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1.
Reprod Domest Anim ; 2021 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-33713470

RESUMO

The bovine IGF2 locus is a genomic region with alternative transcripts controlled by five promoters (P0, P1, P2, P3 and P4). As transcriptional regulation can affect messenger RNA (mRNA) stability and translation, and thus, subsequent biological effects, this study evaluated the bovine IGF2 promoter-specific expression patterns in oocytes and pre-implantation embryos produced in vitro by our standard IVP procedures. Immature and matured oocytes, and pre-implantation embryos at the 1-, 2-, 4-, 8- and 16-cell, and at early morula, compact morula, blastocyst and expanded blastocyst stages were collected in three pools of five structures per stage, in four replicates. Total RNA was extracted and subjected to RT-qPCR, using four sets of IGF2 promoter-specific primers covering transcripts driven by promoters P0/P1, P2, P3 and P4, with fragments sequenced for confirmation. Expression of P2- and P4-derived transcripts showed an initial peak between immature (P4) or matured (P2/P4) oocytes and 2-cell embryos, gradually falling until embryo genome activation (EGA), rising again at compaction and cavitation. P0/P1-derived transcripts were identified after EGA, during compaction, whereas P3 activity was not detected at any stage. Our findings suggest that P0/P1 and P2 likely have secondary roles during early stages, whereas P3 may be more relevant later in development. P4 seems to be the main pathway for bovine IGF2 expression during oocyte maturation and embryo development and, therefore, the main target to influence IVP in modulation of embryo growth and in studies in developmental biology.

2.
Nutrients ; 12(1)2020 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-31968556

RESUMO

The pursuit of cholesterol lowering natural products with less side effects is needed for controlling dyslipidemia and reducing the increasing toll of cardiovascular diseases that are associated with morbidity and mortality worldwide. The present study aimed at the examining effects of p-methoxycinnamic acid diesters (PCO-C) from carnauba (Copernicia prunifera)-derived wax on cytotoxic, genotoxic responses in vitro and on dyslipidemia and liver oxidative stress in vivo, utilizing high-fat diet (HFD) chronically fed Swiss mice. In addition, we evaluated the effect of PCO-C on the expression of key cholesterol metabolism-related genes, as well as the structural interactions between PCO-C and lecithin-cholesterol acyl transferase (LCAT) in silico. Oral treatment with PCO-C was able to reduce total serum cholesterol and low-density lipoprotein (LDL) levels following HFD. In addition, PCO-C reduced excessive weight gain and lipid peroxidation, and increased the gene expression of LCAT following HFD. Furthermore, the high affinity of the studied compound (ΔG: -8.78 Kcal/mol) towards the active sites of mutant LCAT owing to hydrophobic and van der Waals interactions was confirmed using bioinformatics. PCO-C showed no evidence of renal and hepatic toxicity, unlike simvastatin, that elevated aspartate aminotransferase (AST) levels, a marker of liver dysfunction. Finally, PCO-C showed no cytotoxicity or genotoxicity towards human peripheral blood lymphocytes in vitro. Our results suggest that PCO-C exerts hypocholesterolemic effects. The safety of PCO-C in the toxicological tests performed and the reports of its beneficial biological effects render this a promising compound for the development of new cholesterol-lowering therapeutics to control dyslipidemia. More work is needed for further elucidating PCO-C role on lipid metabolism to support future clinical studies.


Assuntos
Antioxidantes/farmacologia , Cinamatos/farmacologia , Dieta Hiperlipídica , Dislipidemias/prevenção & controle , Hipolipemiantes/farmacologia , Lipídeos/sangue , Fígado/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/toxicidade , Biomarcadores/sangue , Células Cultivadas , Cinamatos/toxicidade , Modelos Animais de Doenças , Dislipidemias/sangue , Dislipidemias/etiologia , Humanos , Hipolipemiantes/toxicidade , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Linfócitos/metabolismo , Linfócitos/patologia , Masculino , Camundongos , Fosfatidilcolina-Esterol O-Aciltransferase/metabolismo
3.
Animals (Basel) ; 9(11)2019 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-31717727

