• Animal cells and systems
• /
• 제1권4호
• /
• pp.595-602
• /
• 1997

Treatment of newborn female rats with gonadal steroids induces permanent sterility in adulthood. We investigated the alteration in expression patterns of gonadotropin-releasing hormone (GnRH) and luteinizing hormone (LH) in neonatally estrogenized sterile rats (ESR). Newborn female rats received daily injections of 17${\beta}$-estradiol (E, 10 ${\mu}$g) from the day of birth (day 1) to postnatal day 5. Controls were subjected to vehicles over the same period. All animals were sacrificed on week 7 after birth. Hypothalamic GnRH mANA levels were markedly higher in all ESR than in controls, while hypothalamic GnRH contents in ESR increased in proportion to the frequency of daily administration of E. However, both pituitary LH6 mRNA and serum LH levels were inversely decreased by the same treatment. The data indicate that neonatal exposure of E equally elevates the expression of GnRH gene, but reduces the secretion of GnRH, accordingly leading to attenuation of LH6 gene expression and circulating LH levels. The temporal effect of E and/or progesterone (P) on GnRH and LH6 mRNA levels was also examined in ESR. Newborn female rats were daily injected with E (10 ${\mu}$g) or vehicle for five successive days from day 1 and ovariectomized at week 5. They were implanted with E (235 ${\mu}$g/ml) two days prior to week 7, injected with P (1 mg) 42 h later, and sacrificed 7 h after P administration. In ovariectomized controls, hypothalamic GnRH mRNA levels were dropped to half by treatment of E and restored by subsequent treatment of P. The negative feedback action of E on GnRH mRNA levels observed in ovariectomized rats was completely blocked by neonatal exposure of E. The change in pituitary LH mRNA levels was similar to that in hypothalamic GnRH mRNA levels. Taken together, the results suggest that neonatal treatment of E alters the synthesis and release of GnRH in adulthood and furthermore blocks the negative feedback regulation of E which occurs normally after ovariectomy.

• 한국수정란이식학회지
• /
• 제29권4호
• /
• pp.333-337
• /
• 2014

To compensate for the critical shortage of human organs for allotransplantation, xenotransplantation studies using genetically modified pigs are being performed in Korea. Two types of pigs that are used are ${\alpha}1,3$-galactosyltransferase gene knockout (GalT KO) pigs and GalT KO+hCD46 (human complement regulatory protein) pigs. The present study measured the gestation time, birth weight, daily growth rate, and heart weight of both kinds of transgenic minipigs. The gestation period for both types of pigs was 117~119 days. There was no difference in the body weight of GalT KO (-/+) and GalT KO (-/-) piglets, but GalT KO+hCD46 ($-^{hCD46+}/+$) pigs were significantly heavier at birth than were GalT KO+hCD46 ($-^{hCD46+}/-^{hCD46+}$) pigs. During the first 10 weeks of life, the daily weight gain of GalT KO+hCD46 ($-^{hCD46+}/-^{CD46+}$) piglets, which are considered the optimal type for xenotransplantation, was 0.19 kg. The weight of hearts from GalT KO piglets up to two months of age was affected more by body weight than by age. Transgenic pigs showed no differences in gestation period or reproductive ability compared with normal pigs. These results comprise basic data that may be used in xenotransplantation studies and transgenic animal production in Korea.

• Clinical and Experimental Reproductive Medicine
• /
• 제33권4호
• /
• pp.253-263
• /
• 2006

목 적: 본 연구에서는 EST Clustering 방법을 이용하여 이전의 연구에서 발굴한 B357 EST의 염기서열을 포함하는 유전자를 동정하고, 이 유전자의 발현을 생쥐의 난소와 정소를 포함한 여러 가지 조직들에서 살펴보고자 하였다. 연구방법: EST Clustering으로 얻어진 전체 염기서열을 5-day-ovary-specific gene-1 (5DOS1)이라고 명명하여 GenBank에 등록하였으며 (AY751521), northern blotting, real-time RT-PCR, in situ hybridization, western blotting, immunohistochemistry 등의 방법을 이용하여 생쥐 난소와 고환의 발달단계에 따라 그 발현양상을 관찰하였다. 결 과: 5DOS1의 전사체는 성장한 정소, 뇌, 근육에서 높게 발현하였으며, 난소의 경우에서는 원시난포시기부터 모든 난자에서 발현하였으며 특히 생후 5일째 높게 발현하였고 그 이후로는 점차 감소하였다. 반면 정소의 경우는 발달과 함께 계속 증가하였으며, 정모세포를 제외한 모든 발달단계별 정자에서 발현함을 관찰하였다. 결 론: 본 연구결과는 5DOS1에 대한 발견과 동정에 대한 첫 보고로써, 유전자 및 단백질이 생쥐의 난소 및 정소의 생식세포에서 발현하는 것을 관찰하였다. 앞으로 생식세포 발생 및 분화에 관련된 5DOS1의 기능에 대한 심층 연구가 더 필요하다고 사료된다.

