Although cryopreservation of sperm is routinely used for clinical requirement, it has some problems, such as high generation of reactive oxygen species (ROS) and cold-shock. To reduce the detrimental damage in sperm, anti-oxidants were added to cryoprotectant for sperm. Curcumin is one of anti-oxidants, which are added in cryoprotectants. However, recent studies have demonstrated that curcumin decreases sperm viability and motility. This study was performed to identify the effect of curcumin on hydrogen peroxide (H2O2)-exposed bovine sperm, which were cryopreserved-thawed. In H2O2-exposed bovine sperm, reactive oxygen species (ROS) were significantly reduced by treatment with curcumin in a dose-dependent manner (p < 0.05). Among tested concentrations of curcumin (1 to 50 μM), 30 and 50 μM curcumin showed anti-oxidant effect on H2O2-induced ROS generation. On the other hand, combination of 30 or 50 μM curcumin with anti-oxidant H2O2 increased the percentage of apoptotic sperm compared to only H2O2 treatment. Sperm viability was also decreased in the combination of 30 or 50 μM curcumin with H2O2 as judged by FDA/PI staining. H2O2–induced decrease in sperm progressive motility was recovered by treatment with 1 μM curcumin. These results show that high concentration of curcumin has anti-oxidant effect, but it has also cytotoxic effect on bovine sperm. Sperm viability and motility might be more affected by cytotoxic signals of curcumin compared to antioxidant signals.

The objective of this study was to investigate the result of in vivo embryo collection and pregnancy rate after embryo transfer using sex-sorted sperm of Korean brindle cattle. Donor Korean brindle cattle superovulation treated by decreasing dose of FSH injection. Embryos were recovered on 7 days after the third artificial insemination. Control group semen straw used artificial insemination contained 20 million sperm. Sex-sorted semen straws contained 4 million sperm or 10 million sperm. As for the result of the recovery of the in vivo embryos derived from sex-sorted sperm, the number of transferable embryos was significantly highly recovered to be 6.20±2.28/donor from the control group and was significantly lowly recovered to be 1.57±1.72/donor from the group treated at a sperm concentration of 10×106 (p<0.05). The number of unfertilized embryo was 0.8±1.30/donor in control group which was significantly lower than the group treated at a sperm concentration of 4×106 (p<0.05). However, there was no significant difference in the number of undeveloped ova between control and treatment groups. Pregnancy rate after embryo transfer was shown to be 35.00% in control group and 12.50% in treatment group. The karyotype analysis of the calf derived from sex-sorted sperm resulted in a similar chromosomal distribution pattern (2n=60, XX) compared to those of common Korean native cattle.

Cryopreservation affects osmotic tolerance and intracellular ion concentration through changes in expression levels of water and ion channels. Control of these changes is important for cell survival after cryopreservation. Relatively little is known about changes in K+ channel expression compared to water channel expression. This study was performed to investigate changes in TASK-2 channel (KCNK5: potassium channel, subfamily K, member 5), a member of two-pore domain K+ channel family, in cryopreserved mouse ovaries. Cryopreservation increased TASK-2 mRNA expression in mouse ovaries. In addition, TASK-2 protein expression was upregulated in vitrified and slowly frozen ovaries. TASK-2 protein was expressed in all area of granulosa cells that surround the oocyte within the follicle, except nucleus. Viability of cells overexpressed with TASK-2 was higher than that of vector-transfected cells. Our results found that TASK-2 expression was increased by cryopreservation and overexpression of TASK-2 decreased cryopreservation-induced cell death. These results suggest that TASK-2 upregulation might reduce cryodamage.

Toll-like receptor 7 (TLR7) is critical for the triggering of innate immune response by recognizing the conserved molecular patterns of single-stranded RNA (ssRNA) viruses and mediated antigenic adaptive immunity. To understand how TLR7 distinguish pathogen-derived molecular patterns from the host self, it is essential to be able to identify TLR7 receptor interaction interfaces, such as active sites or R848-agonist binding sites. The functional interfaces of TLR7 can serve as targets for structure-based drug design in studying the TLR7 receptor’s structure-function relationship. In contrast to mammalian TLR7, chicken TLR7 (chTLR7) is unknown for its important biological function. Therefore, it has been targeted to mediate contrasting evolutionary patterns of positive selection into non-synonymous SNPs across eleven species using TLR7 conservation patterns (evolutionary conserved and class-specific trace residues), where protein sequence differences to the TLR7 receptors of interest record mutation that have passed positive section across the species. In this study, we characterized the Lys609 residue on chTLR7-ECD homodimer interfaces to reflect the current tendency of evolving positive selection to be transfer into a stabilization direction of the R848-agonist/ chTLR7-ECDs complex under the phylogenetically variable position across species and we suggest a potential indicator for contrasting evolutionary patterns of both the species TLR-ECDs.

