Thesis--Wisconsin. / Vita. Includes bibliographical references (leaves 131-135).

Thesis (M.S.)--University of Missouri-Columbia, 2007. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on April 3, 2009) Vita. Includes bibliographical references.

Theses (M.S.)--Marshall University, 2006. / Title from document title page. Includes abstract. Document formatted into pages: contains x, 90 p. including illustrations. Bibliography: p. 58-70.

This study was initiated in an attempt toadd the four-striped field mouse (Rhabdomys pumilio) to the small group of mammals that are used to study and develop assisted reproductive technologies. To accomplish this, a complete knowledge of both the male and female gametes was necessary, as well as a repeatable and successful IVF protocol. The gametes of the field mouse, both the falciformspermand the mature oocytes, were shown to be similar in several respects to those of many other rodent species. The induction of both the capacitated state and the acrosome reaction in the sperm were readily achieved. The success of retrieving mature oocytes was very low, with the rare occurrence of polar bodies within ovulated oocytes. The difficulties with oocytes was either related to the lack of a regular oestrous cycle, or to the hormonal regime used to induce superovulation. Although several protocols were used to accomplish superovulation in this species, it was seldom achieved. In in vitro fertilization trials, sperm were found to attach to both the cumulus oophorus and the zona pellucida of the oocytes. However, fertilization was never accomplished. This failure to achieve fertilization was probably related to the immaturity of the oocytes. Without further studies on Rhabdomys pumilio and further attempts at accomplishing in vitro fertilization, it would seem that this species would not easily be introduced as an animalmodelfor assisted reproduction.

