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1.
Artigo em Inglês | LILACS-Express | VETINDEX | ID: biblio-1457665

Resumo

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened prim

2.
Acta sci. vet. (Impr.) ; 41: 01-06, 2013.
Artigo em Inglês | LILACS-Express | VETINDEX | ID: biblio-1457079

Resumo

Background: The main advantage of the cryopreservation of ovarian fragments is a thinner tissue, which facilitates the penetration of cryoprotective agents, but the size of tissue may not be a limiting factor in achieving a successful cryopreservation of the ovarian tissue. This information is highly significant considering that the cryopreservation of hemi-ovary or whole ovary may preserve the entire or major part of the contingent of primordial follicles of ovarian fragments. Therefore, the aim of this study was to evaluate the vitrification of different dimensions goat ovarian tissue on the follicular morphology, viability, diameter, and the stromal cell density. Materials, Methods & Results: The ovarian tissue was vitrified as fragment, hemi-ovary, or whole ovary, and after warming, the preantral follicles were examined by trypan blue dye exclusion test and histological analysis. Preantral follicles incubated with trypan blue were considered viable if the oocyte and granulosa cells remained unstained. Preantral follicles were classified as morphologically normal only when they contained intact oocyte and granulosa cells. The follicular diameter was measured considering the major and minor axes of each follicle; the average of these 2 measurements was used to determine the diameter of each follicle. Ovarian stroma cells density was evaluated by calculating the number of


Background: The main advantage of the cryopreservation of ovarian fragments is a thinner tissue, which facilitates the penetration of cryoprotective agents, but the size of tissue may not be a limiting factor in achieving a successful cryopreservation of the ovarian tissue. This information is highly significant considering that the cryopreservation of hemi-ovary or whole ovary may preserve the entire or major part of the contingent of primordial follicles of ovarian fragments. Therefore, the aim of this study was to evaluate the vitrification of different dimensions goat ovarian tissue on the follicular morphology, viability, diameter, and the stromal cell density. Materials, Methods & Results: The ovarian tissue was vitrified as fragment, hemi-ovary, or whole ovary, and after warming, the preantral follicles were examined by trypan blue dye exclusion test and histological analysis. Preantral follicles incubated with trypan blue were considered viable if the oocyte and granulosa cells remained unstained. Preantral follicles were classified as morphologically normal only when they contained intact oocyte and granulosa cells. The follicular diameter was measured considering the major and minor axes of each follicle; the average of these 2 measurements was used to determine the diameter of each follicle. Ovarian stroma cells density was evaluated by calculating the number of

3.
Acta sci. vet. (Impr.) ; 41: 01-12, 2013.
Artigo em Português | LILACS-Express | VETINDEX | ID: biblio-1457105

Resumo

Background: The anti-müllerian hormone (AMH) is a member of the transforming growth factor (TGF-) superfamily, which exerts important functions on local regulation of folliculogenesis. Although in vivo and in vitro studies suggest that AMH affects the primordial follicle assembly and activation, as well as the responsiveness of growing follicles to folliclestimulating hormone (FSH), the physiological mechanisms involved in these actions remain to be fully elucidated. Given the relevance of AMH in the folliculogenesis, this review aimed to describe the structural features, expression and the main biological effects of AMH on the follicular development. Review: Originally identified as a testicular product, AMH is responsible for regression of the Müllerian ducts during sexual differentiation of male embryos. In females, AMH is produced almost exclusively by granulosa cells of ovarian growing follicles, whose serum levels are positively related to the number of ovarian follicles, making AMH an excellent clinic marker of ovarian reserve. Along this work, it was shown aspects related to the structural characterization of AMH and its specific type II receptor (AMHRII). AMH is a glycoprotein dimer linked by disulfide bonds. The mature protein comprises two unequal domains: a long N-terminal domain (110-kDa) and a short C-terminal domain (25-kDa), responsible for the biological act


A foliculogênese é um evento complexo que envolve o recrutamento do pool de folículos primordiais, seguido por uma fase de crescimento e diferenciação. É sabido que para a sua regulação são requeridos vários fatores parácrinos e autócrinos, especialmente fatores de crescimento produzidos pelas células da granulosa. Dentre esses fatores, destaca-se o hormônio anti-mülleriano (AMH), uma glicoproteína membro da superfamília de fatores de crescimento transformantes (TGF-). Inicialmente o AMH foi estudado por seu papel regulatório no processo de diferenciação sexual masculina. [...]

