Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2018-09-24T15:40:23Z No. of bitstreams: 1 CAROLINE MARTINEZ.pdf: 15483260 bytes, checksum: 336be2ac82e6f35a3e3945f6a2b3b533 (MD5) / Approved for entry into archive by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2018-09-24T15:48:42Z (GMT) No. of bitstreams: 1 CAROLINE MARTINEZ.pdf: 15483260 bytes, checksum: 336be2ac82e6f35a3e3945f6a2b3b533 (MD5) / Made available in DSpace on 2018-09-24T15:48:42Z (GMT). No. of bitstreams: 1 CAROLINE MARTINEZ.pdf: 15483260 bytes, checksum: 336be2ac82e6f35a3e3945f6a2b3b533 (MD5) Previous issue date: 2017-12-13 / O Alumínio (Al) é o metal de maior exposição humana, no entanto os efeitos do metal em nível de exposição humana ainda são pouco conhecidos. Assim, o objetivo desse estudo foi investigar os efeitos da exposição ao Al por 60 dias em dose equivalente a exposição humana ao metal através da dieta sobre o Sistema Nervoso Central (SNC), Sistema Nervoso Periférico (SNP), sistema reprodutor masculino e sistema cardiovascular e, comparar com os efeitos de uma exposição alta ao metal com efeitos tóxicos conhecidos. Para isso, ratos Wistar com 3 meses de idade foram divididos em: 1) Grupo 1: baixas doses de Al, onde durante 60 dias os ratos receberam por água de beber: a) Controle – água ultrapura; b) Al na dose de 1,5 mg/kg de peso corporal e, c) Al na dose de 8,3 mg/kg de peso corporal e, 2) Grupo 2: alta dose de Al, onde durante 42 dias os ratos receberam por gavagem: a) Controle – água ultrapura; b) Al na dose de 100 mg/kg de peso corporal. O tratamento com Al mesmo em baixas doses prejudicou a memória de reconhecimento de objetos e promoveu o desenvolvimento de catalepsia nos ratos. Somado a isso, a exposição ao Al aumentou os níveis de espécies reativas de oxigênio (EROs) e de peroxidação lipídica, reduziu a capacidade antioxidante e inibiu a atividade da acetilcolinesterase no hipocampo dos animais. No SNP, o Al promoveu o desenvolvimento de alodínea mecânica, aumentou o estresse oxidativo sistêmico, induziu inflamação com recrutamento de macrófagos e, o metal foi capaz de depositar-se entre as fibras do nervo ciático. Já no sistema reprodutor masculino, a exposição ao Al reduziu a contagem espermática, a motilidade e a produção diária de espermatozides, aumentou a porcentagem de espermatozoides com anormalidades morfológicas, alterou a estrutura testicular, aumentou os níveis de estresse 14 oxidativo e a inflamação testicular, demonstrando que uma baixa concentração do metal nos testículos (3.35 μg/g) é o suficiente para comprometer a espermatogênese e a qualidade dos gametas masculinos. No sistema cardiovascular, o Al aumentou a pressão arterial sistólica, reduziu a resposta vasodilatadora a acetilcolina, aumentou a resposta vasoconstritora a fenilefrina, reduziu a modulação endotelial na resposta vasoconstritora, reduziu a biodisponibilidade de óxido nítrico, o envolvimento dos canais de potássio nas respostas vasculares e aumentou a produção de EROs principalmente via NAD(P)H oxidase e de prostanóides contráteis da via da COX-2. A exposição ao Al aumentou o estresse oxidativo em artérias aorta e mesentérica, reduziu a expressão de mRNA de eNOS e SOD1 e aumentou a expressão da isoforma da NAD(P)H oxidase 1, COX-2 e a expressão de TXA-2 R. Tomados em conjunto, nossos dados demonstram que a exposição subcrônica ao Al por 60 dias em baixa dose, que reflete a exposição humana ao metal através da dieta, alcança um limiar tóxico suficiente para promover efeitos adversos no SNC, SNP, sistema reprodutor masculino e sistema cardiovascular. Além disso, os efeitos de uma exposição em baixa dose são praticamente os mesmos de uma exposição alta ao metal. / Aluminum (Al) is the most important environmental and human contaminant. While a good deal of research has been conducted on the acute toxic effects of Al, little is known about the effects of longer-term exposure at human dietary Al levels. Therefore, the purpose of this study was to investigate the effects of 60-day Al exposure at low doses on Central Nervous System (CNS), Peripheral Nervous System (PNS), male reproductive system and cardiovascular system for comparison with a model of exposure known to produce toxicity in rats. Three-month-old male Wistar rats were divided into two