본 연구는 현재 국립축산과학원 가축유전자원세터에서 보유 하고 있는 재래닭을 순수화된 품종인 것으로 판단하고, 일반 적으로 이용되고 있는 산란사료 및 사양관리 방법을 적용하여 특히, 동절기에 있어 재래닭의 정자의 보존 기간과 수정률 및 초기배자의 생존율을 각각 비교함으로써 재래닭의 생산성 향 상을 위한 기초자료를 제공하고, 나아가 표준능력을 고찰하고 자 수행하였다. 본 시험에 사용된 공시계는 39주령의 재래닭 6계통 적갈색(R, Red Brown Strain), 황갈색(Y, Yellow Brown Strain), 회갈색(G, Gray Brown Strain), 흑색(L, Black Strain), 백색(W, White Strain) 그리고 오계(O, Ogol Strain)를 대상 으로 하고 대조군으로는 3계통 즉, 외래도입종 중에서 대표적 인 다산종인 White Leghorn (F Strain), Rhode Island (C Strain) 그리고 육용종인 Cornish (H Strain)의 수정률 및 초기배자 생존율을 조사하였다. 단 한번의 인공수정 후, 3 주간 생산된 알의 수정률 확인을 한 결과, 재래닭의 경우, 6 계통간의 유의 적인 차이는 없지만 93.3 ~ 100.0비율로 인공 수정 후, 2일째 부터 수정률이 6일째 동안 최고 높음을 확인했다. 6일째까지 상대적으로 일정하게 높은 수정률을 유지하다가 7일부터 17일 째 까지 점진적으로 감소함을 확인했다. 17일 이후 생산된 알 의 경우 무정란임을 확인 할 수 있었다. 재래닭(R, Y, 그리고 O) 21일간 생산된 알의 배발생정지율의 결과, 인공수정 후, 약 4일째 생산된 알(3 ~ 6일)에서 외래도입종 3품종간의 유의적인 차이는 보이지 않았지만, 배발생정지율이 0%임을 확인하였다. 세 품종 모두 약 7일째 생산된 알부터 배발생정지율이 13.8 ~ 26.7%로 급격히 증가하고 12일부터 감소하는 것을 확 인했다. 재래닭 3 계통의 결과도 인공수정 후, 약 4 일째 생 산된 알에서 외래도입종 3 품종과 유사한 패턴을 나타내면서 배아 사망율이 6 일째까지 0%를 보였다. 금후, 체내 정자보존 기간이 수정률 및 초기배자 생존율에 미치는 영향을 좀더 엄 밀하게 조사하기 위해서는 정액 성상 및 활력 검사와 더불어 암탉의 주령에 따른 변화도 함께 보다 세부적이고 입체적인 방법의 체계적인 조사가 반드시 필요하다고 할 수 있겠다.

Reproductive disorders in cows cause economic loss in livestock farms. Reproductive diseases, such as follicular cyst, luteal cyst, endometritis, pyometra, and repeat breeding cause infertility. Among these diseases, endometritis and pyometra are uterine infections that are leading causes of infertility. This study was performed to investigate the causative agents of uterine diseases using bacterial culture. Bacteria were obtained from the reproductive organs (vagina, uterine cervix, and uterine horn) of dairy cow diagnosed with endometritis or pyometra, and cultured on blood agar. The colonies obtained from cultivation for 24 hours were passaged. To identify the bacteria, the colonies grown in passaged culture Gram stained and applied to an automatic biochemical microbial identification system. Escherichia coli were commonly detected in vagina, uterine cervix, and uterine horn of dairy cows diagnosed to pyometra. The cows having endometritis showed not only Escherichia coli but also Pantoea spp. and Klebsiella spp. strains. Dairy cows that were infected with Escherichia coli in uterus caused mastitis or digestive disease. These results suggest that sanitary feeding and management beforehand are needed to prevent bacterial infections.