4.
Acta sci. vet. (Impr.) ; 40(3): 01-015, 2012.
Artigo em Português | LILACS-Express | VETINDEX | ID: biblio-1457009

Resumo

Background: Cryopreservation is a biotech successfully employed in female gametes and embryos. This technique is of great importance for propagation of genetic material from animals with high-value livestock as well as to preserve the fertility of women undergoing cancer treatments. However, low temperatures can result in damage to different cellular compartments and organelles. This damage culminates in reduced viability, since they affect cell metabolism.Review: Cryopreservation consists of maintenance of biological material at low temperatures, in which chemical reactions are ceased, however, allowing the cells to preserve their viability. However, the decrease of temperature and subsequent warming may result in cellular damage. These damages occur in the cell membrane, cytoplasmic organelles and the cell nucleus. It is believed that the fi rst cell structure undergoing cryoinjury is the plasma membrane, responsible for maintaining homeostasis within the cell, and the loss of plasma membrane during the reduction temperature reported the main damage. The membrane damage due to cryopreservation appears to correlate with the reduction of thermal energy at low temperatures, thus limiting the movement of molecules through the phospholipids of lipid bilayer. Cryopreservation also alters the morphology, structure and cellular distribution of lipid droplets, reducing the survival of


Background: Cryopreservation is a biotech successfully employed in female gametes and embryos. This technique is of great importance for propagation of genetic material from animals with high-value livestock as well as to preserve the fertility of women undergoing cancer treatments. However, low temperatures can result in damage to different cellular compartments and organelles. This damage culminates in reduced viability, since they affect cell metabolism.Review: Cryopreservation consists of maintenance of biological material at low temperatures, in which chemical reactions are ceased, however, allowing the cells to preserve their viability. However, the decrease of temperature and subsequent warming may result in cellular damage. These damages occur in the cell membrane, cytoplasmic organelles and the cell nucleus. It is believed that the fi rst cell structure undergoing cryoinjury is the plasma membrane, responsible for maintaining homeostasis within the cell, and the loss of plasma membrane during the reduction temperature reported the main damage. The membrane damage due to cryopreservation appears to correlate with the reduction of thermal energy at low temperatures, thus limiting the movement of molecules through the phospholipids of lipid bilayer. Cryopreservation also alters the morphology, structure and cellular distribution of lipid droplets, reducing the survival of

5.
Braz. j. vet. res. anim. sci ; 39(5): 254-259, 2002.
Artigo em Inglês | VETINDEX | ID: vti-710483

Resumo

The present work investigated the efficiency of 0.9% saline solution and Phosphate Buffered Saline (PBS) in the preservation of goat preantral follicles in situ at different temperatures and incubation times. The ovarian pair of each animal was divided into 19 fragments. One ovarian fragment was taken randomly and fixed (control). The other 18 fragments were randomly distributed in tubes containing 0.9% saline solution or PBS at 4, 20 or 39 ºC for 4, 12 or 24 h. A total of 5,921 preantral follicles were examined. The quality of preantral follicles was evaluated by classical histology. The storage of ovarian fragments in 0.9% saline solution or PBS at 4 ºC did not reduce significantly the percentage of morphologically normal follicles when compared with the control, except after preservation in 0.9% saline solution for 24 h. The storage of ovarian fragments at 20 or 39C reduced the percentage of normal preantral follicles when compared to the control, except after preservation in PBS at 20C for 4 h. In conclusion, this study showed for the first time that goat preantral follicles can be stored in situ successfully at 4 ºC in 0.9% saline solution for 12 h and in PBS for 24 h, and at 20 ºC in PBS for 4 h.