• Clinical and Experimental Reproductive Medicine
• /
• 제40권1호
• /
• pp.42-46
• /
• 2013

Hypochondroplasia (HCH) is an autosomal dominant inherited skeletal dysplasia, usually caused by a heterozygous mutation in the fibroblast growth factor receptor 3 gene (FGFR3). A 27-year-old HCH woman with a history of two consecutive abortions of HCH-affected fetuses visited our clinic for preimplantation genetic diagnosis (PGD). We confirmed the mutation in the proband (FGFR3:c.1620C>A, p.N540K), and established a nested allele-specific PCR and sequence analysis for PGD using single lymphocyte cells. We performed this molecular genetic analysis to detect the presence of mutation among 20 blastomeres from 18 different embryos, and selected 9 embryos with the wild-type sequence (FGFR3:c.1620C). A successful pregnancy was achieved through a frozen-thawed cycle and resulted in the full-term birth of a normal neonate. To the best of our knowledge, this is the first report of a successful pregnancy and birth using single-cell allele-specific PCR and sequencing for PGD in an HCH patient.

• Animal cells and systems
• /
• 제3권3호
• /
• pp.319-329
• /
• 1999

Onset of female puberty follows a series of prepubertal cellular and molecular events including changes of synaptic plasticity, synthetic and releasing activity and gene expression. Dramatic increase of gonadal steroid level is one of the most prominent changes before the onset of puberty. Based on the importance of steroid feedback upon the hypothalamus, we adopted an estrogen sterilized rat (ESR) model where 100 ng of 17$\eta$-estradiol were administered into neonatal pubs for 7 days after birth. To identify genes involved in the onset of female puberty, we applied PCR differential display using RNA samples derived from ESR and control rat hypothalami. About 100 out of more than 1000 RNA species examined displayed differential expression patterns between a 60-day old control rat and ESR. Sequence analysis of differentially amplified PCR products showed homology with genes such as mouse kinesin superfamily-associated protein 3 (KAP3) and several cDNAs previously described by others in mouse and human tissues. Several gene products such as 2-1 and 8-1 corresponded to novel DNA sequences. We analyzed mRNA levels of KAP3, 2-1 and 8-1 genes in the hypothalami derived from neonatal, 6-, 28-, 31-, and 40-day old rats. Northern blot analysis showed that mRNAs of KAP3, 2-1 and 8-1 genes were markedly increased before the initiation of puberty. Neonatal treatment of estrogen clearly inhibited prepubertal increases in KAP3, 2-1 and 8-1 mRNA levels. Therefore, these genes may play important roles in the initiation of hypothalamic puberty. In addition, intracerebroventricular (icv) injection of antisense KAP3 oligodeoxynucleotide (ODN) clearly delayed puberty initiation determined by vaginal opening, which further confirmed that KAP3 plays an important role in the regulation of puberty initiation.

• Asian-Australasian Journal of Animal Sciences
• /
• 제27권1호
• /
• pp.123-130
• /
• 2014

The A-type lamin deficient mouse line ($Lmna^{-/-}$) has become one of the most frequently used models for providing insights into many different aspects of A-type lamin function. To elucidate the function of Lmna in the growth and metabolism of mice, tissue growth and blood biochemistry were monitored in Lmna-deficient mice, heterozygous ($Lmna^{+/-}$) and wide-type ($Lmna^{+/+}$) backcrossed to C57BL/6 background. At 4 weeks after birth, the weight of various organs of the $Lmna^{-/-}$, $Lmna^{+/-}$ and $Lmna^{+/+}$ mice was measured. A panel of biochemical analyses consisting of 15 serological tests was examined. The results showed that Lmna deficient mice had significantly decreased body weight and increased the ratio of organ to body weight in most of tissues. Compared with $Lmna^{+/+}$ and $Lmna^{+/-}$ mice, $Lmna^{-/-}$ mice exhibited lower levels of ALP (alkaline phosphatase), Chol (cholesterol), CR (creatinine), GLU (glucose), HDL (high-density lipoprotein cholesterol) and higher levels of ALT (alanine aminotransferase) (p<0.05). $Lmna^{-/-}$ mice displayed higher AST (aspartate aminotransferase) values and lower LDL (lowdensity lipoprotein cholesterol), CK-MB (creatine kinase-MB) levels than $Lmna^{+/+}$ mice (p<0.05). There were no significant differences among the three groups of mice with respect to BUN (blood urea nitrogen), CK (creatine kinase), Cyc C (cystatin C), TP (total protein), TG (triacylglycerols) and UA (uric acid) levels (p>0.05). These changes of serological parameters may provide an experimental basis for the elucidation of Lmna gene functions.

