• Animal cells and systems
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• 제3권1호
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• pp.1-7
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• 1999

Gonadotropin-releasing hormone (GnRH) was originally isolated as a hypothalamic peptide that regulates reproduction by stimulating the release of gonadotropins. Using comparative animal models has led to the discovery that GnRH has a more ancient evolutionary origin. Durinq evolution GnRH peptide underwent gene duplication and structural changes to give rise to multiple molecular forms of GnRHs. Mammalian GnRH initially considered to be the sole molecular form, is now grouped as a family of peptides along with GnRH variants determined from representatives in all classes of vertebrates. Vertebrate species including primates and humanshave more than one GnRH variant in individual brains; a unique GnRH form in the forebrain and chicken IIGnRH in the midbrain. Furthermore, several species of bony fish have three molecular variants of GnRH: salmon GnRH sea-bream GnRH and chicken II GnRH. Also, it has been shown that in addition to the olfactory placodes and the midbrain, there is a third embryonic source of GnRH neurons from the basal diencephalon in birds and fish, which might be true for other vertebrates. Therefore, comparative animal models like fish with discrete sites of expression of three molecular variants of GnRH in individual brains, could provide insight into novel functions of GnRH variants, conservation of gene regulation, and mechanisms governing reproduction in vertebrates.

• 한국가축번식학회지
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• 제27권3호
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• pp.207-213
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• 2003

본 연구는 한우에 PGF$_2$$\alpha$와 GnRH+PGF$_2$$\alpha$+GnRH(Ov-synch)를 처리하여 발정 및 배란을 동기화 시켰으며, 2차 GnRH 투여후 배란시간, 2차 GnRH 투여후 시간 경과에 따른 수태율과 발정ㆍ배란 동기화법에 의한 수태율을 조사하고자 실시하였다. 시험축은 총 4개 농가에서 경산우 150두를 무작위로 선발하여 시험에 공시하였으며, 발정ㆍ배란동기화 방법에 따라 발정을 유기한 후 1회 인공수정을 실시하고 수태율에 미치는 영향을 조사하였다. 호르몬처리 방법으로는 GnRH+PCF$_2$$\alpha$+GnRH(Ov-synch)와 PGF$_2$$\alpha$를 이용한 발정동기화 방법을 사용하였다. 2차 GnRH 투여 후 배란시간을 알아보기 GnRH 투여 후 24시간 후부터 32시간까지 2시간 간격으로 초음파 Sonovet-600(Medison. Korea)를 이용하여 난소를 촬영하였다. 1. 호르몬 투여후 발정동기 화율은 PGF$_2$$\alpha$ 투여구에서 40.0%와 GnRH+PGF$_2$$\alpha$+GnRH(Ov-synch)처리구에서 91.3%로 나타났다. 2. 2차 GnRH 주사후 24시간에 배란이 시작되어 32시간에 배란이 종료되었으며, 배란율은 28시간째에 46.6%가 배란되어 가장 높게 나타났다. 3. 2차 GnRH주사후 6∼24시간에 수정한 군이 6시간 이전과 30시간 이후에 수정시킨 군에 비해 높은 수태율을 나타냈다. 4. 호르몬 처리별 수태율은 PGF$_2$$\alpha$, CIDR 및 GnRH+PGF$_2$$\alpha$+GnRH(Ov-synch)에서 각각 50.0, 36.0와 76.9%로 GnRH+PGF$_2$$\alpha$+CnRH(Ov-synch)군에서 가장 높은 수태율을 나타냈다.

• Animal cells and systems
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• 제5권3호
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• pp.217-224
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• 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.

• 한국발생생물학회지:발생과생식
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• 제6권2호
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• pp.131-139
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• 2002