O presente trabalho investigou a eficiência da solução salina 0,9% e tampão fosfato salina (PBS) na conservação de folículos pré-antrais caprinos in situ a diferentes temperaturas e tempos de incubação. O par ovariano de cada animal foi dividido em 19 fragmentos. Um fragmento foi escolhido aleatoriamente e fixado (controle). Os outros 18 fragmentos foram distribuídos aleatoriamente em tubos contendo solução salina 0,9% ou PBS a 4, 20 ou 39 C por 4, 12 ou 24 h. Um total de 5.921 folículos pré-antrais foram analisados. A qualidade dos folículos pré-antrais foi avaliada através de histologia clássica. A incubação de fragmentos ovarianos em solução salina 0,9% ou PBS a 4 ºC não reduziu significativamente a percentagem de folículos morfologicamente normais quando comparados com o controle, exceto após a conservação em solução salina 0,9% por 24 h. A incubação de fragmentos ovarianos a 20 ou 39C reduziu a percentagem de folículos pré-antrais normais quando comparados com o controle, exceto após conservação em PBS a 20C por 4 h. Em conclusão, este estudo mostrou pela primeira vez que folículos pré-antrais caprinos podem ser conservados in situ com sucesso a 4 ºC em solução salina 0,9% por 12 h e em PBS por 24 h, e a 20 ºC em PBS por 4 h.

6.
Acta sci. vet. (Online) ; 38(3): 221-236, 2010.
Artigo em Português | VETINDEX | ID: vti-733071

Resumo

Background:  :  :      : The platelet-derived growth factor (PDGF) is expressed in a wide variety of cell types, exerts a potent mitogenic role and acts on the growth, differentiation and cell chemotaxis. Studies have shown that during folliculogenesis, PDGF and their receptors are expressed in oocytes, granulosa cells and thecal cells of ovarian follicles at different developmental stages in several species. Although exist many information about its expression sites, as well as about its action in different cells types, the role of PDGF on ovarian folliculogenesis remains understudied. Thus, this article aims to review issues related to PDGF, suggesting the involvement of this mitogenic factor during follicular development. Review: Along this work, it was shown aspects related to structural characterization of PDGF and its receptors, as well as PDGF expression in different cells types, emphasizing its importance to follicular development. PDGF family is composed by four polypeptide chains (each encoded by a different gene), which are synthesized in the form of inactive pro-proteins. After a proteolytic processing, these chains undergo homo or heterodimerization, resulting in five isoforms (PDGF-AA, -BB, -AB, -CC e -DD). The cellular effects of these different PDGF isoforms are mediated by binding, with different specificities, to three transmembrane receptors isoforms of type

7.
Acta sci. vet. (Online) ; 38(3): 221-236, 2010.
Artigo em Português | VETINDEX | ID: vti-732366

Resumo

Background:  :  :      : The platelet-derived growth factor (PDGF) is expressed in a wide variety of cell types, exerts a potent mitogenic role and acts on the growth, differentiation and cell chemotaxis. Studies have shown that during folliculogenesis, PDGF and their receptors are expressed in oocytes, granulosa cells and thecal cells of ovarian follicles at different developmental stages in several species. Although exist many information about its expression sites, as well as about its action in different cells types, the role of PDGF on ovarian folliculogenesis remains understudied. Thus, this article aims to review issues related to PDGF, suggesting the involvement of this mitogenic factor during follicular development. Review: Along this work, it was shown aspects related to structural characterization of PDGF and its receptors, as well as PDGF expression in different cells types, emphasizing its importance to follicular development. PDGF family is composed by four polypeptide chains (each encoded by a different gene), which are synthesized in the form of inactive pro-proteins. After a proteolytic processing, these chains undergo homo or heterodimerization, resulting in five isoforms (PDGF-AA, -BB, -AB, -CC e -DD). The cellular effects of these different PDGF isoforms are mediated by binding, with different specificities, to three transmembrane receptors isoforms of type

8.
Artigo em Inglês | VETINDEX | ID: vti-731936

Resumo

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened prim

9.
Artigo em Inglês | VETINDEX | ID: vti-731552

Resumo

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened prim

10.
Artigo em Inglês | VETINDEX | ID: vti-730818

Resumo

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened prim

11.
Artigo em Inglês | VETINDEX | ID: vti-730172

Resumo

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened prim

12.
Acta Vet. bras. ; 5(3): 236-248, 2011.
Artigo em Português | VETINDEX | ID: vti-722245

Resumo

Vitrification is a cryopreservation methods cheap, fast and easy to perform and has been used, with relatively success, for the preservation of embryos and oocytes from preantral and antral follicles. The present review describes the different methods of vitrification, the main results obtained so far as well as its important for the assisted reproduction technologies in human, genetic superior animal and endangered mammalian species. Keywords: Blastocyst, oocyte, preantral follicle, cryopreservation.


