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Ultrastructure of the Testis and Germ Cell Development During Spermatogenesis in Male Crassostrea gigas (Bivalvia: Ostreidae) in Western Korea

  • Kim, Jin-Hee;Chung, Ee-Yung;Choi, Ki-Ho;Lee, Ki-Young;Choi, Moon-Sul
    • The Korean Journal of Malacology
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    • v.26 no.3
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    • pp.235-244
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    • 2010
  • Ultrastructural characteristics of the testis and spermatogenesis of Crassostrea gigas were investigated by Transmission and Scanning Electron microscope observations. The testis is a diffuse organ consisting of branching acini containing differentiating germ cells in a variety of stages. The acinus is surrounded by an intermitent layer of myoepithelial cells andis divided into subcompartments that are partially separated by pleomorphic accessory cells which remain in close contact with germ cells until late stages of development. these accessory cells contain a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in the supplying of the nutrients for germ cell development, while any phenomena associated with phagocytosis of undischarged, residual sperms by lysosomes could be find in the cytoplasm of the accessory cells. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves. Mature spermatozoa consist of broad, cap-shaped acrosomal vesicle, subacrosomal material (containing axial rod embedded in a granular matrix), a oval nucleus showing deeply invaginated anteriorly, two triplet substructure centrioles surrounded by four spherical mitochondria, and satelite fibres appear to the distal centriole and plasma membrane. Spermatozoa of C. gigas resemble to those of other investigated ostreids. In particular, the anterior region of the acrosomal vesicle is transversely banded. It is assumed that differences in this acrosomal substructure are associated with the inability of fertilization between the genus Crassostrea and other genus species in Ostreidae. Therefore, we can use sperm morphology in the resolution of taxonomic relationships within the Ostreidea. The spermatozoon is approximately $42-47{\mu}m$ in length including an oval sperm nucleus (about $0.91{\mu}m$ in length), an acrosome (about $0.42{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$). The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9 + 2 structure. These morphological charateristics of acrosomal vesicle belong to the family Ostreidae in the subclass Pteriomorphia.

The Conservation Treatment of the Bark of Wooden Sculpture (목제 조각품의 수피부 보존처리)

  • Kim, Young Mok;Han, Ye Bin;Shin, Jeong Ah;Cha, Sun Min;Kwon, Hee Hong
    • Journal of Conservation Science
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    • v.37 no.5
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    • pp.516-524
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    • 2021
  • The 'artist's intention' plays an important role in the conservation process of contemporary art. Accordingly, the information on artworks owned by the artist, their bereaved family members, and foundations that have decision-making power is important. The artist's interview is the kind of data that can clear 'artist's intentions' to ensure reliable conservation treatment can be carried out even after the artist's death. Therefore, this study attempted to use information from the artist's interview on the type of wood in the manufacture of the filler required for lifted and separated bark conditions in the conservation process of wooden artwork in the National Museum of Modern and Contemporary Art. While the conservation treatment resulted in the stable preservation of the bark of artwork, an analysis of the wood confirmed that the information in the artist's interview was not true. Consequently, we suggest that attention must be paid toward the information provided by artist, their bereaved families, etc. Based on the result of this study, this is also expected to help preserve upcoming artworks with similar bark conditions.

Development of Individual Residue Analysis Method for Cyanazine in Agricultural Commodities as an Unregistered Herbicide in Korea (국내 미등록 제초제 cyanazine의 농산물 중 개별 잔류분석법 개발)

  • Choung, Myoung-Gun;Im, Moo-Hyeog
    • Journal of the Korean Society of International Agriculture
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    • v.30 no.4
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    • pp.339-346
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    • 2018
  • Cyanazine is a member of the triazine family of herbicides. Cyanazine is used as a pre- and post-emergence herbicide for the control of annual grasses and broadleaf weeds. This experiment was conducted to establish a determination method for cyanazine, as domestic unregistered pesticide, residue in major agricultural commodities using HPLC-DAD/MS. Cyanazine was extracted with acetone from representative samples of five raw products which comprised apple, green pepper, Kimchi cabbage, hulled rice and soybean. The extract was diluted with saline water and partitioned to dichloromethane for remove polar extractive in the aqueous phase. For the hulled rice and soybean samples, n-hexane/acetonitrile partition was additionally employed to remove non-polar lipids. The extract was finally purified by optimized florisil column chromatography. On a $C_{18}$ column in HPLC, cyanazine was successfully separated from co-extractives of sample, and sensitively quantitated by diode array detection at 220 nm. Accuracy and precision of the proposed method was validated by the recovery experiment on every major agricultural commodity samples fortified with cyanazine at 3 concentration levels per agricultural commodity in each triplication. Mean recoveries were ranged from 83.6 to 93.3% in five major representative agricultural commodities. The coefficients of variation were all less than 10%, irrespective of sample types and fortification levels. Limit of quantitation(LOQ) of cyanazine was 0.02 mg/kg as verified by the recovery experiment. A confirmatory method using LC/MS with selected-ion monitoring(SIM) technique was also provided to clearly identify the suspected residue.

