title: High performance collection of cerebrospinal fluid in rats: evaluation of erythropoietin penetration after osmotic opening of the blood-brain barrier.
abstract: An important issue to be considered when studying a new drug for treatment of central nervous system (CNS) diseases is its ability to cross the blood-brain barrier (BBB) and distribute throughout the brain. As cerebrospinal fluid (CSF) has demonstrated to be an invaluable reservoir to study CNS availability of therapeutic proteins, we have developed an improved method for CSF sampling from the cisterna magna of rats. The technique enables the simple and rapid collection of adequate quantities (50-75 μl) of blood-free CSF, rendering a high percentage of animal survival (99%) without clinic or neurological consequences. Its success in avoiding blood contamination of CSF lays in the use of a mixture of lidocaine/ephinephrine topically injected in the rat's suboccipital area and neck. Another relevant feature of the methodology is its low cost, since the puncture device can be easily assembled with cheap and available materials and, more importantly, neither expensive stereotaxic equipment nor frame is required. The present method is demonstrated by studying the CSF pharmacokinetics of recombinant human erythropoietin (rhEPO), a well-studied therapeutic candidate for neurological diseases. Moreover, we applied this technique to evaluate a strategy of osmotic disruption of the BBB to achieve a faster delivery of rhEPO into the CNS.
authors.affiliation: Cell Culture Laboratory, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, Ciudad Universitaria, Paraje "El Pozo", C.C. 242, S3000ZAA Santa Fe, Argentina. nceaglio@fbcb.unl.edu.ar
authors.forename: Natalia
authors.initials: N
authors.lastname: Ceaglio
authors.forename: Gustavo
authors.initials: G
authors.lastname: Orozco
authors.forename: Marina
authors.initials: M
authors.lastname: Etcheverrigaray
authors.forename: Mónica
authors.initials: M
authors.lastname: Mattio
authors.forename: Ricardo
authors.initials: R
authors.lastname: Kratje
authors.forename: Norma
authors.initials: N
authors.lastname: Perotti
authors.forename: Marcos
authors.initials: M
authors.lastname: Oggero
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000502:physiology*; D002452:Cell Count; D002555:Cerebrospinal Fluid / Q000737:chemistry* / Q000166:cytology; D002946:Cisterna Magna / Q000000981:diagnostic imaging / Q000502:physiology; D003627:Data Interpretation, Statistical; D000068817:Epoetin Alfa; D004921:Erythropoietin / Q000134:cerebrospinal fluid* / Q000493:pharmacokinetics; D005260:Female; D005916:Globulins / Q000134:cerebrospinal fluid; D007275:Injections, Intravenous; D008353:Mannitol / Q000494:pharmacology; D009333:Neck / Q000000981:diagnostic imaging; D009995:Osmosis; D011859:Radiography; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000134:cerebrospinal fluid / Q000493:pharmacokinetics; D012886:Skull / Q000000981:diagnostic imaging; D013048:Specimen Handling / Q000379:methods*
chemical_list: D005916:Globulins; D011994:Recombinant Proteins; D004921:Erythropoietin; D008353:Mannitol; D000068817:Epoetin Alfa
keywords: Blood–brain barrier; Cerebrospinal fluid; Osmotic disruption; Sampling method; rhEPO