Corrigendum: Dysregulation of BMP, Wnt, and insulin signaling in fragile X syndrome.

[This corrects the article DOI: 10.3389/fcell.2022.934662.].

Expression of Insl3 Protein in Adult Danio rerio.

Insulin-like peptide 3 (INSL3) is a biomarker for Leydig cells in the testes of vertebrates, and it is principally involved in spermatogenesis through specific binding with the RXFP2 receptor. This study reports the insl3 gene transcript and the Insl3 prepropeptide expression in both non-reproductive and reproductive tissues of Danio rerio. An immunohistochemistry analysis shows that the hormone is present at a low level in the Leydig cells and germ cells at all stages of Danio rerio testis differentiation. Considering that the insl3 gene is transcribed in Leydig cells, our results highlight an autocrine and paracrine function of this hormone in the Danio rerio testis, adding new information on the Insl3 mode of action in reproduction. We also show that Insl3 and Rxfp2 belonging to Danio rerio and other vertebrate species share most of the amino acid residues involved in the ligand-receptor interaction and activation, suggesting a conserved mechanism of action during vertebrate evolution.

Endocrine Regulation of Aging in the Fruit Fly Drosophila melanogaster.

The past few decades have witnessed increasing research clarifying the role of endocrine signaling in the regulation of aging in both vertebrates and invertebrates. Studies using the model organism fruit fly Drosophila melanogaster have largely advanced our understanding of evolutionarily conserved mechanisms in the endocrinology of aging and anti-aging. Mutations in single genes involved in endocrine signaling modify lifespan, as do alterations of endocrine signaling in a tissue- or cell-specific manner, highlighting a central role of endocrine signaling in coordinating the crosstalk between tissues and cells to determine the pace of aging. Here, we review the current landscape of research in D. melanogaster that offers valuable insights into the endocrine-governed mechanisms which influence lifespan and age-related physiology.

Regulation of forager honey bee appetite independent of the glucose-insulin signaling pathway.

Introduction: To maintain energetic homeostasis the energetic state of the individual needs to communicate with appetite regulatory mechanisms on a regular basis. Although hunger levels indicated by the energetic state and appetite levels, the desire for food intake, tend to be correlated, and on their own are well studied, how the two cross-talk and regulate one another is less known. Insects, in contrast to vertebrates, tend to have trehalose as the primary sugar found in the hemolymph, which could possibly serve as an alternative monitor of the energetic state in comparison to the glucose-insulin signaling pathway, found in vertebrates. Methods: We investigate how manipulating hemolymph sugar levels alter the biogenic amines in the honey bee brain, appetite levels, and insulin like peptide gene expression, across three age classes, to determine how the energetic state of the honey bee might be connected to appetite regulation. Results: We found that only in the forager bees, with a lowering of hemolymph trehalose levels, there was an increase in octopamine and a decrease in tyramine levels in the honey bee brain that corresponded with increased appetite levels, while there was no significant changes in Insulin Like Peptide-1 or 2 gene expression. Discussion: Our findings suggest that hemolymph trehalose levels aid in regulating appetite levels, in forager bees, via octopamine and tyramine, and this regulation appears to be functioning independent of the glucose insulin signaling pathway. Whether this potentially more direct and rapid appetite regulatory pathway can be generalized to other insects, which also undergo energy demanding activities, remains to be investigated.

The diverse influences of relaxin-like peptide family on tumor progression: Potential opportunities and emerging challenges.

Relaxin-like peptide family exhibit differential expression patterns in various types of cancers and play a role in cancer development. This family participates in tumorigenic processes encompassing proliferation, migration, invasion, tumor microenvironment, immune microenvironment, and anti-cancer resistance, ultimately influencing patient prognosis. In this review, we explore the mechanisms underlying the interaction between the RLN-like peptide family and tumors and provide an overview of therapeutic approaches utilizing this interaction.

Pathogenicity of enterotoxigenic Escherichia coli in Caenorhabditis elegans as an alternative model host.