RESUMO

The aim of this work was to determine the effect of enalapril maleate administration, during oocyte recovery by serial laparoscopic ovum pick-up (LOPU), on the ovarian response and in vitro embryo production (IVP). Twenty cross-bred goats were allocated equally into two groups: Nulliparous and Multiparous. In each group, five animals were selected to receive daily doses of enalapril maleate during the hormonal protocol. Estrus was synchronized by a PGF2α analog, followed 48 h later by insertion of an intravaginal device with progesterone. Forty-eight hours after, a single dose of FSH/eCG was administered. The FSH/eCG doses were repeated three times, on every four day. Oocytes were recovered by LOPU 24 h after each FSH/eCG dose. Viable oocytes were matured in vitro, to be parthenogenetically activated and cultured for 72 h to the cleavage stage. The drug treatment increased the proportion of total follicles observed at LOPU (p < 0.01) in multiparous goats. In both parity groups, enalapril administration had no effect on the proportion or quality of oocytes recovered. Furthermore, the number of embryos cleaved was similar between the groups. Thus, enalapril maleate affected the ovarian response in multiparous animals only and had no effect on the oocyte quality or IVP.

4.
Reprod Domest Anim ; 54(9): 1206-1216, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31240745

RESUMO

The aim of this study was to verify the effect of the energy source for a short-term diet supplementation on follicular dynamics, ovarian response and oocyte recovery in goats. Thirty Anglo Nubian crossbred does received a diet for 4 weeks to satisfy the nutritional requirements of breeding for adult non-dairy goats. Seven days prior to oocyte recovery (OR), a group of does (n = 10) was supplemented with ground full-fat linseed in the diet (Diet A), whereas a second group of does (n = 10) received crude glycerine in the diet (Diet B). The total mixed ration (TMR) diet was maintained as the Control Diet (n = 10). All animals were oestrous-synchronized by the use of a progesterone insert for 12 days prior to OR. Follicles were stimulated by using pFSH (five 40-mg/ml doses) during the supplementation time. At OR, follicles were counted and recovered oocytes were classified as viable or degenerated. Follicular dynamics was monitored by ultrasonography, and plasma glucose, cholesterol and triglyceride levels were measured during supplementation. Glucose was higher in Diet B and cholesterol in Diet A. Diet B had a lower proportion of small (<3 mm) and large follicles (≥3 mm; p = 0.01). The follicular growth rate was higher in Diet A (p < 0.01), with follicles emerging in the 5th day of supplementation. No differences were observed for follicles counted and oocytes recovered. Thus, the type of energy source supplemented for a short term was capable to alter the follicular dynamics, without affecting the proportion of morphologically viable oocytes upon recovery.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Dieta/veterinária , Oócitos/fisiologia , Folículo Ovariano/crescimento & desenvolvimento , Ração Animal/análise , Animais , Glicemia/análise , Colesterol/sangue , Feminino , Linho , Hormônio Foliculoestimulante/administração & dosagem , Glicerol , Cabras , Recuperação de Oócitos/veterinária , Progesterona/administração & dosagem , Triglicerídeos/sangue
5.
Reprod Domest Anim ; 54(3): 480-485, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30444528

RESUMO

This study aimed to examine the in vitro culture of secondary preantral follicles, using reused ovaries, to compare both the 2D and 3D methods of in vitro culture of preantral follicles, and the system of medium replacement. Twenty-five pairs of ovaries from mixed-breed goats were used for the experiment. Follicular puncture of antral follicles was performed for in vitro production. After this procedure, the secondary preantral follicles were submitted to a microdissection procedure. The isolated preantral follicles were randomly divided into three treatments: (a) Two-dimensional culture with partial replacement of medium during culture (2D PR), (b) Three-dimensional culture with addition of medium during culture (3D AD) and (c) Three-dimensional culture with partial replacement of medium (3D PR). The culture period was 18 days. All treatments at the end of the in vitro culture period (18 days) presented a follicular survival rate which ranged from 59% to 70%, demonstrating that it was possible to perform an experiment with preantral follicles using ovaries that had previously been used in another reproductive biotechnique. The 3D AD treatment showed a survival percentage and follicular diameter higher than the 2D PR treatment, however, it did not differ from the 3D PR treatment. In conclusion, experiments employing the use of preantral follicles can be performed with success after the ovaries have been used for experiments with antral follicles. Moreover, the three-dimensional system with the addition of medium is recommended for in vitro culture of preantral follicles, since this system is more practical and financially feasible.


