Spinal Motor Neuronal Degeneration After Knee Joint Immobilization in the Guinea Pig



      This study used a unilateral knee joint immobilization model in adult guinea pigs to test the hypothesis that retrograde degeneration of motor neurons in the spinal cord is the result of attenuation of knee joint activities.


      A total of 32 adult guinea pigs were used and divided into 8 groups based on the duration of knee joint immobilization. Light microscopic studies of Nissl, nitric oxide synthase immunohistochemistry, horseradish peroxidase, and fast blue were carried out to examine the neurons in the spinal cord. Electron microscopy was also performed to examine the neurons and axons.


      After various periods of knee joint immobilization, a variety of features of motor neuronal degeneration were observed. Specific characteristics included gradual increases in the expressions of neuronal nitric oxide synthase and ultrastructural changes in affected motor neurons including reduction of cell organelles, indentation of the nuclear envelop, and small compact clumps of chromatin in the nuclei. Observation of the peripheral nerve (femoral nerve) also revealed demyelination alterations in some axons innervating the muscles of the knee joint. Interestingly, motor neuronal degenerative changes and demyelination were reversible after the knee joint immobilization was removed and knee joint activity was restored. These findings may assist in further development of models for spinal dysfunction such as the chiropractic subluxation complex.


      We conclude that motor neuronal degeneration in the spinal cord and axons in this study was the result of knee joint immobilization. Increases in motor neuronal nitric oxide-mediated oxidative stress level after reduction of target tissue activity may contribute to the mechanism for degenerative changes in the motor neurons in adult spinal cord of the guinea pig.

