Pub Date : 2022-06-01Epub Date: 2022-06-30DOI: 10.12717/DR.2022.26.2.71
Ji-Eun Park, Seung Gee Lee, Young Hyun Yoo, Jong-Min Kim
In response to luteinizing hormone (LH), a higher concentration of progesterone (P4) is produced in luteal cells of corpus luteum (CL). Mitochondria are an essential cellular organelle in steroidogenesis. The specific engagement of the concept regarding mitochondrial shaping with early stages of steroidogenesis was suggested in reproductive endocrine cells. Although the specific involvement of GTPase dynamin-related protein 1 (Drp1) with steroidogenesis has been demonstrated in luteal cells of bovine CL in vitro, its actual relationship with ovarian steroidogenesis during the estrous cycle remains unknown. In this study, while Fis1 and Opa1 protein levels did not show significant changes during the estrous cycle, Drp1, Mfn1, and Mfn2 proteins exhibited relatively lower levels at proestrus than at estrus or diestrus. 3β-HSD showed higher levels at proestrus than at estrus or diestrus. In addition, Drp1 phosphorylation (s637) was higher in proestrus than in estrus or diestrus. Immune-positive cells for Drp1, pDrp1 (s637), and 3β-HSD were all localized in the cytoplasm of luteal cells in the CL. The immune-positive cells for 3β-HSD were more frequently seen in the CL at proestrus than at estrus or diestrus. Immunoreactivity for Drp1 in luteal cells at proestrus was weaker than that at estrus or diestrus. However, pDrp1 (s637) immune-positive cells were mostly detected in luteal cells at proestrus. These results imply that steroidogenesis (P4 production) in the CL is closely related to phosphorylation of Drp1 at serine 637. Taken together, this study presents evidence that Drp1 phosphorylation at serine 637 is an important step in steroidogenesis in the CL.
{"title":"Drp1 Expression and Phosphorylation in Steroidogenic Corpus Luteum during the Estrous Cycle in Rat Ovaries.","authors":"Ji-Eun Park, Seung Gee Lee, Young Hyun Yoo, Jong-Min Kim","doi":"10.12717/DR.2022.26.2.71","DOIUrl":"https://doi.org/10.12717/DR.2022.26.2.71","url":null,"abstract":"<p><p>In response to luteinizing hormone (LH), a higher concentration of progesterone (P4) is produced in luteal cells of corpus luteum (CL). Mitochondria are an essential cellular organelle in steroidogenesis. The specific engagement of the concept regarding mitochondrial shaping with early stages of steroidogenesis was suggested in reproductive endocrine cells. Although the specific involvement of GTPase dynamin-related protein 1 (Drp1) with steroidogenesis has been demonstrated in luteal cells of bovine CL <i>in vitro</i>, its actual relationship with ovarian steroidogenesis during the estrous cycle remains unknown. In this study, while Fis1 and Opa1 protein levels did not show significant changes during the estrous cycle, Drp1, Mfn1, and Mfn2 proteins exhibited relatively lower levels at proestrus than at estrus or diestrus. 3<i>β</i>-HSD showed higher levels at proestrus than at estrus or diestrus. In addition, Drp1 phosphorylation (s637) was higher in proestrus than in estrus or diestrus. Immune-positive cells for Drp1, pDrp1 (s637), and 3<i>β</i>-HSD were all localized in the cytoplasm of luteal cells in the CL. The immune-positive cells for 3<i>β</i>-HSD were more frequently seen in the CL at proestrus than at estrus or diestrus. Immunoreactivity for Drp1 in luteal cells at proestrus was weaker than that at estrus or diestrus. However, pDrp1 (s637) immune-positive cells were mostly detected in luteal cells at proestrus. These results imply that steroidogenesis (P4 production) in the CL is closely related to phosphorylation of Drp1 at serine 637. Taken together, this study presents evidence that Drp1 phosphorylation at serine 637 is an important step in steroidogenesis in the CL.</p>","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 2","pages":"71-77"},"PeriodicalIF":0.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/a5/4d/dr-26-2-71.PMC9336213.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40709075","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-01Epub Date: 2022-06-30DOI: 10.12717/DR.2022.26.2.91
Jong-Man Yoon