Assuntos
Folículo Ovariano/crescimento & desenvolvimento , Ovário/fisiologia , Técnicas de Cultura de Tecidos/veterinária , Animais , Meios de Cultura , Feminino , Hormônio Foliculoestimulante/farmacologia , Cabras , Oócitos/metabolismo , Reprodução , Técnicas de Cultura de Tecidos/métodos
6.
Ciênc. rural (Online) ; 49(3): e20180340, 2019. tab, graf
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1045314

RESUMO

ABSTRACT: The effect of insulin administration on the productive responses of Saanen goats during early lactation was investigated. Ten of 20 adult females were subjected to subcutaneous administration of intermediate-acting insulin (0.14UI/kg body weight) at 2, 9, and 14 days postpartum. Milk yield was measured twice daily for 13 weeks and milk samples were collected to measure protein and fat contents. Plasma levels of progesterone, insulin, non-esterifies fatty acids, glucose and other metabolites were measured. Results showed a significantly increased effect of insulin treatment on the content of milk fat and protein; moreover, milk production in the first and second postpartum weeks were higher than control group. The peak of lactation in the insulin group was achieved one week earlier in comparison to the control group. In addition, the milk production rate showed lower persistency (milk yield 13 week/milk yield at peak) in the same group. During the first four weeks of postpartum, treated animals showed greater weight loss and higher non-esterified fatty acid concentration, whereas no effect was observed on the concentration of progesterone and other metabolites. The above results indicated that repeated administration of insulin in dairy goats during early lactation increase yield and qualitative components of milk, but has substantial consequences on animal productive rate and metabolic response.


RESUMO: O objetivo do estudo foi avaliar o efeito da administração de insulina sobre a resposta produtiva de cabras Saanen durante a lactação inicial. Dez de vinte fêmeas adultas foram sujeitas à administração subcutânea de repetidas e baixas doses de insulina de liberação intermediária aos 2, 9 e 14 dias pós-parto. A produção de leite foi mensurada duas vezes ao dia, por 13 semanas, e amostras de leite foram coletadas para mensurar teores de proteína e gordura. Os níveis plasmáticos de progesterona, insulina, ácidos graxos não-esterificados (AGNE), glicose e outros metabólitos foram mensurados. Os resultados mostraram um efeito significativamente maior nos animais tratados com insulina sobre o teor de gordura e proteína no leite. Além disso, a produção de leite na primeira e segunda semana pós-parto foi maior no grupo tratado do que no grupo controle. O pico de lactação no grupo insulina foi alcançado uma semana antes em comparação ao grupo controle. Além disso, a taxa de produção de leite nos animais tratados mostrou uma menor persistência de produção de leite durante o período analisado. Durante as primeiras quatro semanas pós-parto, os animais tratados com insulina mostraram maior perda de peso e maior concentração de AGNE, enquanto não se observou nenhum efeito sobre a concentração de progesterona ou outros metabólitos. Os resultados acima indicam que repetidas doses de insulina em cabras leiteiras durante a lactação inicial aumenta o rendimento de produção e concentração de componentes qualitativos do leite, mas apresenta consequências consideráveis sobre taxa de produção animal e resposta metabólica.

