T. Muiño‐Blanco

Universidad de Zaragoza

Author Statistics

Papers

Citation

H-Index

i-10 index

Research Trends

Author Order

Document Type

Co-Authors

J.A. Cebrián-Pérez

Universidad de Zaragoza

126

Rosaura Pérez‐Pé

Universidad de Zaragoza

Adriana Casao

Universidad de Zaragoza

J.A. Abecia

Universidad de Zaragoza

F. Forcada

Universidad de Zaragoza

Carmen Colás

Universidad de Zaragoza

Noelia Mendoza

Universidad de Zaragoza

JÁ Cebrián‐Pérez

Universidad de Zaragoza

Mario Ollero

Inserm

Patricia Grasa

University of Oxford

Cooperative Institutions

Universidad de Zaragoza

Universitat Autònoma de Barcelona

Hospital Universitario Miguel Servet

Consejo Superior de Investigaciones Científicas

Instituto Aragonés de Ciencias de la Salud

Universidade de São Paulo

Universitat de Barcelona

Universidad de Murcia

Instituto de Salud Carlos III

Instituto de Investigación Sanitaria Aragón

Author Statistics

Papers

Citation

H-Index

i-10 index

Research Field

Caffeine induces ram sperm hyperactivation independent of cAMP‐dependent protein kinase

Andrology (2009)

Carmen Colás J.A. Cebrián-Pérez T. Muiño‐Blanco

Summary We have previously shown that a cocktail‐containing phosphodiesterase inhibitors (theophylline and caffeine), a phosphatase inhibitor (okadaic acid) and dibutyryl‐cAMP promoted specific protein tyrosine phosphorylation in ram spermatozoa during incubation in capacitating conditions. Here, we show, for the first time, that this cocktail induced a progressive time‐dependent increase in the capacitated‐sperm subpopulation . The addition of either the analogue of adenosine, 2‐chloro‐2′‐deoxyadenosine (Cl‐Ado) or caffeine provided a significant increase in the proportion of capacitated spermatozoa and total tyrosine phosphorylation. Computer‐assisted semen analysis was used to identify hyperactivated spermatozoa by setting maximum threshold for linearity (≤45%) and minimum for amplitude of lateral head displacement (≥3.5 μm). Our results showed that ram spermatozoa can be capacitated in vitro without displaying hyperactivated movement. Among the above‐mentioned compounds, only caffeine was able to induce hyperactivation that achieved the maximal response at 8 min of incubation, with a significant increase in hyperactivated spermatozoa of 44.4 ± 5.6% related to control samples. Flow cytometry analyses showed that caffeine induced a significant increase in the content of calcium in viable spermatozoa during the time‐course of incubation in capacitating conditions. BAPTA‐AM, a cell‐permeable calcium chelator, did not suppress the caffeine‐dependent hyperactivation. Quantitative analysis revealed that the addition of caffeine or Cl‐Ado accounted for an increase in intracellular cAMP level. However, this increase in cAMP does not seem to be responsible for the caffeine‐induced hyperactivation because the cAMP‐elevating agents (cocktail) did not promote hyperactivation either, although they greatly induced capacitation and protein tyrosine phosphorylation. The inhibition of PKA with H89 reduced both capacitation and protein tyrosine phosphorylation although hyperactivation increased. These results suggest that calcium from internal stores would be enough to initiate the hyperactivated movement, and that protein tyrosine phosphorylation implicated in ram sperm hyperactivation would be regulated by calcium rather than by PKA‐dependent cAMP.

Hyperactivation

10.1111/j.1365-2605.2009.00991.x

Cite

Citations (52)

Effects of 17-β estradiol and progesterone on ram sperm functionality

Animal Reproduction Science (2016)

Sergio Gimeno L. Del Molino Adriana Casao J.A. Cebrián-Pérez T. Muiño‐Blanco

10.1016/j.anireprosci.2016.03.043

Cite

Citations (1)

A New Method for the Treatment of Sperm Samples for Ultrastructural Study Based on the Use of Animal Tissues as Biological Containers

Microscopy Research and Technique (2007)

Concepción Junquera Carmen Colás Carmen Martínez‐Ciriano Pedro Serrano Tomás Castiella

Abstract The study of the ultrastructure of spematozoa by means of transmission electron microscopy often presents with problems of interpretation according to the method employed, depending on whether samples are either centrifuged previously to the fixation or immersed in viscous gels. The major problems of interpretation are: changes in the location of vesicles originated during the maturation process and modifications in the adsorption of seminal plasma proteins to the sperm membrane surface. The aim of our study is to communicate an original new method for the treatment of spermatozoa for ultrastructural study. Our method is based on the use of animal tissues as biological containers, inside which the spermatic suspensions are included. We developed this method using fresh sperm samples taken from mature Rasa aragonesa rams. As biological container, we used 2.5‐cm long segments of the intestine of 1‐week‐old chickens ( Gallus gallus ) (diameter around 4 mm). To avoid any influence of digestive enzymes of the mucosa on the sperm surface, we put each intestine fragment inside out by means of microdissection forceps under bifocal optical microscope and cold light. One of the edges was tied with thin suture silk. The sperm suspension was injected in the optimal experimental condition and amount. Finally, the still open edge of the intestine segment was tied with silk in the same way as the other segment edge. By using this technique, we can perform a suitable morphological study at an ultrastructural level. In addition, the functional relationship of the ultrastructural components of the target cells is correctly preserved. Microsc. Res. Tech., 2007. © 2007 Wiley‐Liss, Inc.

10.1002/jemt.20466

Cite

Citations (1)

Cloning of DNA sequences encoding RSVP14, RSVP20 and RSVP22 proteins from ovine seminal plasma.