Genomic DNA (gDNA) set apart from two populations of Korean Charybdis crab (Charybdis japonica) was augmented by PCR experiments. The five oligonucleotides primers (ONT-primers) were spent to yield the number of unique loci shared to each crab population (ULSECP) and number of loci shared by the two crab populations (LSTCP). 305 fragments (FRAGs) were identified in the Charybdis crab population A (CCPA), and 344 in the Charybdis crab population B (CCPB): 44 number of ULSECP (14.43%) in the CCPA and 110 (31.98%) in the CCPB. 44 number of LSTCP, with an average of 8.8 per primer, were detected in the two crab populations. The bandsharing (BS) value between entity's no. 01 and no. 10 was the lowest (0.371) between the two CCPs. The average bandsharing (ABS) values of individuals in the CCPA (0.575±0.014) were lesser than in those originated from the CCPB (0.705±0.011) (p < 0.05). The polar hierarchical dendrogram (PHD) achieved by the five ONT-primers denotes three genetic clusters (GCs): cluster I (CHARYBCRAB 01, 04, 05, 06, and 08), cluster II (CHARYBCRAB 02, 03, 07, 09, 10, and 11) and cluster III (CHARYBCRAB 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and 22). The shortest genetic distance (GD) displaying significant molecular difference (MD) was between individuals CHARYBCRAB no. 18 and CHARYBCRAB no. 17 (0.055).
{"title":"PCR Analysis for Genetic Distances of Two <i>Charybdis</i> Crab Populations.","authors":"Jong-Man Yoon","doi":"10.12717/DR.2022.26.2.91","DOIUrl":"https://doi.org/10.12717/DR.2022.26.2.91","url":null,"abstract":"<p><p>Genomic DNA (gDNA) set apart from two populations of Korean <i>Charybdis</i> crab (<i>Charybdis japonica</i>) was augmented by PCR experiments. The five oligonucleotides primers (ONT-primers) were spent to yield the number of unique loci shared to each crab population (ULSECP) and number of loci shared by the two crab populations (LSTCP). 305 fragments (FRAGs) were identified in the <i>Charybdis</i> crab population A (CCPA), and 344 in the <i>Charybdis</i> crab population B (CCPB): 44 number of ULSECP (14.43%) in the CCPA and 110 (31.98%) in the CCPB. 44 number of LSTCP, with an average of 8.8 per primer, were detected in the two crab populations. The bandsharing (BS) value between entity's no. 01 and no. 10 was the lowest (0.371) between the two CCPs. The average bandsharing (ABS) values of individuals in the CCPA (0.575±0.014) were lesser than in those originated from the CCPB (0.705±0.011) (<i>p</i> < 0.05). The polar hierarchical dendrogram (PHD) achieved by the five ONT-primers denotes three genetic clusters (GCs): cluster I (CHARYBCRAB 01, 04, 05, 06, and 08), cluster II (CHARYBCRAB 02, 03, 07, 09, 10, and 11) and cluster III (CHARYBCRAB 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, and 22). The shortest genetic distance (GD) displaying significant molecular difference (MD) was between individuals CHARYBCRAB no. 18 and CHARYBCRAB no. 17 (0.055).</p>","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 2","pages":"91-98"},"PeriodicalIF":0.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/b4/11/dr-26-2-91.PMC9336212.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40599997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-01Epub Date: 2022-06-30DOI: 10.12717/DR.2022.26.2.79
Hee Jeong Kong, Jung Jin Lee, Ju-Won Kim, Julan Kim, Young-Ok Kim, Sang-Yeob Yeo
Krüppel-like factor 10 (KLF10) regulates various cellular functions, such as proliferation, differentiation and apoptosis, as well as the homeostasis of several types of tissue. In the present study, we attempted a loss-of-function analysis of zebrafish Klf11a and Klf11b, which constitute human KLF10 homologs. Embryos injected with klf11b-morpholino (MO) showed developmental retardation and cell death, whereas klf11a-MO-injected embryos showed normal development. In klf11b-MO-injected embryos, a dramatic increase in the amount of zebrafish p53 mRNA might be the cause of the increase in that of bax. The degree of apoptosis decreased in the klf11b-MO and p53-MO co-injected embryos. These findings imply that KLF10 is a negative regulator of p53-dependent transcription, suggesting that the KLF10/p53 complex may play an important role in apoptosis for maintenance of tissue homeostasis during embryonic development.