7.
Diabetes Metab Syndr Obes ; 11: 289-301, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29942143

RESUMO

Purpose: Type 1 diabetes mellitus (DM1) is one of the most common chronic diseases observed during childhood. The incidence of DM1 is increasing worldwide, and there is currently no way to prevent or delay the onset or to cure the disease. Most diseases, including diabetes, stem from abnormalities in the functioning of proteins, and some studies have reported the expression of protein variation to be involved in the development of DM1. Thus, the aim of this study was to investigate the differential expression of serum proteins in patients with DM1. Materials and methods: Serum of patients with DM1 (n=30) and healthy controls (n=30) was collected. A proteomic approach was used with depletion of albumin and immunoglobulin G chromatography on serum samples followed by data-independent, label-free mass spectrometric analysis. Results: A total of eight serum proteins were identified as being differentially expressed and involved in the immune system, lipid metabolism, and pathways of coagulation. DM1 was associated with the upregulation of six proteins: alpha-2-macroglobulin, apolipoprotein A-II, ß2 glycoprotein I, Ig alpha-2 chain C region, alpha-1-microglobulin, and prothrombin. A total of two proteins were downregulated, including pregnancy zone protein and complement C4. Conclusion: To the best of our knowledge, these findings show differential expression of proteins revealing new proteins that may be involved in the development and progression of diabetes.

8.
Reprod Domest Anim ; 53(5): 1085-1095, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29885005

RESUMO

This study was conducted to evaluate the effects of different feeding levels on the proteome of oviduct and uterus tissues of hormonally stimulated goats during the periovulatory period. Forty goats were separated into four different diet groups: Diet 1.0 M (n = 11), Diet 1.3 M (n = 10), Diet 1.6 M (n = 9), Diet 1.9 M (n = 10), fed with 1.0, 1.3, 1.6 and 1.9 times live weight maintenance, respectively. After four weeks of treatment, six hormonally stimulated females per treatment group were randomly selected for collection of uterine and the oviduct tissue samples. Samples were collected after animals were slaughtered in a commercial unit. Feeding goats with 1.3 to 1.9 times more nutrients than a control group directly influenced the proteome of the oviduct and uterus, altering the expression of proteins that participate in biological processes such as apoptosis, antioxidant, and immunological activities. These events are crucial for fertilization and early embryonic survival. Expression of oviduct proteins such as Tubulin Beta 2B, Transferrin and Disulphide-isomerase A3 increased in the 1.9 M group in relation to the other feeding levels. Disulphide-isomerase A4 showed higher expression in the 1.0 M group compared to diets with higher energetic levels. As energy intake increased in the diets, there was higher expression of Alpha-1-antitrypsin and downregulation of Profilin-1 in the uterus of the goats. In conclusion, this study showed that specific proteins of the goat oviduct and uterus expressed during the periovulatory period are modified as the result of nutritional balance.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Dieta/veterinária , Oviductos/fisiologia , Ovulação/fisiologia , Proteoma/fisiologia , Ração Animal , Animais , Ingestão de Energia , Feminino , Cabras/fisiologia , Útero/fisiologia
10.
Artigo em Inglês | MEDLINE | ID: mdl-29081820

RESUMO

The present objective of the investigation is to evaluate the antihypercholesterolemic activity of the aqueous fruit pulp extract (APE) of Copernicia prunifera (Miller) H. E. Moore (Arecaceae family). Various chemical characterization methods like thin layer chromatography, Fourier transform infrared spectroscopy, 1H and 13C NMR, and molecular weight by gel permeation chromatography have been employed to characterize the extracted pectin. The present study demonstrated that hypercholesterolemic diet (HD) created hypercholesterolemia, caused significant increases in body weight, total cholesterol, and low-density lipoprotein, and caused decreases in high-density lipoprotein in serum compared with SD group. Two doses (APE 150 and 300 mg/Kg b.w./day) were administered to hyperlipidemic mice for 90 days. APE reversed body weight changes, changed serum lipids to normal values, and significantly inhibited the changes of lipid peroxidation and inflammation in the liver tissues. The renal parameters analyzed (urea and creatinine) altered by diet were reverted to normal values. Our results revealed that aqueous fruit pulp extracts of carnauba reduced hypercholesterolemia showing a potential preventive effect against cardiovascular diseases without side effects cause.

