Copulatory behaviour increases sperm viability in female spiders

One remarkable reproductive feature in animals with internal fertilization is a reduction in sperm viability over time in females. Whether this reduction is driven by male-male competition and/or cryptic female choice is unclear. From the perspective of cryptic female choice, we postulated that sperm viability is affected by a particular male copulatory behaviour. In this study, we investigated the following aspects: (1) sperm viability in mated females vs. males; (2) whether sperm viability varies temporally after mating; and (3) whether male copulatory behaviour covaries positively with sperm viability within females. We used the spider Holocnemus pluchei, whose males use several copulatory behaviours to court females. We found that females that stored sperm for 4 or 15 days showed no difference in sperm viability but had lower sperm viability compared with males, and males that performed a longer post-insemination behaviour had higher sperm viability inside the female. It is unclear how sperm viability is reduced and how male post-insemination behaviour affects this. It is possible that extending copulation allows males to induce females to keep sperm alive for longer. This result is predicted by theory whereby males induce females to facilitate sperm to reach and fertilize eggs based on male postcopulatory behaviour.Fil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Uñates, Diego Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Costa Schmidt, Luiz Ernesto. Universidade Federal de Pelotas; Brasil. Universidade do Vale do Rio dos Sinos, Rio Grande do Sul; BrasilFil: Córdoba-Aguilar, Alex. Universidad Nacional Autónoma de México. Instituto de Ecología; MéxicoFil: Peretti, Alfredo Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin

Human sperm pattern of movement during chemotactic re-orientation towards a progesterone source

Human spermatozoa may chemotactically find out the egg by following an increasing gradient of attractant molecules. Although human spermatozoa have been observed to show several of the physiological characteristics of chemotaxis, the chemotactic pattern of movement has not been easy to describe. However, it is apparent that chemotactic cells may be identified while returning to the attractant source. This study characterizes the pattern of movement of human spermatozoa during chemotactic re-orientation towards a progesterone source, which is a physiological attractant candidate. By means of videomicroscopy and image analysis, a chemotactic pattern of movement was identified as the spermatozoon returned towards the source of a chemotactic concentration of progesterone (10 pmol l 21 ). First, as a continuation of its original path, the spermatozoon swims away from the progesterone source with linear movement and then turns back with a transitional movement that can be characterized by an increased velocity and decreased linearity. This sperm behaviour may help the spermatozoon to re-orient itself towards a progesterone source and may be used to identify the few cells that are undergoing chemotaxis at a given time

Progesterone from the Cumulus Cells Is the Sperm Chemoattractant Secreted by the Rabbit Oocyte Cumulus Complex

Sperm chemotaxis in mammals have been identified towards several female sources as follicular fluid (FF), oviduct fluid, and conditioned medium from the cumulus oophorus (CU) and the oocyte (O). Though several substances were confirmed as sperm chemoattractant, Progesterone (P) seems to be the best chemoattractant candidate, because: 1) spermatozoa express a cell surface P receptor, 2) capacitated spermatozoa are chemotactically attracted in vitro by gradients of low quantities of P; 3) the CU cells produce and secrete P after ovulation; 4) a gradient of P may be kept stable along the CU; and 5) the most probable site for sperm chemotaxis in vivo could be near and/or inside the CU. The aim of this study was to verify whether P is the sperm chemoattractant secreted by the rabbit oocyte-cumulus complex (OCC) in the rabbit, as a mammalian animal model. By means of videomicroscopy and computer image analysis we observed that only the CU are a stable source of sperm attractants. The CU produce and secrete P since the hormone was localized inside these cells by immunocytochemistry and in the conditioned medium by enzyme immunoassay. In addition, rabbit spermatozoa express a cell surface P receptor detected by western blot and localized over the acrosomal region by immunocytochemistry. To confirm that P is the sperm chemoattractant secreted by the CU, the sperm chemotactic response towards the OCC conditioned medium was inhibited by three different approaches: P from the OCC conditioned medium was removed with an anti-P antibody, the attractant gradient of the OCC conditioned medium was disrupted by a P counter gradient, and the sperm P receptor was blocked with a specific antibody. We concluded that only the CU but not the oocyte secretes P, and the latter chemoattract spermatozoa by means of a cell surface receptor. Our findings may be of interest in assisted reproduction procedures in humans, animals of economic importance and endangered species

Sperm parameters in Pristidactylus achalensis (Squamata: Leiosauridae), a lizard endemic to the highest mountain areas in central Argentina

