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International Journal of Molecular... Feb 2024Insulin-like growth factors (IGFs) are hormones that primarily stimulate and regulate animal physiological processes. In this study, we cloned and identified the open...
Insulin-like growth factors (IGFs) are hormones that primarily stimulate and regulate animal physiological processes. In this study, we cloned and identified the open reading frame (ORF) cDNA sequences of IGF family genes: the insulin-like growth factor 1 (IGF1), insulin-like growth factor 2 (IGF2), and insulin-like growth factor 3 (IGF3). We found that IGF1, IGF2, and IGF3 have a total length of 558, 648, and 585 base pairs (bp), which encoded a predicted protein with 185, 215, and 194 amino acids (aa), respectively. Multiple sequences and phylogenetic tree analysis showed that the mature golden pompano IGFs had been conserved and showed high similarities with other teleosts. The tissue distribution experiment showed that IGF1 and IGF2 mRNA levels were highly expressed in the liver of female and male fish. In contrast, IGF3 was highly expressed in the gonads and livers of male and female fish, suggesting a high influence on fish reproduction. The effect of fasting showed that IGF1 and mRNA expression had no significant difference in the liver but significantly decreased after long-term (7 days) fasting in the muscles and started to recover after refeeding. IGF2 mRNA expression showed no significant difference in the liver but had a significant difference in muscles for short-term (2 days) and long-term fasting, which started to recover after refeeding, suggesting muscles are more susceptible to both short-term and long-term fasting. In vitro incubation of 17β-estradiol (E) was observed to decrease the IGF1 and IGF3 mRNA expression level in a dose- (0.1, 1, and 10 μM) and time- (3, 6, and 12 h) dependent manner. In addition, E had no effect on IGF2 mRNA expression levels in a time- and dose-dependent manner. The effect of 17α-methyltestosterone (MT) in vitro incubation was observed to significantly increase the IGF3 mRNA expression level in a time- and dose-dependent manner. MT had no effect on IGF2 mRNA but was observed to decrease the IGF1 mRNA expression in the liver. Taken together, these data indicate that E and MT may either increase or decrease IGF expression in fish; this study provides basic knowledge and understanding of the expression and regulation of IGF family genes in relation to the nutritional status, somatic growth, and reproductive endocrinology of golden pompano for aquaculture development.
Topics: Animals; Insulin-Like Peptides; Phylogeny; Amino Acid Sequence; Fishes; RNA, Messenger; Gene Expression; Cloning, Molecular
PubMed: 38473747
DOI: 10.3390/ijms25052499 -
BMC Clinical Pathology 2014Immunoassays are widely used in clinical laboratories for measurement of plasma/serum concentrations of steroid hormones such as cortisol and testosterone. Immunoassays...
BACKGROUND
Immunoassays are widely used in clinical laboratories for measurement of plasma/serum concentrations of steroid hormones such as cortisol and testosterone. Immunoassays can be performed on a variety of standard clinical chemistry analyzers, thus allowing even small clinical laboratories to do analysis on-site. One limitation of steroid hormone immunoassays is interference caused by compounds with structural similarity to the target steroid of the assay. Interfering molecules include structurally related endogenous compounds and their metabolites as well as drugs such as anabolic steroids and synthetic glucocorticoids.
METHODS
Cross-reactivity of a structurally diverse set of compounds were determined for the Roche Diagnostics Elecsys assays for cortisol, dehydroepiandrosterone (DHEA) sulfate, estradiol, progesterone, and testosterone. These data were compared and contrasted to package insert data and published cross-reactivity studies for other marketed steroid hormone immunoassays. Cross-reactivity was computationally predicted using the technique of two-dimensional molecular similarity.
RESULTS
The Roche Elecsys Cortisol and Testosterone II assays showed a wider range of cross-reactivity than the DHEA sulfate, Estradiol II, and Progesterone II assays. 6-Methylprednisolone and prednisolone showed high cross-reactivity for the cortisol assay, with high likelihood of clinically significant effect for patients administered these drugs. In addition, 21-deoxycortisol likely produces clinically relevant cross-reactivity for cortisol in patients with 21-hydroxylase deficiency, while 11-deoxycortisol may produce clinically relevant cross-reactivity in 11β-hydroxylase deficiency or following metyrapone challenge. Several anabolic steroids may produce clinically significant false positives on the testosterone assay, although interpretation is limited by sparse pharmacokinetic data for some of these drugs. Norethindrone therapy may impact immunoassay measurement of testosterone in women. Using two-dimensional similarity calculations, all compounds with high cross-reactivity also showed a high degree of similarity to the target molecule of the immunoassay.