A vitrificação é um método de criopreservação barato, rápido e fácil de ser realizado e tem sido usado com relativo sucesso para a preservação de embriões e oócitos obtidos a partir de folículos antrais e pré-antrais. A presente revisão descreve os diferentes métodos de vitrificação, os principais resultados obtidos, bem como sua importância para a tecnologia de reprodução assistida em humanos, animais de genética superior e espécies mamíferas ameaçadas de extinção. Palavras-Chave: Blastocisto, oócito, folículo ovariano pré-antral, criopreservação.

13.
Acta Vet. bras. ; 4(4): 215-226, 2010.
Artigo em Português | VETINDEX | ID: vti-722103

Resumo

Several cellular processes, such as cell survival, proliferation, differentiation, adhesion and migration, are controlled by a variety of growth factors. Among these factors it is possible to highlight the epidermal growth factor (EGF) family, which is constituted of at least eight members, described as important mediators of ovarian function. Its action on the follicular and embryo development is mediated by tyrosina kinase transmembrane receptors that are able to interact with at least six different members of this family. This paper aims focus on the role of EGF family on follicular and embryonic development in mammals. Keywords: EGF, tyrosine kinases receptor, ovarian follicle.


Diversos processos celulares, tais como a sobrevivência, proliferação, diferenciação, adesão e migração celular, são controlados por uma variedade de fatores de crescimento. Dentre estes fatores destaca-se a família do fator de crescimento epidermal (EGF), que é constituída de pelo menos oito membros, descritos como importantes mediadores da função ovariana. Sua ação no desenvolvimento folicular e embrionário é mediada por receptores transmembranários do tipo tirosina quinase, capazes de interagir com pelo menos seis diferentes membros desta família. O presente trabalho tem como objetivo abordar o papel da família EGF no desenvolvimento folicular e embrionário em mamíferos. Palavras-Chave: EGF, receptor tirosina quinase, folículo ovariano.

14.
Acta sci. vet. (Online) ; 40(3): 01-015, 2012.
Artigo em Português | VETINDEX | ID: vti-475739

Resumo

Background: Cryopreservation is a biotech successfully employed in female gametes and embryos. This technique is of great importance for propagation of genetic material from animals with high-value livestock as well as to preserve the fertility of women undergoing cancer treatments. However, low temperatures can result in damage to different cellular compartments and organelles. This damage culminates in reduced viability, since they affect cell metabolism.Review: Cryopreservation consists of maintenance of biological material at low temperatures, in which chemical reactions are ceased, however, allowing the cells to preserve their viability. However, the decrease of temperature and subsequent warming may result in cellular damage. These damages occur in the cell membrane, cytoplasmic organelles and the cell nucleus. It is believed that the fi rst cell structure undergoing cryoinjury is the plasma membrane, responsible for maintaining homeostasis within the cell, and the loss of plasma membrane during the reduction temperature reported the main damage. The membrane damage due to cryopreservation appears to correlate with the reduction of thermal energy at low temperatures, thus limiting the movement of molecules through the phospholipids of lipid bilayer. Cryopreservation also alters the morphology, structure and cellular distribution of lipid droplets, reducing the survival of


Background: Cryopreservation is a biotech successfully employed in female gametes and embryos. This technique is of great importance for propagation of genetic material from animals with high-value livestock as well as to preserve the fertility of women undergoing cancer treatments. However, low temperatures can result in damage to different cellular compartments and organelles. This damage culminates in reduced viability, since they affect cell metabolism.Review: Cryopreservation consists of maintenance of biological material at low temperatures, in which chemical reactions are ceased, however, allowing the cells to preserve their viability. However, the decrease of temperature and subsequent warming may result in cellular damage. These damages occur in the cell membrane, cytoplasmic organelles and the cell nucleus. It is believed that the fi rst cell structure undergoing cryoinjury is the plasma membrane, responsible for maintaining homeostasis within the cell, and the loss of plasma membrane during the reduction temperature reported the main damage. The membrane damage due to cryopreservation appears to correlate with the reduction of thermal energy at low temperatures, thus limiting the movement of molecules through the phospholipids of lipid bilayer. Cryopreservation also alters the morphology, structure and cellular distribution of lipid droplets, reducing the survival of

