Criopreservação de sêmen de epidídimo de garanhões

It is a fact that Brasil has a featured position in the equine market due to the size and the high quality of its squad. To keep the competitivity, the reproduction biotechnologies have a big and important role. The cryopreservation of stallion semen, for example, generates innumerous advantages, which includes: larger number of obtained doses, storing genetic material for undetermined time, using the semen of a good stallion even after its dead or any other event that makes both semen collection and mating impossible, etc. In this context, the cryopreservation of epididymal sperm, has been seen as a promising technique in equine reproduction. The epididymal cauda has a significant quantity of fertile spermatozoa and this ensures a huge store of cells in cases of unexpected accidents, which can early interrupt with the reproductive life of a stallion. Lots of studies are being developed using the cryopreservation of stallion epididymal sperm. These studies permit the propagation of high quality genetic material and make possible for the stallion owner to opt for a final semen collect in unexpected circumstances

Principais marcadores utilizados na avaliação da qualidade do sêmen de animais domésticos

Methods of semen cryopreservation allow changes in spermatic cells, such as damage in plasma and acrossomal membrane and modifications in mitochondrial function due to a disorder in the lipidic bilayer. For effective oocyte fertilization, spermatozoa require functional competent membranes, and intact organelles, acrosome and DNA. However, most laboratory methods used to evaluate semen quality are not highly correlated with fertilizing capacity. The discovery of a variety of fluorochromes and compounds conjugated to fluorescent probes has enabled an accurate assessment of the viability, integrity and function of spermatozoa. Among the most used probes that label the various compartments of the sperm cell there are the membrane impermeable fluorescent dyes to test the membrane integrity, as well as acylated dyes that pass the intact membrane. For the acrossomal integrity the most commonly used method is lectins labeled by a fluorescent probe. The acrosome reaction and spermatic capacitation is detected by the evaluation of membrane architecture and disorder of lipids in plasma membrane. Mitochondrial function can be determined using markers for their aerobic activity. The DNA status of spermatozoa has been determined using the metachromatic properties of Acridine Orange, and the DNA fragmentation can also be assessed by TUNEL assay. Finally, DNA condensation is analyzed using a single cell DNA gel electrophoresis assay that indicates DNA compactation. This monograph aims to compile the various tests used to detect damaged spermatozoa under cryopreservation methods, searching for improve the predictive value of semen analysis with the intention of a successful conception

Correlação entre aneuploidia cromossômica em espermatozóides e taxa de implantação embrionária: subtitulo

Infertility is directly related to chromosomal abnormalities in germ cells. Among them, the aneuploidies are the most frequent chromosomal abnormalities and responsible for embryo implantation failures, miscarriages, fetal losses and newborns with congenital malformations, mental disability and neuropsychomotor developmental delay. Male patients with normal somatic karyotype may present different rates of aneuploidies in sperm, resulting in abnormal embryos. This study aimed to correlate the frequency of chromosomal aneuploidies in spermatozoa with embryo implantation rate in couples undergoing assisted reproductive techniques. The methodology has included chromosomal analysis by GTG banding and molecular cytogenetic study using Fluorescent In Situ Hybridization technique for evaluation of chromosomes 9, X and Y in germ cells of 22 patients referred to the Human Reproduction Service of the Clinical Hospital FMRP-USP. Embryo implantation rates were determined by hormonal evaluation in maternal peripheral blood and ultrasound confirmation. Two patients presented abnormal karyotype, characterized by polymorphism of the heterochromatic region of the long arm of chromosome 9 and a satellite in the short arm of chromosome 22. Both alterations, usually considered variants of normality, have been related to infertility phenotype and miscarriages. Significant differences were detected between couples who presented pregnancy (group 1) and couples with embryo implantation failure (group 2), with higher frequency of aneusomy and diploidy of chromosome 9, as well as total aneuploidy in sperm of group 2 patients. Our results suggest a correlation between aneuploidy and embryo implantation rates, since the infertile group with reproductive failure has showed higher frequency of aneuploidy. Screening for aneuploidies detection in male germ cells should be included in order to decrease embryo implantation failures, miscarriages and fetuses with chromosomal ...

Morfometria e compactação da cromatina espermática de touros Nelore de acordo com a idade e sua influência na produção de embriões in vitro

Pós-graduação em Medicina Veterinária - FCAV

Imunocastração e ractopamina na dieta de suínos: implicações na qualidade de carne fresca e melhorada por injeção (enhancement)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Microscopic Evidence of the Opportunistic Reproductive Strategy and Early Sexual Maturation of the Small-Sized Characin Knodus moenkhausii (Characidae, Pisces)

