Rats are one of the most widely used animals in biomedical sciences because their metabolism and physiology are comparable to humans. In recent years, gene-targeted models have been developed using various animal species utilizing engineered nucleases such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated gene (Cas). It has recently become possible to efficiently transfect CRISPR/Cas into embryos via electroporation. However, electroporation can damage fertilized eggs; therefore, it is important to determine the optimal embryo culture conditions. A standardized approach for routine and reproducible rat transgenesis will render rat models more accessible for research. We performed experiments to obtain rat embryos with efficient superovulation and synchronization, and to investigate the appropriate medium conditions for pronuclear stage embryos subjected to electroporation stimulation for the introduction of engineered nuclease.

Lilium lancifolium (LL) is widely cultivated in East Asia and used to attenuate airway diseases. Our current study aimed to investigate the therapeutic effect of LL on pain level and inflammatory response in a rat model of monosodium iodoacetate (MIA)-induced osteoarthritis (OA). We first examined the effect of LL on inflammatory cytokines and inflammatory mediators in IL-1β-treated HTB-94 cells. The LL extract was found to significantly inhibit the levels of Interleukin-6 (IL-6), Interleukin-8 (IL-8), and prostaglandin-E2 (PGE-2) in Interleukin-1 β (IL-1β)-stimulated HTB-94 cells in a dose-dependent manner. Moreover, chronic oral administration of LL effectively restored the weight-bearing distribution in the rat model of MIA-induced OA. In addition, administration of LL inhibited inflammatory cytokines and inflammatory mediators, such as C-reactive protein (CRP), IL-6, leukotriene B4 (LTB-4), PGE-2, and matrix metalloproteinase-9 (MMP-9). Our findings collectively suggested LL as one of the potential therapeutic agents for OA, owing to its properties of reducing pain and inflammatory responses.

In present study, we investigated the antidiabetic effect of Momordica charantia(as well known “bitter melon”). This study was conducted to determine antidiabetic mechanism of Bitter Melon Extract (BME). We measured blood glucose, insulin, glucagon level in a Sprague-Dawley rat model of high-fat diet/streptozotocin(HFD/STZ)-induced diabetes. Five experimental groups were used: normal, HFD/STZ, BME 62.5 mg/kg HFD/STZ, BME 125 mg/kg HFD/STZ and BME 250 mg/kg HFD/STZ. BME was orally administered to the rats every other day for 9 weeks. Results showed that fasting blood glucose levels were significantly lower in the BME 125 mg/kg(150.17 ± 20.22 mg/dL) and 250 mg/kg(124.17 ± 22.17 mg/dL) groups than in the vehicle group(188.83 ± 26.63 mg/dL)(p<0.05). In addition, glucagon levels were lower in the three BME treatment groups than in the vehicle group(p<0.05). Oral glucose tolerance tests revealed that the BME 250 mg/kg group had significantly(p<0.05) reduced 120-minute blood glucose levels and areas under the curve. Our results suggest that BME induces antidiabetic effects via the reduction of glucagon and blood glucose levels.

본 연구에서는 강력한 항산화제로 알려진 셀레늄의 방사선방호효과를 연구하였다. 렛드에 셀레늄을 14 일간 경구투여 후 10 Gy의 방사선을 전신조사하였다. 그리고 1일, 3.5일, 7일, 21일 후에 혈구 성분 및 SOD (Superoxide Dismutase)와 소장의 변화를 관찰하였다. 방사선조사군에 비해 셀레늄 투여 후 방사선조사군에 서 조혈면역계의 손상을 경감시키는 유의한 방호 효과가 있었다(p<0.05). 그리고 셀레늄이 항산화 효소인 Superoxide Dismutase(SOD)의 활성을 증가시키는 유효한 성분이며, 방사선 조사에 의한 소장움 세포의 apop tosis 발현을 억제하는 효과가 있음을 확인하였다. 이 결과를 바탕으로 셀레늄은 방사선으로부터 생명체를 보호하는 필수 성분이라 생각한다.

