• Development and Reproduction
• /
• v.17 no.4
• /
• pp.461-467
• /
• 2013

Nesfatin-1/NUCB2, which is associated with the control of appetite and energy metabolism, was reported for the first time to be expressed in the hypothalamus. However, recent studies have shown that nesfatin-1/NUCB2 was expressed not only in the hypothalamus, but also in various tissues including digestive and reproductive organs. We also demonstrated that nesfatin-1/NUCB2 was expressed in the reproductive organs, pituitary gland, heart, lung, and gastrointestinal tract of the adult mouse. However, little is known about nesfatin-1/NUCB2 expression in fetal and neonatal mice. Therefore, we examined here the distribution of nesfatin-1/NUCB2 in various organs of fetal and neonatal mice and compared them with the distribution in adult mice. As a result of immunohistochemical staining, nesfatin-1/NUCB2 protein was expressed relatively higher in the lung, kidney, heart, and liver compared to other organs in the fetus. Western blot results also showed that nesfatin-1/NUCB2 protein was detected in the lung, kidney, heart, and stomach. Next, we compared the expression levels of nesfatin-1/NUCB2 mRNA in the fetus and neonate with the expression levels in both male and female adult mice. The expression levels in heart, lung, stomach, and kidney were higher compared with other organs in fetal and neonatal mice and in both male and female adult mice. Interestingly, the expression of nesfatin-1/NUCB2 mRNA in the kidney was dramatically increased in male and female adult mice compared to fetal and neonatal mice. These results indicate that nesfatin-1/NUCB2 may regulate the development and physiological function of mouse organs. In the future, we need more study on the function of nesfatin-1/NUCB2, which is highly expressed in the heart, lung, and kidney during mouse development.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.11
• /
• pp.1565-1571
• /
• 2003

Physiological mechanisms, involved in unusual ovulatory sequences in domestic hen are remaining undefined. One hundred individually caged white leghorn birds were divided into two equal groups viz. control and treatment, and 2-bromo-$\alpha$-ergocryptine, was administered to birds in the treatment group to modulate prolactin (PRL) secretion from anterior pituitary gland. The effect of modulation of PRL concentrations on egg production, sequence length and intersequence pause length were studied by analysis of oviposition records of the birds from 24 to 72 weeks of age. The surviving 48 birds in the control and treatment groups averaged $34.58{\pm}1.7$ and $25.67{\pm}1.15$ sequences of oviposition, with a mean sequence length of $9.92{\pm}0.63$ and ${\pm}1.12$ days respectively. Most of the birds had a single characteristically long sequence during the entire reproductive cycle, which averaged $46.04{\pm}3.09$ days in the control birds and $59.33{\pm}4.44$ days in the treated birds. 2-bromo-$\alpha$-ergocriptine treatments had significantly decreased (p$\leq$0.01) the circulating concentrations of PRL compared to the birds of the control group. This resulted in a significant increase (p$\leq$0.01) in the number of laying days in birds of the treatment group with a concomitant decrease in the intersequence pause length. The decreased PRL levels during prime sequences in birds of the both groups, reveals the negative role of the circulating PRL levels on egg production with concomitant shorter intersequence pause length. Hence, modulation of PRL with dopamine agonist may enhance the reproductive efficiency of hens later in life.

• Herbal Formula Science
• /
• v.11 no.2
• /
• pp.111-124
• /
• 2003

Gamiguibitang(GMGBT) is used in amenorrhea and female infertility caused by ovulation disorder. An attempt was made to evaluate the influences of GMGBT on the serum concentrations of FSH, LH, estradiol(E2) and progesterone, the histological and optical changes of ovary of rats. The results of the study were as follows: 1. Blood FSH level significantly increased in experimental group controlled by four times quantity as compared with control group. 2. Blood LH level increased in experimental group controlled by four times quantity as compared with control group, which showed no efficacy. 3. Blood E2 level increased in experimental group controlled by four times quantity as compared with control group, which showed no efficacy. 4. Blood progesterone level significantly decreased in experimental group controlled by double quantity as compared with control group. 5. In optically observations of ovary, weight of ovary significantly increased in experimental group controlled by double quantity as compared with control group 6. In histological observations of ovary, ovulation significantly increased in experimental group controlled by both double and four times quantity as compared with control group. According to these results, it can be concluded that GMGBT influences the pituitary gland and ovary to increase the ovulation of rats.

