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Expression and localization of alpha V and alpha 3 integrins in male germ cells of selected mammalian species
Bajerová, Kateřina ; Frolíková, Michaela (advisor) ; Krejčová, Tereza (referee)
Integrins are transmembrane glycoproteins mediating cell-cell adhesion and ensuring communication between extracellular matrix and cell cytoskeleton. The presence of integrin subunits has already been described on somatic cells, as well as on oocytes and sperm cells. In oocyte plasma membrane, integrins together with tetraspanins are involved in the formation of vast molecular webs. Analogical structures have been recently discovered in sperm plasma membrane as well. While the role of integrins on oocyte has been researched quite extensively, there is much less known about its presence and function on sperm. This Master's thesis focuses on expression and localization of integrins alpha V and alpha 3 on acrosome-intact sperm and its comparison between mouse, pig and human sperm cells. Furthermore, this thesis also describes relocation of the alpha V integrin after the acrosome reaction, which is an essential step of sperm maturation. Using confocal microscopy, the alpha V integrin localization was detected in acrosomal cap area of acrosome-intact sperm of all selected species. After acrosome reaction, the alpha V integrin relocated and was depicted in postacrosomal region and equatorial segment of acrosome-reacted sperm. Immunodetection revealed a decreasing expression of the alpha V integrin in...
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Sperm morphology and motility in estrildid finches of the genus Lonchura
Šárová, Markéta ; Reifová, Radka (advisor) ; Frolíková, Michaela (referee)
Sexual selection plays an important role in the evolution of animals. Today we already know that it takes place not only before copulation (precopulatory sexual selection), but also after copulation. This type of sexual selection is called postcopulatory sexual selection, and occurs mainly in promiscuous species, where females mate with multiple males. In this case, sperm competition occurs in the female reproductive tract. To increase the likelihood of their reproductive success, males began to develop surprisingly diverse sperm adaptations at the morphological, physiological, or behavioural levels. These adaptations often affect sperm velocity (motility), which is a key factor for successful egg fertilization. However, the result of reproductive success can also be influenced by females, who may prefer sperm with a certain phenotype in the process of cryptic female choice, and thus, for example, obtain better genes for offspring. In some species, females even can have the ability to sort and store sperm in specialized organs in which the sperms are nourished for some time, and then used to fertilize the egg. Even in this case, the storage of sperm is often affected by sperm morphology. Due to these mechanisms of postcopulatory sexual selection, sperm are under strong selection pressure, which can...
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The effect of oxidative stress on gametes and early embryos concerning assisted reproductive techniques in human
Kapsdorferová, Viktória ; Krylov, Vladimír (advisor) ; Frolíková, Michaela (referee)
One of the factors influencing the successful outcome of assisted reproduction could be the oxidative stress (OS), which is characterized by a disequilibrium between prooxidants and antioxidants in favour of prooxidants. Important prooxidants are both radical and non-radical reactive oxygen species (ROS) with a high reactivity and strong oxidizing effects on biological substrates. The origins of ROS and OS are assisted reproductive technologies (ART) and several exogenous sources to which gametes and early embryos are exposed during in vitro manipulation. The overproduction of ROS is associated with several negative effects on the cellular and molecular level. However, ROS are a common part of the cellular metabolism and important signaling molecules participating in processes necessary for the physiological development of gametes and early embryos. Hence, strategies to reduce OS may lead to improved results in assisted reproduction. The purpose of this bachelor's thesis is to provide a comprehensive review of literature focused on the identification of sources of ROS and OS, their effect on gametes and early embryos, as well as the possibilities for OS reduction and improvement of cultivation conditions in ART in human. Key words: oxidative stress, reactive oxygen species, gametes, early embryo,...