최근 난포에서 GnRH와 그 수용체의 발현이 확인되면서 GnRH가 국소적으로 난소의 기능을 조절하고,특 히 과립세포의 세포자연사(apoptosis)를 유도하는 것으로 보고되고 있다. 그러나 황체에서 GnRH와 그 수용체의 발현과 기능에 대해서는 잘 알려져 있지 않다. 따라서 본 연구는 임신한 흰쥐의 황체세포에서 GnRH와 그 수용체가 발현되는지를 확인하고, 또한 GnRH가 황체세포의 세포자연사를 직접적으로 유발시킬 수 있는지를 알아보고자 시행하였다. 임신된 흰쥐로부터 황체세포를 획득하여 배양한 후 면역조직화학적 염색과 Western blot 방법으로 GnRH와 그 수용체의 발현을 확인한 결과 배양된 황체세포에서 GnRH와 그 수용체가 강하게 발현되는 것을 관찰할 수 있었다. GnRH가 배양된 황체세포의 세포자연사에 미치는 영향을 조사하기 위하여, $10^{-6}$ GnRH-agonist(GnRH-Ag)를 처리한 후 3, 8, 12시간에 TUNEL 방법과 DNA 분절화 검증 방법으로 세포자연사를 조사하였다. TUNEL 결과 세포자연사를 보이는 황체세포는 처리 후 12 시간에 GnRH-Ag 처리군에서 유의하게 증가하였다(p<0.05). 또한 DNA 분절화를 조사한 결과에서도 TUNEL 결과와 유사하게 GnRH-Ag처리 후 12 시간에 DNA 분절화가 유의하게 증가하였다(p<0.05). 이러한 세포자연사의 증가가 cytochrome c 방출과 연관이 있는지를 알아보고자 미토콘드리아로부터 방출된 cytochrome c를 Western blot 방법으로 정량한 결과, GnRH-Ag 처리 후 12 시간에 cytochrome c가 미토콘드리아로부터 세포질쪽으로 방출된 것을 확인할 수 있었다. 결론적으로 임신된 흰쥐의 황체세포에서 GnRH와 그 수용체 단백질이 발현되며 GnRH-Ag가 GnRH 수용체에 결합함으로써 cytochrome c가 미토콘드리아로부터 방출되고, 이로 인해 황체세포가 세포자연사하는 것을 알 수 있었다. 이러한 결과들은 국소적으로 분비되는 GnRH가 미토콘드리아로부터 cytochrome c의 방출을 유발시켜 황체세포의 세포자연사를 유도할 수 있다는 것을 제시하고 있다.

• 한국임상수의학회지
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• 제19권3호
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• pp.322-327
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• 2002

The effect of GnRH alone and concomitant with PMSG on the prevention of implantation. termination of pregnancy, and concentration of plasma progesterone were studied in pregnant rats. GnRH 50, 100 or 200 ug alone and concomitant with PMSG 25 or 50 IU were administered once on day 2 or 9 of gestation, respectively. Rats were autopsied on days 7 or 20. Administration of GnRH on day 2 did not result in the prevention of implantation and termination of pregnancy but resulted in termination of pregnancy administering on day 9. Administration of GnRM concomitant with PMSG on day 2 or 9 resulted in prevention of implantation and termination of pregnancy, but injection of GnRH 50 ug concomitant with PMSG 25 IU on day 9 had only one live fetus. Administration of GnRH alone and concomitant with PMSG on day 2 had no effect on the concentration of plasma progesterone determining on day 7. Administration of GnRH concomitant with PMSG on day 2 resulted in decrease of progesterone level determining on day 20 but GnRH alone was normal level. Administration of GnRH alone and concomitant with PMSG on day 9 resulted in decrease of the concentration of progesterone but was normal concentration administering GnRH 50 ug concomitant with PMSG 25 IU.

• Clinical and Experimental Reproductive Medicine
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• 제21권1호
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• pp.121-130
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• 1994

Expression of gonadotropin releasing hormone(GnRH) has been described in the rat ovary. It remains, however, unkown whether GnRH is synthesized as a prohormone. Therefore, this study was performed to verify the expression of pro-GnRH by in situ hybridization and further to investigate the effect of gonadotropin on GnRH or GnRH mRNA in rat ovary by immunohistochemical and in situ hybridization techniques. Adult female Sprague-Dawely rats were used and the estrous cycle was synchronized by intraperitoneal injection of pregnant mare's serum gonadotropin(PMSG). Ovaries were fixed with 4% paraformaldehyde and embedded with G.C.T. compound and cut by cryostat. For immunohistochemistry, avidin-biotin peroxidase complex(ABS) method was employed and for in situ hybridization, $^{35}S$-end labeled oligonucleotide was used and followed by autoradiography. By in situ hybridization using GnRH oligomer and GAP(GnRH associated protein) oligomer, GnRH mRNA and GAP mRNA were co-localized in the fullicular cells, luteal cells, interstitial cells and theca cells. GnRH or GnRH mRNA signals in the ovary increased by human chorionic gonadotropin(hCG) injection. At the 3 and 6 hrs after hCG injection, the number of GnRH and GnRH mRNA containing cells increased rapidly and the density of GnRH and GnRH mRHA culminated at 9 hrs after heG injection. With the follicular development, the high expression of GnRH and GnRH mRNA was also observed within the follicles. After ovulation, the density of GnRH or GnRH mRNA decreased in the follicles but increased in the corpus lutea.

