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Fish Follicle-Stimulating Hormone (FSH) ELISA Kit, Species Fish, Sample Type serum, plasma

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[#CSB-E15790Fh] Fish Follicle-Stimulating Hormone (FSH) ELISA Kit, Species Fish, Sample Type serum, plasma

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CSB-E15790Fh | Fish Follicle-Stimulating Hormone (FSH) ELISA Kit, Species Fish, Sample Type serum, plasma, 96T
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(1) Seasonal ovarian development in relation to the gonadotropins, steroids, aromatase and steroidogenic factor 1 (SF-1) in the banded gourami, Trichogaster fasciata.[TOP]

Pubmed ID :30055147
Publication Date : //
The endocrine regulation of gonadal development and annual variation of key sex steroids is the basic knowledge to understand the reproductive cycle of teleost fish. Present study was aimed to investigate the levels of gonadotropins in relation to the follicular development and plasma steroids during the reproductive cycle of female Trichogaster fasciata. Female fish were sampled and ovarian development is described histologically throughout the year in relation to the seasonal variations of gonadosomatic index (GSI); follicle stimulating hormone (FSH) and luteinizing hormone (LH); three key steroids for folliculogenesis and maturation i.e. testosterone (T), 17β-estradiol (E) and 17α20βdihydroxy4pregnen3one (17,20β-P). Relatively higher level of FSH was observed till the ovary reaches in late vitellogenic stage confirms that FSH regulates the early folliculogenesis of the ovary, whereas LH peak was observed in the postvitellogenic stage, which indicates that maturation and ovulation were controlled by LH. Seasonal steroid profiles show that both T and E reach its maximum level prior to the 17,20β-P which attain its peak value in the month of August. Thus, single peak values of LH and 17,20β-P coinciding with GSI peak, clearly indicates that T. fasciata breeds only once in a year. Furthermore, to elucidate the molecular basis of the reproductive cycle, this study analyzes the other key factors of ovarian function such as cyp19a1a gene expression, aromatase activity and SF-1 localization throughout the year. cyp19a1a gene expression and the aromatase activity were highest in vitellogenic stages indicate that relatively higher E production in this stage is regulated by FSH. Immunohistochemical localizations of aromatase and SF-1 in the cellular layer of oocytes demonstrated that aromatase is FSH-dependent and SF-1 could be regulated by both FSH and LH as relatively higher amount of aromatase was localized in the vitellogenic stage oocytes than the postvitellogenic and post germinal vesicle breakdown (post-GVBD) stages; whereas, high amount of SF-1 was observed in vitellogenic, postvitellogenic and post-GVBD stages. These data regarding the reproductive endocrinology of T. fasciata may be useful to understand the interaction between gonadotropins, steroids, aromatase and SF-1 in teleost fishes and may contribute to restoration of the ecologically important fish through artificial reproduction.

Authors : Guchhait Rajkumar, Chatterjee Ankit, Mukherjee Dilip, Pramanick Kousik,



(2) Ctrp9 and adiponectin receptors in Nile tilapia (Oreochromis niloticus): Molecular cloning, tissue distribution and effects on reproductive genes.[TOP]

Pubmed ID :29864417
Publication Date : //
As the close paralog of adiponectin, C1q/TNF-Related Protein 9 (CTRP9) has been reported to be involved in the regulation of glucose and fat metabolism, immunization and endothelial cell functions. However, information regarding the actions of Ctrp9 on reproduction is extremely limited in fish. As a first step, Ctrp9, adiponectin receptor 1 (Adipor1) and Adipor2 were identified from Nile tilapia. The open reading frame (ORF) of ctrp9 was 1020 bp which encoded a 339 amino acids. Moreover, the ORFs of adipor1 and adipor2 were 1131 bp and 1134 bp encoding 376 and 377 amino acids, respectively. Tissue distribution showed that ctrp9 mRNA levels were highest in the kidney in both sexes. And, the expression of adipor1 and adipor2 were widely distributed in all tissues examined, exhibiting high levels in the brain, gonad, gut and stomach. In addition, intraperitoneal (i.p.) injection of gCtrp9 (globular Ctrp9) suppressed the hypothalamic expression of gnrh2 (gonadotropin-releasing hormone 2) and gnrh3, as well as gthα (gonadotropic hormone α), fshβ (follicle-stimulating hormone β), lhβ (luteinizing hormone β), lhr (LH receptor) and fshr (FSH receptor) mRNA levels in the pituitary. The mRNA levels of adipor1, but not adipor2, in the gonads were also inhibited after injection. Moreover, the levels of serum E2 (estrogen) in female and T (testosterone) in male were significantly decreased after injection of gCtrp9. Overall, our data provides novel data indicating, for the first time, a regulatory effect of CTRP9 on teleost reproduction.