major groups: 1) Grou 1, low aluminum levels - rats were treated orally by drinking water for 60 days as follows: a) Control – received ultrapure drinking water; b) Aluminum at 1.5 mg/kg b.w. and c) Aluminum at 8.3 mg/kg b.w. and 2) Group 2, high aluminum level - rats were treated through oral gavages for 42 days as follows: a) Control – received ultrapure water; b) Aluminum at 100 mg/kg b.w. Al treatment even at low doses promoted recognition memory impairment seen in object recognition memory testing and catalepsy behavior in rats. Moreover, Al increased hippocampal reactive oxygen species (ROS) and lipid peroxidation levels, reduced antioxidant capacity and decreased acetylcholinesterase activity. On PNS, Al promoted the development of mechanical allodynia, increased inflammation in the sciatic nerve, systemic oxidative stress and, is able to be retained in the sciatic nerve. Regarding the male reproductive system, Al decreased sperm count, daily sperm production, sperm motility, normal morphological sperm, impaired testis histology; increased oxidative stress in reproductive organs and inflammation in testis, showing that low concentrations of Al in testes (3.35 μg/g) are sufficient to impair 16 spermatogenesis and sperm quality. On cardiovascular system, Al increased systolic blood pressure, decreased acetylcholine-induced relaxation, increased response to phenylephrine, decreased endothelial modulation of vasoconstrictor responses, the bioavailability of nitric oxide, the involvement of potassium channels on vascular responses, as well as increased ROS production from NAD(P)H oxidase and contractile prostanoids mainly from COX-2 in both aorta and mesenteric resistance arteries (MRA). Al exposure increased vascular ROS production and lipid peroxidation as well as altered the antioxidant status in aorta and MRA. Al decreased vascular eNOS and SOD1 mRNA levels and increased the NAD(P)H oxidase 1, COX-2 and TXA-2 R mRNA levels. Taken together, our data demonstrate that 60-day subchronic exposure to low doses of Al from feed and added to the water, which reflect human dietary Al intake, reaches a threshold sufficient to promote adverse effects on SNC, PNS, male reproductive system and cardiovascular system. Moreover, these effects were almost the same as Al exposure at much higher levels.

En aquest treball s'ha dissenyat un mètode ràpid i fiable de tinció amb fluorocroms per a l'anàlisi de la integritat i viabilitat espermàtiques a partir del marcatge de la beina mitocondrial amb MitoTracker®Green FM, de l'acrosoma amb la lectina Trypsin inhibitor from Soybean (SBTI) conjugada amb el fluorocrom Alexa Fluor®488 específic per la proacrosina i del nucli amb els fluorocroms bis-benzimida (específic per a cèl·lules viables) i iodur de propidi (específic per a cèl·lules no viables). També s'ha determinat l'efecte de la filtració de dosis seminals de mascles astentoteratonecrospèrmics en columnes de Sephadex neutre i de dosis de mascles amb baixa qualitat espermàtica per filtració en columnes de Sephadex iònic, llana de vidre i glass beads sobre la qualitat espermàtica dels diferents grups de mascles analitzats. Els resultats obtinguts han mostrat que diversos paràmetres de qualitat espermàtica milloren després de la filtració en les diferents reïnes segons la patologia que presentin. / In this work, a quick and reliable method of staining with fluorochromes was designed for the assessment of the sperm integrity and vitality by marking the mitochondrial sheath with MitoTracker®Green FM, the acrosome with the lectine Trypsin inhibitor from Soybean (SBTI) conjugated with the fluorochrome Alexa Fluor ® 488m, specific for the proacrosine, and the nucleus with the fluorochromes bis-benzimida (specific for viable cells) and propidium iodide (specific for non-viable cells).Moreover, in this work, the effects of filtration of seminal doses from asthenoteratonecrospermic boars through columns of Sephadex neuter and seminal doses from boars with low sperm quality through ionic Sephadex, glass wool and glass beads columns on sperm quality were tested. The results obtained showed that some sperm quality parameters improved after filtration through the different matrixes depending on the pathology that the males presented before filtration.