Reproductive physiology is influenced by environmental pressures, particularly in ectothermic species living in harsh climatic conditions. Studying the period of time in which males exhibit semen availability and their sperm traits constitutes a powerful tool to evaluate the fertility potential of males because sperm size and sperm velocity have been proposed as determinants in fertilization success. Pristidactylus achalensis is a lizard endemic to the highest mountain areas in Central Argentina and shows a pattern of seasonal activity. Here, we study the period of time when males possess sperm during the active season and characterize the spermatozoa based on sperm morphometric and dynamic traits among males and within ejaculate. Males possess sperm in their ejaculates only during the spring months of the active season, suggesting that reproductive activity is restricted to this time. Spermatozoa of Pristidactylus achalensis show intra-specific variation in sperm traits among males and within ejaculate, suggesting different reproductive potential among males. Since this species only occupies the highest areas of the mountains and is categorized as vulnerable, basic information about reproduction and sperm physiology could be useful for potential population conservation strategies.Fil: Blengini, Cecilia Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: López Juri, Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Chiaraviglio, Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Uñates, Diego Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Naretto, Sergio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin

Progesterone sperm chemoattraction may be modulated by its corticosteroid-binding globulin carrier protein

Progesterone, the main steroidal component secreted by the cumulus cells that surround the egg, chemotactically guides human spermatozoa. The aim of this work was to evaluate whether the carrier protein corticosteroid-binding globulin also participates in the sperm P chemotactic response. By means of videomicroscopy and image analysis, we observed that corticosteroid-binding globulin modulates the chemotactic activity of P, when a solution of corticosteroid-binding globulin + P is at the nanomolar range.Fil: Teves, Maria Eugenia. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Guidobaldi, Héctor Alejandro. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Uñates, Diego Rafael. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sanchez, Raúl. Universidad de la Frontera; ChileFil: Miska, Werner. Justus Liebig University; AlemaniaFil: Giojalas, Laura Cecilia. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Cytogenetics of tuna in Argentina (two forms of Opuntia ficus-indica (L.) Mill. and O. robusta J. C. Wendl., Cactaceae)

Opuntia ficus-indica (L.) Mill. f. ficus-indica (yellow tuna, cordobesa orange tuna, italiana orange tuna, salteña tuna) and f. amyclaea (white tuna, reddish tuna) and O. robusta J. C. Wendl. (cuaresma tuna) are extensively cultivated cacti in Argentina, mainly used as fodder and human food. Despite the importance of these resources, no studies of molecular cytogenetics considering the morphological characteristics of the varieties have been carried out. The main goal of this contribution is to determine the chromosome number, physical localization of ribosomal genes and amount of DNA in these taxa, including several populations with different fruit colors. The f. ficus-indica samples were octoploid, f. amyclaea hexaploid, and O. rubusta tetraploid. All taxa exhibited small similar-sized symmetrical chromosomes. Karyotypes were symmetrical with slight variations within each species. The 18S-5.8S-26S sites in all taxa were located on the secondary constrictions and the adjacent satellites at telomeric positions. The f. ficus-indica presented four 18S-5.8S-26S rDNA sites and the f. amyclaea three, whereas O. robusta had two. There were eight and six 5S rDNA sites in f. ficus-indica and f. amyclaea, respectively, and four in O. robusta. They were always proximally located on short arms of different chromosome pairs, always in a pericentromeric position. Both ribosomal genes were always asynthenic. The nuclear DNA in the octoploid f. ficus-indica presented a lower value (1Cx = 0.91 pg) than the hexaploid f.amyclaea (1Cx = 1.06 pg), whereas the tetraploid O. robusta had 1Cx = 0.87 pg. This cytogenetic evaluation is important for developing more efficient and faster selection strategies, contributing to obtaining new and improved varietiesFil: Ahumada, Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina. Universidad Nacional de Córdoba; ArgentinaFil: Montenegro, Gonzalo. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Trillo, Cecilia. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Uñates, Diego Rafael. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Bernardello, Gabriel Luis Mario. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Las Peñas, María Laura. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentin

Versatile action of picomolar gradients of progesterone on different sperm subpopulations.

High step concentrations of progesterone may stimulate various sperm physiological processes, such as priming and the acrosome reaction. However, approaching the egg, spermatozoa face increasing concentrations of the hormone, as it is secreted by the cumulus cells and then passively diffuses along the cumulus matrix and beyond. In this context, several questions arise: are spermatozoa sensitive to the steroid gradients as they undergo priming and the acrosome reaction? If so, what are the functional gradual concentrations of progesterone? Do spermatozoa in different physiological states respond differentially to steroid gradients? To answer these questions, spermatozoa were confronted with progesterone gradients generated by different hormone concentrations (1 pM to 100 µM). Brief exposure to a 10 pM progesterone gradient stimulated priming for the acrosome reaction in one sperm subpopulation, and simultaneously induced the acrosome reaction in a different sperm subpopulation. This effect was not observed in non-capacitated cells or when progesterone was homogeneously distributed. The results suggest a versatile role of the gradual distribution of very low doses of progesterone, which selectively stimulate the priming and the acrosome reaction in different sperm subpopulations