• Development and Reproduction
• /
• v.14 no.4
• /
• pp.261-268
• /
• 2010

As in the O. mykiss electrophoretic profiles of RNA, the signals of each RNA sample from 9 individual tissues such as liver, muscle, brain, heart, pituitary gland, kidney, intestine, spleen and gill similar to positive control were obtained. The tissue distributions of the complimentary DNA (cDNA) of O. mykiss four genes were analyzed using quantitative real-time PCR with primer sets for tissue expression analysis. In this rainbow trout species, author obtained bands of various sizes, ranged from 700 bp to 1,400 bp. A dissociation curve was made at the end of each run to make sure that there was no non-specific amplification. Supplementarily, the Ct of each DNA was compared. The Ct values of vinculin with rainbow trout tissues were determined in a manner similar to those for agouti-related protein (AgRP) and melanocortin receptors (MC4R I and MC4R II). Further, obtained Cts for standard curve of each DNA were affected by specific product (vinculin, AgRP and MC4R II genes). After several experiments with four individual genes of rainbow trout, author estimated a variation ratio of the mean Ct value of the DNA extracted using the comparative CTt method was 37.27, and the standard deviation was 5.33. The correlation coefficient between the Ct values and the concentration of cDNA was -0.98, -0.99, -0.91 and -0.86, respectively (vinculin, AgRP, MC4R I and MC4R II genes). Since this correlation showed high linearity, the straight line obtained was used as a standard for the O. mykiss tissues reared in aquarium. A PCR efficiency of 100% is ideally achieved when the slopes are close to the theoretical value of -3.31. According to quantification method, the results of quantification are strongly affected by the DNA fragmentation. The size of most DNA fragments obtained from various tissues of rainbow trout used in the experiment was approximately 100 bp. According to the four slopes, an efficiency of nearly 100% was estimated for four genes detection methods. Additionally, further analysis with more individuals and primers will be required to fully establish optimization in rainbow trout.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.49-49
• /
• 2003

본 연구에서는 vesicular stomatitis virus G glycoprotein (VSV-G)를 envelope로 가지는 pantropic retrovirus vector system을 이용하여 재조합 human FSH 유전자가 전이된 형질전환 닭을 생산하고자 하였다. Human FSH $\alpha$ 및 $\beta$ 유전자와 CTP linker는 human pituitary gland cDNA library에서 RT-PCR 방법을 이용하여 cloning하였으며, 각각의 fragment는 FSH$\beta$-CTP-FSH$\alpha$ 순서의 단일사슬로 연결하였다. 연결된 FSH$\beta$-CTP-FSH$\alpha$는 retroviral vector 내의 $\beta$-actin promoter의 조절 하에 도입한 후, PT67 packaging cell line에 transfection하여 virus를 생산하였으며 생산된 virus는 pantropic한 virus producing cell인 GP293에 infection하여 FSH 유전자가 도입된 virus를 생산하였다. FSH 유전자의 발현을 in vitro에서 확인하기 위하여 CHO (chinese hamster ovary) 세포에 virus를 감염시킨 후, 세포의 배양액을 취하여 electrochemilumine-scence immunoassay 방법으로 정량하였다. In vitro에서 전이 후 발현이 확인된 FSH 외래유전자의 retroviral vector virus를 초원심분리로 고농축하여 stageX의 계란의 배반엽 층에 주입하였으며, 그 결과 18%의 부화율과 91%의 부화한 닭의 유전자 전이율을 확인할 수 있었다. 전이된 유전자의 확인은 FSH$\beta$와 Neo 유전자에 대한 primer를 이용한 RT-PCR의 방법을 이용하였다. In vitro에서와는 달리 in vivo에서는 FSH 유전자의 전이는 확인되었으나 발현을 확인하지는 못하였는데, 이는 적은 수의 실험군이 형질전환율에 비해 상대적이지 못하였거나, 외래 유전자인 FSH의 발현에 의한 생리적인 부작용이 유발되어 해당개체가 부화되지 못한 것으로 추정된다. 본 문제점을 해결하기 위하여 실험군의 수를 늘리고 외래 유전자에 대한 controllable expression system이 보완될 필요성이 요구되며, 이러한 점이 해결된다면 높은 유전자 전이율에 기인하여 retrovirus를 이용한 형질전환 방법은 형질전환 가금의 생산에 있어서 매우 효율적이고 주목할 만한 방법으로 사료된다.

