Cell proliferation in ovarian follicles fromBos taurus indicus females with different antral follicle count

The aim of this study was to compare the cell proliferation rate of follicles from Bostaurus indicusfemales with different antral follicle counts (AFCs). Thirty pairs of ovaries were classified as having low(≤ 31 follicles), intermediate (≥ 46 and ≤ 76 follicles) or high AFC (≥ 91 follicles). The ovaries were cut into 1 x 1 x 0.3 cm fragments, fixed in 10% buffered formalin solution and subjected to immunohistochemical analysis for proliferating cell nuclear antigen (PCNA). PCNA detection was detected in all stages of follicular development, being 1,388 (82.23%) primordial, 197 (11.67%) primary, 29 (1.72%) secondary and 74 (4.83%) antral follicles. The PCNA detection rate was higher (P ≤ 0.05) for antral and secondary follicles in ovaries with intermediate and low AFCs. This study showed that higher PCNA at early stages of follicular growth is detected in animals with high AFC, whereas a higher PCNA is seen in secondary and antral follicles of animals with low AFC.

Cell proliferation in ovarian follicles fromBos taurus indicus females with different antral follicle count

The aim of this study was to compare the cell proliferation rate of follicles from Bostaurus indicusfemales with different antral follicle counts (AFCs). Thirty pairs of ovaries were classified as having low(≤ 31 follicles), intermediate (≥ 46 and ≤ 76 follicles) or high AFC (≥ 91 follicles). The ovaries were cut into 1 x 1 x 0.3 cm fragments, fixed in 10% buffered formalin solution and subjected to immunohistochemical analysis for proliferating cell nuclear antigen (PCNA). PCNA detection was detected in all stages of follicular development, being 1,388 (82.23%) primordial, 197 (11.67%) primary, 29 (1.72%) secondary and 74 (4.83%) antral follicles. The PCNA detection rate was higher (P ≤ 0.05) for antral and secondary follicles in ovaries with intermediate and low AFCs. This study showed that higher PCNA at early stages of follicular growth is detected in animals with high AFC, whereas a higher PCNA is seen in secondary and antral follicles of animals with low AFC.(AU)

Proliferative activity of oocytes in multi-oocyte follicles of bovine ovary / Atividade proliferativa em folículos multi-oócitos na espécie bovina

We characterized the proliferative activity of multi-oocyte follicles with anti-nuclear antigen of proliferating cells (PCNA). Ovaries (n = 12) from heifers were processed for histology. From 789 multi-oocytes follicles observed, only 11 were considered appropriated for immunostaining, since they presented all nuclei of the oocytes clearly visible. All multi-oocyte follicles were positive for PCNA, but some oocytes showed no proliferative activity. We conclude that oocytes in multi-oocyte follicles seem to be in different stages of the cell cycle.

O objetivo deste trabalho foi caracterizar a atividade proliferativa de folículos multi-oócitos com antígeno marcador de proliferação celular (PCNA). Ovários (n = 12) de novilhas foram processados para histologia. A partir de 789 folículos multi-oócitos identificados, somente 11 foram considerados adequados para imunohistoquímica por apresentarem todos os núcleos claramente visíveis. Todos os folículos multi-oócitos foram positivos para PCNA, mas alguns oócitos não apresentaram marcação. Concluímos que oócitos presentes em folículos multi-oócitos parecem estar em diferentes fases do ciclo celular.

Proliferative activity of oocytes in multi-oocyte follicles of bovine ovary / Atividade proliferativa em folículos multi-oócitos na espécie bovina

We characterized the proliferative activity of multi-oocyte follicles with anti-nuclear antigen of proliferating cells (PCNA). Ovaries (n = 12) from heifers were processed for histology. From 789 multi-oocytes follicles observed, only 11 were considered appropriated for immunostaining, since they presented all nuclei of the oocytes clearly visible. All multi-oocyte follicles were positive for PCNA, but some oocytes showed no proliferative activity. We conclude that oocytes in multi-oocyte follicles seem to be in different stages of the cell cycle.(AU)

