• Title/Summary/Keyword: puberty

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CRISPR/Cas9-mediated knockout of CD47 causes hemolytic anemia with splenomegaly in C57BL/6 mice

  • Kim, Joo-Il;Park, Jin-Sung;Kwak, Jina;Lim, Hyun-Jin;Ryu, Soo-Kyung;Kwon, Euna;Han, Kang-Min;Nam, Ki-Taek;Lee, Han-Woong;Kang, Byeong-Cheol
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.302-310
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    • 2018
  • CD47 (integrin-associated protein), a multi-spanning transmembrane protein expressed in all cells including red blood cells (RBCs) and leukocytes, interacts with signal regulatory protein ${\alpha}$ ($SIRP{\alpha}$) on macrophages and thereby inhibits phagocytosis of RBCs. Recently, we generated a novel C57BL/6J CD47 knockout ($CD47^{-/-}$ hereafter) mouse line by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease, and here report their hematological phenotypes. On monitoring their birth and development, $CD47^{-/-}$ mice were born viable with a natural male-to-female sex ratio and normally developed from birth through puberty to adulthood without noticeable changes in growth, food/water intake compared to their age and sex-matched wild-type littermates up to 26 weeks. Hematological analysis revealed a mild but significant reduction of RBC counts and hemoglobin in 16 week-old male $CD47^{-/-}$ mice which were aggravated at the age of 26 weeks with increased reticulocyte counts and mean corpuscular volume (MCV), suggesting hemolytic anemia. Interestingly, anemia in female $CD47^{-/-}$ mice became evident at 26 weeks, but splenomegaly was identified in both genders of $CD47^{-/-}$ mice from the age of 16 weeks, consistent with development of hemolytic anemia. Additionally, helper and cytotoxic T cell populations were considerably reduced in the spleen, but not in thymus, of $CD47^{-/-}$ mice, suggesting a crucial role of CD47 in proliferation of T cells. Collectively, these findings indicate that our $CD47^{-/-}$ mice have progressive hemolytic anemia and splenic depletion of mature T cell populations and therefore may be useful as an in vivo model to study the function of CD47.

Classification of junior high school boys' body types (남자 중학생의 체형분류에 관한 연구)

  • Shin, Jang-Hee
    • Journal of the Korea Fashion and Costume Design Association
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    • v.21 no.3
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    • pp.13-24
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    • 2019
  • Adolescence is a transitional stage of physical development which occurs during the period from puberty to adulthood. Going through this period, various parts of an adolescent's body grow at different rates, leading to different body shapes and proportions when compared to adults. Therefore, this study aimed to investigate the body sizes and shapes of junior high school boys from ages 13-15 based on body measurement items that are used as the basis for school uniform designs including jackets, shirts, and pants. For this, the study sought the basic data needed to develop body shapes and school uniform patterns for junior high school boys using the data from the 6th Size Korea Survey (2010). Specifically, it provided basic data for the development of school uniform patterns that fit well through the classification of bodies into particular types. After extracting body shape componen a cluster analysis using ANOVA was performed. According to the factor analysis conducted to determine body shape components, 5 factors were obtained as follows: Factor 1: bulk and horizontal size, Factor 2: body height and length, Factor 3: shoulder shape and length, Factor 4: characteristics of horizontal size, Factor 5: shape of the upper body with a variance of 82.62%. To classify junior high school boys' body shape was determined using various characteristics, and a cluster analysis was performed with the variables obtained by the factor analysis. For this, body shapes were classified into 3 different types: Type 1 accounted for 33.4%, with a total of 463 subjects. This type was a tall, long body individual with the smallest bulk and size. Type 2 accounted for 22.7%, with a total of 315 subjects. This type was large in bulk and horizontal size, but the lowest in height and length. Type 3 accounted for 43.9%, with a total of 610 subjects. This type was close to average in terms of horizontal size, length, and height. To develop well-fitting school uniforms for junior high school students, there should be further studies on changes in body shape and their associated causes. The study results will be available as basic data for comparing branded school uniform patterns for junior high school boys and developing school uniform patterns based on body shape, using 3D virtual clothing simulations.

