Long-term non-surgical contraceptive methods for cats, especially community cats, are of global interest for cost-effective and humane reasons. This study aimed to investigate the effectiveness of a gonadotropin-releasing hormone (GnRH)-based vaccine for immunocontraception and to confirm its safety in intact female cats. Recombinant Salmonella typhimurium flagellin fljB (STF2)-GnRH protein was expressed in Escherichia coli. We divided female cats into vehicle control group (n = 4) and two experimental groups (100 μg injection group [n = 7] and 1000 μg injection group [n=7]), and immunized them twice intramuscularly (0.2 mL/cat at zero week and 4 weeks later into the other leg). Breeding trials started on day 120. All control cats (n = 4/4), 71% of the 100 μg injection group (n = 5/7), and 57% of the 1000 μg injection group (n = 4/7) became pregnant within 203 days after the introduction of male cats. The 1000 μg injection group had significantly a longer median time to conception following treatment (166 days) than the control (17 days, p < 0.05). Average litter size was significantly lower in the 1000 μg GnRH-vaccinated cats (2.8 ± 0.7) than in the control cats (4.5 ± 0.5, p < 0.05). Injection site reactions were not observed in any cat. The E. coli-expressed STF2-GnRH vaccine did not provide contraception in a sufficient proportion of the cats. However, it might be effective to suppress fertility through infertility vaccines before inducing permanent infertility through the trap-neuter-return.

Gonadotropin releasing hormone (GnRH) centrally plays a role in control of the hypothalamicpituitary- gonadal axis-related hormone secretions in the reproductive neuroendocrine system. In addition, hormone receptors like luteinizing hormone receptor (LHR) are important element for hormones to take effect in target organ. However, ageing-dependent changes in terms of the distribution of GnRH neurons in the brain and LHR expression in the acyclic ovary have not been fully understood yet. Therefore, we comparatively investigated those ageing-dependent changes using young (1-5 months), middle (11-14 months) and old (21-27 months) aged female mice. Whereas a number of GnRH positive fibers and neurons with monopolar or bipolar morphology were abundantly observed in the brain of the young and middle aged mice, a few GnRH positive neurons with multiple dendrites were observed in the old aged mice. In addition, acyclic ovary without repeated development and degeneration of the follicles was shown in the old aged mice than others. LHR expression was localized in theca cells, granulosa cell, corpora lutea and atretic follicle in the ovaries from young and middle aged mice, in contrast, old aged mice had few positive LHR expression on the follicles due to acyclic ovary. However, the whole protein level of LHR was higher in the ovary of old aged mice than others. These results are expected to be used as an important basis on the relationship between GnRH and LHR in old aged animals as well as in further research for reproduction failure.

On January 6, 2010, two months earlier than normal breeding season, a red fox vixen was implanted with synthetic GnRH analogue, Deslorelin. Blood was sampled every 2~3 days from the day of implant to identifying spermatozoa on stains of epithelial cells. Estradiol and progesterone were examined. Even though the vixen was in non-breeding season, she was mated by a male fox. Pregnancy was confirmed by canine pregnancy detection kit that detect relaxin released from placenta. Four healthy pups were born on March 9, 2010. This is the first report showing synthetic GnRH can activate ovarian function and lead to fertile estrus of red fox in non-breeding season.

본 연구는 한우에 PGF₂α와 GnRH+PGF₂α+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+PCF₂α+GnRH(Ov-synch)와 PGF₂α를 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 PGF₂α 투여구에서 40.0%와 GnRH+PGF₂α+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 PGF₂α, CIDR 및 GnRH+PGF₂α+GnRH(Ov-synch)에서 각각 50.0, 36.0와 76.9%로 GnRH+PGF₂α+CnRH(Ov-synch)군에서 가장 높은 수태율을 나타냈다.

본 연구에서는 GnRH가 과배란 처치된 래트의 초기 난포기와 후기 난포기에서 난소기능에 어떠한 영향을 미치는지를 이해하기 위해서, 30IU PMSG와 10IU hCG로 전처치된 미성숙 래트에 있어서 배란반응, 배란 난자의 형태학적 이상 유무 및 핵 성숙도, 난소 중량, 난소의 조직학적인 변화 및 혈중 스테로이드 호르몬 (17-estradiol, progesterone 및 testosterone) 농도에 대하여 GnRH agonist의 효과를 검사하였다.

From September 1993 to August 1997, we treated ovarian disorders in 1,782 repeat breeder cows after diagnosis by ultrasound on 35 farms in Kyeong-ki do. The rates of ovarian appearance were 59.8% of CL group, 16.7% of ovarian atrophy or hypofunction, 15.4% of luteal cyst, 4.3% of follicular cyst and 3.7% of follicle group in diagnosis with rectal palpation and ultrasound. The results of treatment for ovarian disorders were 1,316 cows(73.8%) in estrus, 348 cows(19.5%) in non-detected and 118 cows(6.6%) in unidentified. The rates of PGF, GnRH and mineral vitamin complex treatment to estrus were 79.6, 69.2 and 50.3%. Two groups were treated with 5 ml PGF intramuscular injection(I.M.) and 1.5 ml PGF intraovarian injection(I.O.), and the results of 1.5ml PGF I.0. were significantly higher than that of 5ml PGF I.M. in inductiom estrus(p<0.05). The pregnant rates were 29.8% in total repeat breeder cows with ovarian disorders following diagnosis and treatment. In summary, rectal palpation and ultrasonography were proven to be useful tools of diagnosis and treatment in ovarian disorders, and it was also suggested that the response to treatment with PGF I.0. was better than PGF I.M.