Many pronuclear stage eggs were used to generate transgenic mice (Tg) by microinjection. In this study, we used in vitro fertilized mouse eggs, followed by ultrarapid freezing to establish a simple procedure for production of Tg mice. We produced in vitro fertilized mouse eggs and cryopreserved them by ultrarapid freezing method. A total of 139 cryopreserved-thawed pronuclear eggs, of which 101 (72.6%) were survived following microinjection of chicken b-actin promoter-driven firefly improved luciferase cDNA (β-act/luc+) and were transferred into 5 recipients. All recipients became pregnant and gave birth to a total of 15 (14.8%) pups. As a control, same DNA construction (β- act/luc+) was also injected into 450 in vitro fertilized eggs, of which 338 (75.1%) were survived and then were transferred into 14 recipients. Eleven (78%) mice became pregnant and littered a total of 54 (19.1%) pups. Southern blotting analysis of Tg mice indicated that one (1/15, 6.6%) and three (3/54, 5.5%) transgenic mice were production from cryopreserved and in vitro fertilized eggs, respectively. All Tg mice produced from both eggs showed the expression of improved luciferase gene. These results indicated that efficiency of produced of Tg mice from cryopreserved eggs was comparable to that from in vitro fertilized eggs. Furthermore, it is suggested that microinjection of transgene into in vitro fertilized eggs cryopreserved by ultrarapid freezing is an easy and conveniently method for production of Tg mice.

Selenium (Se) is an essential trace element for humans and animals, and several findings suggest that dietary Se intake may be necessary for bone health. Accumulating evidence indicates that Se compounds possess anticancer properties. Se is specifically incorporated into proteins in the form of selenocysteine and non-specifically incorporated as selenomethionine in place of methionine. This study evaluated protection by Se in the bone repair process in ovariectomized rats after irradiation. For such purpose, 80 ovariectomized female Sprague-Dawley rats were randomly divided into 4 experimental groups: ovariectomized (Ov), Ov/Se, Ov/irradiated (Irr) and Ov/ Se/Irr. A bone defect was created on the tibia of all animals 40 days after ovariectomy. Two days after surgery, only the Ov/Se and Ov/Se/Irr rats received 0.8 mg Se/kg. Three days after surgery, only the Ov/Irr and Ov/Se/Irr rats received 10 Gy of X-rays on the lower limb region. The animals were euthanized at 7, 15, 22 and 29 days after surgery to assess the repair process, which was evaluated by analysis of trabecular bone number (Masson Trichrome) and birefringence analysis (Picrosirius). It was possible to observe a delay in the bone repair process in the ovariectomized/irradiated group and similarity between the ovariectomized, Ov/ Se and Ov/Se/Irr groups. Our findings suggest that sodium selenite may influence a radioprotective effect in the bone repair of tibia of ovariectomized rats without toxicity.

The objective of this study was to evaluate the toxicities of permeable cryoprotectants and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Toxicities of permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), Glycerol, and 1,2-PROH were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to among DMSO, EG, Glycerol, and 1,2-PROH. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO and Glycerol at the 2-cell stage was significantly lower than that were treated with EG (81.1±15.1), 1,2-PROH (88.0±21.1) or the control (99.9±21.3) (p<0.001). On comparison of four cryoprotectant treated groups, the DMSO and Glycerol treated group showed a decreased cell count compared with the EG and 1,2-PROH treated group (p<0.05). Both DMSO (14.7±1.3), EG (12.1±1.1), Glycerol (15.2±1.8), and 1,2-PROH (11.5±1.3) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control (6.5±0.7, p<0.0001). In addition, the DMSO or Glycerol treated group showed more apoptotic cells than the EG or 1,2-PROH treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to among DMSO, EG, Glycerol, and 1,2-PROH at room temperature. When comparing four permeable cryoprotective agents, EG and 1,2-PROH appeared to be less toxic than DMSO and Glycerol at least in a murine embryo model.

The objective of this study was to monitor health conditions of genetically identical somatic cells cloned Korean white cattle, endangered indigenous cattle (EIC) and indigenous cattle (IC) by analysis of hematologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and one live cattle were delivered by caesarean section. The cloned Korean white cattle were genetically identical to the nuclear donor cattle. As a result, the mean values of RBC and platelet of cloned cattle and white cattle were significantly decreased by age (P<0.05). The mean values of RBC, HCT, MCV and MCHC between cloned cattle and IC of the same age (1∼2 years) showed the statistical significance (P<0.05). Also, in the WBC of Korean white cattle, the estimated values were decreased according to the age from 12.0×103/μl under 1 year to 11.0×103/μl over 1 years respectively. Although clone-cattle had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned cattle were not different when compared to reference range. This study suggests that cloned Korean white cattle derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological parameters. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