Effect of Developmental Disorders of Maxillary First Molars on Orofacial Morphology (악안면 형태에 대한 상악 제1대구치 발육장애의 영향)

  • Park, Soyoung;Jeong, Taesung;Kim, Jiyeon;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.46 no.2
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    • pp.209-218
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    • 2019
  • This study was aimed to evaluate orofacial morphologies on the cases of developmental disorders of maxillary first molars. Panoramic radiographs, lateral cephalographs, and clinical photos of 2983 children who attended the Pediatric Dental Clinic of Pusan National University Dental Hospital from 2006 to August 2017 were assessed retrospectively. 34 patients were selected whose maxillary first molars were missed or developmentally delayed unilaterally or bilaterally. Demirjian's method was used for estimating dental age, then which was compared to chronologic age of children. Parameters expressing skeletal and dentoalveolar disharmony were checked and compared with control. Additionally, occlusion relationship was evaluated. Maxillary dental age was significantly delayed compared to chronologic age. Several parameters which show skeletal open-bite tendency and skeletal class III malocclusion with maxillary retrusion were statistically significant. Anterior crossbite and edge-bite were expected in most of these cases, but compensation by occlusion and soft tissue was also verified which might mask skeletal class III tendency. Congenital missed or developmentally delayed maxillary first molars might be related with declined growth of maxilla. If developmental disorders of maxillary first molars were verified during clinical examination, careful monitoring of orofacial growth was necessary during puberty and timed orthopedic and orthodontic intervention were considered.

Alterations of mRNA and lncRNA profiles associated with the extracellular matrix and spermatogenesis in goats

  • Chen, Haolin;Miao Xiaomeng;Xu, Jinge;Pu, Ling;Li, Liang;Han, Yong;Mao, Fengxian;Ma, Youji
    • Animal Bioscience
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    • v.35 no.4
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    • pp.544-555
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    • 2022
  • Objective: Spermatozoa are produced within the seminiferous tubules after sexual maturity. The expression levels of mRNAs and lncRNAs in testicular tissues are different at each stage of testicular development and are closely related to formation of the extracellular matrix (ECM) and spermatogenesis. Therefore, we set out to study the expression of lncRNAs and mRNAs during the different developmental stages of the goat testis. Methods: We constructed 12 RNA libraries using testicular tissues from goats aged 3, 6, and 12 months, and studied the functions of mRNAs and lncRNAs using the gene ontogeny (GO) and Kyoto encyclopedia of genes and genomes (KEGG) databases. Relationships between differentially expressed genes (DEGs) were analyzed by lncRNA-mRNA co-expression network and protein-protein interaction network (PPI). Finally, the protein expression levels of matrix metalloproteinase 2 (MMP2), insulin-like growth factor 2 (IGF2), and insulin-like growth factor-binding protein 6 (IGFBP6) were detected by western blotting. Results: We found 23, 8, and 135 differentially expressed lncRNAs and 161, 12, and 665 differentially expressed mRNAs that were identified between 3 vs 6, 6 vs 12, and 3 vs 12 months, respectively. GO, KEGG, and PPI analyses showed that the differential genes were mainly related to the ECM. Moreover, MMP2 was a hub gene and co-expressed with the lncRNA TCONS-0002139 and TCONS-00093342. The results of quantitative reverse-transcription polymerase chain reaction verification were consistent with those of RNA-seq sequencing. The expression trends of MMP2, IGF2, and IGFBP6 protein were the same as that of mRNA, which all decreased with age. IGF2 and MMP2 were significantly different in the 3 vs 6-month-old group (p<0.05). Conclusion: These results improve our understanding of the molecular mechanisms involved in sexual maturation of the goat testis.