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The acrosome reaction in mammalian sperm
Picková, Jana ; Frolíková, Michaela (advisor) ; Kuntová, Barbora (referee)
The acrosome reaction (AR) of the sperm is a prerequisite for egg fertilization, which takes place in the female reproductive tract. The AR allows sperm to penetrate extracellular egg coats and fuse with the egg. At first, the sperm must undergo the process called capacitation, then AR is initiated and acrosomal content is released. While it is not clear, what initiates the AR, it is probably the egg's extracellular coats - the zona pellucida and cumulus cells, secreting progesterone and some other substances, which can initiate the AR. Lately, it was demonstrated, that in the mouse the most sperms undergo the AR in the upper isthmus of the oviduct. Only a few sperms reach the ampulla, but all of them can fertilize eggs. During the AR, the acrosomal content is released into the extracellular space. It was discovered, that the release of acrosomal proteins is not synchronous, soluble components are released faster from the acrosome than are acrosomal matrix proteins. Before the acrosomal release, G-coupled receptors and tyrosine kinase receptors activate phospholipases. Protein kinases are also activated, which results in the opening of Ca2+ channels in the acrosome and sperm plasma membrane and the release of Ca2+ . The increase of Ca2+ leads to actin depolymerization, membrane fusion, and finally,...
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Sperm acrosomal reactien in selected species of mammals
Frolíková, Michaela
Mammalian sperm must undergo the process of capacitation - series of physiological and biochemical modifications prior fertilization. In last stage of capacitation sperm undergoes acrosome reaction (AR). During AR the cell membrane of the sperm fuses with the outer acrosomal membrane and the contents of acrosomal vesicle are released into extracellular space. Sperm which did not undergo AR or sperm missing acrosome at all are unable to fertilize. AR results into dramatical changes in the sperm head. Most of the proteins present in plasmatic and outer acrosomal membrane are reorganized or lost. There are also significant changes in cytoskeletal and intraacrosomal proteins are released to extracellular space uncovering new surface domains. Some sperms undergo AR even without presence of inductor of AR during capacitation in vitro. This event is called spontaneous (accelerated) AR. The latest research indicates that spontaneous AR is natural part of the process of fertilization. Field mice (Apodemus) show high level of promiscuity leading to significant risk of sperm competition. Unique reproduction strategy where the sperms form so-called sperm trains was evolved in field mice. Spontaneous AR is probably enabling the dissociation of sperms from the sperm train. The spontaneous AR rate is dependent on...
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Sperm motility and postmating prezygotic isolation in two nightingale species
Baránková, Lucie ; Reifová, Radka (advisor) ; Frolíková, Michaela (referee)
The motility of male gametes (sperm) is one of the important factors influencing the reproductive success of males. Because sperms are often subjected to strong postmating sexual selection and even closely related species often differ in sperm morphology, sperm motility could also differ between species, which may contribute to reproductive isolation between species. As part of my diploma thesis, I studied sperm motility in two closely related species of songbirds, the common nightingale (Luscinia megarhynchos) and the thrush nightingale (Luscinia luscinia). These two species of nightingales are an ideal model system because the areas of these two species overlap in the secondary contact zone across Central and Eastern Europe, where they occasionally hybridize and thus allow the study of speciation mechanisms in the natural environment. Both species also differ greatly in total sperm length. As part of my diploma thesis, I studied the possible influence of different sperm morphology on their motility. I further tested whether the motility of nightingale sperm differs in the fluid from the cloaca of a female of the same species and a different species, which would demonstrate the presence of postmating prezygotic reproductive isolation between species. The results of my work showed that despite the...