• Clinical and Experimental Pediatrics
• /
• v.46 no.4
• /
• pp.400-403
• /
• 2003

Congenital adrenal hyperplasia(CAH) is a recognized cause of precocious pseudopuberty. Some children with CAH also develop true precocious puberty with early maturation of the hypothalamic-pituitary-gonadal axis. We review a case of CAH who eventually developed central precocious puberty nine months after initial treatment with corticosteroid. A 3-year-old boy visited complaining of rapid growth, a large penis and frequent penile erections. This patient was diagnosed with CAH with elevated 17-OH progesterone and cortical hypertrophy of adrenal gland on CT scan. His gonadotropin levels were within the normal prepubertal range. Even on treatment with corticosteroid he grew rapidly and had testicular enlargement, pubic hair development and rapid bone maturation. At second admission, his gonadotropin levels were elevated both basally and in response to LHRH stimulation, suggesting that the CAH led to early activation of pubertal gonadotropin secretion(true precocious puberty). He was treated with monthly depot injections of a LHRH analog in addition to the hydrocortisone. His second sexual characteristics regressed gradually and rate of linear growth and bone maturation decreased.

• Journal of the Korean Institute of Oriental Medical Informatics
• /
• v.15 no.2
• /
• pp.57-76
• /
• 2009

Objective : All kinds of stimulus can be work as a stressor, and too much stress can makes disease and leads to a death. It has studied to know what kinds of study have carried out for controlling stress with acupuncture. Methods : It has searched articles in various web sites with key words of acupuncture, stress, hormone, adrenal, cortisol, catecholamine, epinephrine, and ACTH. Result : 1. It will work in the hypothalamus-pituitary gland-adrenal axis to control stress with acupuncture. 2. It has studied most in the years of 1990's and with acupoint of BL23 (腎兪) about controlling stress with acupuncture in Korea. 3. It has studied most in USA, the years of 2000's and with acupoints of ST36 (足三里), PC6 (內關) about controlling stress with acupuncture in Korea. Conclusion : It can be said that is growing interest about controlling stress with acupuncture in other countries, and it should be studied more various about controlling stress with acupuncture in Korea.