CONCLUSIONS
Compounds producing cross-reactivity in steroid hormone immunoassays generally have a high degree of structural similarity to the target hormone. Clinically significant interactions can occur with structurally similar drugs (e.g., prednisolone and cortisol immunoassays; methyltestosterone and testosterone immunoassays) or with endogenous compounds such as 21-deoxycortisol that can accumulate to very high concentrations in certain disease conditions. Simple similarity calculations can help triage compounds for future testing of assay cross-reactivity.
PubMed: 25071417
DOI: 10.1186/1472-6890-14-33 -
PloS One 2015Androgen administration has been widely used for masculinization in fish. The mechanism of the sex change in sexual fate regulation is not clear. Oral administration or...
Androgen administration has been widely used for masculinization in fish. The mechanism of the sex change in sexual fate regulation is not clear. Oral administration or pellet implantation was applied. We orally applied an aromatase inhibitor (AI, to decrease estrogen levels) and 17α-methyltestosterone (MT, to increase androgen levels) to induce masculinization to clarify the mechanism of the sex change in the protogynous orange-spotted grouper. After 3 mo of AI/MT administration, male characteristics were observed in the female-to-male sex change fish. These male characteristics included increased plasma 11-ketotestosterone (11-KT), decreased estradiol (E2) levels, increased male-related gene (dmrt1, sox9, and cyp11b2) expression, and decreased female-related gene (figla, foxl2, and cyp19a1a) expression. However, the reduced male characteristics and male-to-female sex change occurred after AI/MT-termination in the AI- and MT-induced maleness. Furthermore, the MT-induced oocyte-depleted follicle cells (from MT-implantation) had increased proliferating activity, and the sexual fate in a portion of female gonadal soma cells was altered to male function during the female-to-male sex change. In contrast, the gonadal soma cells were not proliferative during the early process of the male-to-female sex change. Additionally, the male gonadal soma cells did not alter to female function during the male-to-female sex change in the AI/MT-terminated fish. After MT termination in the male-to-female sex-changed fish, the differentiated male germ cells showed increased proliferating activities together with dormancy and did not show characteristics of both sexes in the early germ cells. In conclusion, these findings indicate for the first time in a single species that the mechanism involved in the replacement of soma cells is different between the female-to-male and male-to-female sex change processes in grouper. These results also demonstrate that sexual fate determination (secondary sex determination) is regulated by endogenous sex steroid levels.
Topics: Administration, Oral; Animals; Aromatase Inhibitors; Bass; Cell Proliferation; Female; Gene Expression Regulation; Gonadal Steroid Hormones; Male; Methyltestosterone; Ovum; Sex Determination Processes; Spermatozoa
PubMed: 26714271
DOI: 10.1371/journal.pone.0145438 -
Theriogenology Jan 2018The common snook, Centropomus undecimalis, is an emerging species for intensive fish culture, however, some reproductive aspects of this species, especially the... (Randomized Controlled Trial)
Randomized Controlled Trial
The common snook, Centropomus undecimalis, is an emerging species for intensive fish culture, however, some reproductive aspects of this species, especially the development of the testes and the action of androgen hormones on spermatogenesis have not been studied. The objective of this study was to evaluate the effects of 17α-methyltestosterone (MT) on spermatogenesis and steroidogenesis during the first sexual maturation of the common snook. The fish, which were reproduced in captivity, had a body weight of 305.80 ± 35.60 g and a total length of 34,11 ± 1,08 cm. We used ethylene-vinyl-acetate (EVAc) implants with four concentrations of the hormone MT: T1 (0.3 mg/kg); T2 (3.0 mg/kg); T3 (15.0 mg/kg) and T4 (30.0 mg/kg), and a control group that did not receive the hormone. The gonads increased (P < 0.05) in relation to the concentrations of MT. Histological analysis revealed a progression of spermatogenesis in the MT treatments, especially in T3 and T4. Sperm release was attained in some fish treated with MT. However, there was a partial suppression of the levels of testosterone (T) and 11-ketotestosterone (11-KT) in plasma in the MT treatments, indicating a negative feedback on steroidogenesis. However, this suppression of T and 11-KT in plasma did not prevent an increase in the gonadosomatic index and the progression of gametogenesis. There was also an increase of estradiol (E2) in plasma in the treatments with the highest MT concentrations. The results suggest that the application of EVAc implants with MT at concentrations of 15 and 30 mg/kg stimulates the development and growth of the testes and accelerates spermatogenesis.
