2024-03-29T08:36:12Zhttp://buleria.unileon.es/oai/requestoai:buleria.unileon.es:10612/107492020-12-10T08:56:05Zcom_10612_17col_10612_18
The Acidic Probe LysoSensor™ is not Useful for Acrosome Evaluation of Cryopreserved Ram Spermatozoa
Castro González, David
Álvarez García, Mercedes
Muro, J.
Esteso, Milagros
Paz Cabello, Paulino de
Anel Rodríguez, Luis
Martínez Pastor, Felipe
Biologia Celular
Facultad de Ciencias Biologicas y Ambientales
Veterinaria
Criopreservación
Esperma
Ganado ovino
P. 363-367
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.
SI
2010-04
2019-05-13T17:38:09Z
2019-05-13T17:38:09Z
2019-05-13
info:eu-repo/semantics/article
Reproduction in Domestic Animals, 2010, vol. 45, n. 2
https://onlinelibrary.wiley.com/doi/full/10.1111/j.1439-0531.2008.01333.x
http://hdl.handle.net/10612/10749
eng
John Wiley & Sons
https://buleria.unileon.es/bitstream/10612/10749/4/42%20The%20Acidic%20Probe%20LysoSensorTM%20%20ReprodDomestAnim_045_0363_2010%20El%20que%20facilit%c3%b3n%20PDF.pdf.jpg
Hispana
TEXT
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BULERIA. Repositorio Institucional de la Universidad de León
http://hdl.handle.net/10612/10749