15.
Braz. j. vet. res. anim. sci ; 39(6): 324-330, 2002.
Artigo em Inglês | VETINDEX | ID: vti-710475

Resumo

The present study investigated the efficiency of saline solution and coconut water solution in the preservation of goat preantral follicles enclosed in ovarian tissue, at different temperatures and for different incubation periods. At the slaughterhouse, the ovarian pair was divided into 19 fragments; one ovarian fragment was immediately fixed for histology (control-time zero). The other 18 ovarian fragments were preserved in both solutions at 4ºC, 20ºC or 39ºC for 4 h, 12 h or 24 h. The histological analysis showed that the storage of ovarian fragments in both solutions at 4ºC for up to 24 h kept the percentage of normal preantral follicles similar to the control values. In contrast, preservation at 20C or 39ºC, in either solution, reduced significantly the percentage of normal preantral follicles compared to the control values, except in saline solution at 20ºC for 4 h or in coconut water solution at 20ºC for 4 h and 12 h. In conclusion, this study shows that both solutions can be used with the same efficiency to preserve goat preantral follicles at 4C, irrespective of the incubation time. However, to preserve goat preantral follicles at higher temperatures, coconut water solution is recommended.


O presente estudo investigou a eficiência da solução salina e solução à base de água de coco na preservação de folículos pré-antrais inclusos em tecido ovariano, em diferentes temperaturas e diferentes tempos de incubação. No abatedouro, o par ovariano foi dividido em 19 fragmentos; um fragmento ovariano foi imediatamente fixado para histologia clássica (controle-tempo zero). Os outros 18 fragmentos ovarianos foram conservados em ambas as soluções a 4ºC, 20ºC ou 39ºC por 4 h, 12 h ou 24 h. A análise histológica mostrou que a conservação de fragmentos ovarianos em ambas as soluções a 4ºC por até 24 h mantém a percentagem de folículos pré-antrais normais similar aos valores do controle. Ao contrário, a conservação a 20C ou 39ºC, em ambas as soluções, reduziu significativamente a percentagem de folículos pré-antrais normais comparado aos valores do controle, exceto em solução salina a 20ºC por 4 h ou em solução à base de água de coco a 20ºC por 4 h e 12 h. Em conclusão, esse estudo mostrou que ambas as soluções podem ser usadas com igual eficiência para conservar folículos pré-antrais caprinos a 4C, independente do tempo de incubação. No entanto, para conservar folículos pré-antrais caprinos a altas temperaturas, a solução à base de água de coco é recomendada.

16.
Ci. Rural ; 33(5)2003.
Artigo em Inglês | VETINDEX | ID: vti-704247

Resumo

The present work has investigated the degeneration rate of goat primordial follicles in situ after preservation in PBS or TCM 199 at different temperatures and incubation times. For each animal the ovarian pair was divided into 19 fragments. One ovarian fragment was taken randomly and immediately fixed (control). The other 18 ovarian fragments were randomly distributed in tubes containing PBS or TCM 199 and stored at 4º, 20º or 39ºC for 4, 12 or 24h. The storage of ovarian fragments in PBS or TCM 199 at 20ºC for 12h and 24h or at 39ºC, in all incubation times tested, increased significantly the percentage of degenerated primordial follicles (P 0.05). In contrast, for both media tested the degeneration rate of primordial follicles preserved at 4ºC for up to 24h and at 20ºC for 4h was similar to control values (P>0.05). In conclusion, this study shows that PBS was as efficient as TCM 199 in the preservation of goat primordial follicles in situ, being the best results observed at 4ºC.


O presente trabalho investigou a taxa de degeneração de folículos primordiais caprinos in situ após conservação em PBS ou TCM 199, em diferentes temperaturas e tempos de incubação. Para cada animal, o par ovariano foi dividido em 19 fragmentos. Um fragmento foi escolhido aleatoriamente e imediatamente fixado (controle). Os 18 fragmentos ovarianos restantes foram aleatoriamente distribuídos em tubos contendo PBS ou TCM 199 e conservados a 4, 20 or 39ºC por 4, 12 or 24h. A conservação de fragmentos ovarianos em PBS ou TCM 199 a 20ºC por 12 e 24h ou a 39ºC, em todos os tempos de incubação testados, aumentou significativamente a percentagem de folículos primordiais degenerados (P 0,05). Ao contrário, em ambos os meios testados, a taxa de degeneração de folículos primordiais conservados a 4ºC em todos os tempos testados e a 20ºC por 4h foi similar ao controle (P>0,05). Em conclusão, este estudo mostrou que o PBS foi tão eficiente quanto o TCM 199 na conservação de folículos primordiais caprinos in situ, sendo os melhores resultados observados na temperatura de 4ºC.