GnRH (gonadotropin-releasing hormone) is a supreme hormone regulating reproductive activity in most animals. The sequences of amino acid and nucleic acid of GnRH reported up to now are examined from the evolutionary framework of Chordata. All identified GnRH are classified into GnRH1, GnRH2, or GnRH3. In all three forms of GnRH both N-terminal and C-terminal are conserved, which allows for effective binding to their receptors. The three amino acids in the middle of GnRH1 sequence have altered diversely from the primitive Chordata, which is indicative of the adaptation process to the ambient environment. GnRH2 and GnRH3 sequences are well conserved. There are more diverse modifications in the nucleic acids than in amino acid sequence of GnRH1. These variations can result from meiosis, mutation, or epigenetics and indicate that GnRH is the product of natural selection.

Red spotted grouper, Epinephelus akaara, is a popular aquaculture species and a protogynous hermaphrodite. Induction of artificial sex change at the time of primary sex differentiation is of interest but has not been successful due to the lack of necessary basic information. To find out the potential neuroendocrine influence on the primary sex differentiation, the expression of key genes in the brain was investigated during the formation of ovarian cavity. Expression of cyp19a1b, esr1, gnrhr1, fsh, lh and cga in the brain was positively associated with the formation of ovarian cavity, showing gradual increase as the formation proceeds. However, the expression of gnrh1 was suppressed during the early part of the ovarian cavity formation, signifying potential hypothalamic influence on the primary sex differentiation in this species.

Neurokinin B (NKB) and neurokinin B related peptide (NKBRP) belong to tachykinin peptide family. They act as a neurotransmitter and/or neuromodulator. Mutation of NKB and/or its cognate receptor, NK3R resulted in hypogonadotropic hypogonadism in mammals, implying a strong involvement of NKB/NK3R system in controlling mammalian reproduction. Teleosts possess NKBRP as well as NKB, but their roles in fish reproduction need to be clarified. In this study, NKB and NKBRP coding gene (tac3) was cloned from Nile tilapia and sequenced. Based on the sequence, Nile tilapia NKB and NKBRP peptide were synthesized and their biological potencies were tested in vitro pituitary culture. The synthetic NKBRP showed direct inhibitory effect on the expression of GTH subunits at the pituitary level. This inhibitory effect was confirmed in vivo by means of intraperitoneal (ip) injection of synthetic NKB and NKBRP to mature female tilapia (20 pmol/g body weight [BW]). Both NKB and NKBRP had no effect on the plasma level of sex steroids, E2 and 11-KT. However, NKBRP caused declines of expression level of GnRH I, Kiss2 and tac3 mRNAs in the brain while NKB seemed to have no distinct effect. These results indicate some inhibitory roles of NKBRP in reproduction of mature female Nile tilapia, although their exact functions are not clear at the moment.

