Laboratory Animals > Volume 42, Number 1 > Pp. 19-25

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Goicoechea, M Articles by de Munain, A L. Search for Related Content PubMed PubMed Citation Social Bookmarking                      What's this?

Papers

Minimizing creatine kinase variability in rats for neuromuscular research purposes

M Goicoechea ^* ¶  1, F Cía ^# 1, C San José ^*, A Asensio ^*, J I Emparanza , A G Gil ^#, A López de Cerain ^#, P Aldazabal ^*, M Azpitarte , D Otaegui ^* ¶ and A López de Munain  ^¶

^* Unidad Experimental; Unidad de Epidemiología; Servicio de Neurología; Servicio de Bioquímica, Laboratorio Unificado, Hospital Donostia, San Sebastián, Spain; ^¶ Ilundain Fundazioa, San Sebastián, Spain; ^# Nutrition and Food Science, Physiology and Toxicology, CIFA, Universidad de Navarra, Pamplona, Spain

Correspondence: M Goicoechea. Email: mgoicoechea{at}chdo.osakidetza.net

Rat serum or plasma creatine kinase (CK) activity is widely used to evaluate myopathic processes, to test the myotoxicity of different drugs, or to analyse the benefits of emerging gene therapies in some neuromuscular disorders. However, great variability is found in this determination. The aim of this study has been to control some factors of variation in order to reduce variability and increase the reproducibility of analytical data. 8–10-week-old Wistar-Han rats were used. The study consisted of four sequential phases. Phase I aimed to analyse the effect of ether and isoflurane as anaesthetic drugs. The objective of Phase II was to evaluate bleeding rats via retro-orbital sinus vs. tail vein. Phases III and IV were designed as two separate, repeated measure experiments on two factors: habituation to laboratory handling procedures in Phase III and gender in Phase IV. The repeated factor was the storage temperature of blood sample prior to centrifugation. Ether did not significantly increased the CK value. Using isoflurane, getting rats accustomed to laboratory handling procedures and whole blood refrigeration prior to centrifugation and serum separation resulted in statistically significant reduction in CK value and variability. Male rats showed significantly higher values than female rats. In the light of our findings, CK value and variability in rats may be minimized by choosing tail vein as site of bleeding, getting rats accustomed to laboratory handling procedures and maintaining whole blood refrigerated until centrifugation and serum separation.

Key Words: Creatine kinase • blood sampling • anaesthesia • variability reduction • refrigeration • serum

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?