비알코올성 지방간 질환은 만성 간질환의 가장 흔한 원인이다. 이 연구의 목적은 비알코올성 지방간 질환 동물모델을 확립하고 생체 내 짧은 양성자 자기공명분광법을 이용하여 비침습적으로 간의 대사물질과 지질의 변화를 정량 분석하고자 하였다. 각 실험은 고지방식 먹이를 먹고 나서 0주를 기준선으로 하고, 2, 4, 6, 8주에 쥐의 간 실질 조직으로부터 측정되었다. 0주를 기준선으로 하고 2주와 비교했을 때 0.9, 1.3, 2.3, 2.8 및 5.3 ppm 에서 유의미하게 증가하였다(p<0.01). 따라서 양성자 자기공명분광법은 고지방식 먹이를 먹고 2주 후부터 다양한 쥐의 간 지질 변화를 검출하고 특성화하는데 유용할 것이다.

In the multicellular tissue, cell-cell interaction is important for a precise control of its function. The exchange of signaling molecules between adjacent cells via connexon allows the functional harmony of cells in the tissue. The present research was to determine the presence and expressional patterns of connexin (Cx) isoforms in the rat epididymal fat during postnatal development using quantitative real-time polymerase chain reaction (PCR) analysis. Of 13 Cx isoforms examined, expression of 11 Cx isoforms in the epididymal fat during postnatal development was detected. These Cx isoforms include Cx26, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx43, Cx45, and Cx50. Expressional levels of all Cx isoforms at 1 and 2 years of age were significantly higher than those at the early postnatal ages, such as 7 days, 14 days, and 24 days of ages. Except Cx33 and Cx43, the transcript levels of rest Cx isoforms at 1 year of age were significantly lower than that at 2 years of age. In addition, expressional patterns of Cx isoforms between 7 days and 5 months of ages generally varied according to the isoform. The existence of various Cx isoforms in the rat epididymal fat has been identified and expression of each Cx isoform in the epididymal fat during postnatal development has shown a particular pattern, distinguishable from the others. To our knowledges, this is the first report showing expressional patterns of Cx isoforms at transcript level in the epididymal fat at various postnatal ages.

In the present study, we employed Hershberger assay to determine possible androgenic or antiandrogenic activities of three di-2-ethylhexyl phthalate (DEHP) substitute candidates. The assay was carried out using immature castrated Sprague–Dawley male rats. After 7 days of the surgery, testosterone propionate (TP, 0.4 mg/kg/day) and test materials (low dose, 40 mg/kg/day; high dose, 400 mg/kg/day) were administered for 10 consecutive days by subcutaneous (s.c.) injection and oral gavage, respectively. Test materials were DEHP, 2-ethylhexyl oleate (IOO), 2-ethylhexyl stearate (IOS) and triethyl 2-acetylcitrate (ATEC). The rats were necropsied, and then the weights of five androgen-dependent tissues [ventral prostate, seminal vesicle, coagulating glands, levator ani-bulbocavernosus (LABC) muscle, paired Cowper’s glands, and glans penis] and four androgen-insensitive tissues (kidney, adrenal glands, spleen and liver) were measured. All test materials including DEHP did not exhibit any androgenic activity in the assay. On the contrary, antiandrogen-like activities were found in all test groups, and the order of the intensity was ATEC < DEHP < ISO < IOO in the five androgen-sensitive tissues. There was no statistical difference between low dose treatment and high dose treatment of all replacement candidate groups. In DEHP groups, high dose treatment exhibited significant weight gains in LABC and Glan Penis. There was no statistical difference in androgen-insensitive tissue measurements. Since the effects of ATEC treatment on the accessory sex organs were much less or not present at all when compared to those of DEHP, ATEC could be a strong candidate to replace DEHP. IOO treatment brought most severe weight reduction in all of androgen-sensitive tissues, so this material should be excluded for further screening of DEHP substitute selection.