O objetivo deste trabalho foi caracterizar a atividade proliferativa de folículos multi-oócitos com antígeno marcador de proliferação celular (PCNA). Ovários (n = 12) de novilhas foram processados para histologia. A partir de 789 folículos multi-oócitos identificados, somente 11 foram considerados adequados para imunohistoquímica por apresentarem todos os núcleos claramente visíveis. Todos os folículos multi-oócitos foram positivos para PCNA, mas alguns oócitos não apresentaram marcação. Concluímos que oócitos presentes em folículos multi-oócitos parecem estar em diferentes fases do ciclo celular.(AU)

Recovery of equine oocytes by scraping of the follicular wall with different specifications of needles and morphological analysis of cumulus oophorus / Recuperação de oócitos equinos por raspagem da parede folicular com diferentes especificações de agulhas e análise morfológica do cumulus oophorus

In follicular aspiration, physical aspects are of high significance for the technique to succeed, such as vacuum pressure, caliber of the needle and the way the follicular wall curettage is performed. The aim of this study was to investigate the recovery rate of equine oocytes aspirated by scraping of the follicular wall, testing different calibers of disposable needles, as well as the morphological evaluation of the cumulus oophorus complexes (COCs). Mares ovaries (n=447) obtained at a local slaughterhouse were transported to the laboratory in a thermal container (20 °C) and had the tunica albuginea and connective tissues dissected. The aspirated follicles had 10 to 25 mm in diameter, and 30x8 (21G 1 ») or 40x12 (18G 1 ½) needles were used for the aspiration, forming group A (G-A) and group B (G-B), respectively. In G-A and G-B, 480 and 548 follicles were aspirated, respectively. Under the stereomicroscope, the oocytes were evaluated according to the quality of the ooplasm and characteristics of the cumulus cells (grade I, II, III and denuded). The statistical analysis was performed using the Students t-test, logistic regression and test of proportions, and differences were considered significant when P<0.05. There was no difference between recovery rates of groups G-A (66.5%; 330/496) and G-B (65.5%; 359/548). In the G-A group, grade II oocytes were related to higher recovery rates (46.9%; 145/330) than grade I (23.6%; 72/330), grade III (20.6%; 59/330) and denuded oocytes (8.5%; 24/330; P<0.05).(AU)

Na obtenção de oócitos para a espécie equina, aspectos físicos são de alta significância para o sucesso da técnica, como a pressão de vácuo e o calibre de agulha utilizado, além da forma como é realizada a raspagem da parede folicular. O objetivo deste estudo foi investigar o índice de recuperação de oócitos equinos aspirados pela técnica de raspagem da parede folicular, testando calibres distintos de agulhas descartáveis, assim como avaliação morfológica dos complexos cumulus oophorus (CCOs). Foram utilizados ovários de éguas (n=447), obtidos em abatedouro local, transportados ao laboratório em recipiente térmico (20 ºC) e submetidos à dissecação da túnica albugínea e tecidos conectores. Os folículos aspirados obtinham diâmetro entre 10 mm a 25 mm, e para tanto utilizou-se a agulha 30x8 (21G 1 ») ou 40x12 (18G 1 ½), formando respectivamente, o grupo A (G-A) e grupo B (G-B). No G-A e G-B aspiraram-se 480 e 548 foliculos, respectivamente. Sob lupa estéreo-microscópica avaliou-se os oócitos quanto à qualidade do ooplasma e características das células do cumulus oophorus (grau I, II, III e desnudo). Para análise estatística utilizou-se teste t Student, regressão logistica e teste de proporções, e as diferenças foram consideradas significativas quando P 0,05. A taxa de recuperação entre G-A e G-B não apresentou diferença; 66,5% (330/496) e 65,5% (359/548), respectivamente. Houve diferençaquanto à qualidade dos oócitos no G-A, no qual os oócitos de grau II obtiveram melhor taxa (46,9%; 145/330; P<0,05) frente ao grau I (23,6%; 72/330), grau III (20,6%; 59/330) e desnudo (8,5%; 24/330). Entretanto, no G-B não houve diferença estatística quanto aos graus de qualidade do CCOs recuperados, grau I (23,4%; 77/359), grau II (43,2%; 145/359), grau III (22,5%; 73/359) e desnudo (11,1%; 32/359).(AU)