Functional characterization of naturally-occurring constitutively activating/inactivating mutations in equine follicle-stimulating hormone receptor

  • Byambaragchaa, Munkhzaya;Ahn, Tae-Young;Choi, Seung-Hee;Kang, Myung-Hwa;Min, Kwan-Sik
    • Animal Bioscience
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    • v.35 no.3
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    • pp.399-409
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    • 2022
  • Objective: Follicle-stimulating hormone (FSH) is the central hormone involved in mammalian reproduction, maturation at puberty, and gamete production that mediates its function by control of follicle growth and function. The present study investigated the mutations involved in the regulation of FSH receptor (FSHR) activation. Methods: We analyzed seven naturally-occurring mutations that were previously reported in human FSHR (hFSHR), in the context of equine FSHR (eFSHR); these include one constitutively activation variant, one allelic variant, and five inactivating variants. These mutations were introduced into wild-type eFSHR (eFSHR-wt) sequence to generate mutants that were designated as eFSHR-D566G, -A306T, -A189V, -N191I, -R572C, -A574V, and -R633H. Mutants were transfected into PathHunter EA-parental CHO-K1 cells expressing β-arrestin. The biological function of mutants was analyzed by quantitating cAMP accumulation in cells incubated with increasing concentrations of FSH. Results: Cells expressing eFSHR-D566G exhibited an 8.6-fold increase in basal cAMP response, as compared to that in eFSHR-wt. The allelic variation mutant eFSHR-A306T was not found to affect the basal cAMP response or half maximal effective concentration (EC50) levels. On the other hand, eFSHR-D566G and eFSHR-A306T displayed a 1.5- and 1.4-fold increase in the maximal response, respectively. Signal transduction was found to be completely impaired in case of the inactivating mutants eFSHR-A189V, -R572C, and -A574V. When compared with eFSHR-wt, eFSHR-N191I displayed a 5.4-fold decrease in the EC50 levels (3,910 ng/mL) and a 2.3-fold decrease in the maximal response. In contrast, cells expressing eFSHR-R633H displayed in a similar manner to that of the cells expressing the eFSHR-wt on signal transduction and maximal response. Conclusion: The activating mutant eFSHR-D566G greatly enhanced the signal transduction in response to FSH, in the absence of agonist treatment. We suggest that the state of activation of the eFSHR can modulate its basal cAMP accumulation.

Evaluation of coat color inheritance and production performance for crossbreed from Chinese indigenous Chenghua pig crossbred with Berkshire

  • Li, Yujing;Yuan, Rong;Gong, Zhengyin;Zou, Qin;Wang, Yifei;Tang, Guoqing;Zhu, Li;Li, Xuewei;Jiang, Yanzhi
    • Animal Bioscience
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    • v.35 no.10
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    • pp.1479-1488
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    • 2022
  • Objective: This work was to determine coat inheritance and evaluate production performance for crossbred pigs from Berkshire×Chenghua (BC) compared with Chinese indigenous Chenghua (CH) pigs. Methods: The coat color phenotypes were recorded for more than 16,000 pigs, and the genotypes of melanocortin 1 receptor (MCIR) gene were identified by sequencing. The reproductive performance of 927 crossbred BC F4 gilts and 320 purebred CH gilts was recorded. Sixty pigs of each breed were randomly selected at approximately 60 days of age to determine growth performance during fattening period, which lasted for 150 days for BC pigs and 240 days for CH pigs. At the end of the fattening period, 30 pigs of each breed were slaughtered to determine carcass composition and meat quality. Results: The coat color of BC pigs exhibits a "dominant black" hereditary pattern, and all piglets derived from boars or sows genotyped ED1 ED1 homozygous for MC1R gene showed a uniform black coat phenotype. The BC F4 gilts displayed a good reproductive performance, showing a higher litter and tear size and were heavier at farrowing litter and at weaning litter than the CH gilts, but they reached puberty later than the CH gilts. BC F4 pigs exhibited improved growth and carcass characteristics with a higher average daily live weight gain, lower feed-to-gain ratio, and higher carcass lean meat rate than CH pigs. Like CH pigs, BC F4 pigs produced superior meat-quality characteristics, showing ideal pH and meat-color values, high intramuscular fat content and water-holding capacity, and acceptable muscle-fiber parameters. C18:1, C16:0, C18:0, and C18:2 were the main fatty acids in M. longissimus lumborum in the two breeds, and a remarkably high polyunsaturated/saturated fatty acid ratio of ~0.39 was observed in the BC F4 pigs. Conclusion: The BC F4 pigs exhibit a uniform black coat pattern and acceptable total production performance.

