• 한국동물위생학회지
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• 제41권4호
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• pp.263-269
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• 2018

Developing drugs targeting respiratory pathogen is essential to control respiratory diseases. Many experiments have been performed under in vivo situation. However, in vivo experiments have economical and ethical issues. The objective of this study was to determine the possibility of developing an ex vivo lung culture system with possible application for respiratory infection studies. After isolating lungs from naïve pigs, agarose-inflated lung tissues were prepared and sliced manually. These sliced lung tissues were then subsequently placed on 24-well plates. Eight different combinations of media were used to determine the optimum ex vivo lung culture condition. In addition, lung tissues were infected with porcine reproductive and respiratory syndrome (PRRS) virus at a titer of $1{\times}10^4\;TCID_{50}/mL$. Virus growth was confirmed by titration in MARC-145 cells at 2, 4, 6 days post infection (dpi). We found that ex vivo lung culture in physiological environment was not media specific based on histopathology and cytotoxicity. However, under virus-infected condition, thickened alveolar walls in the lung tissues and stable virus titers at 2, 4, 6 dpi were shown in F12K medium suggesting that it was useful for tissue maintenance and virus infection using PRRS virus infected lung tissues. The present study shows the possibility of using porcine ex vivo lung model for respiratory infection studies.

• Clinical and Experimental Reproductive Medicine
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• 제11권2호
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• pp.41-49
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• 1984

임신직후 자궁조직은 난소 스테로이드 호르몬에 의하여 분화한다. 착상전 여러 단계에 있는 자궁조직내의 cAMP의 함량을 측정함으로서 cAMP와 호르몬과의 관계를 규명할 목적으로 흰쥐를 재료로 해서 본 실험을 행하였다. 임신제 2일에 난소를 제거하고 즉시, 제3, 또는 제 4일에 estradiol과 progesterone 을 각각 혹은 동시에 주사한 뒤 대조군인 정상임신군, 난소제거군, 난소제거후 호르몬주사군의 자궁조직에서 cAMP농도를 competitive protein-binding assay로 측정하여 이를 각각 비교하였다. 본 연구결과는 다음과 같았다. 정상임신군인 경우 자궁조직이 분화해 갈수록 자궁조직내 cAMP의 농도는 낮아지고 있다. 난소제거군에서는 임신기간이 길수록 자궁조직내의 cAMP농도가 높아진다. 난소제거후 progesterone 혹은 progesterone 과 estradiol을 주사한 실험군에서는 임신 제 6일이 되면 cAMP 농도가 정상임신군과 난소제거군의 중간수치를 보여준다. 한편, estradiol 단독주사인 경우는 임신제3, 제6일의 자궁조직내의 cAMP의 농도는 크게 감소한다. 이처럼 estradiol의 영향아래 세포분열등 분화가 왕성한 자궁조직에서는 cAMP 농도가 크게 낮아지지만 자궁조직의 분화가 억제된 상태에서는 cAMP의 농도가 반대로 높아진다. 난자, 배아 또는 여러 형태의 세포에서처럼 cAMP의 농도는 세포의 대사에 크게 영향을 주고 있다는 것을 알 수 있었다.

• 한국패류학회지
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• 제20권1호
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• pp.55-63
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• 2004

Gonadosomatic index, condition index and reproductive cycle with the gonadal development of the female Octopus ocellatus were investigated by histological observations and morphometric data, from January to December, 2000. And changes in biochemical components of the ovary and the trunk tissues including the digestive organ associated with gonadal development were studied by biochemical analysis from January to October, 2001. The specimens were collected at the coastal waters of Buan, Jeollabuk-do, Korea, from January 2000 to October 2001. O. ocellatus is a dioecious organism. The gonad of O. ocellatus locates medially in posterior region of the body. Morphology of the ovary shows round and oval in shape, the average diameter and external colour of ripe ovary was 32 mm and semitransparent light brown in colour. As the ovary was getting mature, transparent elongated eggs covered with chorion were present in the ovarian cavity. Monthly changes in the gonadosomatic index (GSI) showed a similar pattern with those of the condition index. The GSI and condition index began to increase in March and reached the maximum in April. And then, their values decreased from May and reached the minimum in September. Reproductive cycle of O. ocellatus can be categorized into five successive stages: early developing stage (September to December), late developing stage (November to March), ripe stage (March to May), partially spawned stage (April to June), and degenerative/resting stage (June to October). Follicle cells attached to an oocyte were involved in vitellogenesis in the cytoplasm of the vitellogeneic oocyte and formation of chorion (secondary egg membrane) of the ovarian eggs. Spawning occurred between April and June. The spawning period was once a year and the peak took place between May and June. This species belongs to semelparity. According to changes in biochemical contents of the ovary and the digestive organ, monthly variations of moisture, total protein, total lipid and glycogen contents (%) in the ovary showed a negative correlationship with those of the trunk tissues including the digestive organ. Accordingly, it is assumed that the ovary only may be received nutrient supply (total lipid content) for gonadal development from the trunk tissues including the digestive organ (r = -0.55, p < 0.05).