A Qualitative Case Study on the Child-rearing Experiences of a Couple who was Raised in a Child Care Institution (아동양육시설에서 성장한 부부의 자녀양육 경험에 관한 질적 사례연구)

  • Park, Shinae
    • Asia-pacific Journal of Multimedia Services Convergent with Art, Humanities, and Sociology
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    • v.7 no.9
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    • pp.519-527
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    • 2017
  • The purpose of this study is to take a closer look at the child-rearing experiences of a two married individuals who were both raised in a child care institution after they had been separated from their parents. Data was collected from a husband and wife who had grown up together in the same child care institution, and in-depth interviews were conducted on them. The research method was analyzed by a qualitative case study. According to the results, the following main themes were extracted from the experiences of the child-rearing couple : 'A family made to escape loneliness', 'Childbirth that started without preparation and fell into the swamp of child rearing', 'Faced reality, regret, and reflection', and 'Finally regaining strength because of the responsibility to be a parent'. The couple married with vague fantasies and without any preparation to escape from the terrible loneliness. Like the marriage, the couple subsequently did not have a specific plan for childbirth. Later, it proved difficult to raise children who did not receive enough information or education, and the couple became exhausted. After the child entered the day care center, the couple experienced a sense of regret and was forced to reflect as they faced a child who resembled their negative appearance. The couple also reported that they felt deeply responsible for their parents even though they did not grow up with any form of parental care. Based on these findings, this study suggested a social welfare intervention plan which could have helped the married couple grow up in the child care institution.

Effect of Solidago altissima L. Extract on Forage Crop Germination

  • Ho-Jun Gam;Yosep Kang;Eun-Jung Park;Ki-Yong Kim;Sang-Mo Kang;In-Jung Lee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.114-114
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    • 2022
  • There are 28 families and 166 species of exotic weeds on agricultural land and among these, 23 families and 80 species of exotic weeds occur on pastures. Among them, the Solidago altissima is a perennial weed belonging to the asteraceae family and it is an exotic weed that spreads to the surrounding area using methods such as high seed production, vegetative propagation using underground rhizomes and allelochemical. Accordingly, in 2009, the Ministry of Environment designated it as an ecosystem-disrupting species. This study was conducted to obtain basic data about the effects of S.altissima derived allelochemicals on forage crops. The root of S.altissima was separated, dried in the shade and then pulverized to prepare an root powder. Powder was repeatedly extracted with methanol for 3 days and concentrated under reduced pressure to obtain an root methanol extract. Dissolve the extract in distilled water, dispense it in a separate-funnel and proceed with liquid-liquid extraction by adding equal amounts of n-haxane (Hex), chloroform (CHCI3), ethyl acetate (EtoAC), and butanol (BuOH) in order of increasing polarity. A seed-bioassay was performed using fractions for each solvent, followed by separation and purification by silica gel column chromatography. As a result of the fraction germination test for each solvent, the IC50 values using the fresh weight of each fraction were 898.3 mg L-1, 676.3 mg L-1, 1160 mg L-1 and 1360 mg L-1. CA, CB, and CC fractions were obtained through primary silica gel column chromatography that used CHCI3 fraction. As a result of seed-bioassay using each fraction, the IC50 values for the fresh weight of each fraction was 537.3 mg L-1, 1280 mg L-1 and 1947 mg L-1. Based on this, 5 fractions were obtained as a result of secondary silica gel column chromatography using the CA fraction. A seed-bioassay was performed, as a result, the lowest IC50 value was calculated as 226.7 mg L-1 in the CAE fraction. Based on this, the fraction was analyzed by GC-MS. The results of this study can be used as basic research data on the effects of weeds on forage crops and allelochemicals secreted from S. altissima.

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Regeneration of adventitious root from Calystegia soldanella L. in Jeju island and mass proliferation method using bioreactor system (제주지역 갯메꽃(Calystegia soldanella L.) 유래 부정근 재분화 및 생물반응기 시스템 이용 대량증식법)

  • Jong-Du Lee;Eunbi Jang;Weon-Jong Yoon;Yong-Hwan Jung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.37-37
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    • 2021
  • Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L-1, and IBA 1mg·L-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.