Enterotoxigenic Escherichia coli (ETEC), one of the diarrheagenic E. coli, is the most common cause of diarrhea in developing country and in travelers to those areas. In this study, Caenorhabditis elegans was used as an alternative model host to evaluate ETEC infections. The ETEC strain ETEC1, which was isolated from a patient with diarrhea, possessed enterotoxins STh, LT1, and EAST1 and colonization factors CS2 and CS3. Live ETEC1 shortened the life span and body size of C. elegans in association with increased expression of enterotoxin genes and intestinal colonization. In contrast, heat-killed ETEC1 did not affect the life span of C. elegans. Caenorhabditis elegans infected with ETEC1 showed upregulated expression of genes related to insulin-like peptides and host defense responses. These results suggest that ETEC1 exhibits pathogenicity through intestinal colonization and enterotoxin production in C. elegans. This system is useful as an ETEC infection model.

Monosex Populations of the Giant Freshwater Prawn Macrobrachium rosenbergii-From a Pre-Molecular Start to the Next Generation Era.

Sexual manipulation in the giant freshwater prawn Macrobrachium rosenbergii has proven successful in generating monosex (both all-male and all-female) populations for aquaculture using a crustacean-specific endocrine gland, the androgenic gland (AG), which serves as a key masculinizing factor by producing and secreting an insulin-like AG hormone (IAG). Here, we provide a summary of the advancements from the discovery of the AG and IAG in decapods through to the development of monosex populations in M. rosenbergii. We discuss the broader sexual development pathway, which is highly divergent across decapods, and provide our future perspective on the utility of novel genetic and genomic tools in promoting refined approaches towards monosex biotechnology. Finally, the future potential benefits of deploying monosex prawn populations for environmental management are discussed.

Matrix metalloproteinase 2 degrades collagen I to regulate ovarian development by association with an insulin-like peptide.

The ovary generally undergoes tissue remodeling during larval to pupal transition, which includes membrane degeneration and ovariole growth. At the same time, the hormones produced by insects significantly change during metamorphosis. However, the regulatory mechanism for ovarian development and hormones is not fully understood in insects. Herein, we found that matrix metalloproteinase 2 (MMP2) was highly expressed in the ovarian capsules and ovarioles, and the development was abnormal after knocking out MMP2 in Bombyx mori. The process of abnormal degradation of collagen I due to MMP2 deletion, which resulted in abnormal development of ovarioles and eggs, was analyzed in detail. The proteomics of ovaries in the MMP2-knock out and wild type strains showed a critically significant difference in the expression of a protein, insulin-like peptide (ILP). Additional analysis revealed significant alteration of ILP during ovarian development, and abnormal expression of ILP significantly affected ovarian development in vivo and MMP2 expression in vitro and in vivo. These results showed that MMP2 regulation of ovarian tissue remodeling is closely related to ILP expression. Our study provides new insights into the regulatory mechanism of MMP2 and ovarian development in B. mori.

Chemical synthesis of insulin-like peptide 1 and its potential role in vitellogenesis of the kuruma prawn Marsupenaeus japonicus.

The insulin superfamily comprises a group of peptides with diverse physiological functions and is conserved across the animal kingdom. Insulin-like peptides (ILPs) of crustaceans are classified into four major types: insulin, relaxin, gonadulin, and androgenic gland hormone (AGH)/insulin-like androgenic gland factor (IAG). Of these, the physiological functions of AGH/IAG have been clarified to be the regulation of male sex differentiation, but those of the other types have not been uncovered. In this study, we chemically synthesized Maj-ILP1, an ILP identified in the ovary of the kuruma prawn Marsupenaeus japonicus, using a combination of solid-phase peptide synthesis and regioselective disulfide bond formation reactions. As the circular dichroism spectral pattern of synthetic Maj-ILP1 is typical of other ILPs reported thus far, the synthetic peptide likely possessed the proper conformation. Functional analysis using ex vivo tissue incubation revealed that Maj-ILP1 significantly increased the expression of the yolk protein genes Maj-Vg1 and Maj-Vg2 in the hepatopancreas and Maj-Vg1 in the ovary of adolescent prawns. This is the first report on the synthesis of a crustacean ILP other than IAGs and also shows the positive relationship between the reproductive process and female-dominant ILP.

The circadian and photoperiodic clock of the pea aphid.