Topics: Androgens; Animals; Dose-Response Relationship, Drug; Drug Implants; Estradiol; Fishes; Male; Methyltestosterone; Spermatogenesis; Testis; Testosterone
PubMed: 29059600
DOI: 10.1016/j.theriogenology.2017.10.015 -
Zoological Research Mar 2019Genetically improved farmed tilapia (GIFT) and GIFT-derived strains account for the majority of farmed tilapia worldwide. As male tilapias grow much faster than females,...
Genetically improved farmed tilapia (GIFT) and GIFT-derived strains account for the majority of farmed tilapia worldwide. As male tilapias grow much faster than females, they are often considered more desirable in the aquacultural industry. Sex reversal of females to males using the male sex hormone 17-α-methyltestosterone (MT) is generally used to induce phenotypic males during large-scale production of all male fingerlings. However, the widespread use of large quantities of sex reversal hormone in hatcheries may pose a health risk to workers and ecological threats to surrounding environments. Breeding procedures to produce genetically all-male tilapia with limited or no use of sex hormones are therefore urgently needed. In this study, by applying marker-assisted selection (MAS) for the selection of YY supermales from a GIFT-derived strain, we identified 24 XY pseudofemale and 431 YY supermale tilapias. Further performance evaluation on the progenies of the YY supermales resulted in male rates of 94.1%, 99.5% and 99.6%, respectively, in three populations, and a daily increase in body weight of 1.4 g at 3 months (n=997). Our study established a highly effective MAS procedure in the selection of YY supermales from a GIFT-derived strain. Furthermore, the development of MAS-selected YY supermales will help reduce the utilization of hormones for controlling sex in the tilapia aquaculture.
Topics: Animals; Aquaculture; Male; Selection, Genetic; Sex Determination Processes; Sex Ratio; Tilapia; Y Chromosome
PubMed: 30213922
DOI: 10.24272/j.issn.2095-8137.2018.071 -
PloS One 2020Fish oogenesis is characterised by a massive growth of oocytes each reproductive season. This growth requires the stockpiling of certain molecules, such as ribosomal...
Fish oogenesis is characterised by a massive growth of oocytes each reproductive season. This growth requires the stockpiling of certain molecules, such as ribosomal RNAs to assist the rapid ribosomal assembly and protein synthesis required to allow developmental processes in the newly formed embryo. Massive 5S rRNA expression in oocytes, facilitated by transcription factor 3A (Gtf3a), serves as marker of intersex condition in fish exposed to xenoestrogens. Our present work on Gtf3a gene evolution has been analysed in silico in teleost genomes and functionally in the case of the zebrafish Danio rerio. Synteny-analysis of fish genomes has allowed the identification of two gtf3a paralog genes, probably emerged from the teleost specific genome duplication event. Functional analyses demonstrated that gtf3ab has evolved as a gene specially transcribed in oocytes as observed in Danio rerio, and also in Oreochromis niloticus. Instead, gtf3aa was observed to be ubiquitously expressed. In addition, in zebrafish embryos gtf3aa transcription began with the activation of the zygotic genome (~8 hpf), while gtf3ab transcription began only at the onset of oogenesis. Under exposure to 100 ng/L 17β-estradiol, fully feminised 61 dpf zebrafish showed transcription of ovarian gtf3ab, while masculinised (100 ng/L 17α-methyltestosterone treated) zebrafish only transcribed gtf3aa. Sex related transcription of gtf3ab coincided with that of cyp19a1a being opposite to that of amh and dmrt1. Such sex dimorphic pattern of gtf3ab transcription was not observed earlier in larvae that had not yet shown any signs of gonad formation after 26 days of oestradiol exposure. Thus, gtf3ab transcription is a consequence of oocyte differentiation and not a direct result of estrogen exposure, and could constitute a useful marker of gonad feminisation and intersex condition.