• 한국발생생물학회지:발생과생식
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• 제16권3호
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• pp.227-233
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• 2012

Circadian timing system plays a major role in a wide range of reproductive function. However it is plausible idea that other environmental and/or internal cue might be simultaneously participated in the optimal regulation of reproductive system. In the present study we extended the reverse feeding (RF) time regimen up to 8 weeks, then measured the general and reproductive indices of the animals. The animals of ad libitum feeding group (Control, CON) have free access to food for 4, 6 and 8 weeks, respectively. The day feeding animals (reverse feeding, RF group) have restricted access to food during daytime (09:00-18:00) for 4, 6 and 8 weeks, respectively. When the feeding schedules were over, key indices were measured. After 4 weeks and 8 weeks of feeding, body weights of animals were not significantly different. However, body weights of 6 weeks RF animals were significantly smaller than those of control animals (CON : RF = $333.46{\pm}12.71$ g : $289.91{\pm}8.31$ g, p<0.01). The blood glucose levels of 4 weeks RF animals were significantly decreased compared to the levels of control animals (CON : RF = $161.4{\pm}2.7$ mg/dL : $176.7{\pm}5$ mg/dL, p<0.01) while the levels of 6 weeks RF and 8 weeks RF animals were not different form those of control animals. Reproductive and non-reproductive tissue weights from 6 weeks RF group were significantly lowered than those from CON group (testis, CON : RF = $1.4714{\pm}0.0174$ g : $1.3724{\pm}0.0168$ g, p<0.001; epididymis, CON : RF = $0.3574{\pm}0.0059$ g : $0.3243{\pm}0.0068$ g, p<0.001; seminal vesicle, CON : RF = $0.1655{\pm}0.0068$ g : $0.1328{\pm}0.0054$ g, p<0.001; prostate, CON : RF = $0.3350{\pm}0.0231$ g : $0.2528{\pm}0.0143$ g, p<0.01). After 4 weeks and 8 weeks of reverse feeding, sperm counts in RF animals were markedly reduced than those in control animals[CON 4W : RF 4W = $121.17{\pm}9.96\;({\times}10^6)$ : $50.86{\pm}9\;({\times}10^6)$, p<0.001; CON 8W : RF 8W= $138.69{\pm}9.8\;({\times}10^6)$ : $108.94{\pm}4.22\;({\times}10^6)$, p<0.001]. Present study indicates that RF may induce an adaptable metabolic stress and cause impairment of androgen-dependent reproductive tissues. On-going longitudinal studies will allow a better understanding of the how does mealtime shift affect the reproductive function and exact nature of adaptation.

• Clinical and Experimental Reproductive Medicine
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• 제26권2호
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• pp.219-223
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• 1999

Ovarian development of the vitrified neonatal ovaries after orthotopical transplantation into the ovariectomized adult recipient mouse were observed. Ovaries were collected from the neonatal females on day of birth and grouped for fresh, vitrification for 1-minute, and 3-minute. Vitrified and thawed neonatal ovaries were orthotopically transplanted into ovarian bursa of the adult mice from which endogenous ovaries have removed just prior to the transplantation (1 minute: n=25; 3 minutes n=23). Fresh ovarian tissue transplanted (n=25) mice were included as control groups. Returning of the estrus cycles and the survival and development of the transplanted ovaries were evaluated. Intact ovaries from neonatal, and four weeks old mice were used for comparison of the ovarian development as in vivo-developed control. From 2 weeks after transplantation, 64%, 36%, and 75% of the transplanted mice showed return of the estrus cycles in fresh, 1-minute, and 3-minute groups, respectively. Four weeks after transplantation, all mice were sacrificed and ovarian tissues were recovered for histological analysis. 57.1%, 33.3%, and 64.7% mice in fresh, 1-minute, and 3-minute groups, respectively, had survived ovaries with follicles at various stages of growth from primordial to preovulatory follicles. Corpus lutea were also observed. Results of the present study suggest that 1) normal folliculogenesis has initiated in vivo after vitrification, and 2) the vitrification may be used as a preservation method for ovarian tissues for establishment of ovarian tissue bank.