{"title":"Zebrafish Klf11b is Required to Maintain Cell Viability by Inhibiting p53-Mediated Apoptosis.","authors":"Hee Jeong Kong, Jung Jin Lee, Ju-Won Kim, Julan Kim, Young-Ok Kim, Sang-Yeob Yeo","doi":"10.12717/DR.2022.26.2.79","DOIUrl":"https://doi.org/10.12717/DR.2022.26.2.79","url":null,"abstract":"<p><p>Krüppel-like factor 10 (KLF10) regulates various cellular functions, such as proliferation, differentiation and apoptosis, as well as the homeostasis of several types of tissue. In the present study, we attempted a loss-of-function analysis of zebrafish <i>Klf11a</i> and <i>Klf11b</i>, which constitute human KLF10 homologs. Embryos injected with <i>klf11b-morpholino</i> (<i>MO</i>) showed developmental retardation and cell death, whereas <i>klf11a-MO</i>-injected embryos showed normal development. In <i>klf11b-MO</i>-injected embryos, a dramatic increase in the amount of zebrafish <i>p53</i> mRNA might be the cause of the increase in that of <i>bax</i>. The degree of apoptosis decreased in the <i>klf11b-MO</i> and <i>p53-MO</i> co-injected embryos. These findings imply that KLF10 is a negative regulator of p53-dependent transcription, suggesting that the KLF10/p53 complex may play an important role in apoptosis for maintenance of tissue homeostasis during embryonic development.</p>","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 2","pages":"79-90"},"PeriodicalIF":0.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/82/47/dr-26-2-79.PMC9336215.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40599993","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-01Epub Date: 2022-06-30DOI: 10.12717/DR.2022.26.2.59
Elsayed A Mohamed, Ji Woo Im, Dong-Hwan Kim, Hae-Rahn Bae
Many efforts have been made to study the expression of aquaporins (AQP) in the mammalian reproductive system, but there are not enough data available regarding their localized expression to fully understand their specific roles in male reproduction. The present study investigated the expression and localization patterns of different AQP subtypes in the adult mouse testes and testicular spermatozoa using an immunofluorescence assay. All the studied AQPs were expressed in the testes and revealed subtype-specific patterns in the intensity and localization depending on the cell types of the testes. AQP7 was the most abundant and intensive AQP subtype in the seminiferous tubules, expressing in Leydig cells and Sertoli cells as well as all stages of germ cells, especially the spermatids and testicular spermatozoa. The expression pattern of AQP3 was similar to that of AQP7, but with higher expression in the basal and lower adluminal compartments rather than the upper adluminalcompartment. AQP8 expression was limited to the spermatogonia and Leydig cells whereas AQP9 expression was exclusive to tails of the testicular spermatozoa and elongated spermatids. Taken together, the abundance and distribution of the AQPs across the different cell types in the testes indicating to their relavance in spermatogenesis, as well as in sperm maturation, transition, and function.
