Dual effect of adenovirus‐mediated transfer of BMP7 in mixed neuron‐glial cultures: Neuroprotection and cellular differentiation

Abstract

Bone morphogenetic proteins (BMPs), members of the TGF-beta superfamily, have been implicated in nervous system development and in response to injury. Previous studies have shown that recombinant BMP7 can enhance dendritic growth and protect cultured neurons from oxidative stress. Because of the presence of extracellular BMP antagonists, BMP7 seems to act locally. Therefore, the present study uses BMP7 overexpression using adenovirus (Ad)-mediated gene transfer to examine its effect in mixed neuronal cultures. Enhanced BMP7 expression selectively induces neuronal CGRP expression in a time-dependent manner. BMP7 overexpression not only significantly protects cultures from H2O2 toxicity but reduces lipopolysaccharide (LPS) stimulation. Concurrently, it profoundly reduces microglial numbers, but increases oligodendroglial and endothelial cells. Together, low-dose and continuously expressed BMP7 is both neuroprotective and differentiation-inductive.

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

“…12,34,35 Briefly, full-length cDNA of human BMP7 or GFP, as a vector control, was cloned into an Ad shuttle vector. Recombinant adenovirus carrying BMP7 (AdBMP7) or GFP (AdGFP), which contained a human phosphoglycerate kinase promoter and a polyadenylation signal of bovine growth hormone, was constructed by homologous recombination in human embryonic kidney (HEK 293) cells.…”

“…The nucleotide sequences of the primers were based on published cDNA sequences of human BMP7 (forward, BMP7_2F: 5 0 -CATGCTGGA CCTGTACAACGC-3 0 ; reverse, BMP7_2R: 5 0 -CCTCACAG TAGTAGGCGGCG-3 0 ). 12 RT-PCR for BMP7 was performed by reverse transcription for 60 min at 50 1C, heat denaturation at 94 1C for 5 min and 38 cycles of denaturation at 95 1C (15 s), annealing at 60 1C (20 s) and elongation at 68 1C (1 min). RT-PCR for actin, an internal standard, was conducted by reverse transcription for 1 h at 50 1C, heat denaturation at 94 1C for 5 min and 25 cycles of denaturation at 94 1C (15 s), annealing at 60 1C (20 s) and elongation at 68 1C (30 s).…”

“…8 In cultures, BMP7 has been shown to promote the outgrowth of both dendritic and axonal processes from both peripheral and central neurons. [9][10][11][12][13] A previous study from this laboratory has shown dual neuroprotecting/differentiating effects of adenoviral (Ad) BMP7 gene transfer in cortical cultures. 12 The role of BMP7 in peripheral nerve regeneration after injury in vivo is not yet understood.…”

“…[9][10][11][12][13] A previous study from this laboratory has shown dual neuroprotecting/differentiating effects of adenoviral (Ad) BMP7 gene transfer in cortical cultures. 12 The role of BMP7 in peripheral nerve regeneration after injury in vivo is not yet understood. The fact that continuously released BMP7 by gene transfer is beneficial in peripheral nerve regeneration, and can correspond to improved functional recovery, has not been explored.…”

Cannabinoids – Can What Hurts You Make You Stronger?: Commentary on the article by Alvarez et al. on page 653

“…This receptor is present in the brain and when CBD binds to it, evokes the release of calcitonin gene related product (CGRP) in dorsal root ganglia (17). CGRP may protect the brain through its modulation of growth hormones like erythropoietin or BMP7 (18,19). However, CBD has been reported to reduce striatal atrophy caused by 3-nitroproprionic acid but the protective effects were not reversed by CB1, TRPV1 or A2A receptor antagonism (20).…”