• 대한본초학회지
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• 제30권3호
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• pp.55-61
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• 2015

Objectives : This study aimed to investigate the effects of Shingi-whan(SG) on the male reproductive and sexual function, so we measured the spermatogenesis and the testicular toxicity in mice and the vasorelaxation in isolated rabbit corpus cavernosum smooth muscle. Methods : To evaluate effect on the spermatogenesis in mice, we prepared two groups, control group and SG group that was orally administered SG(1,000mg/kg) for 20 days, and compared. To analyze testicular toxicity in mice, we also prepared two groups, doxo group that was injected with doxorubicin (3mg/kg) on three times and doxo + SG group that was injected with doxorubicin and SG for 20 days, and compared. To investigate sexual function of SG in mice, we prepared three groups, normal group and aging elicited group consisting of 18-month-old mice, SG treated aging group that was orally administered SG for 60 days, and compared using histochemical staining on mice corpus cavernous tissues. In order to define the relaxation effects of SG, rabbit corpus cavernous tissues were prepared in $2{\times}2{\times}6mm$ sized strip. Then the dose-dependent relaxation responses of SG at 0.01-3.0 mg/ml in contracted strips induced by phenylephrine were measured. Results : The sperm density in dutus epididymis and the diameter of seminiferous tubules of SG group was significantly increased when compared to control group. The testicular weight and the diameter and height of epithelial layer of seminiferous tubules of doxo + SG group was significantly increased when compared to doxo group. The cavernous strips were significantly relaxed by SG extract In SG treated aging group, ratio of smooth muscles to collagen fibers and red blood cell count in venous sinus was increased as compared to aging elicited group. Conclusions : Our findings have shown that SG extract have effect on spermatogenesis and mitigating effect on doxo-induced testicular toxicity. Further, it also have the vasorelaxant effect on rabbit corpus cavernosum.

• 한국발생생물학회지:발생과생식
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• 제18권4호
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• pp.301-309
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• 2014

Nesfatin-1, an anorexic nucleobindin-2 (NUCB2)-derived hypothalamic peptide, controls appetite and energy metabolism. Recent studies show that nesfatin-1/NUCB2 is expressed not only in the brain but also in gastric and adipose tissues. Thus, we investigated the distributions of nesfatin-1/NUCB2 in various tissues of male and female mice by real-time PCR, western blotting, and immunohistochemical staining. Real-time PCR analyses showed that NUCB2 mRNA was predominantly expressed in the pituitary and at lower levels in the hypothalamus, spleen, thymus, heart, liver, and muscle of both male and female mice. Expression was much higher in reproductive organs, such as the testis, epididymis, ovary, and uterus, than in the hypothalamus. Western blot analysis of the nesfatin-1 protein level showed similar results to the real-time PCR analyses in both male and female mice. These results suggest that nesfatin-1/NUCB2 have widespread physiological effects in endocrine and non-endocrine organs. In addition, immunohistochemical staining revealed that nesfatin-1 was localized in interstitial cells, including Leydig cells and in the columnar epithelium of the epididymis. Nesfatin-1 was also expressed in theca cells and interstitial cells in the ovary and in epithelial cells of the endometrium and uterine glands in the uterus. These results suggest that nesfatin-1 is a novel potent regulator of steroidogenesis and gonadal function in male and female reproductive organs. Further studies are required to elucidate the functions of nesfatin-1 in various organs of male and female mice.