Topics: Animals; Cichlids; Disorders of Sex Development; Evolution, Molecular; Female; Fish Proteins; Gene Duplication; Male; Oogenesis; Ovary; Phylogeny; Sex Characteristics; Sex Differentiation; Synteny; Transcription Factor TFIIIA; Zebrafish; Zebrafish Proteins
PubMed: 31999691
DOI: 10.1371/journal.pone.0227690 -
Journal of the American College of... 2015Current methods for measuring regional body fat are expensive and inconvenient compared to the relative cost-effectiveness and ease of use of a stereovision body imaging...
OBJECTIVE
Current methods for measuring regional body fat are expensive and inconvenient compared to the relative cost-effectiveness and ease of use of a stereovision body imaging (SBI) system. The primary goal of this research is to develop prediction models for android and gynoid fat by body measurements assessed via SBI and dual-energy x-ray absorptiometry (DXA). Subsequently, mathematical equations for prediction of total and regional (trunk, leg) body adiposity were established via parameters measured by SBI and DXA.
METHODS
A total of 121 participants were randomly assigned into primary and cross-validation groups. Body measurements were obtained via traditional anthropometrics, SBI, and DXA. Multiple regression analysis was conducted to develop mathematical equations by demographics and SBI assessed body measurements as independent variables and body adiposity (fat mass and percentage fat) as dependent variables. The validity of the prediction models was evaluated by a split sample method and Bland-Altman analysis.
RESULTS
The R(2) of the prediction equations for fat mass and percentage body fat were 93.2% and 76.4% for android and 91.4% and 66.5% for gynoid, respectively. The limits of agreement for the fat mass and percentage fat were -0.06 ± 0.87 kg and -0.11% ± 1.97% for android and -0.04 ± 1.58 kg and -0.19% ± 4.27% for gynoid. Prediction values for fat mass and percentage fat were 94.6% and 88.9% for total body, 93.9% and 71.0% for trunk, and 92.4% and 64.1% for leg, respectively.
CONCLUSIONS
The three-dimensional (3D) SBI produces reliable parameters that can predict android and gynoid as well as total and regional (trunk, leg) fat mass.
Topics: Absorptiometry, Photon; Adipose Tissue; Adiposity; Adult; Aged; Body Composition; Body Mass Index; Female; Humans; Imaging, Three-Dimensional; Linear Models; Male; Methyltestosterone; Middle Aged; Young Adult
PubMed: 25915106
DOI: 10.1080/07315724.2014.966396 -
PloS One 2016Gastropod mollusks have been proposed as alternative models for male reproductive toxicity testing, due to similarities in their reproductive anatomy compared to...
Gastropod mollusks have been proposed as alternative models for male reproductive toxicity testing, due to similarities in their reproductive anatomy compared to mammals, together with evidence that endocrine disrupting chemicals can cause effects in some mollusks analogous to those seen in mammals. To test this hypothesis, we used the freshwater pulmonate snail, Biomphalaria glabrata, for which various genetic tools and a draft genome have recently become available, to investigate the effects of two steroid androgens on the development of mollusk secondary sexual organs. Here we present the results of exposures to two potent androgens, the vertebrate steroid; 5α-dihydrotestosterone (DHT) and the pharmaceutical anabolic steroid; 17α-methyltestosterone (MT), under continuous flow-through conditions throughout embryonic development and up to sexual maturity. Secondary sexual gland morphology, histopathology and differential gene expression analysis were used to determine whether steroid androgens stimulated or inhibited organ development. No significant differences between tissues from control and exposed snails were identified, suggesting that these androgens elicited no biologically detectable response normally associated with exposure to androgens in vertebrate model systems. Identifying no effect of androgens in this mollusk is significant, not only in the context of the suitability of mollusks as alternative model organisms for testing vertebrate androgen receptor agonists but also, if applicable to other similar mollusks, in terms of the likely impacts of androgens and anti-androgenic pollutants present in the aquatic environment.