• Childhood Kidney Diseases
• /
• 제9권2호
• /
• pp.269-274
• /
• 2005

저자들은 불규칙한 발열은 주소로 내원한 5개월 된 어린 영아에서 유전자 검사를 통하여 선천성 신성 요붕증을 조기 확진하였으며 thiazide 치료에 반응을 보였기에 문헌고찰과 함께 보고하는 바이다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제33권3호
• /
• pp.506-514
• /
• 2020

Objective: We tested the hypothesis that increasing dietary copper (Cu) to gravid ewes would enhance brown adipose tissue (BAT) thermogenesis in their offspring. Methods: Twin-bearing ewes were assigned on d 70 of gestation to diets containing 3, 10, or 20 ppm dietary Cu (n = 8 per group). Twin lambs were assigned at birth to a cold (6℃) or warm (28℃) environmental chamber for 48 h. Blood was collected from ewes and from lambs and perirenal BAT was collected after 48 h in the environmental chambers. Results: Prenatal Cu exposure increased ewe plasma triiodothyronine (T₃) and thyroxine concentration (T₄) (p<0.01) but prenatal Cu exposure had no effect on lamb plasma concentrations of T₃, T₄, glucose, or nonesterified fatty acid concentration (p≥0.08). The high level of prenatal Cu exposure depressed 48-h rectal temperature (p = 0.03). Cold exposure decreased BAT norepinephrine (NE) and increased BAT dopamine (p≤0.01), but prenatal Cu exposure had no effect on BAT cytochrome C oxidase activity or BAT NE or dopamine (p≥0.07). However, BAT of lambs from high-Cu ewes maintained higher uncoupling protein-1 (UCP1) gene expression than BAT of lambs from low- and medium-Cu ewes following warm or cold exposure in environmental chambers (p = 0.02). Cold exposure caused near depletion of BAT lipid by 48 h (p<0.001), increased BAT cytochrome c oxidase activity (p<0.01), and depressed plasma fatty acid concentrations (p<0.001). Conclusion: Although prenatal Cu exposure increased BAT UCP1 expression during warm and cold exposure, prenatal cold Cu exposure depressed 48-h rectal temperature. Cold exposure decreased BAT lipid content by over 80% and decreased lamb plasma fatty acid concentration by over 40%, indicating that fuel reserves for thermogenesis were nearly depleted by 48 h of cold exposure.

• Journal of Genetic Medicine
• /
• 제6권1호
• /
• pp.87-90
• /
• 2009

Cardiofaciocutaneous (CFC) 증후군은 누난-관련질환중 하나로서, 심장기형, 특징적 얼굴형태, 피부이상과 발달지연이 동반되는 질환이다. CFC증후군은 성긴 두피, 옅은 눈썹, 안구 돌출이나 안구 진탕 등의 얼굴 형태와 과각화증, 어린선 등의 피부증상으로 다른누난 관련-질환과 임상적으로 구분을 할 수 있으나, 임상적인 구분이 모호한 경우도 있다. 최근 누난 증후군과 누난-관련 증후군의 원인유전자들이 밝혀짐에 따라 이들의 감별 진단에 도움을 받고 있다. 이에 본 저자들은 유전자 검사를 통하여 진단된 CFC증후군 1례를 보고하는 바이며, 누난 증후군과 누난-관련 질환의 유전형과 표현형의 다양성에 대해 논의하고자 한다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제18권5호
• /
• pp.723-727
• /
• 2005

Ghrelin is a novel growth-hormone-releasing acylated peptide, which has been purified and identified in rat stomach. In the present study, the full-length sequence of bovine ghrelin cDNA was cloned by RT-PCR. The bovine ghrelin cDNA sequence derived in the present study included a 348 bp open reading frame and a 137 bp 3'UTR. The putative amino acid sequence of bovine prepro-ghrelin consisted of 116 amino acids, which contained the 27-amino acid ghrelin. The sequence analysis of the bovine ghrelin gene revealed that an intron existed between Gln$^{13}$ and Arg$^{14}$ of ghrelin. This exon-intron boundary matched the GT-AG rule of the splicing mechanism. Compared with rats, which have two tandem CAG sequences in the 3'end of intron, bovine ghrelin genome has only one CAG sequence. Therefore, although rats can produce 28 amino acid-ghrelin and 27 amino acid-des-Gln$^{14}$-ghrelin by alternative splicing, ruminant species, including bovines, might be able to produce only one type of ghrelin peptide, des-Gln$^{14}$-ghrelin. The influence of aging on plasma ghrelin concentration was also examined. Plasma ghrelin concentration increased after birth to approximately 600 days of age, and then remained constant.