XIV Jordanas Sobre Produccion Animal, Zaragoza, Espana, 17 y 18 de Mayo de 2011. (2011)

Ester Serrano R. Beamonte Nancy Guillén Lucı́a Calleja Rosaura Pérez‐Pé

Cloning (programming)

Source

Cite

Citations (0)

Utilización de Ribosa por Aspergillus Oryzae

Anales de la Estación Experimental de Aula Dei (1982)

T. Muiño‐Blanco J.A. Cebrián-Pérez M. Luisa Peleato Sánchez

Aspergillus oryzae

Source

Cite

Citations (0)

The effects of low concentrations of melatonin on in vitro maturation and fertilization of oocytes and on early embryo development in sheep.

Adriana Casao M. Ait Amer Meziane J.A. Cebrián-Pérez T. Muiño‐Blanco J.A. Abecia

In vitro maturation

Source

Cite

Citations (0)

Survival rate and antioxidant enzyme activity of ram spermatozoa after dilution with different extenders or selection by a dextran swim-up procedure

Theriogenology (2003)

J. I. Martí Eulàlia Martı́J.A. Cebrián-Pérez T. Muiño‐Blanco

Glutathione reductase

10.1016/s0093-691x(03)00105-5

Cite

Citations (51)

Effects of Melatonin Implants During Non‐Breeding Season on Sperm Motility and Reproductive Parameters in Rasa Aragonesa Rams

Reproduction in Domestic Animals (2008)

Adriana Casao Sonia Vega I. Palacín Rosaura Pérez‐Pé Adolfo Laviña

Contents The effect of melatonin implants administered during non‐breeding season in Rasa Aragonesa rams on sperm motility parameters and other reproductive traits was assessed. In a first experiment, two Rasa Aragonesa rams were implanted (with melatonin group M), remaining other two males as control group (C). Semen of each group was collected from 1 May to 23 June, twice or three times a week, and motility parameters were assessed using a computer‐assisted sperm analysis system. Melatonin increased the percentage of progressive motile spermatozoa, particularly during 46–75 days after melatonin implantation (p < 0.01). In experiment 2, M and C in vitro fertilization ability had been determined by zona‐pellucida binding assays, using spermatozoa from experiment 1, obtained 60–70 days after melatonin was implanted. A significantly higher number of spermatozoa attached per oocyte was observed in frozen‐thawed immature ovine oocytes incubated with sperm from M animals than in those incubated with sperm from the C group (p < 0.01). Finally, a field assay (experiment 3) was performed. In this case, five Rasa Aragonesa rams were implanted with melatonin and three remained as control group. Sperm doses from those animals were used for artificial insemination of 2608 Rasa Aragonesa ewes from 39 different farms at non‐breeding season. Fertility, litter size and fecundity were studied. Semen from melatonin implanted rams seemed to increase both fertility and fecundity in ewes inseminated with spermatozoa obtained 46–60 days after implantation (p < 0.1). Thus, melatonin treatment in rams during non‐breeding season modifies sperm motility parameters and seems to improve the fertilization parameters obtained.

10.1111/j.1439-0531.2008.01215.x

Cite

Citations (99)

Short-term inhibition of the energy metabolism affects motility but not surface properties of sperm cells

Bioscience Reports (1996)

Ma Luisa Pascual J.A. Cebrián-Pérez M.J. López-Pérez T. Muiño‐Blanco

Centrifugal countercurrent distribution (CCCD) in aqueous two-phase systems has been proven to be a useful method to study subtle surface properties of spermatozoa. The present work shows that a short-term inhibition of the energy metabolism of sperm cells effected by incubating bovine sperm cells with KCN or ouabain, did not account for changes in the cell surface properties, as assessed either by estimation of the cell viability or by CCCD analysis. However, the short-term inhibition of energy metabolism provoked a clear decrease of cell motility, suggesting that a drop of cellular ATP levels brings about a rapid decrease of motility followed by a very delayed effect on cell surface properties. The relevance of these results on the handling of sperm and on the understanding of the molecular events underlying asthenospermia is discussed.

Sperm cell

10.1007/bf01200999

Cite

Citations (20)

Ultrastructural study of the ability of seminal plasma proteins to protect ram spermatozoa against cold‐shock

Microscopy Research and Technique (2009)

Carmen Colás Concepción Junquera Rosaura Pérez‐Pé J.A. Cebrián-Pérez T. Muiño‐Blanco

Abstract The process of sperm cryopreservation, involving cooling, freezing, and thawing, induces serious detrimental changes in sperm function. The plasma and acrosomal membranes of spermatozoa are considered to be the primary site of these modifications due to thermal, mechanical, chemical, and osmotic stress. In previous studies, we demonstrated the ability of seminal plasma (SP) proteins to protect ram spermatozoa against cold‐shock by using biochemical markers and scanning electron microscopy. In this study, we have attempted to examine the potential protective effect of SP proteins in membrane ultrastructure of ram spermatozoa subjected to cold‐shock, by means of transmission electron microscopy (TEM). All the experiments were carried out with fresh spermatozoa freed from SP by a dextran/swim‐up procedure. The high proportion of viable spermatozoa found in the swim‐up obtained sample decreased drastically after the cold‐shock treatment, and a considerable blebbing and vesiculation of the plasma and acrosomal membranes was found. The addition of SP proteins increased the sperm resistance to damage due to cold‐shock (48% membrane‐intact spermatozoa versus 15% in the control sample), and TEM analysis revealed that membrane alteration was prevented. This protective effect seems to be specific for SP proteins, as the addition of BSA did not provide any protection. Microsc. Res. Tech. 2009. © 2009 Wiley‐Liss, Inc.

Osmotic shock

Thermal shock

10.1002/jemt.20710

Cite

Citations (42)