Standardization of a Sex-Sorting Protocol for Stallion Spermatozoa by Means of Absolute RT-qPCR
| Primer Autor |
Felmer, Ricardo
|
| Co-autores |
Munoz, Erwin
Castro, Macarena
Aguila, Luis
Contreras, Maria Jose
Fuentes, Fernanda
Arias, Maria Elena
|
| Título |
Standardization of a Sex-Sorting Protocol for Stallion Spermatozoa by Means of Absolute RT-qPCR
|
| Editorial |
MDPI
|
| Revista |
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
|
| Lenguaje |
en
|
| Resumen |
Sperm sexing is a technology that can generate great economic benefits in the animal production sector. Techniques such as sex-sorting promise over 90% accuracy in sperm sexing. However, for the correct standardization of the technique, some laboratory methodologies are required. The present manuscript describes in detail a standardized equine sperm sex-sorting protocol using an absolute qPCR-based methodology. Furthermore, the results of absolute qPCR were implemented and validated by generating equine/bovine heterologous embryos by intracytoplasmic sperm injection (ICSI) of presumably sexed equine spermatozoa into bovine oocytes using a piezoelectric system (Piezo-ICSI). Our results indicated that equine sex-sorting spermatozoa had a 97% and 94% certainty for X and Y sperm, respectively, while presumptive female and male equine/bovine hybrid embryos, generated by Piezo-ICSI, had an accuracy of 92% with respect to the desired sex. Therefore, it is concluded that the presented methodology is a reliable, cost-effective, and relatively simple option for standardizing sex-sorting of equine spermatozoa. This is supported by the results of the correct sexing of Piezo-ICSI heterologous embryos generated with the sexed spermatozoa, validating the correct sexing and viability of these gametes.
|
| Fecha Publicación |
2023
|
| Tipo de Recurso |
artículo original
|
| doi |
10.3390/ijms241511947
|
| Formato Recurso |
PDF
|
| Palabras Claves |
absolute RT-qPCR
PIEZO-ICSI
plasmids
sex-sorting
sperm stallion
|
| Ubicación del archivo | |
| Categoría OCDE |
Bioquímica y biología molecular
Química
|
| Materias |
RT-qPCR absoluta
PIEZO-ICSI
plásmidos
clasificación por sexo
semental de esperma
|
| Identificador del recurso (Mandatado-único) |
artículo original
|
| Versión del recurso (Recomendado-único) |
versión publicada
|
| License |
CC BY 4.0
|
| Condición de la licencia (Recomendado-repetible) |
CC BY 4.0
|
| Derechos de acceso |
acceso abierto
|
| Access Rights |
acceso abierto
|
| Id de Web of Science |
WOS:001046253900001
|
| ISSN |
1661-6596
|
| Tipo de ruta |
verde# dorado
|
| Categoría WOS |
Bioquímica y biología molecular
Química
|
| Referencia del Financiador (Mandatado si es aplicable-repetible) |
ANID-FONDECYT 1201166
ANID 21191434
UFRO PDT21-0001
ANID FONDECYT 1201166
|
cienciaabierta.ufro.cl
ciencia.abierta@ufrontera.cl