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        • Sood C
        A study of the effects of experimental immobilisation on rabbit articular cartilage.
        J Anat. 1971; 108: 497-507
        • Finsterbush A
        • Friedman B
        Early changes in immobilized rabbits knee joints: a light and electron microscopic study.
        Clin Orthop. 1973; 92: 305-319
        • Troyer H
        The effect of short-term immobilization on the rabbit knee joint cartilage. A histochemical study.
        Clin Orthop. 1975; 107: 249-257
        • Langenskiod A
        • Michelsson JE
        • Videman T
        Osteoarthritis of the knee in the rabbit produced by immobilization. Attempts to achieve a reproducible model for studies on pathogenesis and therapy.
        Acta Orthop Scand. 1979; 50: 1-14
        • Candolin T
        • Videman T
        Surface changes in the articular cartilage of rabbit knee during immobilization. A scanning electron microscopic study of experimental osteoarthritis.
        Acta Pathol Microbiol Scand. 1980; 88: 291-297
        • Jurvelin J
        • Helminen HJ
        • Lauritsalo S
        • et al.
        Influences of joint immobilization and running exercise on articular cartilage surfaces of young rabbits. A semi quantitative stereomicroscopic and scanning electron microscopic study.
        Acta Anat. 1985; 122: 62-68
        • Paukkonen K
        • Jurvelin J
        • Helminen HJ
        Effects of immobilization on the articular cartilage in young rabbits. A quantitative light microscopic stereological study.
        Clin Orthop. 1986; 206: 270-280
        • Kiviranta I
        • Tammi M
        • Jurvelin J
        • Arokoski J
        • Saamanen AM
        • Helminen HJ
        Articular cartilage thickness and glycosaminoglycan distribution in the young canine knee joint after remobilization of the immobilized limb.
        J Orthop Res. 1994; 12: 161-167
        • Setton LA
        • Mow VC
        • Muller FJ
        • Pita JC
        • Howell DS
        Mechanical behavior and biochemical composition of canine knee cartilage following periods of joint disuse and disuse with remobilization.
        Osteoarthritis Cartilage. 1997; 5: 1-16
        • Fu LL
        • Maffulli N
        • Yip KM
        • Chan KM
        Articular cartilage lesions of the knee following immobilisation or destabilisation for 6 or 12 weeks in rabbits.
        Clin Rheumatol. 1998; 17: 227-233
        • Haapala J
        • Arokoski J
        • Pirttimaki J
        • Lyyra T
        • Jurvelin J
        • Tammi M
        • Helminen HJ
        • Kiviranta I
        Incomplete restoration of immobilization induced softening of young beagle knee articular cartilage after 50-week remobilization.
        Int J Sports Med. 2000; 21: 76-81
        • Gardner E
        The distribution and termination of nerves in the knee joint of the cat.
        J Comp Neurol. 1944; 80: 11-32
        • Freeman MAR
        • Wyke BD
        The innervation of the knee joint. An anatomical and histological study in the cat.
        J Anat. 1967; 101: 505-532
        • Hildebrand C
        • Öqvist G
        • Brax L
        • Tuisku F
        Anatomy of the rat knee joint and fibre composition of a major articular nerve.
        Anat Rec. 1991; 229: 545-555
        • Lee SH
        • Wang SC
        • Shen CL
        Innervation of the knee joint in the rabbit and the Formosan rock-monkey (Macaca cyclopis): a retrograde HRP study.
        Proc Natl Sci Counc Repub China B. 1991; 15: 171-177
        • He XH
        • La Rose J
        • Zhang N
        Extrinsic and intrinsic Innervation of the Guinea-pig knee joint: toward an animal model for manipulative therapy.
        J Manipulative Physiol Ther. 2009; 32: 223-231
        • Michinaka Y
        • Yamamoto H
        • Morisawa T
        • Uemura H
        Effect of immobilization of the knee joint on mechanoreceptors in anterior cruciate ligament of the rabbit.
        J Orthop Sci. 1997; 2: 259-265
        • Kanemura N
        • Kobayashi R
        • Kajihara H
        • et al.
        Changes of mechamoreceptor in anterior cruciate ligament with hindlimb suspension rats.
        J Phys Ther Sci. 2002; 14: 27-32
        • Fuglsang-Frederiksen A
        • Scheel U
        Transient decrease in number of motor units after immobilization in man.
        J Neurol Neurosurg Psychiatry. 1978; 41: 924-929
        • Gonzalez MF
        • Sharp FR
        • Sagar SM
        Axotomy increase NADPH-diaphorase staining in rat vagal motor neurons.
        Brain Res Bull. 1987; 18: 417-427
        • Yu WH
        Nitric oxide synthase in motor neuron after axotomy.
        J Histochem Cytochem. 1994; 42: 451-457
        • Wu W
        • Li L
        Inhibition of nitric oxide synthase reduces motoneuron death due to spinal root avulsion.
        Neurosci Lett. 1993; 153: 121-124
        • Mesulam M-M
        Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction-product with superior sensitive for visualising neural afferent and efferent.
        J Histochem Cytochem. 1978; 26: 106-117
        • Rexed B
        The cytoarchitectonic organization of the spinal cord in the cat.
        J Comp Neurol. 1952; 96: 415-495
        • Iwasaki Y
        • Shiojima T
        • Ikeda K
        • Kinoshita M
        Bromocriptine prevents spinal motor neuron death following sciatic nerve transection in neonatal rats.
        Neurosci Lett. 1996; 214: 139-142
        • Martin LJ
        • Kaiser AI
        • Price AC
        Motor neuron degeneration after sciatic nerve avulsion in adult rat evolves with oxidative stress and is apoptosis.
        J Neurobiol. 1999; 40: 185-201
        • Koliatsos V
        • Clatterbuck RE
        • Winslow JW.
        • Cayouette MH
        • Price DL
        Evidence that brain derived neurotrophic factor is an atrophic factor for motor neurons in vivo.
        Neuron. 1993; 10: 359-367
        • Dasen JS
        • De Camilli A
        • Wang B
        • Tucker PW
        • Jessell TM
        Hox repertoires for motor neuron diversity and connectivity gated by a single accessory factor.
        Cell. 2008; 134: 304-316
        • Oppenheim RW
        Muscle activity and motor neuron death in the spinal cord of the chick embryo.
        Ciba Found Sym. 1987; 126: 96-112
        • Hanson MG
        • Landmesser LT
        Normal patterns of spontaneous activity are required for correct motor axon guidance and the expression of specific guidance molecules.
        Neuron. 2004; 43: 687-701
        • Gómez-Pinilla F
        • Ying Z
        • Roy RR
        • Molteni R
        • Edgerton VR
        Voluntary exercise induces a BDNF-mediated mechanism that promotes neuroplasticity.
        J Neurophysiol. 2002; 88: 2187-2195
        • Miyata Y
        • Kasihara S
        • Homma S
        • Kuno M
        Effects of nerve growth factor on the survival and synaptic function of Ia sensory neurons axotomized in neonatal rats.
        J Neurosci. 1990; 6: 2012-2018
        • Sendtner M
        • Kreutzberg GW
        • Thoenen H
        Ciliary neurotrophic factor prevents the degeneration of motoneurons after axotomy.
        Nature. 1990; 345: 440-441
        • Feng Z
        • Ko CP
        Schwann cell promote synaptogenesis at the neuromuscular junction via transforming growth factor-beta 1.
        J Neurosci. 2008; 28: 9599-9609
        • Pellitteri R
        • Russo A
        • Stanzani S
        Schwann cell: a source of neurotrophic activity on cortical glutamatergic neurons in culture.
        Brain Res. 2006; 1069: 139-144
        • Cheng C
        • Chen M
        • Shi S
        • Gao S
        • Niu S
        • Li X
        • Liu H
        • Qin Y
        • Shen A
        Effect of peripheral axotomy on gene expression of NIDD in rat neural tissue.
        J Mol Neurosci. 2007; 32: 199-206