{"title":"Differential Expressions of Aquaporin Subtypes in the Adult Mouse Testis.","authors":"Elsayed A Mohamed, Ji Woo Im, Dong-Hwan Kim, Hae-Rahn Bae","doi":"10.12717/DR.2022.26.2.59","DOIUrl":"https://doi.org/10.12717/DR.2022.26.2.59","url":null,"abstract":"<p><p>Many efforts have been made to study the expression of aquaporins (AQP) in the mammalian reproductive system, but there are not enough data available regarding their localized expression to fully understand their specific roles in male reproduction. The present study investigated the expression and localization patterns of different AQP subtypes in the adult mouse testes and testicular spermatozoa using an immunofluorescence assay. All the studied AQPs were expressed in the testes and revealed subtype-specific patterns in the intensity and localization depending on the cell types of the testes. AQP7 was the most abundant and intensive AQP subtype in the seminiferous tubules, expressing in Leydig cells and Sertoli cells as well as all stages of germ cells, especially the spermatids and testicular spermatozoa. The expression pattern of AQP3 was similar to that of AQP7, but with higher expression in the basal and lower adluminal compartments rather than the upper adluminalcompartment. AQP8 expression was limited to the spermatogonia and Leydig cells whereas AQP9 expression was exclusive to tails of the testicular spermatozoa and elongated spermatids. Taken together, the abundance and distribution of the AQPs across the different cell types in the testes indicating to their relavance in spermatogenesis, as well as in sperm maturation, transition, and function.</p>","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 2","pages":"59-69"},"PeriodicalIF":0.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/76/13/dr-26-2-59.PMC9336216.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40599995","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-01Epub Date: 2022-06-30DOI: 10.12717/DR.2022.26.2.49
Yong-Seung Lee, Ki-Ho Lee
The differentiation and development of preadipocyte into mature adipocyte are regulated by transcription factors, such as CCAAT enhancer binding protein (Cebp) gene family and sterol regulatory element binding transcription factor 1 (Srebp1). Steroid hormones give influences on the development and function of adipocyte. The present research examined expression patterns of CCAAT enhancer binding protein alpha (Cebpa), CCAAT enhancer binding protein beta (Cebpb), CCAAT enhancer binding protein gamma (Cebpg), sterol regulatory element binding transcription factor 1 (Srebp1), androgen receptor (Ar), and estrogen receptors (Esr) among different epididymal fat parts during postnatal period by quantitative real-time polymerase chain reaction. In the distal epididymal fat, expression of Cebpa, Cebpb, Cebpg, Srebp1, Ar, and Esr2 was increased until 12 months of age, while expression of Esr1 was decreased at 5 months of age and was not detectable after 8 months of age. In the proximal epididymal fat, transcript levels of Cebps and Srebp1 were increased at 8 months of age, followed by decreases of Cebpb and Cebpg transcript levels at 12 months of age. An additional increase of Srebp1 expression was observed at 12 months of age. Expression of Ar and Esr2 were increased until 8 months of age, followed by a drop of Ar expression level at 12 months of age. Expression pattern of Esr1 was similar to that in the distal epididymal fat. In the tail epididymal fat, expression of Cebpa, Cebpg, Srebp1, Ar, and Esr2 was increased with age. Esr1 was not detectable at all. The highest level of Cebpb was observed at 8 months of age. These data suggest the possibility of developmental and functional differentiation among the epididymal fat parts.
{"title":"Expressional Evaluation of C/EBP Family, SREBP1, and Steroid Hormone Receptors in the Epididiymal Fat of Postnatally Developing Mouse.","authors":"Yong-Seung Lee, Ki-Ho Lee","doi":"10.12717/DR.2022.26.2.49","DOIUrl":"https://doi.org/10.12717/DR.2022.26.2.49","url":null,"abstract":"<p><p>The differentiation and development of preadipocyte into mature adipocyte <i>ar</i>e regulated by transcription factors, such as CCAAT enhancer binding protein (<i>Cebp</i>) gene family and sterol regulatory element binding transcription factor 1 (<i>Srebp1</i>). Steroid hormones give influences on the development and function of adipocyte. The present rese<i>ar</i>ch examined expression patterns of CCAAT enhancer binding protein alpha (<i>Cebpa</i>), CCAAT enhancer binding protein beta (<i>Cebpb</i>), CCAAT enhancer binding protein gamma (<i>Cebpg</i>), sterol regulatory element binding transcription factor 1 (<i>Srebp1</i>), androgen receptor (<i>Ar</i>), and estrogen receptors (<i>Esr</i>) among different epididymal fat p<i>ar</i>ts during postnatal period by quantitative real-time polymerase chain reaction. In the distal epididymal fat, expression of <i>Cebpa</i>, <i>Cebpb</i>, <i>Cebpg</i>, <i>Srebp1</i>, <i>Ar</i>, and <i>Esr</i>2 was increased until 12 months of age, while expression of <i>Esr1</i> was decreased at 5 months of age and was not detectable after 8 months of age. In the proximal epididymal fat, transcript levels of <i>Cebps</i> and <i>Srebp1</i> were increased at 8 months of age, followed by decreases of <i>Cebpb</i> and <i>Cebpg</i> transcript levels at 12 months of age. An additional increase of <i>Srebp1</i> expression was observed at 12 months of age. Expression of <i>Ar</i> and <i>Esr</i>2 were increased until 8 months of age, followed by a drop of <i>Ar</i> expression level at 12 months of age. Expression pattern of <i>Esr1</i> was simil<i>ar</i> to that in the distal epididymal fat. In the tail epididymal fat, expression of <i>Cebpa</i>, <i>Cebpg</i>, <i>Srebp1</i>, <i>Ar</i>, and <i>Esr</i>2 was increased with age. <i>Esr1</i> was not detectable at all. The highest level of <i>Cebpb</i> was observed at 8 months of age. These data suggest the possibility of developmental and functional differentiation among the epididymal fat p<i>ar</i>ts.</p>","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 2","pages":"49-58"},"PeriodicalIF":0.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/b7/e8/dr-26-2-49.PMC9336211.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40599994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-06-01Epub Date: 2022-06-30DOI: 10.12717/DR.2022.26.2.37