The pea aphid, Acyrthosiphon pisum, is a paradigmatic photoperiodic species that exhibits a remarkable annual life cycle, which is tightly coupled to the seasonal changes in day length. During spring and summer, characterised by longer days, aphid populations consist exclusively of viviparous females that reproduce parthenogenetically. When autumn comes and the days shorten, aphids switch their reproductive mode and generate males and oviparous sexual females, which mate and produce cold-resistant eggs that overwinter and survive the unfavourable season. While the photoperiodic responses have been well described, the nature of the timing mechanisms which underlie day length discrimination are still not completely understood. Experiments from the 1960's suggested that aphids rely on an 'hourglass' clock measuring the elapsed time during the dark night by accumulating a biochemical factor, which reaches a critical threshold at a certain night length and triggers the switch in reproduction mode. However, the photoperiodic responses of aphids can also be attributed to a strongly dampened circadian clock. Recent studies have uncovered the molecular components and the location of the circadian clock in the brain of the pea aphid and revealed that it is well connected to the neurohormonal system controlling aphid reproduction. We provide an overview of the putative mechanisms of photoperiodic control in aphids, from the photoreceptors involved in this process to the circadian clock and the neuroendocrine system.

Differential expression of some termite neuropeptides and insulin/IGF-related hormones and their plausible functions in growth, reproduction and caste determination.

Background: Insulin-like growth factor (IGF) and other insulin-like peptides (ilps) are important hormones regulating growth and development in animals. Whereas most animals have a single female and male adult phenotype, in some insect species the same genome may lead to different final forms. Perhaps the best known example is the honeybee where females can either develop into queens or workers. More extreme forms of such polyphenism occur in termites, where queens, kings, workers and soldiers coexist. Both juvenile hormone and insulin-like peptides are known to regulate growth and reproduction as well as polyphenism. In termites the role of juvenile hormone in reproduction and the induction of the soldier caste is well known, but the role of IGF and other ilps in these processes remains largely unknown. Here the various termite ilps are identified and hypotheses regarding their functions suggested. Methods: Genome assemblies and transcriptome short read archives (SRAs) were used to identify insulin-like peptides and neuropeptides in termites and to determine their expression in different species, tissues and castes. Results and Discussion: Termites have seven different ilps, i.e. gonadulin, IGF and an ortholog of Drosophila insulin-like peptide 7 (dilp7), which are commonly present in insects, and four smaller peptides, that have collectively been called short IGF-related peptides (sirps) and individually atirpin, birpin, cirpin and brovirpin. Gonadulin is lost from the higher termites which have however amplified the brovirpin gene, of which they often have two or three paralogs. Based on differential expression of these genes it seems likely that IGF is a growth hormone and atirpin an autocrine tissue factor that is released when a tissue faces metabolic stress. Birpin seems to be responsible for growth and in the absence of juvenile hormone this may lead to reproductive adults or, when juvenile hormone is present, to soldiers. Brovirpin is expressed both by the brain and the ovary and likely stimulates vitellogenesis, while the function of cirpin is less clear.

Insulin-like peptide 5 is associated with insulin resistance in women with polycystic ovary syndrome.

AIMS: Insulin-like peptide 5 (INSL5) plays an important part in metabolic processes in vitro and in vivo. We hypothesized that INSL5 levels are associated with the presence of polycystic ovary syndrome (PCOS) and insulin resistance (IR). METHODS: Circulating INSL5 levels were measured by an enzyme-linked immunosorbent assay in the PCOS group (n = 101) and control (n = 78) groups. The relationship between INSL5 and IR was evaluated by using regression models. RESULTS: The levels of circulating INSL5 were elevated in the individuals with PCOS (P < 0.001) and significantly associated with homeostasis model assessment of insulin resistance (HOMA-IR, r = 0.434, P < 0.001; HOMA-IS, r = 0.432, P < 0.001; QUICKI, r = -0.504, P < 0.001). The subjects in the highest tertile of INSL5 levels were more likely to have PCOS (odds ratio: 12.591, 95 % confidence interval 2.616-60.605) as compared with the lowest tertile after adjustment for potential confounders. Furthermore, the multiple linear regression analyses after adjustment for confounders showed an independent association between INSL5 levels and HOMA-IR (ß = 0.024, P < 0.001). CONCLUSIONS: Circulating INSL5 concentration is linked to PCOS, possibly through increased insulin resistance.

