• Asian-Australasian Journal of Animal Sciences
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• v.19 no.12
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• pp.1711-1715
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• 2006

To predict the number of preantral (primordial, primary and secondary) follicles retrieved from bovine ovaries, we examined the relationship between morphological parameters of ovaries and number of preantral follicles retrieved mechanically. The preantral follicles were retrieved mechanically by slicing ovarian tissue and the influences of size of the ovaries, number of antral follicles, and presence of cystic follicle and corpus luteum on the retrieval were evaluated. Total 77 ovaries were used and significant (p<0.05) relationship was detected between the number of antral follicles and the presence of cystic follicles, and the retrieval number. More preantral follicles were retrieved from the ovaries having more than 20 antral follicles than those having less than 20 antral follicles (17,760${\pm}$5,637 vs. 3,689${\pm}$537) in the ovarian cortex. The retrieval number was significantly reduced in cystic ovaries compared with non-cystic ovaries (5,167${\pm}$825 vs. 20,631${\pm}$6,507). However, neither ovary size (<3.5, 3.5 to 4.0, 4.0 to 4.5 and >4.5 cm) nor the presence of corpus luteum affected the follicle retrieval. In conclusion, the number of preantral follicles retrieved from the ovaries can simply be predicted by the number of antral follicles and the presence of cystic follicles in the ovarian cortex.

• Journal of Embryo Transfer
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• v.30 no.1
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• pp.17-21
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• 2015

Alteration in ion channel or transporter expression levels affects cell volume which is produced by movement of water and ion across the plasma membrane. In particular, aquaporin (AQP) channels among ion channels play a crucial role in movement of water across the cell membrane. This study was performed to identify whether AQP expression is changed in bovine follicular cystic follicles using microarray, RT-PCR and Western blotting analyses. In microarray data, AQP4 expression was decreased, whereas AQP7 was increased in cystic follicles. Additional experiments were focused on the AQP7 expression increased in cystic follicles. The microarray data was confirmed by semi-quantitative polymerase chain reaction (PCR) and Western blot analysis. AQP7 mRNA and protein expressions were significantly increased in the cystic follicles (p<0.05). Application of estrogen ($10{\mu}g/ml$) to bovine ovarian cells showed a trend of increase in AQP7 expression. From these results, we suggest that the increase in AQP7 expression in cystic follicles may play an important role in movement of water in bovine ovary. In addition, AQP7, a aquaglyceroporin permeating water and glycerol, could be a good target in development of methods for the cryopreservation of bovine ovary.

• Journal of Embryo Transfer
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• v.25 no.1
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• pp.29-34
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• 2010

Follicular cystic ovary (FCO) is one of the major causes of reproductive failure in cattle. Genetic alterations affect the function of diverse cells and/or tissues, which could be present in cystic ovaries. A microarray analysis was performed to screen differential gene expressions in follicular cystic follicles of cattle. In this study, we hypothesized that follicular cysts may be induced by changes in ion- and transporter-related gene expression. Microarray data showed that fibrinogen-gamma (FGG) and low density lipoprotein receptor-related protein 8 (LRP8) were up-regulated, while choline transporter-like protein 4 (SLC44A4), very long-chain acyl-CoA synthetase homolog 2 (SLC27A5), annexin 8 (ANXA8), and aquaporin 4 were down-regulated in follicular cystic follicles. A semi-quantitative RT-PCR was carried out to validate DEGs altered in follicular cystic follicles. Of six DEGs, three DEGs (FGG, SLC44A4, and aquaporin 4) showed a positive correlation between microarray and semi-quantitative PCR data. We focused on FGG, among three DEGs, which was highly up-regulated in follicular cystic follicles. The FGG mRNA was upregulated by 8.4-fold and by 1.7-fold in the bovine follicular cystic follicles as judged by microarray and RT-PCR analysis, respectively. However, there was no significant changes in the expression level of FGG protein in both follicular cystic follicles and granulosa cells isolated from follicular cystic follicles by Western blot analysis. Although this study does not reveal a positive correlation between the mRNA and protein level, FGG appears to be an important biomarker in the discrimination of follicular cyst from normal ovary.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.5
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• pp.265-272
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• 2010

Follicular cystic ovary (FCO) is one of the most frequently diagnosed ovarian diseases and is a major cause of reproductive failure in mammalian species. However, the mechanism by which FCO is induced remains unclear. Genetic alterations which affect the functioning of many kinds of cells and/or tissues could be present in cystic ovaries. In this study, we performed a comparison analysis of gene expression in order to identify new molecules useful in discrimination of bovine FCO with follicular cystic follicles (FCFs). Normal follicles and FCFs were classified based on their sizes (5 to 10 mm and $\geq25mm$). These follicles had granulosa cell layer and theca interna and the hormone $17{\beta}$-estradiol ($E_2$)/ progesterone ($P_4$) ratio in follicles was greater than one. Perifollicular regions including follicles were used for the preparation of RNA or protein. Differentially expressed genes (DEG) that showed greater than a 2-fold change in expression were screened by the annealing control primer (ACP)-based PCR method using $GeneFishing^{TM}$ DEG kits in bovine normal follicles and FCFs. We identified two DEGs in the FCFs: ribosomal protein L15 (RPL15) and microtubule-associated protein 1B (MAP1B) based on BLAST searches of the NCBI GenBank. Consistent with the ACP analysis, semi-quantitative PCR data and Western blot analyses revealed an up-regulation of RPL15 and a down-regulation of MAP1B in FCFs. These results suggest that RPL15 and MAP1B may be involved in the regulation of pathological processes in bovine FCOs and may help to establish a bovine gene data-base for the discrimination of FCOs from normal ovaries.