Bisphenol-A(BPA) is a member of alkylphenol family, and shows adverse effects including reduced fertility, reproductive tract abnormalities, metabolic disorder, cancer induction, neurotoxicity and immunotoxicity. In the present study, we conducted Hershberger assay to evaluate whether the two candidates to replace BPA have androgenic or antiandrogenic activity. The assay was carried out using immature castrated Sprague–Dawley male rats. After 7 days of the surgery, testosterone propionate (TP, 0.4 mg/kg/day) and test materials (low dose, 40 mg/kg/day; high dose, 400 mg/kg/day) were administered for 10 consecutive days by subcutaneous (s.c.) injection and oral gavage, respectively. Test materials were BPA, isosorbide (ISO) and cyclohexanedimethanol (CHDM). The rats were necropsied, and then the weights of five androgen-dependent tissues [ventral prostate, seminal vesicle, levator ani-bulbocavernosus (LABC) muscle, paired Cowper’s glands, and glans penis] and three androgen-insensitive tissues (kidney, spleen and liver) were measured. All test materials including BPA did not exhibit any androgenic activity in the assay. On the contrary, antiandrogen-like activities were found in all test groups, and the order of the intensity was CHDM > BPA > ISO in the five androgen-sensitive tissues. There was no statistical difference between low dose treatment and high dose treatment of BPA group as well as ISO group. In CHDM group, high dose treatment exhibited most severe weight reduction in all measured tissues. There was no statistical difference in androgen-insensitive tissue measurements, except BPA groups. Since the effects of ISO treatment on the accessory sex organs were much less or not present at all when compared to those of BPA, ISO could be a strong candidate to replace BPA. CHDM treatment brought most severe weight reduction in all of androgen-sensitive tissues, so this material should be excluded for further screening of BPA substitute selection.

Direct communication between neighboring cells through connexin (Cx)-based gap junction is a crucial biolo– gical manner to regulate functions of a tissue consisting of multi-cell types. The present research evaluated expressional changes of Cx isoforms in the caput epididymis of adult rat exposed to estradiol benzoate (EB) or flutamide (Flu) at the early postnatal age. A single subcutaneous administration of EB at a low-dose [0.015 g /kg body weight (BW)] or a high-dose (1.5 g/kg BW) or Flu at a low-dose (500 g/kg BW) or a high-dose (5 mg/kg BW) was performed to an animal at 1 week of age. Quantitative real-time PCR analysis was employed to determine expressional changes of Cx isoforms. The transcript levels of Cxs30.3 and 37 were decreased by a low-dose EB treatment, while decreases of Cxs31, 31.1, 32, 40, and 45 transcript levels were observed with a low-dose EB treatment. The treatment of a high-dose EB resulted in expressional reduction of Cxs30.3, 31, 31.1, 37, 40, 43, and 45. The Flu treatment at a low dose caused increases of Cxs26, 37, and 40 transcript levels but decreases of Cxs31.1, 43, and 45 transcript levels. Increases of Cxs30.3, 31, 37, and 40 mRNA amounts were induced by a high-dose Flu treatment. However, exposure to a high-dose Flu produced expressional decreases of Cxs31.1, 32, and 43 in the adult caput epididymis. These observations suggest that exposure to EB or Flu at the neonatal period could lead to aberrant expression of Cx isoforms in the adult caput epididymis.

Mammalian spermatogenesis occurs in a precise and coordinated manner in the seminiferous tubules. One of the attempts to understand the detailed biological process during mammalian spermatogenesis at the molecular level has been to identify the testis specific genes followed by study of the testicular expression pattern of the genes. From the subtracted cDNA library of rat testis prepared using representational difference analysis (RDA) method, a complimentary DNA clone encoding type III member of a DnaJ family protein, DnaJC18, was cloned (GenBank Accession No. DQ158861). The fulllength DnaJC18 cDNA has the longest open reading frame of 357 amino acids. Tissue and developmental Northern blot analysis revealed that the DnaJC18 gene was expressed specifically in testis and began to express from postnatal week 4 testis, respectively. In situ hybridization studies showed that DnaJC18 mRNA was expressed only during the maturation stages of late pachytene, round and elongated spermatids of adult rat testis. Western blot analysis with DnaJC18 antibody revealed that 41.2 kDa DnaJC18 protein was detected only in adult testis. Immunohistochemistry study further confirmed that DnaJC18 protein, was expressed in developing germ cells and the result was in concert with the in situ hybridization result. Confocal microscopy with GFP tagged DnaJC18 protein revealed that it was localized in the cytoplasm of cells. Taken together, these results suggested that testis specific DnaJC18, a member of the type III DnaJ protein family, might play a role during germ cell maturation in adult rat testis.