We investigated the reproductive biology of Knodus moenkhausii, an abundantsmall-sized characin fish with broad occurrence in the Parana River basin, Bra-zil. Specimens were collected monthly to determine fecundity, length at firstmaturity, reproductive period and spawning type. Gonads were macroscopically classified according to their form, size and texture in three different stages(immature, maturing or mature). Histological procedures were conducted toconfirm gonadal developmental stages, and it was possible to notice thatmaturing females actually presented atretic oocytes, and all males that weremacroscopically classified as immature, maturing and mature actually presentedabundant spermatozoa in their gonads. Because of these discrepancies, a reclas-sification of gonadal maturations stages was needed after histological analysis,reinforcing its importance to studies on the reproduction of small characins.Reproduction occurred throughout the year though with two peaks. The lengthof the smallest mature individuals was 13 mm SL for males and 24 mm SL for females. Despite presenting relatively small batch fecundity, some life historytraits such as early reproduction, multiple spawning throughout the year, inassociation with known opportunistic feeding habits, explain the high abun-dance of this species in locations where it occurs.

Sexual maturation of the Mongolian gerbil (Meriones unguiculatus): a histological, hormonal and spermatic evaluation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Centrifugação de espermatozóides ovinos em gradiente de densidade contínuo: efeito sobre a taxa de prenhez e proporção do sexo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Impaired protamination and sperm DNA damage in a Nellore bull with high percentages of morphological sperm defects in comparison to normospermic bulls

The routine semen evaluation assessing sperm concentration, motility and morphology, does not identify subtle defects in sperm chromatin architecture. Bulls appear to have stable chromatin, with low levels of DNA fragmentation. However, the nature of fragmentation and its impact on fertility remain unclear and there are no detailed reports characterizing the DNA organization and damage in this species. The intensive genetic selection, the use of artificial insemination and in vitro embryo production associated to the cryopreservation process can contribute to the chromatin damage and highlights the importance of sperm DNA integrity for the success of these technologies. Frozen-thawed semen samples from three ejaculates from a Nellore bull showed high levels of morphological sperm abnormalities (55.8±5.1%), and were selected for complementary tests. Damage of acrosomal (76.9±8.9%) and plasma membranes (75.7±9.3%) as well as sperm DNA strand breaks (13.8±9.5%) and protamination deficiency (3.7±0.6%) were significantly higher compared to the values measured in the semen of five Nellore bulls with normospermia (24.3±3.3%; 24.5±6.1%; 0.6±0.5%; 0.4±0.6% for acrosome, plasma membrane, DNA breaks and protamine deficiency, respectively) (P<0.05). Motility and percentage of spermatozoa with low mitochondrial potential showed no differences between groups. This study shows how routine semen analyses (in this case morphology) may point to the length and complexity of sperm cell damage emphasizing the importance of sperm function testing.

Endocrine and paracrine regulation of zebrafish spermatogenesis: the Sertoli cell perspective

Spermatogonial stem cells (SSCs) either self-renew or differentiate into spermatogonia that further develop into spermatozoa. Self-renewal occurs when residing in a specific micro-environment (niche) while displacement from the niche would tip the signalling balance towards differentiation. Considering the cystic type of spermatogenesis in fish, the SSC candidates are single type A undifferentiated (A(und)) spermatogonia, enveloped by mostly one niche-forming Sertoli cell. When going through a self-renewal cell cycle, the resulting new single type Aund spermatogonium would have to recruit another Sertoli cell to expand the niche space, while a differentiating germ cell cyle would result in a pair of spermatogonia that remain in contact with their cyst-forming Sertoli cells. In zebrafish, thyroid hormone stimulates the proliferation of Sertoli cells and of type Aund spermatogonia, involving Igf3, a new member of the Igf family. In cystic spermatogenesis, type Aund spermatogonia usually do not leave the niche, so that supposedly the signalling in the niche changes when switching from self-renewal to differentiation. and rzAmh inhibited differentiation of type A(und) spermatogonia as well as Fsh-stimulated steroidogenesis. Thus, for Fsh to efficiently stimulate testis functions, Amh bioactivity should be dampened. We also discovered that Fsh increased Sertoli cell Igf3 gene and protein expression; rzIgf3 stimulated spermatogonial proliferation and Fsh-stimulated spermatogenesis was significantly impaired by inhibiting Igf receptor signaling. We propose that in zebrafish, Fsh is the major regulator of testis functions and, supported by other endocrine systems (e.g. thyroid hormone), regulates Leydig cell steroidogenesis as well as Sertoli cell number and growth factor production to promote spermatogenesis.

Tests of fed and unfed Amblyomma cajennense ticks (Acari: Ixodidae): first morphological data

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Impact of the processes of total testicular regression and recrudescence on the epididymal physiology of the bat myotis nigricans (chiroptera: vespertilionidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anti-androgenic activities of diuron and its metabolites in male Nile tilapia (Oreochromis niloticus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Spermatozoon ultrastructure and semen parameters of Brycon vermelha (Characiformes, Characidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Sperm production of Steidachneridion parahybae (Steindachner 1877) and the effect of hormonal induction throughout one reproductive cycle

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)