Topics: Androgens; Animals; Biomphalaria; Dihydrotestosterone; Dose-Response Relationship, Drug; Environmental Exposure; Gene Expression Profiling; Gene Expression Regulation; Methyltestosterone; Reproduction
PubMed: 27448327
DOI: 10.1371/journal.pone.0159852 -
Genetics and Molecular Research : GMR Apr 2016Androgen plays critical roles in vertebrate reproductive systems via androgen receptors (ARs). In the present study, the full-length spotted scat (Scatophagus argus)...
Androgen plays critical roles in vertebrate reproductive systems via androgen receptors (ARs). In the present study, the full-length spotted scat (Scatophagus argus) androgen receptor (sAR) cDNA sequence was cloned from testis. The sAR cDNA measured 2448 bp in length with an open-reading frame of 2289 bp, encoding 763 amino acids. Amino acid alignment analyses showed that the sARs exhibited highly evolutionary conserved functional domains. Phylogenetically, the sARs clustered within the ARβ common vertebrate group. Real-time polymerase chain reaction (RT-PCR) revealed that sAR expression varied in level and distribution throughout the tissues of both females and males. sAR expression was detected during testicular development by quantitative RT-PCR. The results showed that the highest transcription of sARs was observed in the mid-testicular stage, and remained at a high expression level until the late-testicular stage. In addition, the effects of 17α-methyltestosterone (MT) and estrogen (E2) on the expression of sARs in ovaries were determined using quantitative RT-PCR. sAR expression increased at 12 and 24 h post-MT treatment and decreased with E2 treatment. The present study provides preliminary evidence indicating gonadal plasticity of spotted scat under exogenous steroidal hormone treatments. It also provides a theoretical basis for sex reversal and production of artificial pseudo-males for female monosex breeding.
Topics: Animals; Cloning, Molecular; Estrogens; Female; Fish Proteins; Gonads; Male; Methyltestosterone; Open Reading Frames; Perciformes; Protein Domains; Receptors, Androgen
PubMed: 27173207
DOI: 10.4238/gmr.15027838 -
BMC Genomics Jan 2020Spermatogenesis is an intricate process regulated by a finely organized network. The orange-spotted grouper (Epinephelus coioides) is a protogynous hermaphroditic fish,...
Transcriptome profiling of laser-captured germ cells and functional characterization of zbtb40 during 17alpha-methyltestosterone-induced spermatogenesis in orange-spotted grouper (Epinephelus coioides).
BACKGROUND
Spermatogenesis is an intricate process regulated by a finely organized network. The orange-spotted grouper (Epinephelus coioides) is a protogynous hermaphroditic fish, but the regulatory mechanism of its spermatogenesis is not well-understood. In the present study, transcriptome sequencing of the male germ cells isolated from orange-spotted grouper was performed to explore the molecular mechanism underlying spermatogenesis.
RESULTS
In this study, the orange-spotted grouper was induced to change sex from female to male by 17alpha-methyltestosterone (MT) implantation. During the spermatogenesis, male germ cells (spermatogonia, spermatocytes, spermatids, and spermatozoa) were isolated by laser capture microdissection. Transcriptomic analysis for the isolated cells was performed. A total of 244,984,338 clean reads were generated from four cDNA libraries. Real-time PCR results of 13 genes related to sex differentiation and hormone metabolism indicated that transcriptome data are reliable. RNA-seq data showed that the female-related genes and genes involved in hormone metabolism were highly expressed in spermatogonia and spermatozoa, suggesting that these genes participate in the spermatogenesis. Interestingly, the expression of zbtb family genes showed significantly changes in the RNA-seq data, and their expression patterns were further examined during spermatogenesis. The analysis of cellular localization of Eczbtb40 and the co-localization of Eczbtb40 and Eccyp17a1 in different gonadal stages suggested that Eczbtb40 might interact with Eccyp17a1 during spermatogenesis.
CONCLUSIONS
Our study, for the first time, investigated the transcriptome of the male germ cells from orange-spotted grouper, and identified functional genes, GO terms, and KEGG pathways involved in spermatogenesis. Furthermore, Eczbtb40 was first characterized and its role during spermatogenesis was predicted. These data will contribute to future studies on the molecular mechanism of spermatogenesis in teleosts.
Topics: Animals; Fishes; Gene Expression Profiling; Germ Cells; Methyltestosterone; Spermatogenesis; Transcriptome
PubMed: 31973692
DOI: 10.1186/s12864-020-6477-4