Equine chorionic gonadotropin (eCG) is a unique molecule that elicits the response characteristics of both follicle- stimulating hormone (FSH) and luteinizing hormone (LH) in other species. Previous studies from this laboratory had demonstrated that recombinant eCG (rec-eCG) from Chinese hamster ovary (CHO-K1) cells exhibited both FSH- and LH-like activity in rat granulosa and Leydig cells. In this study, we analyzed receptor internalization through rec-eCGs, wild type eCG (eCGβ/α) and mutant eCG (eCGβ/αΔ56) with an N-linked oligosaccharide at Asn56 of the α-subunit. Both the rec-eCGs were obtained from CHO-K1 cells. The agonist activation of receptors was analyzed by measuring stimulation time and concentrations of rec-eCGs. Internalization values in the stably selected rat follicle-stimulating hormone receptor (rFSHR) and rat luteinizing/ chorionic gonadotropin receptor (rLH/CGR) were highest at 50 min after stimulation with 10 ng of rec-eCGβ/α. The dose-dependent response was highest when 10 ng of rec-eCGβ/α was used. The deglycosylated eCGβ/αΔ56 mutant did not enhance the agonist-stimulated internalization. We concluded that the state of activation of rFSHR and rLH/CGR could be modulated through agonist-stimulated internalization. Our results suggested that the eLH/CGRs are mostly internalized within 60 min by agonist-stimulation by rec-eCG. We also suggested that the lack of responsiveness of the deglycosylated eCGβ/ αΔ56 was likely because the site of glycosylation played a pivotal role in agonist-stimulated internalization in cells expressing rFSHR and rLH/CGR.

The present research was chiefly designed to determine the effect of the treatment of estrogenic agonist, estradiol benzoate (EB), or antiandrogenic compound, flutamide (Flu), at the weaning age on the expression of connexin (Cx) isoforms in the caudal epididymis of adult male rat. Animals were subcutaneously administrated with a single shot of either EB at a low-dose (0.015 mg of EB/kg body weight (BW)) or a high-dose (1.5 mg of EB/kg BW) or Flu at a low-dose (500 mg of EB/kg BW) or a high-dose (5 mg of EB/kg BW). Expressional changes of Cx isoforms in the adult caudal epididymis were examined by quantitative real-time PCR analysis. The treatment of a low-dose EB caused significant increases of Cx30.3, Cx31, Cx32, and Cx43 transcript levels but reduction of Cx31.1, Cx37, and Cx45 expression. Exposure to a high-dose EB resulted in very close responses observed in a low-dose EB treatment, except no significant expressional change of Cx37 and a significant induction of Cx40. Expression of all Cx isoforms, except Cx45, was significantly increased by a low-dose Flu treatment. Expressional increases of all Cx isoforms were detected by a high-dose Flu treatment. The current study demonstrates that a single exposure to estrogenic or antiandrogenic compound during the early postnatal developmental period is sufficient to disrupt normal expression of Cx isoforms in the adult caudal epididymis.