11.
Zygote ; 25(5): 567-574, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28835303

RESUMO

The objective of this study was to assess the influence of nutritional regimens such as adequate feeding, restricted feeding, and underfeeding-refeeding on the follicle growth and development from caprine ovaries. Goats were divided into three different groups (n = 5 per group). For 24 weeks, goats received elephant grass plus concentrate to provide 1.5 (n = 5) and 0.72 (n = 10) times the energy requirements for maintenance of live weight. Underfed goats were subsequently refed for 6 weeks with the diet of the nourished group (1.5 times the energetic requirements of maintenance). Follicular morphology and morphometry, as well as granulosa cells mitotic index were assessed. Ovarian follicles were classified as small or large preantral follicles, or as small or large antral follicles. Ovarian volume was smaller in animals from both underfed and refed groups than in those animals from fed group. Although no difference in the total number of normal follicles was observed among the nutritional groups, underfed animals presented higher percentages of atretic preantral and small antral follicles when compared with fed animals. Large antral follicles from underfed and refed goats presented a lower mitotic index when compared with fed ones. In conclusion, ovaries from goats challenged with prolonged undernutrition will be functionally compromised, which is characterized by atresia of preantral and small antral follicles and decreased mitotic index of large antral follicles. Refeeding those animals will not recover ovarian function to a same level experienced by goats fed a diet with adequate energy requirements.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal/fisiologia , Cabras/fisiologia , Índice Mitótico , Folículo Ovariano/fisiologia , Ração Animal , Animais , Contagem de Células , Feminino , Células da Granulosa/fisiologia , Folículo Ovariano/citologia , Ovário/citologia , Ovário/fisiologia
12.
Cell Reprogram ; 18(4): 264-79, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27362734

RESUMO

Cloning by somatic cell nuclear transfer (SCNT) is characterized by low efficiency and the occurrence of developmental abnormalities, which are rather poorly studied phenomena in goats. This study aimed at comparing overall SCNT efficiency in goats by using in vitro-matured (IVM) or in vivo-matured oocytes and fibroblast donor cells (mock transfected, transgenic, or wild type), also characterizing symptoms of the Abnormal Offspring Syndrome (AOS) in development, comparing results with pregnancies produced by artificial insemination (AI) and in vivo-derived (IVD) embryos. The SCNT group had lower pregnancy rate (18.3%, 11/60), total number of concepti (20.0%, 12/60), term births (3.3%, 2/60), and live births (1.7%, 1/60) than both the IVD (77.8%, 7/9; 155.5%, 14/9; 122.2%, 11/9; 88.8%, 8/9) and the AI (71.4%, 10/14; 121.4%, 17/14; 100%, 14/14; 78.5%, 11/14) groups, respectively (p < 0.05). No SCNT pregnancies reached term using IVM oocytes, but in vivo-matured oocytes resulted in two term transgenic cloned kids. The proportion fetal membrane (FM) weight/birth weight reflected an increase in FM size and cotyledonary enlargement in clones, for disproportionally bigger newborns in relation to cotyledonary numbers. Overall, goat cloning showed losses and abnormality patterns similar to the AOS in cloned cattle and sheep, which have not been previously well recognized in goats.


Assuntos
Animais Geneticamente Modificados/crescimento & desenvolvimento , Transferência Embrionária/veterinária , Desenvolvimento Embrionário , Fibroblastos/citologia , Técnicas de Transferência Nuclear/veterinária , Oócitos/citologia , Animais , Animais Geneticamente Modificados/genética , Animais Recém-Nascidos , Feminino , Fibroblastos/metabolismo , Cabras , Oócitos/metabolismo , Gravidez , Taxa de Gravidez , Nascimento a Termo
13.
Int J Biometeorol ; 58(7): 1559-67, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24240541