17.
Ci. Rural ; 28(3)1998.
Artigo em Português | VETINDEX | ID: vti-703401

Resumo

For mechanical isolation of caprine preantral follicles, ovaries from 52 undefinined breed type goats were collected at a local slaughterhouse or cirurgically extracted. The method used for preantral follicle isolation was a direct mechanical procedure (DMP). Briefly, ovaries were cut into small fragments using a tissue chopper adjusted to 500mum. The ovarian fragments were suspended 80 times with a pasteur pipette and then, the suspension was filtered successively through 500 and 100mum nylon mesh filters. The DMP resulted in a large number of isolated preantral follicles. On average, 1,067 ± 160 (mean number ± SEM) preantral follicles were isolated per ovary. To increase the number of preantral follicles recovered per ovary, the ovarian fragments remaining in the 500 and 100mum nylon mesh filters were transferred to the tissue chopper, and the procedure was repeated (complementary mechanical procedure - CMP). The use of CM P resulted in a mean increment of 5.2% of the total preantral follicles isolated per ovary. The mean number (mean ± SEM) of follicles isolated per ovary using the mechanical procedure (DPM + CPM) was 1,126 + 163. Irrespective of the procedure used, the number of preantral follicles isolated was characterized by extreme individual variation. The limits minimum and maximum of this variation were 53 and 7,482, respectively. The diameter of follicles isolated showed a range from l5.0mum to 92.7 mum. In conclusion, these results showed, for the first time, that preantral follicles can be successfully isolated from caprine ovary by applying a mechanical procedure.


Com o objetivo de isolar mecanicamente folículos pré-antrais caprinos, 52 ovários de cabras sem raça definida (SRD) foram coletados em matadouros ou extraídos por ovariectomia. O protocolo utilizado para o isolamento dos folículos, denominado procedimento mecânico direto (PMD), consistiu no corte do tecido ovariano em pequenos fragmentos, utilizando-se um cortador de tecidos. Em seguida, os fragmentos obtidos foram dissociados mecanicamente com pipetas de pasteur (1600mim e 600mim de diâmetro) e a suspensão resultante, contendo os fragmentos ovarianos, foi sucessivamente filtrada em malhas de náilon (500mim e 100mim). A realização deste procedimento permitiu isolar um número médio (± EP) de 1.067 ± 160 folículos por ovário. Com a finalidade de aumentar o número de folículos pré-antrais isolados por ovário, foi utilizado um procedimento mecânico complementar (PMC), que consistiu em submeter os fragmentos retidos nas malhas de 500mim e 100mim às etapas descritas no PMD. A utilização desses fragmentos resultou em um incremento médio de 5,2% no total de folículos isolados por ovário. O número médio (± EP) de folículos isolados por ovário, utilizando-se o procedimento mecânico (PMD + PMC), foi 1.126 + 163. Independentemente do procedimento utilizado, observou-se uma grande variação individual no número de folículos isolados por ovário. Os limites mínimo e máximo desta variação foram 53 e 7.482 folículos, respectivamente. O tamanho dos folículos isolados variou de 15,0mim a 92,7mim. É viável a utilização do procedimento mecânico, descrito pela primeira vez, para o isolamento de folículos ovarianos pré-antrais caprinos.