Made available in DSpace on 2014-08-20T14:38:01Z (GMT). No. of bitstreams: 1 dissertacao_liziane_vianna.pdf: 321138 bytes, checksum: e8e76002b27c6cb6d710d3cd63a4c088 (MD5) Previous issue date: 2011-02-28 / The objective of this study was to evaluate alternative methods to the gradient of Percoll® for sperm selection used in bovine in vitro production systems. We evaluated the following parameters: sperm quality, embryo development, the viability after cryopreservation and sex ratio of bovine embryos produced in vitro. Four treatments were performed: percoll group (90 and 45%) 4 mL, centrifuged at 700xg for 20 min; minipercoll group (90 and 45%) 800 &#956;L, centrifuged at 700xg for 5 min; miniisolate group (90 and 45%) 800 &#956;L, centrifuged at 700xg for 5 min; and minioptiprep group (30,28 and 26%) 1,2 mL, centrifuged at 900xg for 15 min. Different gradients did not affect the semen characteristics compared with the evaluation immediately after thawing (P>0,05). It was only observed a decrease in total motility in minioptiprep group (P<0,05). The cleavage rates were similar among the percoll and miniisolate groups (P>0,05). Minipercoll and minioptiprep groups were lower to percoll (P<0,05) and similar to isolate (P>0,05). At Day 7 the developmental rates were higher in miniisolate group than other groups (P<0,05). But at D8 the developmental rates of minigradients were similar to percoll (P>0,05). Among the minigradientes, the miniisolate was higher to minipercoll and minioptiprep (P<0,05). The treatments did not affect the sex ratio expected of 50:50% (P.0,05) and there was no difference among the four groups (P>0,05). Also, there was no difference in survival rates after vitrification (P>0,05). In conclusion, the miniisolate and the minioptiprep groups, in the concentration and volume used in our experiment, reached similar results to the percoll group. So, they are alternatives to the gradient of Percoll® in the routine of IVP bovine embryos. / O objetivo do presente estudo foi avaliar possíveis alternativas ao gradiente de Percoll®, para seleção espermática, no sistema de produção in vitro de embriões bovinos. Foram avaliados os seguintes parâmetros: qualidade espermática, desenvolvimento embrionário, viabilidade após a criopreservação e a proporção sexual dos embriões bovinos produzidos in vitro. Foram realizados quatro tratamentos: grupo percoll (90 e 45%) - 4 mL, centrifugado 700xg por 20 min; grupo minipercoll (90 e 45%) 800 &#956;L, centrifugado a 700xg por 5 min; gupo miniisolate (90 e 45%) 800 &#956;L, centrifugado a 700xg por 5 min; e grupo minioptiprep (30, 28 e 26%) - 1,2 mL, centrifugado a 900xg por 15 min. A passagem pelos gradientes não afetou as características do sêmen em comparação com a avaliação realizada logo após o descongelamento (P>0,05), sendo somente observada uma diminuição da motilidade total para o grupo minioptiprep (P<0,05). Quanto a produção in vitro de embriões, não foi observada diferença na taxa de clivagem entre os grupos percoll e miniisolate (P>0,05), sendo o minipercoll e o minioptiprep inferiores ao percoll (P<0,05) e semelhantes ao miniisolate (P>0,05). No desenvolvimento embrionário, o miniisolate foi superior aos demais grupos em D7 (P<0,05). Em D8, os três minigradientes foram semelhantes ao percoll (P>0,05). Entre os minigradientes, o miniisolate foi superior ao minipercoll e ao minioptiprep (P<0,05). A passagem pelos gradientes não afetou a proporção sexual esperada de 50:50%, além disso, não houve diferença entre os grupos (P>0,05). Não houve diferença quanto à taxa de sobrevivência a vitrificação nos quatro grupos em 48 h (P>0,05). Concluímos que o miniisolate e o minioptiprep, nas concentrações e volumes estudados neste trabalho, atingiram resultados semelhantes ao percoll. Portanto, são alternativas a utilização do gradiente de Percoll®, na rotina de produção de embriões bovinos in vitro.