Fas ligand (FasL) and its receptor Fas have been implicated in granulosa cell apoptosis during follicular atresia. Although interferon-gamma (IFN-γ) is believed to be involved in the regulation Fas expression in differentiated granulosa or granulosa-luteal cells, the expression of this cytokine and its role in the regulation of the granulosa cell Fas/FasL system and apoptosis during follicular maturation have not been thoroughly investigated. In the present study, we have examined the presence of IFN-γ in ovarian follicles at different stage of development by immunohistochemistry and related their relative intensities with follicular expression of Fas and FasL, and with differences in granulosa cell sensitivity to Fas activation by exogenous agonistic Anti-Fas monoclonal antibody (Fas mAb). Although IFN-γ immunostaining was detectable in oocyte and granulosa cells in antral follicles, most intense immunoreactivity for the cytokine was observed in these cells of preantral follicles. Intense immunoreactivity for IFN-γ was most evident in granulosa cells of atretic early antral follicles where increased Fas and FasL expression and apoptosis were also observed. Whereas low concentrations of IFN-γ (10-100 U/mL) significantly increased Fas expression in undifferentiated granulosa cells (from preantral or very early antral follicles) in vitro, very higher concentrations (≥ 1,000 U/mL) were required to up-regulate of Fas in differentiated cells isolated from eCGprimed (antral) follicles. Addition of agonistic Fas mAb to cultures of granulosa cells at the two stages of differentiation and pretreated with IFN-γ (100 U/mL) elicited morphological and biochemical apoptotic features which were more prominent in cells not previously exposed to the gonadotropin in vivo. These findings suggested that IFN-γ is an important physiologic intra-ovarian regulator of follicular atresia and plays a pivotal role in regulation of expression of Fas receptor and subsequent apoptotic response in undifferentiated (or poorly differentiated) granulosa cells at an early (penultimate) stage of follicular development.

방사선장해 극복을 위한 조직공학실험에서는 다양한 실험방법이 존재한다. 그중에서도 방사선 장해모델을 구현함에 있어서 쥐는 가장 많이 사용되는 실험동물이다. 이번 실험에서는 쥐 피부를 벗긴 후 일정한 두께로 만든 후에 피부 두께에 따른 방사선량을 측정하였다. 또한 쥐의 피부 두께에 따른 방사선 흡수선량을 유추해 날수 있는 수식을 도식화 하였다. 결론적으로 이러한 수식을 적용한다면 방사선장해극복을 위한 조직공학 연구에서 사용하는 다양한 지지체삽입시에 지지체가 받을 수 있는 방사선량의 정도를 유추해 낼 수 있으며 이것을 통하여 쥐 피부두께 피하에 관한 체내 (in-vitro)실험모델 구현 시 방사선량의 정보를 제공하고자 한다. 따라서 이번 연구의 결과를 기반으로 사용한다면 방사선이 노출된 쥐의 피하조직 체내 (in-vitro) 조직공학 실험에서 효과적으로 사용 될 수 있을 것이다.

Background: Many menopausal women suffer from health problems including metabolic diseases such as dyslipidemia and osteoporosis. Thus they need natural products and functional foods particularly highly nutritional food products, that can help alleviate these diseases. This study was carried out to determine the effect of Drynariae Rhizoma water extract on the lipid and bone metabolism of ovariectomized Sprague-Dawley rats.Methods and Results: The animals were randomly divided into six dietary groups comprising SHAM-operated rats, OVX rats (normal diet), and OVX-DR rats (Drynariae Rhizoma extract). After 8 weeks, plasma, liver, and fat samples were collected to analyze the lipid metabolism, plasma Ca, alkaline phosphatase (ALP), osteocalcin and C-terminal telopeptide (CTx) concentrations, which are biochemical makers of bone metabolism. The left femurs of rats were also collected for histological analyses. OVX counteracted menopause induced body weight gain, as well as increases in triglycerides, total cholesterol, and free fatty acids. The Drynariae Rhizoma group showed low levels of triglycerides, high HDL-cholesterol, and decreased lipogenesis based on activity of the lipid-regulating enzymes (fatty acid synthase and malic enzyme). Decreased serum levels of ALP and osteocalcin were observed in Drynariae Rhizoma group.Conclusions: The results of this study show that Drynariae Rhizoma extract may effectively regulate hyperlipidemia and improve bone density.