• Journal of Embryo Transfer
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• v.25 no.4
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• pp.273-279
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• 2010

Follicular cystic follicles (FCFs) show delayed regression with persistent follicle growth. However, the mechanism by which follicles are persistently grown remains unclear. Glutathione S-transferases (GSTs) are drug-metabolizing and detoxification enzymes that are involved in the intracellular transport and metabolism of steroid hormones. In this study, a proteomic analysis was performed to identify whether GST expression is changed in bovine FCFs and to predict the interactions between GST and other proteins. Normal follicles and FCFs were classified based on their sizes (5 to 10 mm and 25 mm). In bovine follicles, GST mu 1 (GSTM1) was detected as a differentially expressed protein (DEP) and significantly up-regulated in FCFs compared to normal follicles (p<0.05). Consistent with the proteomic results, semi-quantitative PCR data and western blot analysis revealed an up-regulation of GSTM1 in FCFs. Expression levels of aromatase and dehydrogenase proteins were changed in FCFs. These results show that the up-regulation of GSTM1 that is observed in bovine FCFs is likely to be responsible for the persistent follicle growth in FCFs as the activity of aromatase and the dehydrogenases.

• Korean Journal of Veterinary Research
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• v.27 no.1
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• pp.141-151
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• 1987

A total of 1200 Korean native cow and 240 dairy cow genitalia were collected during the slaughtering process in Seoul and Kwang Ju abattoir and were examined from July 1985 to March 1986. Ovarian follicles were classified as cystic if the diameter was greater than 2.5cm or if follicles were multiple. In order to investigate the ovarian cysts, anatomical and histological examinations were performed. In addition progesterone and estrogen level in different types of cystic follicular fluid and serum were measured by radioimmunoassay. The results were summerized as follows: 1. The incidences of ovarian cysts were 2.0% in Korean native cow and 7.9% in dairy cow. 2. In distribution of cysts in the left, right and both ovaries, the most encountered ovary was right one. The frequency was 45.8% in right ovaries, 33.4% in left ovaries and 20.8% in both ovaries in Korean native cow. On the contrary the frequency was 42.1% in right ovaries, 31.8% in both ovaries and 26.3% in left ovaries in dairy cow. 3. Six speciemens (25.0%) of Korean native cow and six specimens (31.6%) of dairy cow were associated with corpora lutes in both ovaries. 4. The luteinization of theca layer was most significant in the group 2Aa (71.4%) and 2Ba (38.5%) which associated with no granulosa cell and corpora lutea in the same cystic ovaries. 5. Correlation of progesterone concentration between cystic fluid and serum was found only in the group 2Aa and 2Ab (r=0.86). Progesterone and estrogen concentrations in cystic fluid were closely related to the degree of degeneration of granulosa cell layer. The cystic follicles that consist of thickened theca and degenerated granulosa cell layers contained a large amount of progesterone, and small amount of estrogen. In conclusion, various types of ovarian cysts with various levels of progesterone and estrogen were observed in Korean native cow.

• Korean Journal of Veterinary Pathology
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• v.2 no.1
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• pp.1-8
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• 1998

Chickens at 3-weeks of age were inoculated with a highly virulent strain (SH/92) of Infectious Bursal Disease Virus(IBDV) through ocular and cloacal routes. The infected chickens were killed at 6, 12, 24, 48, 72, and 96 hrs post inoculation (PI) and Bursa of Fabricius(BF) were collected. The sizes of bursal follicules in infected chickens decreased at 48 to 96 hrs PI. Histologically the cellular changes were first evident at 12 hrs PI and characterized by condensation of nuclear chromatin of bursal lymphocytes indicating apoptosis. By 24 hrs PI apoptotic lymphocytes dramatically increased. In addition infiltration of heterophils were also seen in the follicles and in the interfollicular connective tissues. At 48 hrs PI, cystic cavities were observed in the follicles. As the infection advanced the bursal follicles showed atrophy accompanied by disappearance of heterophils and reduction in number of lymphocytes in the cystic cavities which was replaced by proteineous materials. The nuclei of most affected lymphocyte stained positively with the in situ end labeling for apoptosis. Electron microscopy showed viral particles with crystalline array in the lymphocytes of BF infected with IBOV. These results indicated that SH/92 IBDV infection in chickens caused increased apoptosis in the BF.