Authors : Yang Guokun, Song Qinqin, Sun Caiyun, Qin Jingkai, Jia Jirong, Yuan Xi, Zhang Yazhou, Li Wensheng,



(3) Complex chromosomal aberrations in a fetus originating from oocytes with smooth endoplasmic reticulum (SER) aggregates.[TOP]

Pubmed ID :29718716
Publication Date : //
The presence of smooth endoplasmic reticulum aggregates (SERa) in the ooplasm is considered as the most severe oocyte dysmorphism due to its serious and potentially lethal outcomes in offspring. In the present case report, a couple underwent their first intracytoplasmic sperm injection (ICSI) cycle using a gonadotrophin releasing hormone (GnRH) antagonist protocol, followed by fetal ultrasound scanning and amniocentesis. SERa were observed in all oocytes retrieved. A singleton pregnancy was established. The second trimester fetal ultrasound scan revealed a female fetus with overlapping fingers in both hands, and amniocentesis was performed for the detection of chromosomal abnormalities. Comprehensive genetic analysis with the combined use of array-comparative genomic hybridization (CGH), fluoresence in situ hybridization (FISH) and conventional cytogenetics revealed a complex chromosome rearrangement (CCR) involving three break points on two chromosomes, resulting in a reciprocal translocation with a cryptic 2q31 deletion. A week following amniocentesis, there was rupture of amniotic membranes and a stillborn was delivered. This is the first case in the literature to report a CCR with concomitant 2q31 deletion resulting in a well-defined and clinically recognizable contiguous gene syndrome with an abnormal phenotype in a fetus arising from a cohort of oocytes affected by SERa. It is suggested that fertilization and transfer of oocytes with SERa should be avoided, until further research establishes whether there is a causal relationship between the presence of SERa and chromosomal abnormalities in the resulting fetus.

Authors : Sfontouris Ioannis A, Lainas George T, Lainas Tryfon G, Faros Efthimios, Banti Maria, Kardara Katerina, Anagnostopoulou Katerina, Kontos Harris, Petsas George K, Kolibianakis Efstratios M,



(4) Evolution of gonadotropin signaling on gonad development: insights from gene knockout studies in zebrafish.[TOP]

Pubmed ID :29718109
Publication Date : //
Gonadal development is precisely regulated by the two gonadotropins luteinizing hormone (LH) and follicle stimulating hormone (FSH). Much progress on understanding the functions of LH and FSH signaling on gonad development has been achieved in the past decades, mostly from studies in mammals especially genetic studies in both mouse and human. The functions of both LH and FSH signaling in non-mammalian species are still largely unknown. In recent years, using zebrafish, a teleost phylogenetically distant from mammals, we and others have genetically analyzed the functions of gonadotropins and their receptors through gene knockout studies. In this review, we will summarize the pertinent findings and discuss how the actions of gonadotropin signaling on gonad development have evolved during evolution from fish to mammals.

Authors : Li Jianzhen, Cheng Christopher H K,



(5) Cloning and characterization of a second lamprey pituitary glycoprotein hormone, thyrostimulin (GpA2/GpB5).[TOP]

