A real-time quantitative PCR comparative study between rat optic and sciatic nerves: determination of neuregulin-1 mRNA levels.
Injured axons from peripheral nervous system (PNS) possess the ability to regenerate. In contrast, regeneration of injured axons does not occur in the central nervous system (CNS) or occurs to a limited extent. Previous works have shown that rat sciatic nerve conditioned medium (CM) produced PC12 cells neuronal-like differentiation and neurite outgrowth. In the present work, we compared the expression of neuregulin-1s (NRG-1s) from rat sciatic and optic nerves as members of the PNS and CNS, respectively. Sciatic nerve CM showed a higher neurotrophic activity on PC12 cells than rat optic nerve CM. RT-PCR analysis verified the presence of all three types of NRG-1 mRNAs and their receptors in both types of nerves. Real-time quantitative PCR (QPCR) assays showed that the relative expression levels of all three types of NRG-1 mRNAs were higher in optic nerves than in sciatic nerves. Eleven-day cultured optic nerves showed an increased in NDF and SMDF when compared to freshly isolated optic nerves, whereas GGF decreased. However, 11-day-cultured sciatic nerves only showed an increase in SMDF mRNA. Western blots corroborated the differences in NRG-1 expression profile for both types of nerves and their CMs. Incubation of both CMs with the anti-pan-NRG-1 antibody showed that the neurotrophic activity of the optic nerve CM increased, whereas the sciatic nerve CM remained unchanged. These results indicated that different NRG-1 levels are expressed upon nerve degeneration and the balance between those levels and other neurotrophic factors could have an important role on nerve regeneration.
Martínez
J C
JC
Instituto de Estudios Avanzados (IDEA), Apartado 17606, Caracas 1015-A, Miranda 1080, Venezuela. jmartien@reaccium.ve
Malavé
C
C
Bosch
I
I
Castillo
C
C
Núñez
J
J
Villegas
G M
GM
Villegas
R
R
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006023:Glycoproteins / Q000145:classification / Q000235:genetics / Q000378:metabolism; D020890:Neuregulin-1 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D009900:Optic Nerve / Q000378:metabolism*; D009924:Organ Culture Techniques; D016716:PC12 Cells; D020033:Protein Isoforms / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D018719:Erb-b2 Receptor Tyrosine Kinases; D049349:Receptors, Lysosphingolipid / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012584:Sciatic Nerve / Q000378:metabolism*
D000906:Antibodies; D017077:Culture Media, Conditioned; D006023:Glycoproteins; D020890:Neuregulin-1; D020033:Protein Isoforms; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; D049349:Receptors, Lysosphingolipid; C483804:Erbb2 protein, rat