This thesis aimed to study three main aspects: (i) the mechanisms underlying unreduced pollen gamete formation in the diploid `CSO¿ tangor hybrid used as male parent in 4x x 2x triploid breeding programs, (ii) the frequencies and mechanisms involved in the unreduced 2n female gametes production for `Eureka Frost¿ and `Fino¿ lemon genotypes, and (iii) the interspecific recombination and the resulting diploid gamete structures of doubled-diploid `Mexican¿ lime to evaluate the possibility that natural interploid hybridization maybe the origin of C. latifolia (`Tahiti¿ lime type) and C. aurantifolia (`Tanepao¿ lime type) triploid varieties. The production of 54 tetraploid hybrids from 4x x 2x sexual hybridizations allowed the analysis of the mechanisms underlying unreduced pollen gamete formation. SSR and SNP molecular markers revealed that the majority of these plants were obtained from 2n pollen of the diploid tangor parent. Then, the maximum-likelihood method based on parental heterozygosity restitution (PHR) of centromeric loci revealed that both FDR (First Division Restitution ) with predominant occurrence and to a lower extend SDR (Second Division Restitution) were the mechanisms leading to unreduced male gamete formation in the tangor studied. These observations were confirmed by the analysis of PHR pattern along the linkage group (LG) 2. To our knowledge, this is the first report of tetraploid citrus progenies arising from unreduced pollen and the first description of the coexistence of two meiotic restitution mechanisms (SDR and FDR) producing unreduced pollen in citrus. In order to study the frequencies and the mechanisms involved in the unreduced 2n female gametes production in two different genotypes of lemon, we produced 43 triploid and tetraploid hybrids from 2x x 2x and 2x x 4x sexual hybridizations using `Eureka Frost¿ and `Fino¿ as female parents. The frequencies of 2n gamete production were respectively 4.9% and 8.3%. The maximum-likelihood analysis and pattern of PHR along LG1 revealed that SDR is the main mechanism of unreduced female lemon gametes (88%), followed by FDR or pre-meiotic doubling (PRD) (7%) and post-meiotic genome doubling (PMD) mechanisms (5%). This is the first report of the production of a large number of lemon progenies from 2n gametes and the first identification of a new mechanism, PMD that has never been observed in citrus and has rarely been described in other herbaceous or woody species. Across both studies, we demonstrated at the methodological level the effectiveness of using two complementary approaches, the analysis of the PHR pattern in one LG and the maximum-likelihood method based on centromeric loci for distinguishing between the different mechanisms of unreduced gamete production. We analyzed the meiotic mechanisms of a doubled diploid `Mexican¿ lime, the interspecific recombination and the resulting diploid gamete structures combining a segregation analysis of SSR and SNPs markers, a cytogenetic study and pollen viability evaluation. We concluded that the doubled-diploid `Mexican¿ lime had a predominantly disomic segregation for three LGs, intermediate inheritance with disomic tendency was found for five LGs and intermediate models for one LG. The resulting interspecific diploid gamete structures displayed high C. medica / C. micrantha heterozygosity. The revealed genetic structures of the diploid gametes produced by the doubled-diploid `Mexican¿ lime are compatible with the hypothesis that `Tahiti¿ and `Tanepao¿ triploid varieties results from interploid hybridization involving a doubled-diploid `Mexican¿ like lime. This disomic tendency limits the recombination and the diversity of the diploid gamete population; however the observed pollen viability restoration at tetraploid level could be advantageous for intensive breeding projects. / Esta tesis estudia tres aspectos principales: (i) los mecanismos de la formación de gametos 2n de polen originados por un híbrido diploide (tangor `CSO¿) utilizado como parental masculino en hibridaciones sexuales 4x x 2x, (ii) las frecuencias y los mecanismos de la producción de gametos 2n femeninos en dos genotipos de limón, `Eureka Frost¿ y `Fino¿, y (iii) el análisis de la recombinación interespecífica y las estructuras de los gametos diploides originados por la lima `Mejicana¿ doble diploide para evaluar el origen de las las variedades triploides de lima C. latifolia (`Tahiti¿) y C. aurantifolia (`Tanepao¿). La obtención de 54 híbridos tetraploides a partir de hibridaciones 4x x 2x permitió analizar los mecanismos de la formación de gametos 2n de polen. El análisis molecular reveló que la mayoría de estas plantas se obtuvieron a partir de gametos 2n de polen del parental `CSO¿. A continuación, el análisis mediante el método de máxima verosimilitud indicó que FDR (Restitución de la Primera División meiótica) y SDR (Restitución de la Segunda División meiótica) son los mecanismos implicados con una mayor dominancia de FDR respecto SDR. Estos resultados se confirmaron mediante el análisis de la restitución de la heterocigosidad en el grupo de ligamiento (LG) 2. Con los datos publicados hasta la fecha, es la primera vez que se han obtenido progenies tetraploides de cítricos mediante gametos 2n de polen y es la primera descripción en cítricos de la coexistencia de dos mecanismos SDR y FDR. Para estudiar las frecuencias y los mecanismos de la producción de gametos 2n en dos genotipos de limón, se obtuvieron 43 híbridos triploides y tetraploides a partir de hibridaciones sexuales 2x x 2x y 2x x 4x utilizando los limones `Eureka Frost¿ y `Fino¿ como parentales femeninos. Las frecuencias de producción de gametos 2n fueron respectivamente 4,9% y 8,3%. El análisis de máxima verosimilitud y el patrón de PHR a lo largo del LG1 reveló que SDR es el mecanismo principal de formación de gametos 2n femeninos (88%), seguido por