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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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CLINICAL CHARACTERISTICS AND TREATMENT COURSES OF THE CHILDREN WITH SELECTIVE MUTISM (선택적 함구증 아동의 임상특성 및 치료경과)

  • Chung, Sun-Ju;Hong, Kang-E
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.6 no.1
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    • pp.74-89
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    • 1995
  • Selective mutism is a childhood condition defined by persisten failure to speak in specific social situation when speaking is expected, dispite preserved ability to comprehend spoken language and speak. Present study is to investigate clinical characteristics, treatment method and outcome of 23 children who were diagnosed as selective mutism by DSM-IV criteria at the child psychiatry ouptatient department of SNUH. The results were as follows : 1) The Sex ratio was 1: 4.8, female dominant Mear age of onset was 33 years old and mean age of first referral was 7.7 years old. 2) 22% of subjects had perinatal problem such as low birth weight, preterm birth, 26% of the subjects have history of delayed language development. There are subjects who had been separated with mam caretaker before 3 years old(26%) and who experienced physical or psychological trauma before 3 years old(26%). A few subjects had enurests(30%) and encoprests(4%). 3) Many subjects(65%) had symbiotic relationship with their mother. These families consist of dominant, verbally aggressive mother and passive father. Parents of 39% of all subjects were judged to have definite psychopathology(social phobic, depression, hysterical trait or alcohol problem) 26% of all subject, were reported physically abused. 4) The personality trait of the subjects were frequently described as follows(in order of frequency) ; Shy(100%), anxious(83%), stubborn(83%)m rigid and tense posture(78%), immature(65%) overdependent(65%), irritable(52%), manipulative(39%), depressive(39%). 5) The mean performance IQ of 16 subjects by KEDI-WISC was 88.3 Among them, the subjects with IQ below 69 were seven and those with IQ above 70 were nine. When comparing these two group(Mental retardation group vs Normal IQ group), we could find some difference in language development, personality trait, family dynamics and treatment outcome. 6) Among several treatment methods for selective mutism, play therapy was the most frequently used method(65%). Other commonly used treatment methods were pharmacotherapy(21%), behavioral therapy(8%), combined therapy(play therapy+pharmacotherapy+family therapy+behavioral therapy)(12%), 7) Regarding the outcome of treatment 8.6% was evaluated as Excellent, 30.4% as Good, 52% as Fair, 8.7% as Poor at the tinic of treatment. At follow up interview 21.7% was evaluated Excellent, 13% as Good, 21.7% as Fair, 34.8% as Poor. 8) We classified all subjects by Havden's 4 subtype. Symbiotic mutism was most common(65%) and other subtypes are Speech phobic mutism(8.6), Reactive mutism(13%) and Passive-aggressive mutism(30%).

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Characterization of Mutations in Bruton's Tyrosine Kinase(Btk) Gene from Unrelated 3 X-linked Agammaglobulinemia(XLA) Families in Korea (국내 X-관련성 범저감마글로불린혈증 세가족에 대한 Bruton's Tyrosine Kinase 단백질 발현 및 유전자 변이 분석)

  • Song, Chang-Hwa;Jo, Eun-Kyeong;Park, Jeong-Kyu;Kim, Jung-Soo;Hong, Soo-Jong;Lee, Jae-Ho
    • Clinical and Experimental Pediatrics
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    • v.45 no.3
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    • pp.302-310
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    • 2002
  • Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells(PBMC) from three XLA families in Korea. Methods : Heparinized venous blood samples were collected from four XLA patients and additional family members in three unrelated XLA families. Mononuclear cells were separated from their blood and the intracellular Btk protein was characterized by a flow cytometry. The mutation analysis was performed using direct sequencing. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in the patients with XLA. We observed a novel deletion and two point mutations within introns(intron 1 and intron 18) resulting in alternative splicings. In XLA family 2, a 980 bp deletion(from intron 9+191 T to intron 10-215 C) including exon 10 was found in patient P2. He was the only sporadic case in this study, because his mother and brother showed a normal Btk expression by flow cytometry. Conclusion : These identified genetic alterations support the molecular heterogeneity of Btk gene in XLA disease. Additionally, by means of flow cytometric analysis, we diagnosed three hypogammaglobulinemia patients as XLA. Advancements in diagnostic methods has facilitated a prompt and definite diagnosis of this disease.

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

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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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 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-t631 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 agoinst-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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