Donchan Choi
Photoperiod has well been established to regulate testicular activities in golden hamsters. These animals breed actively around summer but become infertile in winter. In males, testicles are full of multistep germ cells including spermatozoa in summer. But in winter only fundamental cells consisting of the testicles are detected. The testicular degeneration is accompanied by the reduced levels of blood gonadotropins and testosterone. In this study, the expressions of gonadotropin subunit genes were investigated in the reproductive active and inactive testicles. And parts of sequences of the gonadotropin subunits were identified and compared with those of other rodents. As results, common gonadotropin alpha (CGa), follicle-stimulating hormone (FSH) β, and luteinizing hormone (LH) β genes were equivalently detected in pituitaries of both sexually active and inactive animals. In considering low concentrations of gonadotropin hormones determined in pituitary, the present findings imply that the processes involved in translation and/or formation of functional hormones could be impeded in the sexually inactive hamsters. All the nucleotide sequences of gonadotropin subunits identified in this study were same as those reported previously except for one base in CGa. An unsure amino acid deduced from the CGa sequence was confirmed from mRNA sequencing. The outcomes mentioned above suggest that animals with regressed testes prepare for the sexually active period forthcoming in the future.
{"title":"Expressions of Gonadotropin Subunit Genes in the Reproductively Inactive Golden Hamsters.","authors":"Donchan Choi","doi":"10.12717/DR.2022.26.2.37","DOIUrl":"https://doi.org/10.12717/DR.2022.26.2.37","url":null,"abstract":"<p><p>Photoperiod has well been established to regulate testicular activities in golden hamsters. These animals breed actively around summer but become infertile in winter. In males, testicles are full of multistep germ cells including spermatozoa in summer. But in winter only fundamental cells consisting of the testicles are detected. The testicular degeneration is accompanied by the reduced levels of blood gonadotropins and testosterone. In this study, the expressions of gonadotropin subunit genes were investigated in the reproductive active and inactive testicles. And parts of sequences of the gonadotropin subunits were identified and compared with those of other rodents. As results, common gonadotropin alpha (CGa), follicle-stimulating hormone (FSH) β, and luteinizing hormone (LH) β genes were equivalently detected in pituitaries of both sexually active and inactive animals. In considering low concentrations of gonadotropin hormones determined in pituitary, the present findings imply that the processes involved in translation and/or formation of functional hormones could be impeded in the sexually inactive hamsters. All the nucleotide sequences of gonadotropin subunits identified in this study were same as those reported previously except for one base in CGa. An unsure amino acid deduced from the CGa sequence was confirmed from mRNA sequencing. The outcomes mentioned above suggest that animals with regressed testes prepare for the sexually active period forthcoming in the future.</p>","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 2","pages":"37-47"},"PeriodicalIF":0.0,"publicationDate":"2022-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/bf/7f/dr-26-2-37.PMC9336214.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40599996","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Munkhzaya Byambaragchaa, Hyo-Eun Joo, Sang-Gwon Kim, Yean-Ji Kim, Gyeong-Eun Park, K. Min
Abstract This study aimed to investigate the signal transduction of phosphorylation sites at the carboxyl (C)-terminal region of equine luteinizing hormone/chorionic gonadotropin receptor (eLH/ CGR). The eLH/CGR has a large extracellular domain of glycoprotein hormone receptors within the G protein-coupled receptors. We constructed a mutant (eLH/CGR-t656) of eLH/ CGR, in which the C-terminal cytoplasmic tail was truncated at the Phe656 residue, through polymerase chain reaction. The eLH/CGR-t656 removed 14 potential phosphorylation sites in the intracellular C-terminal region. The plasmids were transfected into Chinese hamster ovary (CHO)-K1 and PathHunter Parental cells expressing β-arrestin, and agonist-induced cAMP responsiveness was analyzed. In CHO-K1 cells, those expressing eLH/CGR-t656 were lower than those expressing eLH/CGR wild-type (eLH/CGR-wt). The EC50 of the eLH/ CGR-t656 mutant was approximately 72.2% of the expression observed in eLH/CGR-wt. The maximal response in eLH/CGR-t656 also decreased to approximately 43% of that observed in eLH/CGR-wt. However, in PathHunter Parental cells, cAMP activity and maximal response of the eLH/CGR-t656 mutant were approximately 173.5% and 100.8%, respectively, of that of eLH/CGR-wt. These results provide evidence that the signal transduction of C-terminal phosphorylation in eLH/CGR plays a pivotal role in CHO-K1 cells. The cAMP level was recovered in PathHunter Parental cells expressing β-arrestin. We suggest that the signal transduction of the C-terminal region phosphorylation sites is remarkably different depending on the cells expressing β-arrestin in CHO-K1 cells.