FDR o duplicación del genoma pre-meiosis (PRD) (7%) y se identificó un nuevo mecanismo originado a partir de la duplicación del genoma post-meiosis (PMD) (5%). En este trabajo se describe por primera vez en cítricos la producción de un elevado número de híbridos de limón a partir de gametos 2n y es la primera vez que se identifica un nuevo mecanismo PMD que nunca se ha observado en cítricos. En ambos estudios se demostró la efectividad del uso de dos métodos complementarios, el análisis del patrón de PHR a lo largo de un LG y el método de máxima verosimilitud para distinguir entre los diferentes mecanismos implicados en la formación de gametos 2n. También se ha analizado el modelo de segregación cromosómica de la lima `Mejicana¿ doble diploide (DD) así como la recombinación interespecífica y las estructuras de los gametos diploides resultantes. Se ha realizado análisis de la viabilidad del polen junto con un análisis citogenético y con marcadores SSRs y SNPs. Estos trabajos nos han permitido concluir que la lima `Mejicana¿ DD presenta una segregación predominantemente disómica para tres LGs, herencia intermedia con tendencia disómica para cinco LGs y un tipo de segregación intermedia para un LG. Las estructuras de los gametos diploides mostraron una alta heterocigosis en C. medica/C. micrantha, parentales de la lima `Mejicana¿. Las estructuras genéticas de los gametos diploides de la lima `Mejicana¿ DD son compatibles con la hipótesis de que las variedades triploides `Tahiti¿ y `Tanepao¿ se obtuvieran a partir de una hibridación interploide a partir de la lima `Mejicana¿ DD. El tipo de segregación disómico conlleva una limitación de la recombinación y la diversidad genética de la población de gametos 2n. Sin embargo la viabilidad del polen de la lima `Mejicana¿ DD en comparación con la lima `Mejicana¿ diploide permite / Aquesta tesi ha tingut com a objectius l'estudi de (i) els mecanismes subjacents a la formació de pol·len de gàmetes 2n al tangor híbrid diploide `CSO', utilitzat com a progenitor masculí en els programes de millora de triploides 4x x 2x (ii) les freqüències i els mecanismes implicats en la producció gàmetes 2n en la femella als genotips de llimona `Eureka Frost¿ i Fina' (iii) la recombinació interespecífica i les estructures resultants de gàmetes diploides del doble-diploide (DD) de `llima Mèxicana' per avaluar la possibilitat que la hibridació interploid natural potser l'origen de les varietats triploides C. latifolia (llima tipus `Tahiti') i C. aurantifolia (llima tipus `Tanepao'). La producció de 54 híbrids tetraploides obtinguts d¿hibridacions sexuals 4x x 2x va permetre l'anàlisi dels mecanismes de formació de gàmetes 2n de pol·len. Els marcadors moleculars SSR i SNP van revelar que la majoria d'aquestes plantes es van obtenir de pol·len 2n del parental diploide tangor. Llavors, el mètode de màxima probabilitat va revelar que tant FDR (Restitució en la Primera Divisió), amb ocurrència predominant, com SDR (Restitució en la Segona Divisió) van ser els mecanismes que condueixen a la formació de gàmetes masculins 2n en aquest tangor. Aquestes observacions van ser confirmades per l'anàlisi de patró de RHP al llarg del cromosoma 2. Des del nostre coneixement, aquest és el primer estudi de progènies de cítrics tetraploides derivats de pol·len no reduït i la primera descripció de la coexistència de dos mecanismes de restitució meiòtiques (FDR i SDR) produint pol·len no reduït en els cítrics. Per tal d'estudiar les freqüències i els mecanismes implicats en la producció de gàmetes 2n sense reduir de la femella, en dos genotips diferents de llimona, vam obtenir 43 híbrids triploides i tetraploides d¿hibridacions sexuals 2x x 2x i 4x x 2x utilitzant `Eureka Frost ' i `Fino' com progenitors femenins. Les freqüències de la producció de gàmetes 2n van ser, respectivament, 4,9% i 8,3%. L'anàlisi de màxima probabilitat i el patró de RHP al llarg del cromosoma 1 van revelar que SDR és el principal mecanisme de gàmetes 2n en la llimona utilitzada com parent femení (88%), seguit pels mecanismes FDR o duplicació pre-meiòtica (PRD) (7%) i la duplicació del genoma post-meiòtica (PMD) (5%). Per primera volta en els cítrics s¿ha obtingut un gran nombre de progènie de llimona a partir de gàmetes 2n i s¿ha identificat un nou mecanisme, el PMD que poques vegades s'ha descrit en altres espècies herbàcies o llenyoses. A través dels dos treballs, hem demostrat, a nivell metodològic, l'eficàcia d'utilitzar dos enfocaments complementaris, és a dir, l'anàlisi del patró de RHP en un cromosoma amb el mètode de màxima probabilitat basat en loci centromèrics per distingir entre els diferents mecanismes de la producció de gàmetes 2n . Es van analitzar els mecanismes meiòtics d'un DD `llima Mèxicana', la recombinació interespecífica i les estructures resultants de gàmetes diploides combinant una anàlisi de segregació de marcadors SSR i SNP, un estudi citogenètic i l'avaluació de la viabilitat del pol·len. Hem arribat a la conclusió que el DD de `llima Mèxicana' tenia una segregació predominantment disómica en tres cromosomes, herència intermèdia amb tendència disómica en cinc cromosomes i els models intermedis per a un altre. Les estructures resultants de gàmetes diploides interespecífiques mostren alta heterozigositat C. medica / C. micrantha. Les estructures genètiques revelades dels gàmetes diploides produïts pel DD de `llima Mèxicana' són compatibles amb la hipòtesi que les varietats triploides `Tahiti¿ i `Tanepao¿ resulten de la hibridació interploide que impliquen un DD de tipus `llima Mèxicana'. Aquesta tendència disómica limita la recombinació i la diversitat de la població de gàmetes diploides, però, la restauració de la viabilitat / Houssem, R. (2017). Genetic Structure of Diploid Gametes for the Production of Triploid Citrus Hybrids [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/90634 / TESIS

