Assessing the role of dopamine in limb and cranial-oromotor control in a rat model of Parkinson's disease.

UNLABELLED: Parkinson's disease (PD) is a neurodegenerative disorder primarily characterized by sensorimotor dysfunction. The neuropathology of PD includes a loss of dopamine (DA) neurons of the nigrostriatal pathway. Classic signs of the disease include rigidity, bradykinesia, and postural instability. However, as many as 90% of patients also experience significant deficits in speech, swallowing (including mastication), and respiratory control. Oromotor deficits such as these are underappreciated, frequently emerging during the early, often hemi-Parkinson, stage of the disease. In this paper, we review tests commonly used in our labs to model early and hemi-Parkinson deficits in rodents. We have recently expanded our tests to include sensitive models of oromotor deficits. This paper discusses the most commonly used tests in our lab to model both limb and oromotor deficits, including tests of forelimb-use asymmetry, postural instability, vibrissae-evoked forelimb placing, single limb akinesia, dry pasta handling, sunflower seed shelling, and acoustic analyses of ultrasonic vocalizations and pasta biting strength. In particular, we lay new groundwork for developing methods for measuring abnormalities in the acoustic patterns during eating that indicate decreased biting strength and irregular intervals between bites in the hemi-Parkinson rat. Similar to limb motor deficits, oromotor deficits, at least to some degree, appear to be modulated by nigrostriatal DA. Finally, we briefly review the literature on targeted motor rehabilitation effects in PD models. LEARNING OUTCOMES: Readers will: (a) understand how a unilateral lesion to the nigrostriatal pathway affects limb use, (b) understand how a unilateral lesion to the nigrostriatal pathway affects oromotor function, and (c) gain an understanding of how limb motor deficits and oromotor deficits appear to involve dopamine and are modulated by training.

Polyethylene glycol rapidly restores axonal integrity and improves the rate of motor behavior recovery after sciatic nerve crush injury.

The inability to rapidly (within minutes to hours) improve behavioral function after severance of peripheral nervous system axons is an ongoing clinical problem. We have previously reported that polyethylene glycol (PEG) can rapidly restore axonal integrity (PEG-fusion) between proximal and distal segments of cut- and crush-severed rat axons in vitro and in vivo. We now report that PEG-fusion not only reestablishes the integrity of crush-severed rat sciatic axons as measured by the restored conduction of compound action potentials (CAPs) and the intraaxonal diffusion of fluorescent dye across the lesion site, but also produces more rapid recovery of appropriate hindlimb motor behaviors. Improvement in recovery occurred during the first few postoperative weeks for the foot fault (FF) asymmetry test and between week 2 and week 3 for the Sciatic Functional Index (SFI) based on analysis of footprints. That is, the FF test was the more sensitive indicator of early behavioral recovery, showing significant postoperative improvement of motor behavior in PEG-treated animals at 24-48 h. In contrast, the SFI more sensitively measured longer-term postoperative behavioral recovery and deficits at 4-8 wk, perhaps reflecting the development of fine (distal) motor control. These and other data show that PEG-fusion not only rapidly restores physiological and morphological axonal continuity, but also more quickly improves behavioral recovery.

Melatonin enhances the in vitro and in vivo repair of severed rat sciatic axons.

This study examines the effects of several experimental compounds [melatonin (MEL), cyclosporin A (CsA), glial-derived neurotrophic factor (GDNF), and methylprednisolone (MP)] on polyethylene glycol (PEG)-induced repair in vitro and/or in vivo by plasmalemmal fusion (PEG-fusion) of sciatic axons severed by crushing. As measured by conduction of compound action potentials (CAPs) through the lesion site, a significantly (p<0.025) higher percentage (75%) of crushed rat sciatic axons can be repaired in vitro by PEG-fusion following exposure to MEL compared to PEG-fusion of severed sciatic axons in control Krebs saline that contains calcium (CTL=20%). In contrast, no other experimental compound (GDNF: 45%; MP: 42%; CsA: 24%) produces a significant improvement in PEG-fusion success compared to CTL. Further, MEL produces significantly (p<0.001) larger peak CAP amplitudes conducted through the lesion site following PEG-fusion compared to CTL or any other experimental compound in vitro. Additionally, MEL significantly (p<0.025) increases the ability to PEG-fuse sciatic axons in vivo, compared to CTL. Finally, PEG-fusion success in vivo is significantly (p<0.01) greater in calcium-free CTL (CTL-Ca) compared to CTL.

Degeneration of mammalian PNS and CNS axons is accelerated by incubation with protein synthesis inhibitors.

Protein synthesis inhibitors (PSIs) increase the rate of degeneration, as measured by compound action potential (CAP) conduction, in segments of rat PNS and CNS axons. Sciatic axonal segments maintained in vitro in Krebs at 37-38 degrees C generate CAPs for 24 h compared to 8 h for axons exposed to Krebs containing two PSIs, 100 microM anisomycin and/or 35 microM cycloheximide. Spinal axonal segments at 37-38 degrees C generate CAPs for 3 h compared to 2 h for axons exposed to Krebs containing PSIs. While cooling (6-9 degrees C) slows degeneration rate, cooled sciatic axons exposed to PSIs exhibit lower peak CAPs compared to cooled control segments (P<0.005).

Cooling enhances in vitro survival and fusion-repair of severed axons taken from the peripheral and central nervous systems of rats.

Severed segments of rat peripheral (PNS; sciatic) and central nervous system (CNS; spinal) axons continue to conduct action potentials when maintained in vitro at 6-9 degrees C for up to 7 (sciatic axons) and 2 days (spinal axons), compared with only 36 h at 37-38 degrees C for sciatic axons and 6 h for spinal axons. These PNS and CNS axonal segments can be crushed and then treated with polyethylene glycol (PEG), resulting in a rapid reconnection (fusion) of the surviving axons at the crush site, as assessed by conduction of action potentials through the crush site within minutes after PEG administration. Severed PNS or CNS axons maintained in vitro at 6-9 degrees C prior to crushing can be successfully PEG-fused for up to 4 and 1.5 days, respectively, compared with only 24 (sciatic) and 3 h (spinal) at 37-38 degrees C. These data demonstrate that cooling significantly increases both the survival time of severed mammalian PNS and CNS axons and the time that severed axons can still be PEG-fused (rejoined) to rapidly re-establish axonal continuity in vitro.