Effects of electroacupuncture at ST36 and BL20 on the diabetic rat testis.

OBJECTIVE: We aimed to evaluate the effects of electroacupuncture (EA) at ST36 and BL20 on the testicular tissues in a rat model of diabetes and to explore the mechanisms of action. METHODS: A total of 34 male Sprague-Dawley rats were allocated to a control group (n = 10), diabetes (D) group (n = 12) or diabetes + acupuncture (DA) group (n = 12). To model diabetes, rats in groups D and DA received an intraperitoneal injection of a single dose of 35 mg/kg streptozotocin (STZ) dissolved in citrate buffer (pH = 4.5; 0.1 M) after 2 weeks of high-fat diet administration. Under xylazine/ketamine anesthesia, stainless steel needles (30 mm × 0.25 mm) were inserted bilaterally at ST36 and BL20. The needles were connected to an EA device via cables, and EA was applied for 30 min (15 Hz frequency and 0.2-1 mA intensity) twice a week for 5 weeks. RESULTS: The effects of EA at ST36 and BL20 on blood glucose levels and body weight, biochemical parameters, histopathological, morphometric and immunohistochemical findings, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) analysis were evaluated. A significant decrease was detected in DA versus D groups in blood glucose levels, basement membrane thickness and apoptotic cell/tubule indices. In addition, there was a significant increase in the Johnsen scores, seminiferous tubule diameters, serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone, proliferation indices, and sex hormone-binding globulin (SHBG) and insulin-like peptide 3 (INSL3) immunoreactivities. CONCLUSION: EA had multiple positive effects on blood glucose homeostasis and testicular structure/function in this rat model of diabetes. EA may be effective at preventing or eliminating histopathological damage in the diabetic testis.

Insulin-like Peptides of the Western Flower Thrips Frankliniella occidentalis and Their Mediation of Immature Development.

Insulin-like peptides (ILPs) mediate various physiological processes in insects. Specifically, ILP expression is required for immature development in different insects. The western flower thrips, Frankliniella occidentalis, is polyphagous, but its occurrence and population density vary among different hosts. This study assesses the developmental variations in the thrips through quantitative analysis of their ILP expressions. Two types of ILPs (Fo-ILP1 and Fo-ILP2) were identified from the genome of F. occidentalis, and both ILPs were predicted to have the characteristics of signal peptides and B-C-A chains linked by cysteines. A phylogenetic analysis indicates that these two ILPs in the thrips are clustered with the ILP1 of Drosophila melanogaster, suggesting their physiological roles in growth. In addition, the two ILP genes were relatively highly expressed at all feeding stages, but their expression was reduced during the nonfeeding prepupal and pupal stages. Furthermore, RNA interference of each ILP expression led to significant developmental retardation. In validating the ILP expression in the thrips' development, five different varieties of host hot peppers were assessed in a choice test, along with the immature development of F. occidentalis. The expression levels of the two ILP genes were highly correlated with variations in the immature developmental rates of different hot pepper varieties. These suggest that Fo-ILP1 and Fo-ILP2 mediate the immature development of F. occidentalis by sensing different nutritional values of hot peppers. This study is the first report on ILPs in thysanopteran insects.

Insulin peptides and their receptors regulate ovarian development and oviposition behavior in Diaphorina citri.

Diaphorina citri is an important vector of Citrus Huanglongbing (HLB) disease. After feeding on young host plant shoots, the population of D. citri can increase significantly. Females also only lay eggs on young shoots. However, there are few studies on the mechanism of this phenomenon. Exogenous nutrient signals can affect the insulin signaling system of D. citri after feeding on young shoots. In this study, the expression of upstream factors DcILP1, DcILP2, and DcIR in the insulin signaling system of D. citri was upregulated after feeding on young shoots. After being silenced by RNA interference technology, the results showed that the number of oviposited eggs of D. citri was significantly decreased and the ovarian development was inhibited with severe vacuolation. In addition, detection using quantitative reverse transcription-polymerase chain reaction showed that the upstream regulatory gene DcRheb of the target of rapamycin (TOR) pathway and the downstream reproduction-related DcVg gene were also significantly downregulated. These results suggest that feeding upon young shoots may upregulate the expression levels of upstream factors DcILP1, DcILP2, and DcIR in the insulin signaling system. The signal will be through upregulating the expression of DcRheb, an upstream gene of the TOR signaling pathway. This in turn influences yolk metabolism, which eventually causes the ovaries of female D. citri to mature and therefore initiate oviposition behavior.