Thesis (Ph. D.)--UCLA, 2008. / Vita. Includes bibliographical references (leaves 208-222).

Thesis (MSc.(Anatomy)--Faculty of Health Sciences)-University of Pretoria, 2006. / Includes bibliographical references.

Basement of healthy embryo development comes from quality of oocytes and spermatozoa. Today, when percentage of couples suffering infertility together with assisted reproductive therapy (ART) is increasing, understanding to gamete biology and heritable epigenetic code is crucial. The study is focused on promising epigenome based markers that could serve as indicators of gamete quality for either their screening or selection for ART. Accordingly selected markers were used for the investigation of environmental pollutant bisphenol S (BPS) effect on gametes quality. To obtain these aims, we have used human semen samples, boar semen samples and ICR mice gametes. Samples were analyzed by flow cytometry, immunocytochemistry and western blot analysis. All experimental work was in accordance with Ethics committee University Hospital in Pilsen and approved experimental designs for appropriate experimental animal project. In the study, we detected the dimethylation of histone H3 on lysine K4 (H3K4me2) as potential epigenetic marker of sperm quality and chromatin immaturity. Secondly, we observed the role of the gasotransmitter hydrogen sulphide (H2S) as anti-capacitating agents, slowing down capacitation possibly through post-translational modification of proteins. Thirdly, SIRT1 histone deacetylase was...