{"title":"Signal Transduction of C-Terminal Phosphorylation Regions for Equine Luteinizing Hormone/Chorionic Gonadotropin Receptor (eLH/CGR)","authors":"Munkhzaya Byambaragchaa, Hyo-Eun Joo, Sang-Gwon Kim, Yean-Ji Kim, Gyeong-Eun Park, K. Min","doi":"10.12717/DR.2022.26.1.1","DOIUrl":"https://doi.org/10.12717/DR.2022.26.1.1","url":null,"abstract":"Abstract This study aimed to investigate the signal transduction of phosphorylation sites at the carboxyl (C)-terminal region of equine luteinizing hormone/chorionic gonadotropin receptor (eLH/ CGR). The eLH/CGR has a large extracellular domain of glycoprotein hormone receptors within the G protein-coupled receptors. We constructed a mutant (eLH/CGR-t656) of eLH/ CGR, in which the C-terminal cytoplasmic tail was truncated at the Phe656 residue, through polymerase chain reaction. The eLH/CGR-t656 removed 14 potential phosphorylation sites in the intracellular C-terminal region. The plasmids were transfected into Chinese hamster ovary (CHO)-K1 and PathHunter Parental cells expressing β-arrestin, and agonist-induced cAMP responsiveness was analyzed. In CHO-K1 cells, those expressing eLH/CGR-t656 were lower than those expressing eLH/CGR wild-type (eLH/CGR-wt). The EC50 of the eLH/ CGR-t656 mutant was approximately 72.2% of the expression observed in eLH/CGR-wt. The maximal response in eLH/CGR-t656 also decreased to approximately 43% of that observed in eLH/CGR-wt. However, in PathHunter Parental cells, cAMP activity and maximal response of the eLH/CGR-t656 mutant were approximately 173.5% and 100.8%, respectively, of that of eLH/CGR-wt. These results provide evidence that the signal transduction of C-terminal phosphorylation in eLH/CGR plays a pivotal role in CHO-K1 cells. The cAMP level was recovered in PathHunter Parental cells expressing β-arrestin. We suggest that the signal transduction of the C-terminal region phosphorylation sites is remarkably different depending on the cells expressing β-arrestin in CHO-K1 cells.","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 1","pages":"1 - 12"},"PeriodicalIF":0.0,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44871051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-03-01DOI: 10.12717/DR.2022.26.1.23
Sil Jin, Haewon Jeon, Chong Pyo Choe
Abstract Pharyngeal pouches, a series of outgrowths of the pharyngeal endoderm, are a key epithelial structure governing facial skeleton development in vertebrates. Pouch formation is achieved through collective cell migration and rearrangement of pouch-forming cells controlled by actin cytoskeleton dynamics. While essential transcription factors and signaling molecules have been identified in pouch formation, regulators of actin cytoskeleton dynamics have not been reported yet in any vertebrates. Cofilin1-like (Cfl1l) is a fish-specific member of the Actin-depolymerizing factor (ADF)/Cofilin family, a critical regulator of actin cytoskeleton dynamics in eukaryotic cells. Here, we report the expression and function of cfl1l in pouch development in zebrafish. We first showed that fish cfl1l might be an ortholog of vertebrate adf, based on phylogenetic analysis of vertebrate adf and cfl genes. During pouch formation, cfl1l was expressed sequentially in the developing pouches but not in the posterior cell mass in which future pouch-forming cells are present. However, pouches, as well as facial cartilages whose development is dependent upon pouch formation, were unaffected by loss-of-function mutations in cfl1l. Although it could not be completely ruled out a possibility of a genetic redundancy of Cfl1l with other Cfls, our results suggest that the cfl1l expression in the developing pouches might be dispensable for regulating actin cytoskeleton dynamics in pouch-forming cells.