• Journal of Nutrition and Health
• /
• v.1 no.3_4
• /
• pp.197-200
• /
• 1968

Using the $Methionine-2-C^{14}$, the metabolism of methionine to the experimental rats on rice diet was studied comparing with that to the rats on stock diet in this paper. The National Institute of Health strain of weaning albino rats were housed into the individual cages deviding into 2 groups, the rice diet (RD) group and the stock diet (SD) group, and fed on rice diet and stock diet respectively for 10 weeks. On the day of experiment, the rats were parenterally administered. the $methionine-2-C^{14}$ solution after fasting over night. And then the rats were sacrificed by ether anesthesia by time being of one, three, six, and twelve hours each and the organs, pituitary gland, pancreas, spleen, liver, and kidney, were taken out for the determinations of radioactivities. And also the excretion of radioactivities through urine were determined by time being. The radioactivities were determined by Autoscaler SC-51 using the planchets. The results of radioactivities of urine excretion were shown at table 3 and the results of radioactivities distibutions in the organs by time being after parenteral administration of $methionine-2-C^{14}$ were shown table 4 in the original paper. According to the results, the following are summarized; 1. The growth experiment result of rats on. rice diet and stock diet were same as shown by the previous workers indicating significant growth inhibition at the rice diet group. 2. Due to the result of radioactivity excretion through urine after administration of $methionine-2-C^{14}$, it might he considered that methionine in the rice diet seems to be limited. However, it seems to be not 주 mostly limited. 3. And due to the results of radioactivity distribution in the organs by time being, the radioactivity in the liver tissue showed appearently higher readings at this methionine study compared with the results at the lysine study shown by HAW and his co-worker. This might be interpreted, though it is not clear, that liver might require methionine as a deficient amino acid at the tissue because methionine is limited at the rice diet.

• Molecules and Cells
• /
• v.22 no.1
• /
• pp.119-125
• /
• 2006

The rat is an accepted model for studying human psychiatric/neurological disorders. We provide a protocol for total soluble protein extraction using trichloroacetic acid/acetone (TCA/A) from rat (female) whole brain, 10 brain regions and the pituitary gland, and show that two-dimensional gel electrophoresis (2-DGE) using precast immobilized pH (4-7) gradient (IPG) strip gels (13 cm) in the first dimension yields clean silver nitrate stained protein profiles. Though TCA/A precipitation may not be "ideal", the important choice here is the selection of an appropriate lysis buffer (LB) for solubilizing precipitated proteins. Our results reveal enrichment of protein spots by use of individual brain regions rather than whole brain, as well as the presence of differentially expressed spots in their proteomes. Thus individual brain regions provide improved protein coverage and are better suited for differential protein detection. Moreover, using a phosphoprotein-specific dye, ingel detection of phosphoproteins was demonstrated. Representative high-resolution silver nitrate stained proteome profiles of rat whole brain total soluble protein are presented. Shortcomings apart (failure to separate membrane proteins), gel-based proteomics remains a viable option, and 2-DGE is the method of choice for generating high-resolution proteome maps of rat brain and brain regions.

• Animal cells and systems
• /
• v.5 no.3
• /
• pp.217-224
• /
• 2001

The present study examines the expression and regulation of gonadotropin-releasing hormone (GnRH) and its receptor (GnRH-R) mRNA levels during mouse ovarian development. A fully processed, mature GnRH mRNA together with intron-containing primary transcripts was expressed in the immature mouse ovary as determined by Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR). The size of ovarian GnRH mRNA was similar to that of hypothalamus, but its amount was much lower than that in the hypothalamus. Quantitative RT-PCR procedure also revealed the expression of GnRH-R mRNA in the ovary, but the estimated amount was a thousand-fold lower than that in the pituitary gland. We also examined the regulation of ovarian GnRH and GnRH-R mRNA levels during the follicular development induced by pregnant mare's serum gonadotropin (PMSG) and/or human chorionic gonadotropin (hCG). Ovarian luteinizing hormone receptor (LH-R) mRNA was abruptly increased st 48 h after the PMSG administration and rapidly decreased to the basal level thereafter. Ovarian GnRH mRNA level was slightly decreased at 48 h after the PMSG administration, and then returned to the basal value. GnRH-R mRNA level began to increase at 24 h after the PMSG treatment, decreased below the uninduced basal level at 48 h, and gradually increased thereafter. HCG administration did not alter ovarian GnRH mRNA level, while it blocked the PMSG-induced increase in GnRH mRNA level. Taken together, the present study demonstrates that the expression of GnRH and GnRH-R mRNA are regulated by gonadotropin during follicular development, suggesting possible intragonadal paracrine roles of GnRH and GnRH-R in the mouse ovarian development.