18.
Ci. Rural ; 28(3)1998.
Artigo em Português | VETINDEX | ID: vti-703400

Resumo

The aïm of this study was to describe the effects of reproductive stages (pre-pubertal, non-pregnant and pregnant adult animals), ovarian anatomic position (left or right), presence of functional corpus luteum, ovarian weight and number of superficial antral follicles on the number of preantral follicles mechanically isolated. The mean number (mean ± SEM) of isolated preantral follicles per ovary was 1,324 ± 193, 866 ±170 and 779 ± 139, respectively for prepubertal, non-pregnant and pregnant adult animals. Anatomic position (except for nonpregnant animals) and the presence or absence of corpus luteum dia nof affect the number and distribution of isolated preantral follicles. In general, the number of primary and secondary follicles was negatively correlated with the ovarian weight. The numbers of primordial and secondary follicles were, respectively, positively and negatively correlated with the number of superficial antral follicles. Analyzing animal categories separately, it was observed that in prepurbetal animals, the number of primordial follicles was positively correlated with the number of superficial antral follicles. However the numbers of primary and secondary follicles were negatively correlated with the number of superficial antral follicles. In the category of pregnant animals, the number of primary follicles was negatively correlated with the ovarian weight. According to ovary category analyzed, were observed significant correlation between the number of primordial, primary and secondary isolated follicles.


O presente trabalho descreve os efeitos do estágio reprodutivo (animais impúberes, adultos não-gestantes e gestantes), posição anatômica do ovário (direito e esquerdo) presença de corpo lúteo funcional, peso do ovário e número de folículos antrais superficiais (FAS) sobre o número de folículos pré-antrais (FPA) isolados mecanicamente. Os FPA isolados foram divididos em 3 classes a saber: primordiais, primários e secundários. O número médio (± EP) de FPA isolados por ovário foi de 1.324 ± 193; 866 + 170 e 779 ± 139, respectivamente para ovários oriundos de animais impúberes, não-gestantes e gestantes, sendo observada diferença significativa somente entre animais impúberes e gestantes. A posição anatômica do ovário (exceto para os animais não-gestantes) e a presença ou ausência de corpo lúteo não exerceram nenhum efeito sobre o número e distribuição dos FPA isolados. De um modo geral, o número de folículos primários e secundários foi negativamente correlacionado com o peso ovariano. Já o número de folículos primordiais e secundários foi, respectivamente, positiva e negativamente correlacionado com o número de FAS. Analisando as categorias de animais isoladamente, observou-se que, nos animais impúberes, o número de folículos primordiais foi positivamente correlacionado com o número de FAZ, enquanto que o número de folículos primários e secundários foi negativamente correlacionado com o número de FAS. No tocante à categoria de animais gestantes, o número de folículos primários foi negativamente correlacionado com o peso ovariano. Dependendo da categoria de ovários estudada, foram observadas associações significativas entre o número de folículos primordiais, primários e secundários.

19.
Artigo em Português | LILACS-Express | VETINDEX | ID: biblio-1475149

Resumo

The aïm of this study was to describe the effects of reproductive stages (pre-pubertal, non-pregnant and pregnant adult animals), ovarian anatomic position (left or right), presence of functional corpus luteum, ovarian weight and number of superficial antral follicles on the number of preantral follicles mechanically isolated. The mean number (mean ± SEM) of isolated preantral follicles per ovary was 1,324 ± 193, 866 ±170 and 779 ± 139, respectively for prepubertal, non-pregnant and pregnant adult animals. Anatomic position (except for nonpregnant animals) and the presence or absence of corpus luteum dia nof affect the number and distribution of isolated preantral follicles. In general, the number of primary and secondary follicles was negatively correlated with the ovarian weight. The numbers of primordial and secondary follicles were, respectively, positively and negatively correlated with the number of superficial antral follicles. Analyzing animal categories separately, it was observed that in prepurbetal animals, the number of primordial follicles was positively correlated with the number of superficial antral follicles. However the numbers of primary and secondary follicles were negatively correlated with the number of superficial antral follicles. In the category of pregnant animals, the number of primary follicles was negatively correlated with the ovarian weight. According to ovary category analyzed, were observed significant correlation between the number of primordial, primary and secondary isolated follicles.