[ES] La selección de gametos de alta calidad es un requisito indispensable a tener en cuenta en los programas de reproduccion asistida. El desarrollo de herramientas biotecnológicas como la criopreservación de gametos, juega un papel importante en la producción acuicola y en la formación de bancos de germoplasma, que contribuiran luego en la mejora genética de las poblaciónes de peces, principalmente aquellas en peligro y que pudieran estar más afectadas ante futuros cambios climáticos. En la primera fase de esta tesis realizada en la Unesp, se trabajó con una especie neotropical de elevada importancia económica para la region Suramericana. La segunda fase realizada en la UPV se trabajó con la Anguila europea (Anguilla anguilla), una especie clasificada en la Lista Roja de la (UICN) como especie "en peligro crítico de extinción". En la primera fase se buscó caracterizar la composición bioquímica del plasma y las características seminales de la especie para evaluar las posibles relaciones entre estos parámetros. El plasma seminal estuvo principalmente compuesto por iones de sodio (Na+) y dentro de los componentes orgánicos sobresalieron las proteínas totales y la glucosa. A través del análisis de componentes principales (PCA) se observó que la motilidad tenía una fuerte correlación positiva con el tiempo de motilidad, la concentración de espermatozoides y las proteínas totales. Estos análisis sirvieron de base para la creación de una solución diluyente utilizada posteriormente en la sustancia crioprotectora. Luego se determinó la influencia del plasma seminal como constituyente de la solución crioprotectora en la criopreservación de semen de P. reticulatum. Se utilizaron tres tratamientos: glucosa 5% + metanol 10% (T1), a este crioprotector se le agregó 30% de plasma seminal natural (T2) y 30% de plasma seminal artificial en base a los resultados de los componentes bioquímicos del plasma determinados para la especie en el experimento anterior (T3). Se evaluaron parámetros de motilidad espermática, capacidad fecundante del semen criopreservado, así como fragmentación del ADN. El tratamiento T1 resultó con los mejores valores de motilidad seguido del T2, y la capacidad fertilizante de estos dos tratamientos fue similar al control, sin embargo, el tratamiento T2 mostró menos daño en el ADN. Mediante el PCA se demostró que T1 tenía una mejor relación positiva con la fertilidad y la motilidad total y progresiva. Finalmente, evaluamos las estructuras de las subpoblaciones espermáticas en cada uno de los tratamientos utilizados. Mediante análisis multivariado en dos etapas, fue posible determinar tres subpoblaciones espermáticas en el semen crioconservado de la especie, SP1 (rápido-lineal), SP2 (rápido-no lineal) y SP3 (lento-lineal). T1 presentó el mayor porcentaje de SP1, siendo confirmado por la efectividad en la protección de las células de este tratamiento en el proceso de criopreservación de la especie. En la segunda fase que se está llevó a cabo en la UPV, el objetivo general fue determinar el efecto de la temperatura y el pH del agua de mar sobre la motilidad de los espermatozoides en la Anguila europea. Se determinó que el bajo pH del agua de mar (6.5-7.4) disminuyó la motilidad de los espermatozoides de anguila en comparación con el control (pH= 8.2). Cuando estudiamos el efecto combinado del pH del plasma seminal artificial y el pH de ASW (7.8 y 8.2), no encontramos diferencias estadísticas en la motilidad y cinética de los espermatozoides en relación con el pH del plasma seminal artificial, pero sí el pH del agua de mar. Se encontraron valores más altos de motilidad total (MOT), FA(rápidos) y ME (médios) con un pH de 8.2 que con un pH de 7.8. En contraste, la temperatura del agua de mar no afectó los parámetros de motilidad de los espermatozoides o la longevidad de los espermatozoides en el contexto del cambio climático. / [CA] La selecció de gàmetes d'alta qualitat és un requisit indispensable a tenir en compte en els programes de reproducció assistida. El desenvolupament d'eines biotecnològiques com la criopreservació de gàmetes, juga un paper important en la producció aqüicola i en la formació de bancs de germoplasma, que contribuiran després a la millora genètica de les població de peixos, principalment aquelles en perill i que poguessin estar més afectades davant de futurs canvis climàtics. A la primera fase d'aquesta tesi realitzada a la Unesp, es va treballar amb una espècie neotropical d'elevada importància econòmica per a la regió sud-americana. La segona fase realitzada a la UPV es va treballar amb l'Anguilla europea (Anguilla anguilla), una espècie classificada a la Llista Vermella de la (UICN) com a espècie "en perill crític d'extinció". A la primera fase es va buscar caracteritzar la composició bioquímica del plasma i les característiques seminals de l'espècie per avaluar les possibles relacions entre aquests paràmetres. El plasma seminal va estar principalment compost per ions de sodi (Na+) i