RESUMO

The aim of this work was to determine the adaptability of Saanen and ½Saanen × ½Anglo-Nubian (½S½AN) goats bred in tropical climates. The study included 30 goats, 15 Saanen and 15 ½S½AN. The data was collected during the rainy and dry seasons. During the whole experimental period, the environment variables were recorded, as well as rectal temperature (RT), superficial temperature (ST), respiratory rate (RR) and heart rate (HR) and milk production (MP). The adaptability coefficient (AC) was calculated for both genotypes. The averages were evaluated by ANOVA at 5% probability. There was a genotype and period of year effect, as well as the interaction genotype × period of year. Pearson's simple correlation analysis was then carried out between milk production and physiological and environment variables. There was a statistical difference (p < 0.05) between the seasons for RT, ST and RR. RT, RR and HR were lower for ½S½AN than Saanen goats, regardless of the season. MP was greater in the dry season (p < 0.05) (2.52 ± 0.50 kg/day for ½S½AN and 2.41 ± 0.38 kg/day for Saanen) than the rainy season (2.17 ± 0.27 kg/day for ½S½AN and 2.28 ± 0.53 kg/day for Saanen). The MP correlations were very significant (p < 0.05), however low and negative, where it was higher when correlated with RR in Saanen goats. Under the conditions of the present study, it is concluded that the goats were influenced by climatic factors, where the rainy period was more likely to cause thermal stress in the animals.


Assuntos
Cabras/fisiologia , Lactação , Adaptação Fisiológica , Animais , Temperatura Corporal , Brasil , Feminino , Frequência Cardíaca , Umidade , Chuva , Taxa Respiratória , Temperatura , Clima Tropical
14.
J Vet Sci ; 12(2): 127-32, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21586871

RESUMO

The present study aimed at analyzing the genetic variability of indigenous goat breeds (Capra hircus) using the MHC-associated microsatellite markers BF1, BM1818, BM1258, DYMS1, and SMHCC1. The following breeds were included: Chinese Xuhuai, Indian Changthangi and Pashmina, Kenyan Small East African (SEA) and Galla, and Albanian Vendi. To examine genetic variability, the levels of heterozigosity, degrees of inbreeding, and genetic differences among the breeds were analyzed. The mean number of alleles ranged from nine in the Galla to 14.5 in the Vendi breed. The mean observed heterozygosity and mean expected heterozygosity varied from 0.483 in the Vendi to 0.577 in the Galla breed, and from 0.767 in the SEA to 0.879 in the Vendi breed, respectively. Significant loss of heterozygosity (p < 0.01) indicated that these loci were not in Hardy-Weinberg equilibrium. The mean F(IS) values ranged from 0.3299 in the SEA to 0.4605 in the Vendi breed with a mean value of 0.3623 in all breeds (p < 0.001). Analysis of molecular variance indicated that 7.14% and 4.74% genetic variation existed among the different breeds and geographic groups, whereas 92.86% and 95.26% existed in the breeds and the geographic groups, respectively (p < 0.001). The microsatellite marker analysis disclosed a high degree of genetic polymorphism. Loss of heterozygosity could be due to genetic drift and endogamy. The genetic variation among populations and geographic groups does not indicate a correlation of genetic differences with geographic distance.


Assuntos
Cabras/genética , Complexo Principal de Histocompatibilidade/genética , Repetições de Microssatélites , Alelos , Animais , DNA/genética , Feminino , Variação Genética , Masculino , Reação em Cadeia da Polimerase/veterinária , Polimorfismo Genético
15.
Zygote ; 18(1): 89-92, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19586559

RESUMO

Ovarian cortical fragments from five adult ewes were in vitro cultured for 1, 3 or 5 days in the presence of minimum essential medium either supplemented or not by follicle-stimulating hormone (FSH) (100 ng/ml) or indole-3-acetic acid (IAA) (10, 20, 40 or 100 ng/ml), alone or in combination. After in vitro culture, ovarian fragments were submitted to follicular isolation and viability test was performed using trypan blue. Addition of IAA (10 ng/ml) to a free-FSH medium resulted in the highest percentages of viable follicles, but was progressively deleterious in higher concentrations (20, 40 and 100 ng/ml) if in absence of FSH. Follicular development was observed only when FSH was added to an IAA-free medium. In conclusion, IAA at a concentration of 10 ng/ml increases follicular survival in vitro. However, at high concentrations (20, 40 or 100 ng/ml), this auxin may be deleterious to preantral follicles, the addition of FSH to the medium being necessary.