Pubmed ID :29678725
Publication Date : //
A novel heterodimeric glycoprotein hormone (GpH) comprised of alpha (GpA2) and beta (GpB5) subunits was discovered in 2002 and called thyrostimulin for its ability to activate the TSH receptor in mammals, but its central function in vertebrates has not been firmly established. We report here the cloning and expression of lamprey (l)GpB5, and its ability to heterodimerize with lGpA2 to form a functional l-thyrostimulin. The full-length cDNA of lGpB5 encodes 174 amino acids with ten conserved cysteine residues and one glycosylation site that is conserved with other vertebrate GpB5 sequences. Phylogenetic and synteny analyses support that lGpB5 belongs to the vertebrate GpB5 clade. Heterodimerization of lGpB5 and lGpA2 was shown by nickel pull-down of histidine-tagged recombinant subunits. RNA transcripts of lGpB5 were detected in the pituitary of lampreys during both parasitic and adult life stages. Intraperitoneal injection with lGnRH-III (100 μg/kg) increased pituitary lGpA2, lGpB5, and lGpHβ mRNA expression in sexually mature, adult female lampreys. A recombinant l-thyrostimulin produced by expression of a fusion gene in Pichia pastoris activated lamprey GpH receptors I and II as measured by cAMP enzymeimmunoassay. In contrast to jawed vertebrates that have pituitary LH, FSH, and TSH, our data support that lampreys only have two functional pituitary GpHs, lGpH and l-thyrostimulin, which consist of lGpA2 and unique beta subunits. It is hypothesized that lGpH and l-thyrostimulin differentially regulate reproductive and thyroid activities in some unknown way(s) in lampreys.

Authors : Hausken Krist N, Tizon Belen, Shpilman Michal, Barton Shannon, Decatur Wayne, Plachetzki David, Kavanaugh Scott, Ul-Hasan Sabah, Levavi-Sivan Berta, Sower Stacia A,



(6) Gonadotropin receptors of Labeo rohita: Cloning and characterization of full-length cDNAs and their expression analysis during annual reproductive cycle.[TOP]

Pubmed ID :29660307
Publication Date : //
Follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh), secreted from pituitary, stimulate gonadal function by binding to their cognate receptors FSH receptor (FSHR), and LH/choriogonadotropin receptor (LHCGR). Rohu (Labeo rohita) is a commercially important seasonal breeder freshwater fish species, but till date, the regulation of expression of gonadotropins and their receptors gene during different phases of annual reproductive cycle has not been investigated. We envisaged the critical role of these molecules during seasonal gonadal development in this carp species. We cloned full- length cDNAs of fshra and lhcgrba from rohu testis using RACE (Rapid amplification of cDNA ends) and analyzed their expression along with fsh and lh by quantitative real time PCR (qRT-PCR) assay at various gonadal developmental stages of the annual reproductive cycle. Full-length rohu fshra and lhcgrba cDNA encodes 670 and 716 amino acids respectively, and in adult fish, they were widely expressed in brain, pituitary, gonad, liver, kidney, head kidney, heart, muscle, gill, fin, eye and intestine. In male, both fsh and fshra transcripts showed high level of expression during spermatogenesis, however, in female, expression level was found to be higher in the fully grown oocyte stages. The expression of rohu lh and lhcgrba mRNA increased with increment of gonadosomatic index and showed highest level during spermiation stage in male and fully matured oocyte stage in female. These results together may suggest the involvement of fshra and lhcgrba in regulating function of seasonal gonadal development in rohu.

Authors : Pradhan Avinash, Nayak Madhusmita, Samanta Mrinal, Panda Rudra Prasanna, Rath Suresh Chandra, Giri Shiba Shankar, Saha Ashis,



(7) Effects of recombinant vertebrate ancient long opsin on reproduction in goldfish, Carassius auratus: profiling green-wavelength light.[TOP]

Pubmed ID :29542047
Publication Date : //
This study was conducted to identify the possible effect of recombinant vertebrate ancient long (VAL) opsin as a non-visual "photoreceptor" in the deep brain of goldfish, Carassius auratus. In addition, we investigated the effects of green-wavelength light on the predictable reproductive function of VAL-opsin as a green-sensitive pigment in the deep brain. To determine this, we quantified changes in gonadotropin hormone (GTH) [GTHα, follicle stimulating hormone (FSH) and luteinizing hormone (LH)] and estrogen receptor (ER; ERα and ERβ) mRNA expression levels associated with goldfish reproduction as well as changes in plasma FSH, LH, and 17β-estradiol (E) activities after injection of recombinant VAL-opsin protein in two concentrations (0.1 or 0.5 μg/g body mass) for 4 weeks (injection once weekly) and examined the possible impact of green-wavelength light (500, 520, and 540 nm) on the function of VAL-opsin. As a result, all parameters associated with reproduction significantly increased with time and light-emitting diode (LED) exposure. Based on these results, we suggested that VAL-opsin in the deep brain is involved in goldfish maturation, and it is possible that green-wavelength light improves the ability of VAL-opsin to promote maturation by increasing VAL-opsin expression.

