TY - JOUR
T1 - Sex Genotyping of Archival Fixed and Immunolabeled Guinea Pig Cochleas
AU - Depreux, Frédéric F.
AU - Czech, Lyubov
AU - Whitlon, Donna S.
N1 - Funding Information:
We thank Hunter Young and Claus Peter Richter for fresh and fixed guinea pig tissue, Jaime Garcia-Anoveros for comments on the manuscript and Mary Grover for researching Guinea pig sex reporting. Supported by Grant # ONR N00014-16-1-2508-P00004 and the Department of Otolaryngology, Feinberg School of Medicine, Northwestern University. This work was made possible in part by the services of the NUSeq Core Facility, which is supported by the Northwestern University Center for Genetic Medicine, Feinberg School of Medicine, and Shared and Core Facilities of the University’s Office for Research.
Publisher Copyright:
© 2018 The Author(s).
PY - 2018/12/1
Y1 - 2018/12/1
N2 - For decades, outbred Guinea pigs (GP) have been used as research models. Various past research studies using Guinea pigs used measures that, unknown at the time, may be sex-dependent, but from which today, archival tissues may be all that remain. We aimed to provide a protocol for sex-typing archival Guinea pig tissue, whereby past experiments could be re-evaluated for sex effects. No PCR sex-genotyping protocols existed for GP. We found that published sequence of the GP Sry gene differed from that in two separate GP stocks. We used sequences from other species to deduce PCR primers for Sry. After developing a genomic DNA extraction for archival, fixed, decalcified, immunolabeled, Guinea pig cochlear half-turns, we used a multiplex assay (Y-specific Sry; X-specific Dystrophin) to assign sex to tissue as old as 3 years. This procedure should allow reevaluation of prior Guinea pig studies in various research areas for the effects of sex on experimental outcomes.
AB - For decades, outbred Guinea pigs (GP) have been used as research models. Various past research studies using Guinea pigs used measures that, unknown at the time, may be sex-dependent, but from which today, archival tissues may be all that remain. We aimed to provide a protocol for sex-typing archival Guinea pig tissue, whereby past experiments could be re-evaluated for sex effects. No PCR sex-genotyping protocols existed for GP. We found that published sequence of the GP Sry gene differed from that in two separate GP stocks. We used sequences from other species to deduce PCR primers for Sry. After developing a genomic DNA extraction for archival, fixed, decalcified, immunolabeled, Guinea pig cochlear half-turns, we used a multiplex assay (Y-specific Sry; X-specific Dystrophin) to assign sex to tissue as old as 3 years. This procedure should allow reevaluation of prior Guinea pig studies in various research areas for the effects of sex on experimental outcomes.
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U2 - 10.1038/s41598-018-23491-3
DO - 10.1038/s41598-018-23491-3
M3 - Article
C2 - 29581456
AN - SCOPUS:85044508603
SN - 2045-2322
VL - 8
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 5156
ER -