Recovery of equine oocytes by scraping of the follicular wall with different specifications of needles and morphological analysis of cumulus oophorus / Recuperação de oócitos equinos por raspagem da parede folicular com diferentes especificações de agulhas e análise morfológica do cumulus oophorus

In follicular aspiration, physical aspects are of high significance for the technique to succeed, such as vacuum pressure, caliber of the needle and the way the follicular wall curettage is performed. The aim of this study was to investigate the recovery rate of equine oocytes aspirated by scraping of the follicular wall, testing different calibers of disposable needles, as well as the morphological evaluation of the cumulus oophorus complexes (COCs). Mares ovaries (n=447) obtained at a local slaughterhouse were transported to the laboratory in a thermal container (20 °C) and had the tunica albuginea and connective tissues dissected. The aspirated follicles had 10 to 25 mm in diameter, and 30x8 (21G 1 ») or 40x12 (18G 1 ½) needles were used for the aspiration, forming group A (G-A) and group B (G-B), respectively. In G-A and G-B, 480 and 548 follicles were aspirated, respectively. Under the stereomicroscope, the oocytes were evaluated according to the quality of the ooplasm and characteristics of the cumulus cells (grade I, II, III and denuded). The statistical analysis was performed using the Student’s t-test, logistic regression and test of proportions, and differences were considered significant when P<0.05. There was no difference between recovery rates of groups G-A (66.5%; 330/496) and G-B (65.5%; 359/548). In the G-A group, grade II oocytes were related to higher recovery rates (46.9%; 145/330) than grade I (23.6%; 72/330), grade III (20.6%; 59/330) and denuded oocytes (8.5%; 24/330; P<0.05).

Na obtenção de oócitos para a espécie equina, aspectos físicos são de alta significância para o sucesso da técnica, como a pressão de vácuo e o calibre de agulha utilizado, além da forma como é realizada a raspagem da parede folicular. O objetivo deste estudo foi investigar o índice de recuperação de oócitos equinos aspirados pela técnica de raspagem da parede folicular, testando calibres distintos de agulhas descartáveis, assim como avaliação morfológica dos complexos cumulus oophorus (CCOs). Foram utilizados ovários de éguas (n=447), obtidos em abatedouro local, transportados ao laboratório em recipiente térmico (20 ºC) e submetidos à dissecação da túnica albugínea e tecidos conectores. Os folículos aspirados obtinham diâmetro entre 10 mm a 25 mm, e para tanto utilizou-se a agulha 30x8 (21G 1 ») ou 40x12 (18G 1 ½), formando respectivamente, o grupo A (G-A) e grupo B (G-B). No G-A e G-B aspiraram-se 480 e 548 foliculos, respectivamente. Sob lupa estéreo-microscópica avaliou-se os oócitos quanto à qualidade do ooplasma e características das células do cumulus oophorus (grau I, II, III e desnudo). Para análise estatística utilizou-se teste t Student, regressão logistica e teste de proporções, e as diferenças foram consideradas significativas quando P 0,05. A taxa de recuperação entre G-A e G-B não apresentou diferença; 66,5% (330/496) e 65,5% (359/548), respectivamente. Houve diferençaquanto à qualidade dos oócitos no G-A, no qual os oócitos de grau II obtiveram melhor taxa (46,9%; 145/330; P<0,05) frente ao grau I (23,6%; 72/330), grau III (20,6%; 59/330) e desnudo (8,5%; 24/330). Entretanto, no G-B não houve diferença estatística quanto aos graus de qualidade do CCOs recuperados, grau I (23,4%; 77/359), grau II (43,2%; 145/359), grau III (22,5%; 73/359) e desnudo (11,1%; 32/359).