Accepted Manuscript Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by antioxidant supplementation and choice of extender María Mata-Campuzano, Manuel Álvarez-Rodríguez, Mercedes Álvarez, Julio Tamayo-Canul, Luis Anel, Paulino de Paz, Felipe Martínez-Pastor PII:
S0093-691X(14)00570-6
DOI:
10.1016/j.theriogenology.2014.10.018
Reference:
THE 12969
To appear in:
Theriogenology
Received Date: 29 June 2014 Revised Date:
9 October 2014
Accepted Date: 11 October 2014
Please cite this article as: Mata-Campuzano M, Álvarez-Rodríguez M, Álvarez M, Tamayo-Canul J, Anel L, de Paz P, Martínez-Pastor F, Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by antioxidant supplementation and choice of extender, Theriogenology (2014), doi: 10.1016/j.theriogenology.2014.10.018. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT 1
revised Title: Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by
3
antioxidant supplementation and choice of extender
4
María Mata-Campuzano1,2, Manuel Álvarez-Rodríguez1, Mercedes Álvarez1,3 Julio Tamayo-Canul1,4,
5
Luis Anel1,3, Paulino de Paz1,2, Felipe Martínez-Pastor1,2,
6
1
7
of León, 24071 León, Spain. 3 Department of Medicine, Surgery and Veterinary Anatomy, University of
8
León, 24071 León, Spain. 4 DEPI, Tecnological Institute of Conkal, 97345, Conkal, Yucatán, México.
9
Corresponding author:
10
Felipe Martínez-Pastor
11
INDEGSAL
12
University of León,
13
24071, León, Spain
14
e-mail: [email protected]
RI PT
2
AC C
EP
TE D
M AN U
SC
INDEGSAL, University of León, 24071 León, Spain. 2 Department of Molecular Biology, University
1
ACCEPTED MANUSCRIPT Abstract
16
The performance of cryopreserved semen in ovine artificial insemination still needs improvement. Some
17
antioxidants have been tested, with variable success. We cryopreserved semen from Churra rams using
18
TES-Tris-fructose with 4% glycerol and 10% egg-yolk (EY) or 3.5% soybean lecithin (SL), with 1 mM
19
of reduced glutathione (GSH), Trolox, crocin or cysteamine. Samples were analyzed post-thawing and
20
post-incubation (6 h, 38 ◦C) for motility (CASA), viability, acrosomal integrity, apoptosis, mitochondrial
21
activity, chromatin status and lipoperoxidation (malondialdehyde —MDA— production). Interactions
22
(antioxidant/extender/incubation) were significant for most variables. Extenders yielded similar results,
23
although SL depressed mitochondrial activity and linearity (P
S0093-691X(14)00570-6
DOI:
10.1016/j.theriogenology.2014.10.018
Reference:
THE 12969
To appear in:
Theriogenology
Received Date: 29 June 2014 Revised Date:
9 October 2014
Accepted Date: 11 October 2014
Please cite this article as: Mata-Campuzano M, Álvarez-Rodríguez M, Álvarez M, Tamayo-Canul J, Anel L, de Paz P, Martínez-Pastor F, Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by antioxidant supplementation and choice of extender, Theriogenology (2014), doi: 10.1016/j.theriogenology.2014.10.018. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT 1
revised Title: Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by
3
antioxidant supplementation and choice of extender
4
María Mata-Campuzano1,2, Manuel Álvarez-Rodríguez1, Mercedes Álvarez1,3 Julio Tamayo-Canul1,4,
5
Luis Anel1,3, Paulino de Paz1,2, Felipe Martínez-Pastor1,2,
6
1
7
of León, 24071 León, Spain. 3 Department of Medicine, Surgery and Veterinary Anatomy, University of
8
León, 24071 León, Spain. 4 DEPI, Tecnological Institute of Conkal, 97345, Conkal, Yucatán, México.
9
Corresponding author:
10
Felipe Martínez-Pastor
11
INDEGSAL
12
University of León,
13
24071, León, Spain
14
e-mail: [email protected]
RI PT
2
AC C
EP
TE D
M AN U
SC
INDEGSAL, University of León, 24071 León, Spain. 2 Department of Molecular Biology, University
1
ACCEPTED MANUSCRIPT Abstract
16
The performance of cryopreserved semen in ovine artificial insemination still needs improvement. Some
17
antioxidants have been tested, with variable success. We cryopreserved semen from Churra rams using
18
TES-Tris-fructose with 4% glycerol and 10% egg-yolk (EY) or 3.5% soybean lecithin (SL), with 1 mM
19
of reduced glutathione (GSH), Trolox, crocin or cysteamine. Samples were analyzed post-thawing and
20
post-incubation (6 h, 38 ◦C) for motility (CASA), viability, acrosomal integrity, apoptosis, mitochondrial
21
activity, chromatin status and lipoperoxidation (malondialdehyde —MDA— production). Interactions
22
(antioxidant/extender/incubation) were significant for most variables. Extenders yielded similar results,
23
although SL depressed mitochondrial activity and linearity (P