Sperm viability in spiders: A first approach using Holocnemus pluchei (Scopoli, 1763) (Synspermiata: Pholcidae)

Several studies have shown that sperm viability (SV) is a key trait during sexual competition. However, this has not yet been tested in spiders as no protocol has been developed to quantify SV. Here, we describe a methodology for estimating SV using the pholcid spider Holocnemus pluchei (Scopoli, 1763). In this method, male spermatozoa were released from the sperm ducts of copulatory bulbs in 50 μl saline solution and mixed with a vortexer. Finally, 10 μl of the sperm solution were combined with 1 μl of GelRed stain. We counted a total of 200 cells and calculated the percentage that was not stained. Males showed almost 100% of live sperm inside their genital bulbs. GelRed stain demonstrated a high effectiveness to distinguish between dead and living sperm cells in spiders and it is thus a more reliable option compared to the dyes typically used, such as propidium iodide and sybr14. The high SV in H. pluchei may be explained by the multiple mating nature of females, which suggests a selection for enhanced effectiveness during sperm competition within the female genital tract.Fil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Uñates, Diego Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Vrech, David Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Bollatti, Fedra Ariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Córdoba Aguilar, Alex. Universidad Nacional Autónoma de México; MéxicoFil: Peretti, Alfredo Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin

Picomolar gradients of progesterone select functional human sperm even in subfertile samples

More than 1 million infertility treatments are practiced around the world per year, but only 30% of the couples succeed in taking a baby home. Reproductive technology depends in part on sperm quality, which influences not only fertilization but also embryo development and implantation. In order to provide a better quality sperm subpopulation, innovative sperm selection techniques based on physiological sperm features are needed. Spermatozoa at an optimum state may be selected by following an increasing concentration gradient of picomolar progesterone, a steroid secreted by the cumulus cells at the time of ovulation. In this study we developed a method to recruit spermatozoa at the best functional state, based on sperm guidance toward progesterone. The sperm selection assay (SSA) consists of a device with two wells connected by a tube.One well was filled with the sperm suspension and the other with picomolar progesterone, which diffused inside the connecting tube as a gradient. The sperm quality after the SSA was analyzed in normal and subfertile semen samples. Several sperm parameters indicative of sperm physiological state were determined before and after the SSA: capacitation, DNA integrity and oxidative stress. After the SSA, the mean level of capacitated spermatozoa increased three times in normal and in subfertile samples. The level of sperm with intact DNA was significantly increased, while sperm oxidative stress was decreased after sperm selection. Interestingly, the exposure to a progesterone gradient stimulated the completion of capacitation in some spermatozoa that could not do it by themselves. Thus, the SSA supplies a sperm population enriched with spermatozoa at an optimum physiological state that may improve the assisted reproductive technology outcome.Fil: Gatica, Laura Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Guidobaldi, Héctor Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Montesinos, Maria del Mar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Teves, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Moreno, Ignacio Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Uñates, Diego Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Molina, R. I.. Laboratorio de Andrología y Reproducción; ArgentinaFil: Giojalas, Laura Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentin

Picomolar gradients of progesterone generated in the chemotaxis chamber stimulate either priming or the acrosome reaction in different sperm subpopulations.

<p>Percentage of spermatozoa primed (A) or acrosome-reacted (B) after a 15 min exposure to gradients of different progesterone concentrations (1 pM to 100 µM). Percentage of spermatozoa-primed (C) or acrosome-reacted (D), after a 15 min exposure to a homogeneous distribution of different progesterone concentrations (1 pM to 100 µM). Data are shown as means ± s.e.m. calculated from 7 independent experiments, counting 200–600 cells per treatment. ^a, significant differences vs. without progesterone (<i>p</i><0.05). Inset: Percentage of acrosome-reacted spermatozoa after a 30 min exposure to a homogeneous 10 µM progesterone concentration. Data are shown as means ± s.e.m. calculated from 3 independent experiments, counting 200-600 cells per treatment. ^a, significant differences vs. without progesterone (<i>p</i><0.05). Percentage of spermatozoa-primed (E) or acrosome-reacted (F) in capacitated (+cap) or non-capacitated (-cap) populations, after a 15 min exposure to a gradient of 10 pM progesterone. Data are shown as means ± s.e.m. calculated from 4 independent experiments, counting 200–600 cells per treatment. ^a, significant differences vs. without progesterone (<i>p</i><0.05). Percentage of spermatozoa-primed (G) or acrosome-reacted (H) in the presence or absence of progesterone, located in different longitudinal areas of the connection between wells (A, close to the progesterone well; B, in the middle; and C, close to the culture medium well).</p