개체 사이의 사회적 상호작용은 인간을 포함한 대부분의 척추동물의 적절한 성성숙 및 번식활동을 위하여 중요한 과정이므로, 행동양식을 결정하는 조절인자들은 뇌를 포함한 중추신경계에 분포하고 있으며, 이 신호들을 조절하는 신경내분비계는 척추동물 내에서 잘 보존되어 있다. 척추동물 경골어류의 번식현상은 다양한 환경적, 계절적, 사회적 요소들에 의해 많은 영향을 받지만, 궁극적으로는 이러한 요소들의 자극이 뇌에서 통합되며, 뇌-뇌하수체-생식소 축에 의해서 번식활동이 제어된다. 이러한 신호 전달은 뇌의 시상하부 preoptic 지역에 위치한 생식선자극호르몬방출호르몬(GnRH1) 함유 뉴런에 의해 시작되며, 축삭이 뇌하수체 생식선자극호르몬 생산세포로 투사되어 1차적으로 생식선의 성장과 스테로이드 호르몬 생산을 조절한다. 경골어류의 일부 종은 성적 가소성이 현저하며, 사회적 계급 사회를 형성하는 흥미로운 지견을 제공한다. 아프리칸 시크리드 Astatotilapia burtoni 수컷 성어들의 소수 지배계급층(Territorial dominant, T)은 번식능력이 있으며, 공격적이고 고유 수역에 대한 보호본능이 있지만, 나머지 피지배계급층(Non-territorial subordinate, NT) 수컷들은 번식능력도 없으며, 보호본능을 상실한다. 그러나, NT와 T는 수 시간~수 일 이내에 변화되며, 충분히 가역적이다. 가장 현저한 변화는 4일 이내에 GnRH1 뉴런 세포체의 체적이 크게 변화하는 것이다. 그러나, 이 세포크기의 변화를 유도하며, 결과적으로 GnRH1 뉴런의 신경내분비학적 활성을 조절하는 분자는 현재 알려져 있지 않지만, GnRH1 세포 크기를 조절하는 후보의 하나로서 세포체 크기, 수상돌기의 국소적 단백질 합성, 축삭의 성장, 시냅스 가소성을 결정하는 역할을 담당한다고 알려진 serine-threonine protein kinase에 속하는 TOR(target of rapamycin)가 추정된다. 포유동물의 mTOR는 세포 성장과 시냅스 가소성을 통제하는 기능이 시사되었지만, 포유동물 이외에는 그 기능이 잘 알려져 있지 않다. 또한, GnRH 뉴런과 같은 신경내분비세포의 세포크기를 조절하는지는 불명확하다. 본 연구에서는 GnRH1 뉴런의 세포의 크기에 영향을 미치는 분자로 추정되는 mTOR 억제자로 알려진 rapamycin을 T 수컷의 제3뇌실에 주사하여 9개의 대표적인 번식관련 행동, GnRH1 뉴런의 세포형태, 정소의 생식세포에 미치는 영향을 분석하였다. Rapamycin 주사 14시간 후에는 대조군에 비해 상대적으로 번식관련 행동의 빈도수가 낮게 관찰되는 경향을 보였다. 한편, 36시간 후에는 번식행동의 패턴은 주사하기 전의 패턴과 유사한 상태로 회복되는 현상이 관찰되었다. H/E염색과 광학현미경으로 정소를 관찰한 결과, 정소세포 및 정자의 분포와 발달상에는 큰 차이가 없었다. 현재, Rapamycin 주입에 의한 GnRH1 뉴런의 변화를 관찰하고 있으며, T 수컷의 번식행동 패턴의 변화와의 상관관계를 조사하고 있다.

Kisspeptin has been implicated in the process of puberty onset in various animal groups. This peptide is encoded by a gene, Kiss1 in avian and mammalian species. Contrary to these higher vertebrates, however, fish appeared to have another gene, Kiss2 that also codes for the precursor peptide of kisspeptin. To figure out biological significance of this gene during the puberty onset in fish, the expression profile of Kiss2 gene was investigated in the brain of Nile tilapia together with genes of GPR54, GnRH receptorI (rGnRHI) and GTH subunits ( and ). Expression of Kiss2 mRNA significantly increased at 2 weeks post hatch (wph) and 13 wph (<0.05). This increase coincided with the increases of GPR54 and rGnRH I gene expression. Detection of and subunit gene expression was possible later than 13 wph, indicating the activation of gonadotrophs in the pituitary. Data obtained from this study strongly suggest that, in addition to Kiss1 gene, Kiss2 gene is deeply associated with the onset of puberty by the activation of hypothalamus pituitary gonadal axis in Nile tilapia.

In mammals, puberty is a dynamic transition process from infertile immature state to fertile adult state. The neuroendocrine aspect of puberty is started with functional activation of hypothalamus-pituitary-gonadal hormone axis. The timing of puberty can be altered by many factors including hormones and/or hormone-like materials, social cues and metabolic signals. For a long time, attainment of a particular body weight or percentage of body fat has been thought as crucial determinant of puberty onset. However, the precise effect of high-fat (HF) diet on the regulation of hypothalamic GnRH neuron during prepubertal period has not been fully elucidated yet. The present study was undertaken to test the effect of a HF diet on the puberty onset and hypothalamic gene expressions in immature female rats. The HF diet (45% energy from fat, HF group) was applied to female rats from weaning to around puberty onset (postnatal days, PND 22-40). Body weight and vaginal opening (VO) were checked daily during the entire feeding period. In the second experiment, all animals were sacrificed on PND 36 to measure the weights of reproductive tissues. Histological studies were performed to assess the effect of HF diet feeding on the structural alterations in the reproductive tissues. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Body weights of HF group animals tend to be higher than those of control animals between PND 22 and PND 31, and significant differences were observed PND 32, PND 34, PND 35 and PND 36 (p<0.05). Advanced VO was shown in the HF group (PND p<0.001) compared to the control (PND ). The weight of ovaries (p<0.01) and uteri (p<0.05) from HF group animals significantly increased when compared to those from control animals. Corpora lutea were observed in the ovaries from the HF group animals but not in control ovaries. Similarly, hypertrophy of luminal and glandular uterine epithelia was found only in the HF group animals. In the semi-quantitative RT-PCR studies, the transcriptional activities of KiSS-1 in HF group animals were significantly higher than those from the control animals (p<0.001). Likewise, the mRNA levels of GnRH (p<0.05) were significantly elevated in HF group animals. The present study indicated that the feeding HF diet during the post-weaning period activates the upstream modulators of gonadotropin such as GnRH and KiSS-1 in hypothalamus, resulting early onset of puberty in immature female rats.