title: Transient transfection of human CDNF gene reduces the 6-hydroxydopamine-induced neuroinflammation in the rat substantia nigra.
abstract: BACKGROUND
  The anti-inflammatory effect of the cerebral dopamine neurotrophic factor (CDNF) was shown recently in primary glial cell cultures, yet such effect remains unknown both in vivo and in 6-hydroxydopamine (6-OHDA) models of Parkinson's disease (PD). We addressed this issue by performing an intranigral transfection of the human CDNF (hCDNF) gene in the critical period of inflammation after a single intrastriatal 6-OHDA injection in the rat.
  
  
  METHODS
  At day 15 after lesion, the plasmids p3xNBRE-hCDNF or p3xNBRE-EGFP, coding for enhanced green florescent protein (EGFP), were transfected into the rat substantia nigra (SN) using neurotensin (NTS)-polyplex. At day 15 post-transfection, we measured nitrite and lipoperoxide levels in the SN. We used ELISA to quantify the levels of TNF-α, IL-1β, IL-6, endogenous rat CDNF (rCDNF) and hCDNF. We also used qRT-PCR to measure rCDNF and hCDNF transcripts, and immunofluorescence assays to evaluate iNOS, CDNF and glial cells (microglia, astrocytes and Neuron/Glial type 2 (NG2) cells). Intact SNs were additional controls.
  
  
  RESULTS
  In the SN, 6-OHDA triggered nitrosative stress, increased inflammatory cytokines levels, and activated the multipotent progenitor NG2 cells, which convert into astrocytes to produce rCDNF. In comparison with the hemiparkinsonian rats that were transfected with the EGFP gene or without transfection, 6-OHDA treatment and p3xNBRE-hCDNF transfection increased the conversion of NG2 cells into astrocytes resulting in 4-fold increase in the rCDNF protein levels. The overexpressed CDNF reduced nitrosative stress, glial markers and IL-6 levels in the SN, but not TNF-α and IL-1β levels.
  
  
  CONCLUSIONS
  Our results show the anti-inflammatory effect of CDNF in a 6-OHDA rat of Parkinson's disease. Our results also suggest the possible participation of TNF-α, IL-1β and IL-6 in rCDNF production by astrocytes, supporting their anti-inflammatory role.
authors.affiliation: Programa de Doctorado en Nanociencias y Nanotecnología; CINVESTAV, Av. Instituto Politécnico Nacional # 2508, San Pedro Zacatenco, CP 07360, México, DF, México. rasagna.s@gmail.com.|Departamento de Fisiología, Biofísica y Neurociencias; CINVESTAV, Av. Instituto Politécnico Nacional # 2508, San Pedro Zacatenco, CP 07360, México, DF, México. rasagna.s@gmail.com.
authors.forename: Rasajna
authors.initials: R
authors.lastname: Nadella
authors.affiliation: Institute of Biotechnology, PO Box 56, Viikki Biocenter, University of Helsinki, FI-00014, Helsinki, Finland. merja.h.voutilainen@helsinki.fi.
authors.forename: Merja H
authors.initials: MH
authors.lastname: Voutilainen
authors.affiliation: Institute of Biotechnology, PO Box 56, Viikki Biocenter, University of Helsinki, FI-00014, Helsinki, Finland. mart.saarma@helsinki.fi.
authors.forename: Mart
authors.initials: M
authors.lastname: Saarma
authors.affiliation: Laboratorio de Medicina Genómica, Hospital Regional '1° de Octubre', ISSSTE, Av. Instituto Politécnico Nacional # 1667, Magdalena de las Salinas, CP 02800, México, DF, México. jantgonzalez69@gmail.com.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Gonzalez-Barrios
authors.affiliation: Facultad de Ciencias Químicas, Benemérita Universidad Autónoma de Puebla, Avenida San Claudio S/N, Ciudad Universitaria Edif. 105A, CP 72570, Puebla, PUE, México. alileonch@gmail.com.
authors.forename: Bertha A
authors.initials: BA
authors.lastname: Leon-Chavez
authors.affiliation: Laboratorio de Neurofisiología, Departamento de Fisiología, Centro Universitario de Ciencias de la Salud, Universidad de Guadalajara, Av. Juárez 976, Colonia Centro, CP 44100, Guadalajara, Jalisco, México. mduenas@cucs.udg.mx.
authors.forename: Judith M Dueñas
authors.initials: JM
authors.lastname: Jiménez
authors.affiliation: Departamento de Neurociencias, Centro Universitario de Ciencias de la Salud, Universidad de Guadalajara, Av. Juárez 976, Colonia Centro, CP 44100, Guadalajara, Jalisco, México. sduenas@cucs.udg.mx.
authors.forename: Sergio H Dueñas
authors.initials: SH
authors.lastname: Jiménez
authors.affiliation: Departamento de Fisiología, Biofísica y Neurociencias; CINVESTAV, Av. Instituto Politécnico Nacional # 2508, San Pedro Zacatenco, CP 07360, México, DF, México. escobedo@fisio.cinvestav.mx.
authors.forename: Lourdes
authors.initials: L
authors.lastname: Escobedo
authors.affiliation: Programa de Doctorado en Nanociencias y Nanotecnología; CINVESTAV, Av. Instituto Politécnico Nacional # 2508, San Pedro Zacatenco, CP 07360, México, DF, México. martinez.fong@gmail.com.|Departamento de Fisiología, Biofísica y Neurociencias; CINVESTAV, Av. Instituto Politécnico Nacional # 2508, San Pedro Zacatenco, CP 07360, México, DF, México. martinez.fong@gmail.com.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martinez-Fong
mesh_terms: D000818:Animals; D006801:Humans; D007249:Inflammation / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism; D007276:Injections, Intraventricular; D008297:Male; D009414:Nerve Growth Factors / Q000096:biosynthesis* / Q000235:genetics*; D016627:Oxidopamine / Q000008:administration & dosage / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar; D013378:Substantia Nigra / Q000187:drug effects* / Q000378:metabolism*; D014162:Transfection
chemical_list: C522217:CDNF protein, human; D009414:Nerve Growth Factors; D016627:Oxidopamine