• Korean Journal of Animal Reproduction
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• v.13 no.2
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• pp.85-92
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• 1989

The study was carried out to find out the changes of the sex hormone concentrations in the blood plasma and antrum of follicular and lutein cystic ovaries of Holstein cows. The progesterone, estradiol-17$\beta$, testosterone, FSH and LH from samples of the blood plasma and antrum of cystic ovaries of cows assayed by radioimmunoassay method. The results of this study were summarized as follows : 1. The concentrations fo progesterone and estradiol-17$\beta$ in the blood plasmaat estrous and luteal phase were 0.95$\pm$0.18ng/ml, 11.45$\pm$3.12pg/ml and 4.25$\pm$0.27ng/ml, 6.27$\pm$0.82pg/ml respectively. The concentrations of progesterone and estradiol-17$\beta$ in the antrum fluid of follicles at estrous and luteal phase were 24.8$\pm$4.12ng/ml, 54.3$\pm$7.25pg/ml and 21.7$\pm$3.79ng/ml, 14.3$\pm$2.72pg/ml respectively, and showed significant changes among the estrous and luteal phase and blood plasma and antrum fluid of follicles. 2. The concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the blood plasma of follicular cystic ovareis of cows were 0.85$\pm$0.25ng/ml, 9.23$\pm$2.72pg/ml, 17.12$\pm$3.26pg/ml and 3.78$\pm$1.02mIU/ml respectively. And the concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the antrum fluid of follicular cystic ovareis of cows were 284$\pm$48.21ng/ml, 389$\pm$67.23ng/ml, 12.84$\pm$0.29ng/ml and 1.84$\pm$0.17mIU/ml respectively, and showed significant changes between in the blood plasma and antrum fluid of cystic ovaries. 3. The concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the blood plasma of lutein cystic ovaries of cows were 3.40$\pm$0.78ng/ml, 4.02$\pm$0.42pg/ml, 10.72$\pm$2.74pg/ml and 0.76$\pm$0.12mIU/ml respectively. And the concentrations of progesterone, estradiol-17$\beta$, testosterone and LH in the antrum fluid of lutein cystic ovareis of cows were 427$\pm$35.79ng/ml, 0.76$\pm$0.07ng/ml, 3.45$\pm$0.57ng/ml and 0.29$\pm$0.07mIU/ml respectively, and showed significant changes between the blood plasma and antrum fluid of cystic ovaries. 4. Accordingly, the diagnosis of follicular and lutein cystic ovareis of cows from progesterone, estradiol-17$\beta$ and LH levels in the blood plasma and antrum of cystic ovaries of cows is thought to be possible a diagnostic means.

• Journal of Embryo Transfer
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• v.28 no.1
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• pp.57-61
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• 2013

Microtubule-associated protein 1B (MAP1B), a member of MAP1 family, plays a key role in neuronal development. MAP1B binds to many kinds of proteins directly or indirectly. This study was performed to investigate whether MAP1B interacts with GAPDH in bovine follicles using immunoprecipitation (IP) with Western blot analysis and immunohistochemisty. The mRNA expressions of MAP1B and glyceraldehydes 3-phosphate dehydrogenase (GAPDH) were down-regulated in bovine follicular cystic follicles (FCF). In parallel with the mRNA levels, their protein levels were also down-regulated in FCFs. In addition, MAP1B and GAPDH were co-localized at the cytoplasm of follicles. IP with Western blot analysis showed that MAP1B bound to GAPDH in normal follicles, but their binding was absent in FCFs, suggesting a low level of MAP1B and/or GAPDH expressions in FCFs. Taken together, these results suggest that MAP1B interacted with GAPDH may play a role in bovine follicle development, and that GAPDH does not function always as a loading control in bovine follicles.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.4
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• pp.315-322
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• 2020

Aquaporin channels (AQPs) are known to play an important role in the development of ovarian follicles through their function in water transport pathways. Compared to other AQPs, research on the role of AQP4 in female reproductive physiology, particularly in cattle, remains limited. In our previous study, gene chip microarray data showed a downregulation of AQP4 in bovine cystic follicles. This study was performed to validate the AQP4 expression level at the protein level in bovine follicles using immunohistochemistry, Western blotting, and immunoprecipitation assays. Immunostaining data showed that AQP4 was expressed in granulosa and theca cells of bovine ovarian follicles. The ovarian follicles were classified according to size as small (< 10 mm) or large (> 25 mm) in diameter. Consistent with earlier microarray data, semi-quantitative PCR data showed a decrease in AQP4 mRNA expression in large follicles. Western blot analysis showed a downregulation of the AQP4 protein in large follicles. In addition, AQP4 was immunoprecipitated and blotted with anti-AQP4 antibody in small and large follicles. Accordingly, AQP4 exhibited a low expression in large follicles. These results show that AQP4 is downregulated in bovine ovarian large follicles, suggesting that the downregulation of AQP4 expression may interfere with follicular water transport, leading to bovine follicular cysts.