Studies on the Utilization of Woodland for Livestock Farming II. Problem and Its Improvement Followed by the Join Cattle Grazing in king Won Do (임지의 축산적 이용에 관한 연구 제2보. 강원도의 새마을 "소" 임간공동방목사업의 문제점과 개선책)

  • 맹원재;윤익석;유제창;정승헌
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.3 no.2
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    • pp.100-111
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    • 1983
  • The research results reported herein had the objectives to understand and analyze the present problems of saemaeul woodland joint cattle grazing system in Kang Won Do and to take steps of improvement. The study results on actual management conditions, problems analyzed and improvement plan of total 208 joint cattle grazing area which was established 105 area in 1981 and 103 area in 1982 were summarized as follows: 1. the effectiveness of joint cattle grazing projects 1) Average daily weight gain of cattle during joint cattle grazing period was 0.4kg, showing higher daily than the conventional feeding of 0.33kg. 2) Increase of total farm income over the conventional feeding system were \1,031,357,320 during the grazing period from May to October in 1982 by adapting the 208 joint cattle grazing system, of which effectiveness of weight gain was \293,075,300 and labor saving was \543,838,750. 3) According to the results of questionaire investigation from 208 joint cattle grazing area, effectiveness of joint cattle grazing system over the conventional system were (1) labor saving, (2) feed cost saving, (3) reduced diseases, (4) increase of number of feeding, (5) inspiration of joint endeavor, (6) effect of more gain, (7) easiness of feeding and feed cost savings. 2. Problems of joint cattle grazing system. 1) Shortages of grass were a problem at second year of joint cattle grazing period due to the low regrowth rate of wild grass. 2) Proper land for woodland joint cattle grazing is belonging to land of Government ownership and it is very hard to get the permission from office of forestry for cattle grazing purpose. 3) It is also difficult to find a proper time of breeding in grazing area by the difficulty of estrus detection. 4) There are a difficulty to give a proper vaccination and medical examination for the grazing cattle. 3. Improvement plans for woodland joint cattle grazing projects. 1) Obtain sufficient roughages by hoof cultivation and oversowing pasture from the second year of joint cattle grazing period. 2) In order to increase the beef production and to use for a calf production area, Government should arrange that all proper grazing land of Government owned in Kang Won Do convert into woodland joint cattle grazing area. 3) Make a good reproductive record by mixed grazing with a excellent breeding cow in a remote area. And carry out the collective artificial insemination with synchronous puberty induced by injection of puberty stimulation hormone. 4) Make a preventive injection for blackleg, twice medication of fasciola hepatica in a year, and spray and medication of tick insecticide. 4. A policy towards upbringing of woodland joint cattle grazing area. 1) Government should thoroughly investigate about a proper land for woodland joint cattle grazing from all forests. 2) When the area is suitable for the woodland joint cattle grazing, though it is national forest or restricted area, government should make it possible to establish a grazing area. 3) On the proper land foe joint cattle grazing in the remote place, Government should support for the road construction and electric fence equipments by using of national funds. 4) There should be an administrative consideration for well promotion of the project that make woodland joint cattle grazing suitable to the characteristics of Kang Won Do. 5) In order to improve the reproduction record, Government should reform the insufficiency of artificial insemination in the joint cattle grazing area. 6) In order to maintain a proper price of cow, Government should carry out the price plan. 7) When there is any request for grassland formation in the woodland joint cattle grazing area, Government should permit it with preference.

