Adenoviral gene transfer of bone morphogenetic protein-7 enhances functional recovery after sciatic nerve injury in rats

Abstract

Bone morphogenetic proteins (BMPs), members of the transforming growth factor-b subfamily, function as instructive signals for neuronal lineage commitment and promote neuronal differentiation. However, the mechanism of BMP7 action in vivo after peripheral nerve injury is poorly understood. This study examines the efficacy of gene transfer of adenoviral (Ad) BMP7 on peripheral neuropathy. Transgene expression was found in both Ad-infected sciatic nerves and their respective remote neurons, indicating Ad transduction by a retrograde transport. After AdBMP7 infection to nerves, the sciatic nerves were crushed or transected. Hind limb functional behavior, including rotarod test and sciatic functional index, were conducted in rats weekly after nerve injury. Interestingly, enhanced BMP7 expression significantly improved hind limb functional recovery in AdBMP7-transduced rats when compared with AdGFPtransduced nerve-crushed or transected rats. Furthermore, AdBMP7 transduction reduced injury-induced macrophage activation, nerve demyelination and axonal degeneration. By contrast, AdBMP7 infection did not affect the hyperalgesia paw-withdrawal latency after nerve injury. We further examined the effect of AdBMP7 infection on sciatic nerve explant and Schwann cell cultures. Enhanced cell proliferation was significantly increased by AdBMP7 transduction in both cultures. Taken together, BMP7 overexpression by Ad gene transfer was beneficial in both nerves and Schwann cells on functional recovery after sciatic nerve injury in rats.

Enhanced Prostacyclin Synthesis by Adenoviral Gene Transfer Reduced Glial Activation and Ameliorated Dopaminergic Dysfunction in Hemiparkinsonian Rats

“…Recombinant adenoviral vectors target gene expression to the nervous system and offer prolonged expression of foreign proteins [23, 33, 36]. Here, we present evidence that glial cultures, mesencephalic neuron/glial cultures, or nigral dopaminergic neurons (SNpc) could effectively be infected by recombinant adenoviruses and thereby expressing transgenes.…”

“…Protein concentration of the resultant lysate was determined using Bio-Rad protein assay (Bio-Rad, Hercules, CA). Equal amounts of proteins were loaded and separated using 8%–12% gels (SDS-PAGE) as described [33]. After electrophoresis, proteins in the gels were transferred to PVDF membranes (Millipore Corp., USA) and incubated overnight at 4°C with antibodies against PGIS (rabbit, dilution 1/3000), COX-2 (rabbit, dilution 1/5000, Cayman), inducible nitric oxide synthetase (iNOS, mouse, dilution 1/5000, BD Bioscience, USA), or β -actin (goat, dilution 1/5000, Santa Cruz Biotech, USA) followed by a horseradish peroxidase-conjugated secondary antibody (dilution 1/2000, Jackson Lab) for 1 hr at room temperature.…”

Transforming Growth Factor-β in Normal Nociceptive Processing and Pathological Pain Models

“…By contrast, intrathecal delivery of BMP4 cDNA incorporated into an adenovirus vector does not modify somatosensory perceptions in mice subjected to spinal cord injury, while it promotes sensory axon regeneration [40]. Similarly, BMP7 overexpression in the sciatic nerve, DRG and spinal cord neurons by adenoviral gene transfer improves the functional recovery after injury but does not affect the development of neuropathic pain in rats [41].…”

Acid Fibroblast Growth Factor and Peripheral Nerve Grafts Regulate Th2 Cytokine Expression, Macrophage Activation, Polyamine Synthesis, and Neurotrophin Expression in Transected Rat Spinal Cords

“…Therefore, recombinant adenovirus carrying green fluorescence protein (Ad-GFP) was used to trace neurons, as described previously (Tsai et al, 2010a). Two weeks after repair surgery, each rat was anesthetized and received a laminectomy below the injury site.…”