dins dels components orgànics van sobresortir les proteïnes totals i la glucosa. A través de l'anàlisi de components principals (PCA), es va observar que la motilitat tenia una forta correlació positiva amb el temps de motilitat, la concentració d'espermatozoides i les proteïnes totals. Aquestes anàlisis van servir de base per a la creació d'una solució diluent utilitzada posteriorment a la substància crioprotectora. Després es va determinar la influència del plasma seminal com a constituent de la solució crioprotectora en la criopreservació de semen de P. reticulatum. Es van utilitzar tres tractaments: glucosa 5% + metanol 10% (T1), a aquest crioprotector se li va afegir 30% de plasma seminal natural (T2) i 30% de plasma seminal artificial sobre la base dels resultats dels components bioquímics del plasma determinats per a l'espècie a l'experiment anterior (T3). S'avaluaren paràmetres de motilitat espermàtica, capacitat fecundant del semen criopreservat, així com fragmentació de l'ADN. El tractament T1 va resultar amb els millors valors de motilitat seguit del T2, i la capacitat fertilitzant d'aquests dos tractaments va ser similar al control, però el tractament T2 va mostrar menys mal a l'ADN. Mitjançant el PCA es va demostrar que T1 tenia una millor relació positiva amb la fertilitat i la motilitat total i progressiva. Finalment, avaluem les estructures de les subpoblacions espermàtiques a cadascun dels tractaments utilitzats. Mitjançant anàlisi multivariada en dues etapes, va ser possible determinar tres subpoblacions espermàtiques en el semen crioconservat de l'espècie, SP1 (ràpid-lineal), SP2 (ràpid-no lineal) i SP3 (lent-lineal). T1 va presentar el percentatge més gran de SP1, i va ser confirmat per l'efectivitat en la protecció de les cèl·lules d'aquest tractament en el procés de criopreservació de l'espècie. A la segona fase que es va dur a terme a la UPV, l'objectiu general va ser determinar l'efecte de la temperatura i el pH de l'aigua de mar sobre la motilitat dels espermatozoides a l'Anguila europea. Es va determinar que el pH baix de l'aigua de mar (6.5-7.4) va disminuir la motilitat dels espermatozoides d'anguila en comparació del control (pH= 8.2). Quan estudiem l'efecte combinat del pH del plasma seminal artificial i el pH d'ASW (7.8 i 8.2), no trobem diferències estadístiques en la motilitat i la cinètica dels espermatozoides en relació amb el pH del plasma seminal artificial, però sí el pH de l'aigua de mar. Es van trobar valors més alts de motilitat total (MOT), FA(ràpids) i ME (metges) amb un pH de 8.2 que amb un pH de 7.8. En contrast, la temperatura de l'aigua de mar no va afectar els paràmetres de motilitat dels espermatozous o la longevitat dels espermatozous en el context del canvi climàtic. / [EN] The selection of high-quality gametes is an essential requirement to take into account in assisted reproduction programs. The development of biotechnological tools such as cryopreservation of gametes plays an important role in aquaculture production and in the formation of germplasm banks, which will later contribute to the genetic improvement of fish populations, mainly those in danger and that could be more affected by future climate changes. In the first phase of this thesis carried out at Unesp, we worked with a neotropical species of high economic importance for the South American region. The second phase carried out at the UPV worked with the European Eel (Anguilla anguilla), a species classified on the (IUCN) Red List as a "critically endangered" species. In the first phase, we sought to characterize the biochemical composition of the plasma and the seminal characteristics of the species to evaluate the possible relationships between these parameters. The seminal plasma was mainly composed of sodium ions (Na+) and within the organic components, total proteins and glucose stood out. Through principal component analysis (PCA) it was observed that motility had a strong positive correlation with motility time, sperm concentration and total proteins. These analyzes served as the basis for the creation of a diluent solution later used in the cryoprotective substance. Then the influence of seminal plasma as a constituent of the cryoprotectant solution in the cryopreservation of P. reticulatum semen was determined. Three treatments were used: 5% glucose + 10% methanol (T1), 30% natural seminal plasma (T2) and 30% artificial seminal plasma were added to this cryoprotectant based on the results of the determined biochemical components of the plasma. for the species in the previous experiment (T3). Parameters of sperm motility, fertilizing capacity of cryopreserved semen, as well as DNA fragmentation were evaluated. Treatment T1 resulted with the best motility values followed by T2, and the fertilizing capacity of these two treatments was similar to the control, however, treatment T2 showed less DNA damage. Using PCA, it was shown that T1 had a better positive relationship with fertility and total