{"title":"Expression and Functional Analysis of cofilin1-like in Craniofacial Development in Zebrafish","authors":"Sil Jin, Haewon Jeon, Chong Pyo Choe","doi":"10.12717/DR.2022.26.1.23","DOIUrl":"https://doi.org/10.12717/DR.2022.26.1.23","url":null,"abstract":"Abstract Pharyngeal pouches, a series of outgrowths of the pharyngeal endoderm, are a key epithelial structure governing facial skeleton development in vertebrates. Pouch formation is achieved through collective cell migration and rearrangement of pouch-forming cells controlled by actin cytoskeleton dynamics. While essential transcription factors and signaling molecules have been identified in pouch formation, regulators of actin cytoskeleton dynamics have not been reported yet in any vertebrates. Cofilin1-like (Cfl1l) is a fish-specific member of the Actin-depolymerizing factor (ADF)/Cofilin family, a critical regulator of actin cytoskeleton dynamics in eukaryotic cells. Here, we report the expression and function of cfl1l in pouch development in zebrafish. We first showed that fish cfl1l might be an ortholog of vertebrate adf, based on phylogenetic analysis of vertebrate adf and cfl genes. During pouch formation, cfl1l was expressed sequentially in the developing pouches but not in the posterior cell mass in which future pouch-forming cells are present. However, pouches, as well as facial cartilages whose development is dependent upon pouch formation, were unaffected by loss-of-function mutations in cfl1l. Although it could not be completely ruled out a possibility of a genetic redundancy of Cfl1l with other Cfls, our results suggest that the cfl1l expression in the developing pouches might be dispensable for regulating actin cytoskeleton dynamics in pouch-forming cells.","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 1","pages":"23 - 36"},"PeriodicalIF":0.0,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45624002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-03-01DOI: 10.12717/DR.2022.26.1.13
Seong Hee Mun, H. J. Oh, J. Kwon
Abstract Neurokinin B (NKB) is a neuropeptide involved in the regulation of reproductive endocrine system of vertebrate animals, including fish. However, the pathway of NKB action in fish has not been clearly elucidated. In order to clarify the effect of NKB and NKF (neurokinin F) on gonadotropic hormone (GTH) gene expression in the pituitary, we studied the changes of LHβ and FSHβ gene expressions by using two different pituitary culture methods (whole pituitary culture or dispersed pituitary cell culture). Pituitaries were removed from mature female and male Nile tilapia. Changes of LHβ and FSHβ gene expressions were measured and compared after the treatment with NKB or NKF peptides at concentrations 0 to 1,000 nM. Expression of GTH genes in the whole pituitary cultures treated with NKB or NKF peptides did not show significant difference except in female at one concentration when treated with NKF. On the contrary, there were significant changes of GTH gene expressions in the dispersed pituitary cell cultures when treated with NKB and NKF peptides. These results suggest that dispersed pituitary cell culture is more relevant than whole pituitary culture in studying the function of pituitary, and that NKB and NKF could act directly on the pituitary to regulate the expression of GTH genes.