Authors : Choi Ji Yong, Choi Cheol Young,



(8) Characterization of carp gonadotropins: Structure, annual profile, and carp and zebrafish pituitary topographic organization.[TOP]

Pubmed ID :29183794
Publication Date : //
Two gonadotropins, follicle stimulating hormone (FSH) and luteinizing hormone (LH), are important players in the hypothalamic-pituitary-gonadal axis of vertebrates. In the present work, we describe the construction of recombinant (r) common carp (Cyprinus carpio; c) FSH (rcFSH) and LH (rcLH) using the Pichia pastoris system, the generation of specific antibodies against their respective β subunits, and their use in the development and validation of specific ELISAs. We produced carp rLH and rFSH as single-chain polypeptides, wherein the GTH subunit α was joined with either cLHβ or cFSHβ mature protein-coding sequences to form a fusion gene that encodes a yoked polypeptide, in which the GTH β-subunit forms the N-terminal part and the α-subunit forms the C-terminal part. Competitive ELISAs were developed, using primary antibodies against rcLHβ or rcFSHβ, respectively, and rcLHβα or rcFSHβα for the standard curves. The standard curves for cLH paralleled those of pituitary extracts of the homologous fish and also those of other cyprinids species like the black carp (Mylopharyngodon piceus), goldfish (Carassius auratus), silver carp (Hypophthalmichthys molitrix), and grass carp (Ctenopharyngodon idella). We used the specific antibodies raised against cFSH and cLH to study the specific localization of the different GTH cells in the pituitary of carp and its taxonomic relative species - the zebrafish. Both FSH and LH cells are localized in the center of the proximal pars distalis enveloping both sides of the neurohypophysis. LH cells form a continuous population throughout the PPD, while FSH cells are more loosely distributed throughout the same area and form small aggregations. Marked annual changes were encountered in gonadosomatic index (GSI), follicle diameter, mRNA levels and protein levels of FSH and LH. From September to November, all fish had low GSI, and the ovary contained previtellogenic follicles. From December, the GSI level increased and remained high until March, the follicular diameter reached its maximum in January, where the ovary contained large fully grown follicles. Thereafter, spawning occurred through March and April and ended in May, and GSI level and follicle diameter increased again; and the ovary contained mid-vitellogenic follicles. LH pituitary content and mRNA levels were low at pre- and early vitellogenesis, increasing gradually during this process to reach a peak of LH mRNA levels in mid vitellogenic ovary and a peak of LH content in fully grown ovarian follicles. However, no significant change occurred in FSH pituitary content and mRNA levels in vitellogenic fish and in fish during final maturation stages. A dramatic difference was found in the total content of each gonadotropin in the pituitary, with higher LH than FSH. Moreover, follicle diameter was positively and significantly correlated with LH pituitary content and its transcript levels - but not with the pituitary content or mRNA levels of FSH. Taken together, these results indicate that in carp, LH alone is sufficient to regulate both vitellogenesis and final oocyte maturation while FSH may have another, yet undefined role.

Authors : Hollander-Cohen Lian, Golan Matan, Aizen Joseph, Shpilman Michal, Levavi-Sivan Berta,



(9) Expression of two glycoprotein hormone receptors in larval, parasitic phase, and adult sea lampreys.[TOP]

