RT info:eu-repo/semantics/article
T1 The Acidic Probe LysoSensor™ is not Useful for Acrosome Evaluation of Cryopreserved Ram Spermatozoa
A1 Castro González, David
A1 Álvarez García, Mercedes
A1 Muro, J.
A1 Esteso, Milagros
A1 Paz Cabello, Paulino de
A1 Anel Rodríguez, Luis
A1 Martínez Pastor, Felipe
A2 Biologia Celular
K1 Veterinaria
K1 Criopreservación
K1 Esperma
K1 Ganado ovino
AB To try new acrosomal probes for assessing ram spermatozoa, we compared the LysoSensor™ probe, which labels acidic organelles, with the frequently used peanut agglutinin acrosomal probe (PNA‐PE; phycoerythrin as fluorescent moiety). The previous microscopic observations showed a lack of relationship of LysoSensor™ with acrosomal status. Semen was obtained from five rams and frozen in four pools. Each pool was analysed carrying out a triple staining propidium ioide/PNA‐PE/LysoSensor™ Green DND‐189 to test acrosome labelling, and a double staining SYBR‐14/PI, to assess sperm viability. Stained samples were analysed by flow cytometry. All measurements were replicated. Data were processed using agreement and repeatability tests. LysoSensor™ labelling did not agree with PNA (mean of differences: 30.8%; coefficient of agreement: 22.6%), confirming microscopic observations. Nevertheless, when LysoSensor™ was compared with SYBR‐14/PI, the agreement was high (mean of differences: −0.05%; coefficient of agreement: 5.07%). Repeatability of both methods was high and similar. LysoSensor™ did not seem to specifically stain the acrosome, but it may accumulate in the cytoplasm and label viable spermatozoa. Therefore, LysoSensor™ might not be used as an acrosomal probe in ram spermatozoa, but it could be used in other kind of studies, taking advantage of its pH sensitivity.
PB John Wiley & Sons
YR 2019
FD 2019-05-13
LK http://hdl.handle.net/10612/10749
UL http://hdl.handle.net/10612/10749
NO Reproduction in Domestic Animals, 2010, vol. 45, n. 2
NO P. 363-367
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 19-may-2024