and progressive motility. Finally, we evaluated the structures of the sperm subpopulations in each of the treatments used. Through two-stage multivariate analysis, it was possible to determine three sperm subpopulations in the cryopreserved semen of the species, SP1 (fast-linear), SP2 (fast-nonlinear) and SP3 (slow-linear). T1 presented the highest percentage of SP1, being confirmed by the effectiveness in protecting the cells of this treatment in the cryopreservation process of the species. In the second phase that is being carried out at the UPV, the general objective was to determine the effect of temperature and pH of seawater on sperm motility in the European Eel. It was determined that the low pH of seawater (6.5-7.4) decreased the motility of eel sperm compared to the control (pH= 8.2). When we studied the combined effect of the pH of the artificial seminal plasma and the pH of ASW (7.8 and 8.2), we did not find statistical differences in the motility and kinetics of the sperm in relation to the pH of the artificial seminal plasma, but we did find the pH of the water of sea Higher values of total motility (MOT), FA (fast) and ME (medium) were found at a pH of 8.2 than at a pH of 7.8. In contrast, seawater temperature did not affect sperm motility parameters or sperm longevity in the context of climate change. / I also thank the Coordination for the Improvement of Higher Education Personnel (CAPES/PROEX) (N° 88887.302629/2018-00), National Council for Scientific, Technological Development CNPq (N° 200452/2022-3) and the Brazilian fostering agencies Foundation for Research Support of the State of Sao Paulo FAPESP (N° 2020/15020-0), for its financial support in Brazil. In Spain, the ThinkInAzul programme, supported by the Spanish Ministry of Science and Innovation (MCIN) with funding from the European Union NextGenerationEU (PRTR-C17.I1) and the Generalitat Valenciana (THINKINAZUL/2021/012) to SEASPERM, which has made it possible the preparation of this work. To the AUIP (Ibero-American Postgraduate University Association) for the Academic Mobility scholarship between Institutions Associated with the AUIP 2022 / Padilla Sánchez, M. (2023). Sperm Quality and Cryopreservation in Teleost: Effect of Seminal Plasma Component and Climate Change [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/201549

Infertility affects 15% of reproductive-aged couples. The milieu surrounding the growing embryo is of outmost importance, and should be optimised during in vitro fertilisation (IVF). Many biological processes, such as angiogenesis, coagulation, and immune processes need to be well regulated for a pregnancy to occur and progress normally. Histidine-rich glycoprotein (HRG) is a plasma protein that regulates components of these systems by building complexes with various ligands. A single nucleotide polymorphism (SNP) in HRG, denoted HRG C633T, seem to be of importance for IVF treatment outcomes. The aim of this thesis was to further investigate the proposed human fertility effects of the HRG C633T SNP. According to the findings of this thesis, the HRG C633T genotype is associated with primary recurrent miscarriage. Male HRG C633T genotype is associated with semen characteristics in infertile men, and pregnancy rates following IVF. However, the distribution of the HRG C633T SNP does not differ between infertile and fertile couples. We further examined the role of the region surrounding the HRG C633T SNP for regulation of endometrial angiogenesis and human embryo development. The region affects primary endometrial endothelial cell migration, proliferation and tube-formation in vitro but does not appear to affect human embryo development. No effect of the HRG peptide was noted on the secretome of human embryos. However, early embryos secrete proteins into the surrounding culture media and the level of secretion of VEGF-A, IL-6, EMMPRIN and PlGF is greater in embryos of higher developmental stages. In conclusion, the HRG C633T genotype appears to play a role only if infertility is established. The region surrounding HRG C633T SNP is of relevance in vitro for regulation of human endometrial endothelial cell angiogenesis. To predict which embryos to transfer in IVF, we have highlighted a number of proteins of interest for further investigation.

The aim of this diploma thesis is to compare quality of northern pike sperm collected by different methods. First method is collection of stripped sperm by abdominal massage of the belly. Second method is collection of stripped sperm with special catheter to eliminate sperm contamination by urine. The last method is collection of testicular sperm. Differently collected sperm was evaluated and compared its quality. The main observed parameters were sperm volume, spermatozoa concentration, spermatozoa motility and velocity and osmolality of seminal fluid. Sperm samples were used for eggs fertilization. In fertilized eggs, the fertility of eggs and larvae hatching rate were observed.