ANÁLISIS COMPARATIVO DE PROTOCOLOS DE CRIOPRESERVACIÓN DE SEMEN EN XENOPUS TROPICALIS
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2019-07-02
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Jaén: Universidad de Jaén
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[ES]Actualmente, la disminución y extinción de anfibios es una problemática que busca resolverse
por medio de programas de conservación que utilizan herramientas como la investigación,
desarrollo e implementación de biotecnologías de reproducción, las cuales incluyen la
criopreservación de semen, para mitigar el daño y prevenir futuras pérdidas en la biodiversidad
de especies. En este trabajo se plantea comparar un método de criopreservación utilizado en la
especie Xenopus laevis (X. laevis) en el que se emplea solución salina inhibidora de la motilidad
(MIS por Motility Inhibiting Saline solution) + dimetilsulfóxido (DMSO) + sacarosa como
criopreservante, con el método tradicionalmente usado en Xenopus tropicalis (X. tropicalis) en el
que se emplea una solución hecha a partir de sacarosa y yema de huevo. Se propone analizar
de forma comparada estos protocolos y evaluar la viabilidad de los espermatozoides tras la
criopreservación. Se realizó un ensayo donde se utilizaron machos de X. tropicalis, de los cuales
se obtuvieron los testículos para recolectar el semen, que fue sometido a los dos métodos de
criopreservación planteados y se realizó un análisis de fertilidad de cada muestra, que consistió
en un recuento espermático, una medición de la viabilidad espermática con solución de diacetato
de 6-carboxifluoresceína (CFDA) y yoduro de propidio (IP) y un índice de fertilidad mediante
inseminación de huevos. Se obtuvieron resultados que sugieren que el protocolo de MIS + DMSO
+ sacarosa tiene un índice de fertilidad mayor al protocolo convencional con yema de huevo, sin
embargo, se requiere realizar más investigaciones que sostengan esta afirmación.2. Palabras clave
Criopreservación, criopreservante, esperma, espermatozoides, fecundación, fertilidad, semen,
Xenopus tropicalis
[EN]Currently, the decline and extinction of amphibians is a problem that seems to be solved through conservation programs that use tools such as research, development, and implementation of reproductive biotechnologies, which include the cryopreservation of semen, to mitigate the damage and prevent future losses in the biodiversity of species. In this work we propose to compare a cryopreservation method used in the species Xenopus laevis (X. laevis) in which motility inhibiting saline solution (MIS) + dimethylsulfoxide (DMSO) + sucrose is used as cryopreserved, with the method traditionally used in Xenopus tropicalis (X. tropicalis) in which a solution made from sucrose and egg yolk is used. It is proposed to analyze these protocols in a comparative way and evaluate the viability of spermatozoa after cryopreservation. An experiment was carried out where males of X. tropicalis were used, from which the testicles were obtained to collect the semen, which was subjected to the two methods of cryopreservation proposed and a fertility analysis of each sample was carried out, which consisted of a spermatozoa count, a measurement of sperm viability with 6-carboxyfluorescein diacetate solution (CFDA) and propidium iodide (PI) and a fertility index by insemination of eggs. Results were obtained that suggest that the protocol of MIS + DMSO + sucrose has a fertility index higher than the conventional protocol with egg yolk, however, more research is required to support this assertion. 4. Keywords Cryopreservation, cryopreserved, sperm, spermatozoa, fertilization, fertility, semen, Xenopus tropicalis.
[EN]Currently, the decline and extinction of amphibians is a problem that seems to be solved through conservation programs that use tools such as research, development, and implementation of reproductive biotechnologies, which include the cryopreservation of semen, to mitigate the damage and prevent future losses in the biodiversity of species. In this work we propose to compare a cryopreservation method used in the species Xenopus laevis (X. laevis) in which motility inhibiting saline solution (MIS) + dimethylsulfoxide (DMSO) + sucrose is used as cryopreserved, with the method traditionally used in Xenopus tropicalis (X. tropicalis) in which a solution made from sucrose and egg yolk is used. It is proposed to analyze these protocols in a comparative way and evaluate the viability of spermatozoa after cryopreservation. An experiment was carried out where males of X. tropicalis were used, from which the testicles were obtained to collect the semen, which was subjected to the two methods of cryopreservation proposed and a fertility analysis of each sample was carried out, which consisted of a spermatozoa count, a measurement of sperm viability with 6-carboxyfluorescein diacetate solution (CFDA) and propidium iodide (PI) and a fertility index by insemination of eggs. Results were obtained that suggest that the protocol of MIS + DMSO + sucrose has a fertility index higher than the conventional protocol with egg yolk, however, more research is required to support this assertion. 4. Keywords Cryopreservation, cryopreserved, sperm, spermatozoa, fertilization, fertility, semen, Xenopus tropicalis.
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Biología