Assuntos
Hormônio Foliculoestimulante/farmacologia , Ácidos Indolacéticos/farmacologia , Folículo Ovariano/efeitos dos fármacos , Animais , Sobrevivência Celular , Feminino , Folículo Ovariano/citologia , Ovinos , Técnicas de Cultura de Tecidos
16.
Fertil Steril ; 85 Suppl 1: 1077-81, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16616077

RESUMO

OBJECTIVE: To determine the behavior of isolated primordial follicles that were exposed to different concentrations of dimethyl sulfoxide (DMSO), ethylene glycol (EG), propylene glycol (PROH), and glycerol (GLY). DESIGN: Isolated primordial follicles were exposed to the cryoprotectant (CPA) solution and photographed to calculate their volume at different periods of exposure. SETTING: Laboratorio Renzo Giuliani, University of Florence, Italy. ANIMAL(S): Lambs, 30-40 days old. INTERVENTION(S): Isolation of primordial follicles and subsequent exposure to CPA. MAIN OUTCOME MEASURE(S): Follicular volume. RESULT(S): At 2 minutes of CPA exposure, all follicles appeared to be shrunken. At approximately 5 minutes, shrinkage ceased, and follicles started to swell, absorbing the CPA and water to maintain osmotic equilibrium. When DMSO was tested, follicular dehydration in all concentrations did not exceed 17%; with PROH and EG, it reached 33% and 27%, respectively. The highest degree of dehydration (48%) was seen with GLY. In almost all tested concentrations, follicular shrinkage occurred up to 5 minutes. CONCLUSION(S): Volume changes in isolated primordial follicles can fluctuate according to the CPA used and its concentration.


Assuntos
Permeabilidade da Membrana Celular , Crioprotetores/farmacocinética , Soluções para Preservação de Órgãos/farmacocinética , Folículo Ovariano/fisiologia , Animais , Permeabilidade da Membrana Celular/efeitos dos fármacos , Crioprotetores/administração & dosagem , Citoproteção/fisiologia , Relação Dose-Resposta a Droga , Feminino , Soluções para Preservação de Órgãos/administração & dosagem , Permeabilidade/efeitos dos fármacos , Ovinos
17.
Fertil Steril ; 81 Suppl 1: 735-40, 2004 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15019803

RESUMO

OBJECTIVE: To verify the viability of isolated primordial follicles to different propylene glycol (PROH) and glycerol (GLY) concentrations before and after cryopreservation. DESIGN: Isolated primordial follicles were stained with trypan blue to evaluate the effect of different PROH and GLY concentrations before and after cryopreservation. SETTING: Laboratorio Renzo Giuliani, University of Florence, Italy. PATIENT(S): Thirty- to forty-day-old lambs. INTERVENTION(S): : Isolation of primordial follicles with subsequent exposure to cryoprotectant and freezing. MAIN OUTCOME MEASURE(S): Histologic structure and follicular mortality. RESULT(S): After the isolation procedure (control), the mean number of live primordial follicles/mL was 2,688 and 4,452 in the GLY and PROH groups, respectively. When GLY was used, the number of live follicles before cryopreservation was 820, 756, 640, 524, 564, and 460 follicles/mL with concentrations of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 mol/L, respectively. After cryopreservation, this number decreased to 0, 12, 36, 100, 84, and 68 follicles/mL, respectively, with the same concentrations. When PROH was used, the number of live follicles before cryopreservation was 4,216, 3,880, 3,560, 1,812, 704, and 568 follicles/mL with concentrations of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 mol/L, respectively. After cryopreservation, this number decreased to 0, 116, 336, 472, 360, and 244 follicles/mL, respectively, with the same concentrations. CONCLUSION(S): Both cryoprotectants were shown to preserve isolated primordial follicles after cryopreservation.