Multiple endocrine factors regulate nutrient mobilization and storage in Aedes aegypti during a gonadotrophic cycle.

Anautogenous mosquitoes must blood feed on a vertebrate host to produce eggs. Each gonadotrophic cycle is subdivided into a sugar-feeding previtellogenic phase that produces primary follicles and a blood meal-activated vitellogenic phase in which large numbers of eggs synchronously mature and are laid. Multiple endocrine factors including juvenile hormone (JH), insulin-like peptides (ILPs), ovary ecdysteroidogenic hormone (OEH), and 20-hydroxyecdysone (20E) coordinate each gonadotrophic cycle. Egg formation also requires nutrients from feeding that are stored in the fat body. Regulation of egg formation is best understood in Aedes aegypti but the role different endocrine factors play in regulating nutrient mobilization and storage remains unclear. In this study, we report that adult female Ae. aegypti maintained triacylglycerol (TAG) stores during the previtellogenic phase of the first gonadotrophic cycle while glycogen stores declined. In contrast, TAG and glycogen stores were rapidly mobilized during the vitellogenic phase and then replenishment. Several genes encoding enzymes with functions in TAG and glycogen metabolism were differentially expressed in the fat body, which suggested regulation was mediated in part at the transcriptional level. Gain of function assays indicated that stored nutrients were primarily mobilized by adipokinetic hormone (AKH) while juvenoids and OEH regulated replenishment. ILP3 further showed evidence of negatively regulating certain lipolytic enzymes. Loss of function assays indicated AKH depends on the AKH receptor (AKHR) for function. Altogether, our results indicate that the opposing activities of different hormones regulate nutrient stores during a gonadotrophic cycle in Ae. aegypti.

The FMRFamide-like peptide FLP-1 modulates larval development by regulating the production and secretion of the insulin-like peptide DAF-28 in Caenorhabditis elegans.

The FMRFamide-like peptides (FLPs) are conserved in both free-living and parasitic nematodes. This molecular genetic study verified the relevance of the flp-1 gene, which is conserved in many nematode species, to the larval development of the free-living soil nematode Caenorhabditis elegans. Using C. elegans as a model, we found that: (1) FLP-1 suppressed larval development, resulting in diapause; (2) the secretion of FLP-1, which is produced in AVK head neurons, was suppressed by the presence of food (Escherichia coli) as an environmental factor to continue larval development; (3) the FLP-1 reduced the production and secretion of DAF-28, which is produced in ASI head neurons and is the predominant insulin-like peptide (INS) present. FLP-1 is conserved in many species of plant-parasitic root-knot nematodes that cause severe damage to crops. Therefore, our findings may provide insight into the development of new nematicides that can disturb their infection and development.

Effects of insulin-like peptide 7 in Bemisia tabaci MED on tomato chlorosis virus transmission.

BACKGROUND: Tomato chlorosis virus (ToCV) is a semi-persistent plant virus that is primarily transmitted by the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae). It causes a serious disease that lowers tomato yield. Insulin-like peptide (ILP), an insulin homolog, regulates trehalose metabolism in a variety of insects. In a previous study, we discovered that trehalose metabolism is required for whiteflies to transmit ToCV effectively. Furthermore, transcriptome sequencing revealed that the BtILP7 gene was highly expressed in B. tabaci infected with ToCV. Therefore, the whitefly ILP7 gene may facilitate the transmission of ToCV and be an attractive target for the control of whiteflies and subsequently ToCV. RESULTS: The ToCV content in B. tabaci MED was found to be correlated with BtILP7 gene expression. Subsequent RNA interference (RNAi) of the BtILP7 gene had a significant impact on B. tabaci MED's trehalose metabolism and reproductive capacity, as well as ability to transmit ToCV. CONCLUSIONS: These results indicate that the BtILP7 gene was closely related to ToCV transmission by regulating trehalose metabolism and reproduction behavior, thus providing a secure and environmentally friendly management strategy for the control of whiteflies and ToCV-caused disease. © 2022 Society of Chemical Industry.