O presente trabalho descreve os efeitos do estágio reprodutivo (animais impúberes, adultos não-gestantes e gestantes), posição anatômica do ovário (direito e esquerdo) presença de corpo lúteo funcional, peso do ovário e número de folículos antrais superficiais (FAS) sobre o número de folículos pré-antrais (FPA) isolados mecanicamente. Os FPA isolados foram divididos em 3 classes a saber: primordiais, primários e secundários. O número médio (± EP) de FPA isolados por ovário foi de 1.324 ± 193; 866 + 170 e 779 ± 139, respectivamente para ovários oriundos de animais impúberes, não-gestantes e gestantes, sendo observada diferença significativa somente entre animais impúberes e gestantes. A posição anatômica do ovário (exceto para os animais não-gestantes) e a presença ou ausência de corpo lúteo não exerceram nenhum efeito sobre o número e distribuição dos FPA isolados. De um modo geral, o número de folículos primários e secundários foi negativamente correlacionado com o peso ovariano. Já o número de folículos primordiais e secundários foi, respectivamente, positiva e negativamente correlacionado com o número de FAS. Analisando as categorias de animais isoladamente, observou-se que, nos animais impúberes, o número de folículos primordiais foi positivamente correlacionado com o número de FAZ, enquanto que o número de folículos primários e secundários foi negativamente correlacionado com o número de FAS. No tocante à categoria de animais gestantes, o número de folículos primários foi negativamente correlacionado com o peso ovariano. Dependendo da categoria de ovários estudada, foram observadas associações significativas entre o número de folículos primordiais, primários e secundários.

20.
Artigo em Português | LILACS-Express | VETINDEX | ID: biblio-1475150

Resumo

For mechanical isolation of caprine preantral follicles, ovaries from 52 undefinined breed type goats were collected at a local slaughterhouse or cirurgically extracted. The method used for preantral follicle isolation was a direct mechanical procedure (DMP). Briefly, ovaries were cut into small fragments using a tissue chopper adjusted to 500mum. The ovarian fragments were suspended 80 times with a pasteur pipette and then, the suspension was filtered successively through 500 and 100mum nylon mesh filters. The DMP resulted in a large number of isolated preantral follicles. On average, 1,067 ± 160 (mean number ± SEM) preantral follicles were isolated per ovary. To increase the number of preantral follicles recovered per ovary, the ovarian fragments remaining in the 500 and 100mum nylon mesh filters were transferred to the tissue chopper, and the procedure was repeated (complementary mechanical procedure - CMP). The use of CM P resulted in a mean increment of 5.2% of the total preantral follicles isolated per ovary. The mean number (mean ± SEM) of follicles isolated per ovary using the mechanical procedure (DPM + CPM) was 1,126 + 163. Irrespective of the procedure used, the number of preantral follicles isolated was characterized by extreme individual variation. The limits minimum and maximum of this variation were 53 and 7,482, respectively. The diameter of follicles isolated showed a range from l5.0mum to 92.7 mum. In conclusion, these results showed, for the first time, that preantral follicles can be successfully isolated from caprine ovary by applying a mechanical procedure.


Com o objetivo de isolar mecanicamente folículos pré-antrais caprinos, 52 ovários de cabras sem raça definida (SRD) foram coletados em matadouros ou extraídos por ovariectomia. O protocolo utilizado para o isolamento dos folículos, denominado procedimento mecânico direto (PMD), consistiu no corte do tecido ovariano em pequenos fragmentos, utilizando-se um cortador de tecidos. Em seguida, os fragmentos obtidos foram dissociados mecanicamente com pipetas de pasteur (1600mim e 600mim de diâmetro) e a suspensão resultante, contendo os fragmentos ovarianos, foi sucessivamente filtrada em malhas de náilon (500mim e 100mim). A realização deste procedimento permitiu isolar um número médio (± EP) de 1.067 ± 160 folículos por ovário. Com a finalidade de aumentar o número de folículos pré-antrais isolados por ovário, foi utilizado um procedimento mecânico complementar (PMC), que consistiu em submeter os fragmentos retidos nas malhas de 500mim e 100mim às etapas descritas no PMD. A utilização desses fragmentos resultou em um incremento médio de 5,2% no total de folículos isolados por ovário. O número médio (± EP) de folículos isolados por ovário, utilizando-se o procedimento mecânico (PMD + PMC), foi 1.126 + 163. Independentemente do procedimento utilizado, observou-se uma grande variação individual no número de folículos isolados por ovário. Os limites mínimo e máximo desta variação foram 53 e 7.482 folículos, respectivamente. O tamanho dos folículos isolados variou de 15,0mim a 92,7mim. É viável a utilização do procedimento mecânico, descrito pela primeira vez, para o isolamento de folículos ovarianos pré-antrais caprinos.

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