이 연구에서는 정소가 제거된 흰쥐에 비타민 E와 셀레늄(Selevit)을 5주간 투여 후 체중, 장기 무게, 혈액학적 그리고 생화학적 변화를 관찰하였다. 체중의 변화에서는 모든 실험군에서 증가가 나타냈다. Orch+Selevit군의 체중 증가는 11.2±10.25 g으로 가장 낮았으며, Intact군, Sham군, Orch군과 비교 시 유의적으로 감소되었다. 장기 무게의 변화에서는 Orch+Selevit군의 심장과 간장 무게는 Intact군, Sham군과 비교 시 유의적으로 감소되었다. Intact군, Sham군, Orch군의 신장 무게는 Orch+Selevit군 비교 시 유의적인 증가를 확인하였다. 백혈구 수의 혈액학적 변화에서는 Orch+Selevit군은 다른 모든 군과 비교해 유의적인 증가를 확인하였으며, 적혈구수, 평균적혈구용적, 평균적혈구혈색소량, 평균적혈구색소농도 등과 같은 혈액학적 측정치에서는 유의성은 인정되지 않았다. 생화학적 변화에서는 Orch+Selevit군의 혈청총단백질, 알부민은 Orch군과 비교 시 유의적으로 증가하였으며, 알부민은 Intact군, Sham군 그리고 Orch군과 비교 시 유의적으로 감소했다. AST와 ALT는 모든 실험군에서 유의성이 없었다.

본 연구에서 배아의 생식세포 동결에 가장 흔히 쓰이고 있는 두 가지 동결 보호제, 즉 DMSO와 EG의 독성을 비교하고자 생쥐 수정란 모델을 이용한 실험을 하였다. 생후 6주령의 암컷 생쥐 F1 hybrid mice에 10 IU의 PMSG를 복강 주사하여 과배란을 유도하고, 2-세포기 배아를 획득하고 DMSO와 EG 각각 노출시킨 후, 배양을 하였다. 배반포의 전체 세포수는 2-세포기 단계에서 DMSO(68.1±24.1)로 EG(81.2±27.0) 혹은 control(99.0±18.3)(p<0.001) 처리구에 비해서 유의적으로 낮았다. DMSO 처리구가 EG 처리구에 비해 세포수가 적었다. DMSO(15.4±1.5)와 EG(10.2±1.4) 두 처리구는 대조구(6.1±0.9, p<0.0001)와 비교해서 배반포에서 세포사 비율이 더 높음을 확인했다. 또한, DMSO 처리구는 EG 처리구(p<0.001)보다 더 많은 세포사멸된 세포가 확인되었다. DMSO 또는 EG 처리군과 대조군 사이에는 배아 부화율에 있어서 차이가 있었으며, 이는 배아에 대한 동결 보호제의 잠재적인 독성을 확인한 결과였다. 이번 연구에서 장기간 처리했을 때 EG 처리군보다 DMSO 처리군에서 배아발달과 세포수가 저하된 것은 DMSO의 독성이 더 높을 것으로 사료된다.

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 μg/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.

The importance of genetic resource preservation has been highlighted in the literature as a means of maintaining genetic diversity. Investigations for hematologic values and the differential count of white blood cell count (WBC) for Korean indigenous cattle (KIC) and endangered indigenous cattle (EIC) are rarely performed. Therefore, the objective of this study was to investigate the hematologic values of total 40 EIC (White, Black, Mini cattle) and 35 KIC as control by analysis of hematologic characteristics. As a result, the mean values of RBC and platelet of EIC were significantly decreased by age (p<0.05). The mean values of RBC, HCT, MCV and MCHC between EIC and KIC of the same age (2 ～3 years) showed the statistical significance (p<0.05). Also, in the WBC of EIC, the mean values were decreased according to the age from 13.9×103/μL～12.7×103/μL under 1 year to 9.1×103/μL～11.5×103/μL over 2 years respectively. In the differential count of WBC of EIC (White, Black, Mini cattle), it showed generally the rates of 40.2%, 52.2%, 49.0% lymphocyte and 27.2%, 33.9%, 32.0% segmented neutrophil from 2～3 years respectively. Result of this study will be used for establishing reference range for blood analysis in EIC such as white, black and mini cattle. This study reported hematological values which could serve as baseline information for comparison in conditions of nutrient deficiency, physiological and health status of endangered Korean native cattle. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