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Serum levels of free insulin-like growth factor-I and clinical value in healthy children (한국 소아 및 청소년에서 혈중 유리 insulin-like growth factor-I 농도치와 임상적 의의)

  • Chung, Young Hee;Chung, Woo Yeong
    • Clinical and Experimental Pediatrics
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    • v.51 no.1
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    • pp.47-53
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    • 2008
  • Purpose : The serum levels of total insulin-like growth factor (IGF)-I and IGF binding protein (IGFBP)-3 reflect endogenous growth hormone (GH) secretion in healthy children. Free form of IGF-I which is suggested to have more potent biological action than complex form of IGF-I. The aim of this study is to investigate the serum levels of free IGF-I and its clinical value in healthy children. Methods : Serum levels of total IGF-I and IGFBP-3 were determined in 494 healthy children (248 boys and 246 girls) by RIA and IRMA. Serum level of free IGF-I was determined in 206 healthy children (103 boys and 103 girls) by IRMA. Results : The free IGF-I level increased with age in both sex. The free IGF-I level increased continuously between 7 and 15 years of age in boys, but decrement was noted after 14 years of age in girls. Serum total IGF-I level also increased with age in similar pattern of that of free IGF-I. There were no significant differences of mean values of the ratio of free IGF-I/total IGF-I in relation to age in both sex. And there were significant correlations between the level of free IGF-I and total IGF-I and the ratio of total IGF-I/IGFBP-3, respectively. Conclusion : In healthy children, serum free IGF-I increased with age in both sex and high free IGF-I level may play an important role in pubertal growth spurt. Our results suggest that the increased serum free IGF-I level in puberty may reflect changes in total IGF-I rather than IGFBP-3. But free IGF-I does not have more clinical value than total IGF-I because of no significant differences of mean values of the ratio of free IGF-I/total IGF-I in relation to age.

Ultrasensitive Enzymeimmunoassay for Testosterone in Human Saliva (사람 타액내 Testosterone의 초감도 효소면역측정법)

  • 윤용달;전은현;이창주;도병록;이준영
    • Development and Reproduction
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    • v.4 no.1
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    • pp.115-123
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    • 2000
  • A few enzymeimmunoassay (EfA) for testosterone (T) have been reported but was not suitable for all biological samples. The present study was designed to develop a rapid, ultrasensitive EIA and to apply this technique for study the physiological changes of T in biological samples. Saliva samples were collected at 06:00~09:00 hour during one menstrual cycle from 18 normally menstruating women and on 09:00~10:00 hour from 20 normal men. The present study shows an established EIA for testosterone, using horseradish peroxidase (HRP), which was covalently bonded to testosterone-3-carboxymethyloxime (T-3-CMO). One batch of T-antisera was also covalently linked to microcrystalline cellulose particles by a mixed anhydride method in order to facilitate separation of bound and free steroids. The established EIA was validated in terms of sensitivity, accuracy, specificity, precisions etc., comparing with conventional radioimmunoassay. The sensitivity of the established EIA was less than 25 pg/tube. The correlation coefficients between the expected T-values and observed T-values measured by EIA or RIA were r=0.985 and r=0.941 respectively. The cross reactivity of antiserum in EIA was a little higher than that of RIA, especially by 5 ${\alpha}$-DHT. The intra- and inter-assay precisions of the present EIA were similar to those of RIA. The present study also demonstrates that the normal T-values in saliva of Korean male & female samples are 265.65${\pm}$15.80 pmol/l and 109.74${\pm}$ 12.01 pmol/l, respectively. The present EIA seems to be established and suitable for use in the endocrinological studies. The advantages of this EIA system also might make the present T-EIA an ideal procedure for use in a routine assay of ordinary laboratory with a conventional spectrophotometer.

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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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