Insulin-like peptide 3 (INSL3) in congenital hypogonadotrophic hypogonadism (CHH) in boys with delayed puberty and adult men.

Background: Delayed puberty in males is almost invariably associated with constitutional delay of growth and puberty (CDGP) or congenital hypogonadotrophic hypogonadism (CHH). Establishing the cause at presentation is challenging, with "red flag" features of CHH commonly overlooked. Thus, several markers have been evaluated in both the basal state or after stimulation e.g. with gonadotrophin releasing hormone agonist (GnRHa).Insulin-like peptide 3 (INSL3) is a constitutive secretory product of Leydig cells and thus a possible candidate marker, but there have been limited data examining its role in distinguishing CDGP from CHH. In this manuscript, we assess INSL3 and inhibin B (INB) in two cohorts: 1. Adolescent boys with delayed puberty due to CDGP or CHH and 2. Adult men, both eugonadal and having CHH. Materials and methods: Retrospective cohort studies of 60 boys with CDGP or CHH, as well as 44 adult men who were either eugonadal or had CHH, in whom INSL3, INB, testosterone and gonadotrophins were measured. Cohort 1: Boys with delayed puberty aged 13-17 years (51 with CDGP and 9 with CHH) who had GnRHa stimulation (subcutaneous triptorelin 100mcg), previously reported with respect to INB. Cohort 2: Adult cohort of 44 men (22 eugonadal men and 22 men with CHH), previously reported with respect to gonadotrophin responses to kisspeptin-54. Results: Median INSL3 was higher in boys with CDGP than CHH (0.35 vs 0.15 ng/ml; p=0.0002). Similarly, in adult men, median INSL3 was higher in eugonadal men than CHH (1.08 vs 0.05 ng/ml; p<0.0001). However, INSL3 more accurately differentiated CHH in adult men than in boys with delayed puberty (auROC with 95% CI in adult men: 100%, 100-100%; boys with delayed puberty: 86.7%, 77.7-95.7%).Median INB was higher in boys with CDGP than CHH (182 vs 59 pg/ml; p<0.0001). Likewise, in adult men, median INB was higher in eugonadal men than CHH (170 vs 36.5 pg/ml; p<0.0001). INB performed better than INSL3 in differentiating CHH in boys with delayed puberty (auROC 98.5%, 95.9-100%), than in adult men (auROC 93.9%, 87.2-100%). Conclusion: INSL3 better identifies CHH in adult men, whereas INB better identifies CHH in boys with delayed puberty.

Insulin-like peptide 3 in domestic animals with normal and abnormal reproductive functions, in comparison to rodents and humans.

Background: Insulin-like peptide 3 (INSL3) is a circulating hormone secreted from only testis and ovaries in mammals. Findings on INSL3 have been gathered from subjects with normal and abnormal reproductive statuses, especially rodents and humans. However, little to no review articles focusing on INSL3 in domestic animals exist. Methods: The author reviewed the past and recent literature regarding the structure, expression, roles of INSL3 in the reproductive organs, and its circulation under normal and aberrant reproductive conditions in domestic animals in comparison with rodents and humans. Main findings: As with humans and rodents, blood INSL3 concentrations rise around puberty in normal male domestic animals and are associated with testicular size. INSL3 levels are acutely upregulated by luteinizing hormone (LH), and the increase is smaller than that of testosterone in male ruminants, whereas the acute regulation of INSL3 by LH does not occur in human men. Dogs with cryptorchidism and bulls with abnormal semen have lowered INSL3 levels. Conclusion: The findings regarding INSL3 secretions in male domestic animals with normal and aberrant reproductive functions illustrate similar or dissimilar points to humans and rodents. Data on blood INSL3 levels in normal and abnormal female domestic species are still limited and require further investigation.