Numerical analysis of the lengths and positions of the two basal bodies (BBs), lamellar strip (LS) and anterior mitochondrion (AM) relative to each other in mid‐ and late‐stage spermatids of mosses and liverworts reveals the existence of several well denned, but previously unrecognized, features which clearly distinguish the blepharoplasts of the two groups. The ten possible quotients were calculated from measurements of anterior BB lengths, posterior BB lengths, LS lengths, distances between the anterior tips of the BBs and distances between the transition regions of the BBs in mid‐stage spermatids of 9 mosses and 16 hepatics. These critical data may be quickly compiled from a small number of electron micrographs. A Mann‐Whitney rank order t test showed highly significant differences in 6 of the 10 quotients between the moss and liverwort taxa. The primary data for late‐stage spermatids (4 mosses, 6 liverworts) also included the length of the AM. A Wilcoxon signed rank procedure revealed that the relationship between the AM and other blepharoplast components changed significantly between mid‐ and late‐stage spermatids in mosses but not in liverworts. The clear‐cut numerical differences between the blepharoplast components in each group are related to different patterns of development namely (1) bidirectional assembly of the LS in young spermatids of liverworts versus unidirectional (anterior) elongation at the same stage in mosses (2) elongation of the posterior BB over the nucleus in mid‐stage spermatids of mosses and (3) maturational elongation of the AM in mosses. Since the differences between the blepharoplasts of mosses and liverworts become apparent only during the later stages in ontogeny and since the mode of development of basal body stagger, involving the same precisely defined patterns of proximal triplet microtubule extension, is unique to mosses and liverworts, we suggest that the two groups share a common ancestry. The blepharoplasts of all the taxa used in the calculations are illustrated in a simplified form and the ‘average’ blepharoplast for mid‐ and late‐stage spermatids of both mosses and liverworts is reconstructed from all the data presently available on the two groups. The same analysis of the blepharoplasts of hornworts, birlagellate lycopods, and charophytes highlights the differences between these groups and mosses and liverworts. Most striking is the side‐by‐side orientation of the basal bodies in hornworts and charophytes compared with the staggered arrangement in mosses, liverworts and the lycopods.

Estudos taxonômicos foram conduzidos em onufídeos coletados na Baía do Araçá, em São Sebastião, litoral norte do estado de São Paulo, utilizando dados morfológicos e moleculares, e se confirmou que os espécimes coletados pertencem à espécie Diopatra neapolitana, originalmente descrita para o Golfo de Nápoles (Mar Mediterrâneo). Com a identidade específica dos animais confirmada, foi conduzido um estudo anual, com coletas mensais visando entender o ciclo reprodutivo a partir dos gametas femininos presentes no celoma, concluindo que, os indivíduos brasileiros possuem reprodução assíncrona, com o ciclo reprodutivo regido mais pelo tamanho de cada animal, do que pelos meses do ano. O estudo anual focou também na espermatogênese destes animais, que é caracterizada pela presença do citóforo, comum a outras espécies de poliquetas. Observou-se, entretanto, que a espermiogênese possui dois caminhos possíveis: formação de mórulas espermáticas e maturação do espermatozoide na própria estrutura unida pelo citóforo. Foi estudado o hermafroditismo sequencial desta espécie, caracterizado pelas papilas espermoducais, que para os indivíduos da Baía do Araçá, apresentam diferentes estágios de maturação, podendo representar as passagens de um gênero sexual para outro. Por fim foi estudado o desenvolvimento larval e dispersão de gametas, a partir de fertilizações in vitro, com registro de todos os estágios larvais para a espécie estudada, além da construção de protocolos importantes para a biologia do desenvolvimento / Taxonomic studies were conducted with onuphids collected at Araçá Bay, located in São Sebastião, in the northern coast of the state of São Paulo, using morphological and molecular data, and it is confirmed that these specimens belong to the species Diopatra neapolitana, which type locality is the Gulf of Napoli. After confirming the species identity, an annual study was conducted, with mensal collects of specimens, aiming to understand the reproductive cycle, based on feminine gametes present on each celom, and the conclusion is that for the Brazilian specimen, the reproduction is asynchronous, with the reproductive cycle guided rather by the size of each animal, than by the months. The annual study also focused on the spermatogenesis, which is characterized by the presence of the cytophore, common on other polychaete species, however, the spermiogenesis can be accomplished by two paths: formation of spermatic morulae or maturation of spermatozoa linked in the cytophore structure. The sequential hermaphroditism, characterized by the presence of spermoducal papillae, was also studied, and, for the specimen of the Araçá Bay, there are different maturation stages for this feature, probably related to the transition from one gender to another. Lastly, the larval development and gametes dispersion were studied, using in vitro> fertilizations, registering all larval stages for the present species, as well as the origination of development biology protocols