

[31]  P. Thompson Electromyography for Experimentalists , 1987 .

[32]  S. Amado,et al.  The sensitivity of two-dimensional hindlimb joint kinematics analysis in assessing functional recovery in rats after sciatic nerve crush , 2011, Behavioural Brain Research.

[33]  B. Meyerson,et al.  Spinal cord stimulation in animal models of mononeuropathy: effects on the withdrawal response and the flexor reflex , 1995, Pain.

[34]  S. Mackinnon,et al.  Lack of association between outcome measures of nerve regeneration , 1998, Muscle & nerve.

[35]  T Gordon,et al.  Contributing factors to poor functional recovery after delayed nerve repair: prolonged axotomy , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[36]  R. Midha,et al.  Dose and duration of nerve growth factor (NGF) administration determine the extent of behavioral recovery following peripheral nerve injury in the rat , 2011, Experimental Neurology.

[37]  J. Crabbe,et al.  Influence of task parameters on rotarod performance and sensitivity to ethanol in mice , 2003, Behavioural Brain Research.

[38]  X. Navarro,et al.  Neural plasticity after peripheral nerve injury and regeneration , 2007, Progress in Neurobiology.

[39]  R. Berenberg,et al.  Recovery of peripheral nerve function after axotomy: Effect of triiodothyronine , 1977, Experimental Neurology.

[40]  R. Cuppini Age-related changes in maturation of regenerated motor innervation , 1993, Journal of the Neurological Sciences.

[41]  A. Irintchev,et al.  Opposite impacts of tenascin‐C and tenascin‐R deficiency in mice on the functional outcome of facial nerve repair , 2005, The European journal of neuroscience.

[42]  K. Rodenacker,et al.  Temporal progression and extent of the return of sensation in the foot provided by the saphenous nerve after sciatic nerve transection and repair in the rat—implications for nociceptive assessments , 2007, Somatosensory & motor research.

[43]  O. Guntinas-Lichius,et al.  Transplantation of Olfactory Mucosa Minimizes Axonal Branching and Promotes the Recovery of Vibrissae Motor Performance after Facial Nerve Repair in Rats , 2002, The Journal of Neuroscience.

[44]  X. Navarro,et al.  Changes in crossed spinal reflexes after peripheral nerve injury and repair. , 2002, Journal of neurophysiology.

[45]  Z. Wiesenfeld‐Hallin,et al.  Expansion of innervation territory by afferents involved in plasma extravasation after nerve regeneration in adult and neonatal rats , 2004, Experimental Brain Research.

[46]  Paul L. Evans,et al.  Walking Track Analysis: A Long‐Term Assessment of Peripheral Nerve Recovery , 1992, Plastic and reconstructive surgery.

[47]  F. Biedermann,et al.  A surface EMG multi-electrode technique for characterizing muscle activation patterns in mice during treadmill locomotion , 2005, Journal of Neuroscience Methods.

[48]  X. Navarro,et al.  Assessment of sensory thresholds and nociceptive fiber growth after sciatic nerve injury reveals the differential contribution of collateral reinnervation and nerve regeneration to neuropathic pain , 2014, Experimental Neurology.

[49]  Jonathan R Wolpaw,et al.  Recovery of electromyographic activity after transection and surgical repair of the rat sciatic nerve. , 2007, Journal of neurophysiology.

[50]  D Schonfeld,et al.  Two modes of cutaneous reinnervation following peripheral nerve injury , 1979, The Journal of comparative neurology.

[51]  M. Marcus,et al.  The CatWalk gait analysis in assessment of both dynamic and static gait changes after adult rat sciatic nerve resection , 2007, Journal of Neuroscience Methods.

[52]  G. Loeb,et al.  Conduction studies in peripheral cat nerve using implanted electrodes: II. The effects of prolonged constriction on regeneration of crushed nerve fibers , 1988, Muscle & nerve.

[53]  J. Peyronnard,et al.  A comparative study of the effects of chronic axotomy, crush lesion and re-anastomosis of the rat sural nerve on horseradish peroxidase labelling of primary sensory neurons , 1988, Brain Research.

[54]  Natalie L. M. Cappaert,et al.  The functional and morphological characteristics of sciatic nerve degeneration and regeneration after crush injury in rats , 2014, Journal of Neuroscience Methods.

[55]  M. Meek,et al.  Functional evaluation of peripheral nerve regeneration in the rat: walking track analysis , 2001, Journal of Neuroscience Methods.

[56]  W. Kennedy,et al.  Sweat gland reinnervation by sudomotor regeneration after different types of lesions and graft repairs , 1989, Experimental Neurology.

[57]  R. L. Holland,et al.  An assessment of the spread of the signal for terminal sprouting within and between muscles , 1981, Brain Research.

[58]  X. Navarro,et al.  Comparison of Regenerative and Reinnervating Capabilities of Different Functional Types of Nerve Fibers , 1994, Experimental Neurology.

[59]  T. Nichols,et al.  Reinnervated muscles fail to produce stretch reflexes. , 1994, Journal of neurophysiology.