Assuntos
Criopreservação , Crioprotetores/administração & dosagem , Glicerol/administração & dosagem , Folículo Ovariano/efeitos dos fármacos , Propilenoglicol/administração & dosagem , Animais , Relação Dose-Resposta a Droga , Feminino , Folículo Ovariano/anatomia & histologia , Folículo Ovariano/fisiologia , Ovinos , Sobrevivência de Tecidos/efeitos dos fármacos , Testes de Toxicidade
18.
Theriogenology ; 60(4): 735-42, 2003 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-12832021

RESUMO

Cryopreservation of primordial follicles represents an opportunity to preserve female gametes, and consequently to protect the reproductive capacity of humans and animals, as well as to safeguard genetic material from endangered animal species or rare breeds. The aim of this work was to assess the toxicity of different concentrations of ethylene glycol (EG) to primordial follicles, and verify the viability of these follicles after the freezing-thawing procedure. Primordial follicles were isolated from ovine ovaries and exposed to different EG concentrations to evaluate the cryoprotectant (CPA) toxicity before and after cryopreservation. After isolation of primordial follicle (control), the number (mean+/-S.E.M.) of viable primordial follicles/ml was 3764+/-795.21. The number of viable follicles in the toxicity test using EG at 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M was 1096+/-447.9, 960+/-446.67, 948+/-366.14, 832+/-313.59, 856+/-280.67, and 700+/-255.02, respectively. The number of viable follicles at concentrations of 2.5 M was less than for controls. After cryopreservation, the numbers decreased to 0+/-0, 148+/-85.46, 764+/-246.69, 824+/-291.9, 844+/-296.27, and 588+/-200.65, respectively for 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M EG. The number of viable follicles at 0, 0.5, and 2.5 M was less than for controls. In conclusion, after the freezing and thawing procedure, concentrations of 1.0, 1.5, and 2.0 M EG can be successfully used for the cryopreservation of isolated follicles in sheep.


Assuntos
Criopreservação/veterinária , Crioprotetores/administração & dosagem , Etilenoglicol/administração & dosagem , Folículo Ovariano/fisiologia , Ovinos , Animais , Crioprotetores/toxicidade , Etilenoglicol/toxicidade , Feminino , Folículo Ovariano/anatomia & histologia
19.
Fertil Steril ; 79 Suppl 1: 682-6, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12620477

RESUMO

OBJECTIVE: To verify the viability of isolated primordial follicles after exposure to different concentrations of dimethyl sulfoxide (DMSO) and after cryopreservation. DESIGN: Randomized control trial. SETTING: Laboratorio Renzo Giuliani, University of Florence, Italy. ANIMAL(S): Thirty- to 40-day-old lambs. MAIN OUTCOME MEASURE(S): Isolated primordial follicles were stained with trypan blue to evaluate the effect of different DMSO concentrations before and after the cryopreservation. Histological structure and follicular mortality were evaluated. RESULT(S): After the isolation procedure (control), a mean (+/-SE) of 800 +/- 203.86 live primordial follicles/mL were obtained. The number of live follicles in the toxicity test using the DMSO at 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M was 782 +/- 193.96, 754 +/- 172.03, 778 +/- 191.58, 736 +/- 191.92, 476 +/- 122.9, and 316 +/- 83.52, respectively. The number of live follicles at 2.5 M was lower than that in the control procedure. After cryopreservation, the numbers decreased to 0 +/- 0, 232 +/- 44.20, 636 +/- 161.82, 628 +/- 181.28, 208 +/- 11.57, and 184 +/- 47.07, respectively at 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M. The number of live follicles at 0, 0.5, 2.0, and 2.5 M were lower than that in the control procedure. CONCLUSION(S): After cryopreservation, only DMSO concentrations of 1.0 and 1.5 M showed a number of live follicles similar to that of the control procedure.


Assuntos
Criopreservação/veterinária , Crioprotetores/toxicidade , Dimetil Sulfóxido/toxicidade , Folículo Ovariano/efeitos dos fármacos , Ovinos/fisiologia , Animais , Sobrevivência Celular/fisiologia , Corantes/metabolismo , Criopreservação/métodos , Feminino , Azul Tripano/metabolismo
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