The occurrence of mastitis in diary cattle has been caused by genetic, physiological, managemental and environment factors accounted for the highest percentage of worldwide disease in dairy cattle. The purpose of this study was to analyze the occurrences and causative bacteria of subclinical mastitis in milking cows and also examine the distribution of bacteria in milk by isolating and identifying bacteria both in whole milk and quarter milk. 31.4% of the milking cows suffered subclinical mastitis, and 9.5% had it in terms of quarter milk. According to the results of analyzing bacteria in quarter milk of which somatic cell count (SCC) was over 500 thousand, 15 kinds of bacteria were isolated, and among them, Pantoea spp. formed the biggest part as 15.8%. From whole milk, 37 kinds of bacteria were identified, and among them, Klebsiella oxytoca showed the highest identification rate as 30.1%. According to the results of bacteria analyzed from the quarter milk of entire milking cows, 52 kinds of bacteria were identified. Among them, 17 kinds of Staphylococci were isolated, and CNS (Coagulase-Negative Staphylococci) formed a large part as 44.9%. The findings of this study showed that various kinds of bacteria were isolated from cows having subclinical mastitis; therefore, when managing specifications about milking or such, dairy farm will have to take proper action like performing sanitary control or counting somatic cells regularly in order to do their best for reducing mastitis.

The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Holstein Donor Cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with PGF2α and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. Seventy two point seven percentage of the cows(16/22) had normal resumption of ovarian cyclicity(resumption within 40 days after superovulation), and 27.3%(6/22) had delayed resumption(resumption did not occur until＞40 days after superovulation). Delayed resumption Type Ⅱ(first ovulation did not occur until ≥40 days after superovulation, i.e. delayed first ovulation 13.6%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and 10≤ of corpora lutea(CL) was 7.8±1.8 and 12.7±2.7, respectively. The number of transferable embryos differed between < 10 and 10≤ CL was 5.4±1.3 and 8.1±3.4, respectively. Four point five percentage of the cows(1/22) did not resumption their ovarian cyclicity until 60 days after superovulation treatment. Diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.

The objective of this work was to analyze the concentrations of progesterone (P4) and estrogen (E2) hormones changed during estrus synchronization in dairy heifers. Estrus synchronization was carried out with CIDRⓇ (Controlled Intravaginal Drug Release) devices. Corpus luteum (CL) was classified into three grades based on its size and palpable characteristics. The concentrations of P4 and E2 were measured by enzyme-amplified chemiluminescence. Serum P4 concentration was markedly low at the estrus stage (36 hrs after removal of CIDR) compared to other stages, while E2 concentration was kept high during estrus stage. The serum P4 concentration was highest in the CL classified into gradeⅠ. These results indicate that P4 concentration could be used as a criteria for determining recipients for artificial insemination or embryo transfer in dairy cattle.

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.

This study was performed to analyze the causative agents of disease occurred in Korean black goat. Bovine viral diarrhea (BVD) and infectious bovine rhinotracheitis (IBR) known as main causative agents of diarrhea and abortion in cattle were founded in Korean black goats. Escherichia coli and Clostridium perfringens are the main causes underlying diarrhea in Korean black goats as well as other animals. In addition, the orf virus was found as a causative agent of contagious ecthyma that shows frequent occurrence in Korean black goats. Leptospirosis was found in Korean black goats with low occurrence rate. However, unlike the reports from many other countries, we could not detect the antibodies for Johne’s disease and Neospora caninum in Korean black goats. These results are likely to contribute to improving the productivity of raising black goats as a field of livestock farming.

Microtubule-associated protein 1B (MAP1B), a member of MAP1 family, plays a key role in the brain development. MAP1B binds to many kinds of proteins directly or indirectly. In our previous studies, MAP1B and glyceraldehydes 3-phosphate dehydrogenase (GAPDH) were down-regulated in bovine follicular cystic follicles (FCF). This study was performed to examine interaction between MAP1B and GAPDH in bovine follicles using immunoprecipitation (IP) with western blot analysis and immunohistochemisty. MAP1B and GAPDH mRNA expression levels were down-regulated in bovine FCFs. Consistent with the semi-quantitative PCR data, their protein expressions were also down-regulated in FCFs. IP data showed that MAP1B bound to GAPDH in normal follicles, but their binding was absent in FCFs, suggesting that these data might be resulted from a low level of MAP1B and/or GAPDH expression. These results suggest that GAPDH does not as always function as a loading control in bovine follicles.