{"title":"Response of Pituitary Cells and Tissues to Neurokinin B and F in the Nile tilapia","authors":"Seong Hee Mun, H. J. Oh, J. Kwon","doi":"10.12717/DR.2022.26.1.13","DOIUrl":"https://doi.org/10.12717/DR.2022.26.1.13","url":null,"abstract":"Abstract Neurokinin B (NKB) is a neuropeptide involved in the regulation of reproductive endocrine system of vertebrate animals, including fish. However, the pathway of NKB action in fish has not been clearly elucidated. In order to clarify the effect of NKB and NKF (neurokinin F) on gonadotropic hormone (GTH) gene expression in the pituitary, we studied the changes of LHβ and FSHβ gene expressions by using two different pituitary culture methods (whole pituitary culture or dispersed pituitary cell culture). Pituitaries were removed from mature female and male Nile tilapia. Changes of LHβ and FSHβ gene expressions were measured and compared after the treatment with NKB or NKF peptides at concentrations 0 to 1,000 nM. Expression of GTH genes in the whole pituitary cultures treated with NKB or NKF peptides did not show significant difference except in female at one concentration when treated with NKF. On the contrary, there were significant changes of GTH gene expressions in the dispersed pituitary cell cultures when treated with NKB and NKF peptides. These results suggest that dispersed pituitary cell culture is more relevant than whole pituitary culture in studying the function of pituitary, and that NKB and NKF could act directly on the pituitary to regulate the expression of GTH genes.","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"26 1","pages":"13 - 21"},"PeriodicalIF":0.0,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47001329","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-12-01Epub Date: 2021-12-31DOI: 10.12717/DR.2021.25.4.257
Bo Young Lee, Jeong Bin Jo, Donchan Choi, Sung-Ho Lee, Yong-Pil Cheon
Phthalates and their metabolites are well-known endocrine disrupting chemicals. Di-(2-ethylhexyl) phthalate (DEHP) has been widely used in industry and the exposing possibility to adult is high. In this study, DEHP was treated (133 μg/L and 1,330 μg/L in drinking water) according to the OECD test guideline 443 to mature female mice and their adrenal gland were examined for histological characteristics and steroidogenic gene expression. The wet weight of the adrenal gland was increased in all administrated groups compared to control. The diameter of zona fasciculata (ZF) was increased by DEHP in both outer ZF and inner ZF but there was no difference in morphology of the cells and arrangements into zona between groups. In addition, the arrangement of extracellular matrix was not different between control and DEHP groups. CYP11B1 was mainly localized at ZF and the intensity was not different between groups. DAX1 was localized in zona glomerulosa (ZG) and ZF, and its expression levels were decreased by DEHP administration. Its level was lower in DEHP133 group than DEHP1330 group. On the other hand, CYP17A1 was localized in ZG of DEHP1330 group. These results suggest that chronic low-dose DEHP exposing may modify the microstructure and function of the adrenal cortical cortex.
{"title":"A Chronic-Low-Dose Exposing of DEHP with OECD TG 443 Altered the Histological Characteristics and Steroidogeic Gene Expression of Adrenal Gland in Female Mice.","authors":"Bo Young Lee, Jeong Bin Jo, Donchan Choi, Sung-Ho Lee, Yong-Pil Cheon","doi":"10.12717/DR.2021.25.4.257","DOIUrl":"https://doi.org/10.12717/DR.2021.25.4.257","url":null,"abstract":"<p><p>Phthalates and their metabolites are well-known endocrine disrupting chemicals. Di-(2-ethylhexyl) phthalate (DEHP) has been widely used in industry and the exposing possibility to adult is high. In this study, DEHP was treated (133 μg/L and 1,330 μg/L in drinking water) according to the OECD test guideline 443 to mature female mice and their adrenal gland were examined for histological characteristics and steroidogenic gene expression. The wet weight of the adrenal gland was increased in all administrated groups compared to control. The diameter of zona fasciculata (ZF) was increased by DEHP in both outer ZF and inner ZF but there was no difference in morphology of the cells and arrangements into zona between groups. In addition, the arrangement of extracellular matrix was not different between control and DEHP groups. CYP11B1 was mainly localized at ZF and the intensity was not different between groups. DAX1 was localized in zona glomerulosa (ZG) and ZF, and its expression levels were decreased by DEHP administration. Its level was lower in DEHP133 group than DEHP1330 group. On the other hand, CYP17A1 was localized in ZG of DEHP1330 group. These results suggest that chronic low-dose DEHP exposing may modify the microstructure and function of the adrenal cortical cortex.</p>","PeriodicalId":72791,"journal":{"name":"Development & reproduction","volume":"25 4","pages":"257-268"},"PeriodicalIF":0.0,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/01/05/dr-25-4-257.PMC8807134.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39767381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}