[60]  A Gramsbergen,et al.  Biodegradable p(DLLA‐ϵ‐CL) nerve guides versus autologous nerve grafts: Electromyographic and video analysis , 2001, Muscle & nerve.

[61]  J L Walker,et al.  Improved footprint analysis using video recording to assess functional recovery following injury to the rat sciatic nerve. , 1994, Restorative neurology and neuroscience.

[62]  A. Gramsbergen,et al.  Toe out angle: a functional index for the evaluation of sciatic nerve recovery in the rat model , 2003, Experimental Neurology.

[63]  M. Fischer,et al.  Surface EMG-recordings using a miniaturised matrix electrode: a new technique for small animals , 2000, Journal of Neuroscience Methods.

[64]  G. Brook,et al.  CatWalk gait analysis in assessment of functional recovery after sciatic nerve injury , 2008, Journal of Neuroscience Methods.

[65]  A. Irintchev,et al.  Impacts of lesion severity and tyrosine kinase receptor B deficiency on functional outcome of femoral nerve injury assessed by a novel single‐frame motion analysis in mice , 2005, The European journal of neuroscience.

[66]  H. Tilson,et al.  A method for the routine assessment of fore- and hindlimb grip strength of rats and mice. , 1979, Neurobehavioral toxicology.

[67]  J. Young,et al.  THE RATE OF REGENERATION OF NERVE , 1942 .

[68]  R. Hopf,et al.  Serial recording of sensory, corticomotor, and brainstem-derived motor evoked potentials in the rat. , 2001, Somatosensory & motor research.

[69]  F. Bajrović,et al.  Efficacy of fluorescent tracers in retrograde labeling of cutaneous afferent neurons in the rat , 2010, Journal of Neuroscience Methods.

[70]  D. Ceballos,et al.  FK506 enhances reinnervation by regeneration and by collateral sprouting of peripheral nerve fibers , 2003, Experimental Neurology.

[71]  S. Mackinnon,et al.  Functional Evaluation of Complete Sciatic, Peroneal, and Posterior Tibial Nerve Lesions in the Rat , 1989, Plastic and reconstructive surgery.

[72]  A. Brenan,et al.  The demonstration of the cutaneous distribution of saphenous nerve C-fibres using a plasma extravasation technique in the normal rat and following nerve injury. , 1988, Journal of anatomy.

[73]  Xavier Navarro,et al.  Specificity of peripheral nerve regeneration: Interactions at the axon level , 2012, Progress in Neurobiology.

[74]  W. Gispen,et al.  Methods for producing a reproducible crush in the sciatic and tibial nerve of the rat and rapid and precise testing of return of sensory function Beneficial effects of melanocortins , 1986, Journal of the Neurological Sciences.

[75]  T Gordon,et al.  Increased Neuromuscular Activity Reduces Sprouting in Partially Denervated Muscles , 2001, The Journal of Neuroscience.

[76]  I. Mcquarrie,et al.  Axonal regeneration in the rat sciatic nerve: Effect of a conditioning lesion and of dbcAMP , 1977, Brain Research.

[77]  S. Lisney,et al.  Reinnervation of sweat glands in the rat hind paw following peripheral nerve injury. , 1988, Journal of the autonomic nervous system.

[78]  M. Malessy,et al.  Electrophysiology and morphometry of the Aα- and Aβ-fiber populations in the normal and regenerating rat sciatic nerve , 2004, Experimental Neurology.

[79]  H. Kimura,et al.  Brain-Derived Neurotrophic Factor from Bone Marrow-Derived Cells Promotes Post-Injury Repair of Peripheral Nerve , 2012, PloS one.

[80]  Paul L. Evans,et al.  Walking Track Analysis: Utilization of Individual Footprint Parameters , 1993, Annals of plastic surgery.

[81]  E. Stennert,et al.  Contralateral trigeminal nerve lesion reduces polyneuronal muscle innervation after facial nerve repair in rats , 1999, The European journal of neuroscience.

[82]  P. Whelan,et al.  Electromyogram recordings from freely moving animals. , 2003, Methods.

[83]  R. Cuppini,et al.  Enlargement of motoneuron peripheral field following partial denervation with or without dorsal rhizotomy , 1998, Neuroscience.

[84]  Sameer B. Shah,et al.  Phosphorylation of Highly Conserved Neurofilament Medium KSP Repeats Is Not Required for Myelin-Dependent Radial Axonal Growth , 2009, The Journal of Neuroscience.

[85]  C. Krarup,et al.  Post reinnervation maturation of myelinated nerve fibers in the cat tibial nerve: chronic electrophysiological and morphometric studies , 2000, Journal of the peripheral nervous system : JPNS.

[86]  S. Mackinnon,et al.  Sciatic nerve regeneration in the rat. Validity of walking track assessment in the presence of chronic contractures , 1989, Microsurgery.

[87]  S. Shenaq,et al.  Reliability of sciatic function index in assessing nerve regeneration across a 1 cm gap , 1989, Microsurgery.