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Role of importins in reproduction
Mertová, Irem ; Frolíková, Michaela (advisor) ; Bubeníčková, Filipa (referee)
Importins are proteins from a group of karyopherins, which provide transport of proteins into the nucleus both in somatic cells and gametes. Transport of specific transcription factors plays an important role in gametes and affects key events of reproduction. In mammals, reproduction is mediated by the process of sexual reproduction. In this event, a haploid gamete is formed in each parent, which in later stages of fertilization combine and make a diploid zygote. Gene expression in gametes differs dramatically from somatic cells. In gametes, translation is suspended and specific proteins are expressed only at certain stages of cell development. Proteins determined for transport into the nucleus contain various nuclear localization signals, which are detected by different types of importins. Based on studying differences in expression of individual types of importins in various phases of gametogenesis, we can deduce their roles in these processes. Role of importins was proven in gametogenesis, differentiation of stem cells and maturation of germ cells. On top of that, importins appear to play a role in defense of male germ cells against oxidative stress.
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Sperm centrioles and their role in reproduction
Vlčková, Monika ; Frolíková, Michaela (advisor) ; Liška, František (referee)
Centrioles are evolutionarily conserved protein structures composed of microtubules. In somatic cells, centrioles serve as the basal body of cilia and flagella and allow the assembly of pericentriolar material, thereby creating the centrosome. Without centrosome, animal cells are not capable of nuclear division. Centrioles do not arise de novo and their formation always requires the presence of a preexisting centriole. Since there are no centrioles in the egg at the time of fertilization, unlike spermatozoa, sperm is the carrier of centrioles and therefore all of the centrioles of the emerging organism are of paternal origin. There are two centrioles in the sperm - a cylindrical shape proximal and a distal one, which is perpendicular to the proximal. The sperm centrioles are the basis for the formation of the sperm flagellum and after fertilization form the mitotic spindle of the zygote, necessary for equal DNA and cell distribution. It follows from the above that the presence of centrioles in sperm is essential in mammals and defects in their structure may lead to male sterility or embryo development disorders. However, sperm centrioles differ from somatic centrioles in their structure and behavior and understanding these differences is one of the important tasks of reproductive biology.
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Sperm cell penetration and membrane fusion with oocyte during fertilization in mammals
Klimková, Veronika ; Krylov, Vladimír (advisor) ; Frolíková, Michaela (referee)
Fertilization is a process involving multiple steps, which are in continuity and complement each other. Spermatozoa become competent to fertilize after capacitation, hyperactivation and acrosome reaction. Oocytes are surrounded by a layer of cumulus cells and give a signals for spermatozoa to activation and orientation. This study focuses on the latest knowledges about a roles of cumulus cells, production of chemoattractants and the possibility that cumulus cells induce the acrosome reaction after binding a novel sperm protein NYD-SP8 to the cumulus. It seems that progesterone induce the acrosome reaction and also is the best chemoattractant secreted by cumulus cells. Next I am focus on gamete fusion including the role of surface proteins such as Juno and IZUMO1. Key words: membrane fusion, cumulus cells, acrosome reaction, chemotaxis sperm, oocyte, fertilization
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Sperm acrosomal reactien in selected species of mammals
Frolíková, Michaela
Mammalian sperm must undergo the process of capacitation - series of physiological and biochemical modifications prior fertilization. In last stage of capacitation sperm undergoes acrosome reaction (AR). During AR the cell membrane of the sperm fuses with the outer acrosomal membrane and the contents of acrosomal vesicle are released into extracellular space. Sperm which did not undergo AR or sperm missing acrosome at all are unable to fertilize. AR results into dramatical changes in the sperm head. Most of the proteins present in plasmatic and outer acrosomal membrane are reorganized or lost. There are also significant changes in cytoskeletal and intraacrosomal proteins are released to extracellular space uncovering new surface domains. Some sperms undergo AR even without presence of inductor of AR during capacitation in vitro. This event is called spontaneous (accelerated) AR. The latest research indicates that spontaneous AR is natural part of the process of fertilization. Field mice (Apodemus) show high level of promiscuity leading to significant risk of sperm competition. Unique reproduction strategy where the sperms form so-called sperm trains was evolved in field mice. Spontaneous AR is probably enabling the dissociation of sperms from the sperm train. The spontaneous AR rate is dependent on...
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