• Fisheries and Aquatic Sciences
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• 제14권4호
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• pp.379-388
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• 2011

The mechanism by which gonadotropin-releasing hormone (GnRH) and dopamine (DA) control gonadotropin (GTH) release was studied in male and female rainbow trout using cultured pituitary cells obtained at different reproductive stages. The mechanisms of follicle-stimulating hormone (FSH) release by GnRH and DA could not be determined yet. However, basal and salmon-type GnRH (sGnRH)- or chicken-II-type GnRH (cGnRH-II)- induced luteinizing hormone (LH) release increased with gonadal maturation in both sexes. LH release activity was higher after sGnRH stimulation than cGnRH-II stimulation at maturing stages in both sexes. The GnRH antagonist ([Ac-3, 4-dehydro-$Pro^1$, D-p-F-$Phe^2$, D-$Trp^{3,6}$] GnRH) suppressed LH release by sGnRH stimulation in a dose-dependent manner, although the effect was weak in maturing fish. The role of DA as a GTH-release inhibitory factor differs during the reproductive cycle: the inhibition of sGnRH-stimulated LH release by DA was stronger in immature fish than in maturing, ovulating, or spermiated fish. DA did not completely inhibit sGnRH-stimulated LH release, and DA alone did not alter basal LH release. Relatively high doses ($10^{-6}$ or $10^{-5}M$) of domperidone (DOM, a DA D2 antagonist) increased LH release, which did not change with reproductive stage in either sex. The potency of DOM to enhance sGnRH-stimulated LH release was higher in maturing and ovulated fish than in immature fish. These data suggest that LH release from the pituitary gland is controlled by dual neuroendocrine mechanisms by GnRH and DA in rainbow trout, as has been reported in other teleosts. The mechanism of control of FSH release, however, remains unknown.

• 한국동물학회:뉴스레터
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• 제16권1호
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• pp.17-50
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• 1999

시상하부에 극히 적은 수로 존재하는 신경분비세포인 성선자극호르몬-방출호르몬(gonadotropin-releasing hormone; GnRH) 뉴런은인간을 포함한 포유동물의 생식과 발생 과정에 있어 중요한 역할을 담당하고 있다. GnRH 뉴런은 배아 발생과정 중에 후판에서 유래하여 시상하부의 여러 영역으로 이동하며, 생후와 사춘기를 거치면서 분화를 계속한다. GnRH 뉴런에서 합성, 분비되는 10개의 아미노산으로 이루어진 작은 신경호르몬인 GnRH는 맥동적으로 분비되어 뇌하수체 성선자극 세포막에 존재하는 GnRH 수용체와 결합한 후 일련의 신호전달과정을 거쳐 성선자극호르몬의 합성과 분비를 제어하게 된다. GnRH의 합성과 분비는 글루탐산, 노르에피네프린, GABA와 같은 각종 신경입력과 스테로이드 호르몬에 의한 액성 피드백 신호에 의해 조절되나 이들의 GnRH 유전자 발현에 미치는 영향은 최근에 연구되고 있는 실정이다. GnRH 뉴런의 분화와 발생에는 다양한 신경영양인자들이 영향을 미치나 그 분자생물학적 기작은 아직 밝혀져 있지 않다. 본 논단에서는 신경호르몬인 GnRH와 그 수용체에 관하여 최근 연구성과를 중심으로 살펴보고자 한다.

• 한국동물학회지
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• 제34권4호
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• pp.491-499
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• 1991

The present experiment was carried out 1) to study the developmental topography of GnRH neuronal system and 2) to characterize the cellular localization of GnRH neurons in the prenatal brain development of the rat. At embryonic day (I) 14.5, immunoreactive cell bodies of GnRH were first seen in the nasal septum and in the ganglion terminate located in the ventral protion of the caudal olfactory bulb. Two days later (E 16.5), GnRH-containing neurons were observed at the level of olfactory tubercle and diagonal band of Broca, which is the first appearance in the intracerebral region. From 118.5, the topographic pattern of immunoreactive GnRH perikarya was similar to that of adult rats. The present data suggest that GnRH neurons were originated from the nasal septum and gradually extended to the hvpothalamic regions with increasing fetal age.

• 한국동물학회지
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• 제37권3호
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• pp.304-310
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• 1994

Using in situ hybridization, we have mapped the anatomical localization of perikarya containing myNA that codes for sonadotropin releasing hormone (GnRH) in the brains of female frogs, R. dybowskii. DNA olisomers, with sequences complementary to the GnRH portion of pro-GnRH myNA sequence, were synthesized and hybridized to paraformaldehvde-fixed, sagittal sections of the whole brain stem. The distribution of the GnRH mRNA containing cell bodies was similar to that described for GnRH peptide by immunohistochemistrv. That is, cells containing GnRH mRNA were observed in the medial septal area, anterior preoptic area, ventromedial hvpothalamus and infundibular regions. However, another cell groups which contains GnRH mRNAs were also detected by in situ hybridization in the bed nucleus of hippocampal commissure, preoptic area, nucleus infundibularis dorsalis, mesencephalic nuclei and intermediolateral cell column of spinal cord areas. These results demonstrate the feasibility of using in situ hybridization as a strategy to study the distribution of GnRH neurons and the detection of GnRH gene expression in the vertebrates.