• 대한한방부인과학회지
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• 제19권1호
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• pp.111-124
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• 2006

Purpose : This study was conducted to investigate the dose dependent effects of Panax Ginseng on the reproductive functions in mice. Methods : We used the 8-week-old mice, and administered 0.2ml extract solution of Panax Ginseng in the different concentration once a day for 60 days. The control group was administered 0.2ml normal saline in the same way and duration. We counted the total, motile and normal sperm number of the cauda epididymis and measured the activities of sperm hyaluronidase, peroxidase and catalase of the isolated testis tissues. And we observed histological changes of surgically isolated testis by histochemical methods. Results : All Panax Ginseng extract solution groups showed significantly dose dependent differences in the total number, the motility and normality of sperms compared with the control group, respectively. In the histological analysis of the testicular tissues, all Panax Ginseng extract solution groups showed the enlargement of testicular lobe diameter and apparent angiogenesis between seminiferous tubules. And the activity of typical sperm enzyme, hyaluronidase, was significantly increased in the Panax Ginseng extract solution groups compared to the control group. In the antioxidant activity analysis, the activities of peroxidase and catalase were significantly increased in the Panax Ginseng extract solution groups compared to the control group. Conclusion : This study shows that Panax Giseng has the beneficial effects on the concentration, morphology and motility of sperm, the activities of sperm hyaluronidase, testicular peroxidase and catalase. We can suggest that Panax Ginseng be useful for the treatment of male infertility.

• 한국발생생물학회지:발생과생식
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• 제20권2호
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• pp.91-99
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• 2016

Lipopolysaccharide (LPS), an endotoxin, elicits strong immune responses in mammals. Several lines of evidence demonstrate that LPS challenge profoundly affects female reproductive function. For example, LPS exposure affects steroidogenesis and folliculogenesis, resulting in delayed puberty onset. The present study was conducted to clarify the mechanism underlying the adverse effect of LPS on the delayed puberty in female rats. LPS was daily injected for 5 days ($50{\mu}g/kg$, PND 25-29) to treated animals and the date at VO was evaluated through daily visual examination. At PND 39, animals were sacrificed, and the tissues were immediately removed and weighed. Among the reproductive organs, the weights of the ovaries and oviduct from LPS-treated animals were significantly lower than those of control animals. There were no changes in the weights of uterus and vagina between the LPS-treated and their control animals. immunological challenge by LPS delayed VO. Multiple corpora lutea were found in the control ovaries, indicating ovulations were occurred. However, none of corpus luteum was present in the LPS-treated ovary. The transcription level of steroidogenic acute regulatory protein (StAR), CYP11A1, CYP17A1 and CYP19 were significantly increased by LPS treatment. On the other hand, the levels of $3{\beta}$-HSD, $17{\beta}$-HSD and LH receptor were not changed by LPS challenge. In conclusion, the present study demonstrated that the repeated LPS exposure during the prepubertal period could induce multiple alterations in the steroidogenic machinery in ovary, and in turn, delayed puberty onset. The prepubertal LPS challenge model used in our study is useful to understand the reciprocal regulation of immune (stress) - reproductive function in early life.

• 한국동물위생학회지
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• 제37권3호
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• pp.143-150
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• 2014

Porcine reproductive and respiratory syndrome virus (PRRSV) is the etiological agent of PRRS characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRSV is a small enveloped virus containing a positive-sense, single-stranded RNA genome and divided into two genotype, type 1 (European) and type 2 (North American), respectively, by nucleotide identity. In this study, ORF7 gene of the type 1 and type 2 PRRSV was cloned and expressed in Baculovirus expression system. Also, monoclonal antibodies (MAbs) against ORF7 were produced and characterized. The expressed ORF7 proteins in the recombinant virus were confirmed by indirect fluorescence antibody (IFA) test using His6 and PRRSV-specific antiserum. A total of eight MAbs were produced and characterized. One (3G12) MAb was type 1 PRRSV ORF7-specific and two (6B10 and 16H8) were type 2 PRRSV ORF7-specific. Other five (1A1, 2A4, 4B4, 12C4 and 13F11) MAbs reacted with both type 1 and type 2 PRRSV. Some PRRSV ORF7-specific MAbs recognized the porcine tissues infected with PRRSV by IFA or immunohistochemistry (IHC) assay. From this experiment, it was confirmed that MAbs produced in this study were PRRSV ORF7-specific and could be used as reliable reagents for type 1/type 2 PRRSV detection.