[88]  M. Beattie,et al.  An Analysis of Changes in Sensory Thresholds to Mild Tactile and Cold Stimuli after Experimental Spinal Cord Injury in the Rat , 2000, Neurorehabilitation and neural repair.

[89]  X. Navarro,et al.  Quantitative assessment of locomotion and interlimb coordination in rats after different spinal cord injuries , 2013, Journal of Neuroscience Methods.

[90]  R. Cuppini,et al.  Motor activity affects adult skeletal muscle re‐innervation acting via tyrosine kinase receptors , 2013, The European journal of neuroscience.

[91]  P. Yu,et al.  Gait analysis in rats with peripheral nerve injury , 2001, Muscle & nerve.

[92]  N. Lago,et al.  Effects of motor and sensory nerve transplants on amount and specificity of sciatic nerve regeneration , 2007, Journal of neuroscience research.

[93]  A. Baptista,et al.  A new approach to assess function after sciatic nerve lesion in the mouse—Adaptation of the sciatic static index , 2007, Journal of Neuroscience Methods.

[94]  Albert Gramsbergen,et al.  Methods to evaluate functional nerve recovery in adult rats: walking track analysis, video analysis and the withdrawal reflex , 2000, Journal of Neuroscience Methods.

[95]  Xavier Navarro,et al.  H reflex restitution and facilitation after different types of peripheral nerve injury and repair , 2001, Brain Research.

[96]  A. Irintchev,et al.  Mechanical stimulation of paralyzed vibrissal muscles following facial nerve injury in adult rat promotes full recovery of whisking , 2007, Neurobiology of Disease.

[97]  Tessa Gordon,et al.  Electrical Stimulation Promotes Motoneuron Regeneration without Increasing Its Speed or Conditioning the Neuron , 2002, The Journal of Neuroscience.

[98]  M Holmes,et al.  Endogenous NGF and nerve impulses regulate the collateral sprouting of sensory axons in the skin of the adult rat , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[99]  L. Guth,et al.  AN ELECTROPHYSIOLOGICAL STUDY OF THE EARLY STAGES OF PERIPHERAL NERVE REGENERATION. , 1965, Experimental neurology.

[100]  A. Gramsbergen,et al.  Sciatic Nerve Transection in the Adult Rat: Abnormal EMG Patterns during Locomotion by Aberrant Innervation of Hindleg Muscles , 2000, Experimental Neurology.

[101]  C. Rohde,et al.  Pathway sampling by regenerating peripheral axons , 2005, The Journal of comparative neurology.

[102]  M. Sabbahi,et al.  Ankle Stance Angle: A Functional Index for the Evaluation of Sciatic Nerve Recovery after Complete Transection , 1996, Journal of reconstructive microsurgery.

[103]  R. Dubner,et al.  A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia , 1987, Pain.

[104]  P. Dyck,et al.  Number, size, and class of peripheral nerve fibers regenerating after crush, multiple crush, and graft , 1989, Brain Research.

[105]  S. Mackinnon,et al.  Choosing the correct functional assay: A comprehensive assessment of functional tests in the rat , 2005, Behavioural Brain Research.

[106]  C. Krarup,et al.  Factors that influence peripheral nerve regeneration: An electrophysiological study of the monkey median nerve , 2002, Annals of neurology.

[107]  M. Bervar Video analysis of standing — an alternative footprint analysis to assess functional loss following injury to the rat sciatic nerve , 2000, Journal of Neuroscience Methods.

[108]  S. Archibald,et al.  Reinnervation Accuracy of the Rat Femoral Nerve by Motor and Sensory Neurons , 1996, The Journal of Neuroscience.

[109]  D. Nicholls,et al.  Protein kinase C modulates field‐evoked transmitter release from cultured rat cerebellar granule cells via a dendrotoxin‐sensitive K+ channel , 1999, The European journal of neuroscience.

[110]  J. Arezzo,et al.  Nerve growth factor prevents experimental cisplatin neuropathy , 1992, Annals of neurology.

[111]  Jeremy M. Shefner,et al.  Motor unit number estimation in human neurological diseases and animal models , 2001, Clinical Neurophysiology.

[112]  T. Yaksh,et al.  Quantitative assessment of tactile allodynia in the rat paw , 1994, Journal of Neuroscience Methods.

[113]  J. Bertelli,et al.  The grasping test: a simple behavioral method for objective quantitative assessment of peripheral nerve regeneration in the rat , 1995, Journal of Neuroscience Methods.

[114]  J. Kerns,et al.  A twitch tension method to assess motor nerve function in rat , 1987, Journal of Neuroscience Methods.

[115]  Daniel A. Bloch,et al.  Functional indices for sciatic, peroneal, and posterior tibial nerve lesions in the mouse , 1998, Microsurgery.

[116]  Xavier Navarro,et al.  Functional impact of axonal misdirection after peripheral nerve injuries followed by graft or tube repair. , 2002, Journal of neurotrauma.