Pubmed ID :29157942
Publication Date : //
All jawed vertebrates have three canonical glycoprotein hormones (GpHs: luteinizing hormone, LH; follicle stimulating hormone, FSH; and thyroid stimulating hormone, TSH) with three corresponding GpH receptors (GpH-Rs: LH-R, FSH-R, and TSH-R). In contrast, we propose that the jawless vertebrate, the sea lamprey (Petromyzon marinus), only has two pituitary glycoprotein hormones, lamprey (l)GpH and l-thyrostimulin, and two functional glycoprotein receptors, lGpH-R I and II. It is not known at this time whether there is a specific receptor for lGpH and l-thyrostimulin, or if both GpHs can differentially activate the lGpH-Rs. In this report, we determined the RNA expression of lGpH-R I and II in the gonads and thyroids of larval, parasitic phase, and adult lampreys. A highly sensitive dual-label fluorescent in situ hybridization technique (RNAScope™) showed lGpH-R I expression in the ovaries of larval lamprey, and co-localization and co-expression of lGpH-R I and II in the ovaries of parasitic phase and adult lampreys. Both receptors were also highly co-localized and co-expressed in the endostyle of larval lamprey and thyroid follicles of parasitic and adult lampreys. In addition, we performed in vivo studies to determine the actions of lamprey gonadotropin releasing hormones (lGnRHs) on lGpH-R I and II expression by real time PCR, and determined plasma concentrations of estradiol and thyroxine. Administration of lGnRH-III significantly (p ≤ 0.01) increased lGpHR II expression in the thyroid follicles of adult female lampreys but did not cause a significant increase in RNA expression of lGpH-R I and II in ovaries. Concomitantly, there was a significant increase (p ≤ 0.01) of plasma estradiol without any significant changes of plasma thyroxine concentrations in response to treatment to lGnRH-I, -II, or -III. In summary, our results provide supporting evidence that the lamprey pituitary glycoprotein hormones may differentially activate the lamprey GpH-Rs in regulating both thyroid and gonadal activities during each of the three life stages of the sea lamprey.

Authors : Hausken Krist N, Marquis Timothy J, Sower Stacia A,



(10) Landmark discoveries in elucidating the origins of the hypothalamic-pituitary system from the perspective of a basal vertebrate, sea lamprey.[TOP]

Pubmed ID :29111305
Publication Date : //
The hypothalamic-pituitary (HP) system, which is specific to vertebrates, is considered to be an evolutionary innovation that emerged prior to or during the differentiation of the ancestral jawless vertebrates (agnathans) leading to the neuroendocrine control of many complex functions. Along with hagfish, lampreys represent the oldest lineage of vertebrates, agnathans (jawless fish). This review will highlight our discoveries of the major components of the lamprey HP axis. Generally, gnathostomes (jawed vertebrates) have one or two hypothalamic gonadotropin-releasing hormones (GnRH) while lampreys have three hypothalamic GnRHs. GnRH(s) regulate reproduction in all vertebrates via the pituitary. In gnathostomes, there are three classical pituitary glycoprotein hormones (luteinizing hormone, LH; follicle stimulating hormone, FSH; and thyrotropin, TSH) interacting specifically with three receptors, LH-R, FSH-R, and TSH-R, respectively. In general, FSH and LH regulate gonadal activity and TSH regulates thyroidal activity. In contrast to gnathostomes, we propose that lampreys only have two heterodimeric pituitary glycoprotein hormones, lamprey glycoprotein hormone (lGpH) and thyrostimulin, and two lamprey glycoprotein hormone receptors (lGpH-R I and -R II). Our existing data also suggest the existence of a primitive, overlapping yet functional hypothalamic-pituitary-gonadal (HPG) and HP-thyroidal (HPT) endocrine systems in lampreys. The study of basal vertebrates provides promising models for understanding the evolution of the hypothalamic-pituitary-thyroidal and gonadal axes in vertebrates. We hypothesize that the glycoprotein hormone/glycoprotein hormone receptor systems emerged as a link between the neuroendocrine and peripheral control levels during the early stages of gnathostome divergence. Our discovery of a functional HPG axis in lamprey has provided important clues for understanding the forces that ensured a common organization of the hypothalamus and pituitary as essential regulatory systems in all vertebrates. This paper will provide a brief snapshot of my discoveries, collaborations and latest findings including phylogenomic analyses on the origins, co-evolution and divergence of ligand and receptor protein families from the perspective of the lamprey hypothalamic-pituitary system.

Authors : Sower Stacia A,