Imprinting Mechanism Research Articles

Функциональная роль гипоталамо- гипофизарной нейросекреторной системы в миграциях и нересте рыб

The results of the ecological-histophysiological study of the hypothalamo-hypophysial neurosecretory system (HHNS) by use of light- and electron-microscopical morphometric methods and comparative analysis of the results are presented. At the beginning of migrations a state of “HHNS mobilization” has been established. It is expressed in the form of the strong synthesis activation of neurosecretary products in neurosecretory cells (NSC) and their transport in dendrites and axons to neurohypophysis (NH), where, their mass accumulation, however, occurs. This completion of moderate “normal” level of excretion of nonapeptide neurohormones (NP-NH) into the bloodstream disrupts the body’s water-salt homeostasis. The synchronized transventricular direction of the NP-NH excretion from dendrites and axons into the brain liquor of III ventricle provides their neurotropic effect in the behavioral centers of Central Nervous System (CNS). As a result, HHNS has a decisive double synchronous effect, which disrupts the long-adapted “pasture” type of osmoregulation (hyper- or hypoosmotic), which is probably the main physiological stimulus of habitat change and, at the same time, it includes the behavioral centers of the CNS, which causes a dominant state, originally defined as a “migration impulse.”The interactions of nonapeptide- and luliberinergic centers of the hypothalamus in the navigational mechanisms of homing and imprinting are discussed. At the beginning of spawning, regardless of its season a strong activation of HHNS, followed by a decrease in its functional activity, was discovered in fish of continuous spawning. The detected two-phase reaction of the HHNS seems to be a reflection of its participation in the body’s protective and adaptive responses to physiological stress. The key role of the HHNS in integrating migration and spawning processes is discussed.

Late manifested sequelae of medications in the critical periods of development. The widening of the faulty hormonal imprinting conception

Hormonal imprinting is a physiological process, which is a part of the receptor-hormone complex development. It determines the binding capacity of the receptors across the lifespan. It takes place perinatally in the critical period of hormone receptor development, when the developmental window for imprinting is open and permits the binding of hormone-like molecules (related or synthetic hormones, endocrine disruptors etc.) causing disturbances of the endocrine system, and the systems- influenced organs by it, for life. This is the faulty hormonal imprinting. However, studying the medical database, PubMed, a lot of data can be found on the harmful late (adult age) effects of medication in the critical period of development with non-hormonal molecules, which are manifested later in functional alterations or diseases. This could mean that in the process of faulty imprinting, the openness of the developmental window could be more important than the structural similarity of a molecule to hormones. As developmentally critical period for faulty imprinting by hormone-like molecules is not exclusively the perinatal one (this is justified in the case of faulty hormonal imprinting), the pubertal period was also studied from this aspect and similarities to the impact of perinatal use have been found (this could be called "Pubertal Origin of Health and Disease = POHaD). While in the case of hormonal faulty imprinting, the mechanism seems to be clear (considering the role of receptors), the mechanism of drug-provoked imprinting is presently uncleared (considering the variety of medications which cause late-manifested alterations). The medicaments-caused faulty imprinting conception calls attention to the dangers of medication in the perinatal as well as the pubertal periods. Orv Hetil. 2020; 161(2): 43-49.

КОМПЕТЕНТНОСТЬ К ИНДУЦИРОВАННОМУ ПАРТЕНОГЕНЕЗУ ООЦИТОВ BOS TAURUS В ЗАВИСИМОСТИ ОТ ФУНКЦИОНАЛЬНОГО СТАТУСА ГАМЕТЫ И ЕЕ ПРОИСХОЖДЕНИЯ (ДИАМЕТРА ФОЛЛИКУЛА)

The development of effective methods for obtaining parthenogenetic embryos is currently being considered in a new perspective as a promising technology for creating homozygous lines of embryonic stem cells, genetically identical animals. Parthenogenetic embryos of animals of different species are used as a model for studying imprinting mechanisms. In the present study, Bos Taurus parthenogenetic embryos were obtained by cold shock induction (exposure for 20 minutes at a temperature of -0 - (-4 o ) C) of after oocytes culture in TC-119 medium with 10% fetal bovine serum, 50 ng / ml prolactin, 50 μg / ml of gentamicin together with 1 × 10 6 granulosa cells per ml of medium for 24 hours. Previously, the initial population of donor oocytes from the ovaries was evaluated with BCB test to assess functional status. For this, the oocyte-cumulus complexes were placed in a solution of 13 Mm brilliant crystalline blue - BCB (B-5388, Sigma) for 60 minutes at a temperature of 38.5 0 C in an atmosphere of 90% humidity. After the expiration of the BCB exposure time, the oocytes were ranked as follows: oocytes with a blue cytoplasm - BCB + oocytes (oocytes that completed their growth phase) and oocytes with unpainted ooplasm - BCB- oocytes (growing oocytes). It was found that the functional status (growing oocytes or oocytes that have finished growth phase) of oocytes and their origin (the diameter of the follicles from which the gametes were isolated) determine their competence for induced parthenogenesis. Oocytes that completed the growth phase have high potentials to cold shock-induced parthenogenesis, which is expressed in a significant increase in the proportion of embryos developed from them at all stages of preimplantation development (percentage of cleavage: 87% vs 11%, P 5 mm-8 mm) revealed low competence to cold shock-induced parthenogenesis of oocytes isolated from follicles less than 3 mm in diameter ( the percentage of cleavage: 20% vs 67% and 87%, P <0.001, respectively; level of late morula, blastocysts: 9% vs 22% and 37%, P <0.001, respectively).

Effects of Nutrient Intake during Pregnancy and Lactation on the Endocrine Pancreas of the Offspring

The pancreas has an essential role in the regulation of glucose homeostasis by secreting insulin, the only hormone with a blood glucose lowering effect in mammals. Several circulating molecules are able to positively or negatively influence insulin secretion. Among them, nutrients such as fatty acids or amino acids can directly act on specific receptors present on pancreatic beta cells. Dietary intake, especially excessive nutrient intake, is known to modify energy balance in adults, resulting in pancreatic dysfunction. However, gestation and lactation are critical periods for fetal development and pup growth and specific dietary nutrients are required for optimal growth. Feeding alterations during these periods will impact offspring development and increase the risk of developing metabolic disorders in adulthood, leading to metabolic programming. This review will focus on the influence of nutrient intake during gestation and lactation periods on pancreas development and function in offspring, highlighting the molecular mechanism of imprinting on this organ.

From Mother or Father: Uniparental Embryos Uncover Parent-of-Origin Effects in Humans.

In mammals, both parents make unique contributions to the offspring and maternal and paternal genomes are required for development. Two recent papers in Cell Stem Cell (Leng etal., 2019; Sagi etal., 2019) study uniparental embryos and uniparental embryonic stem cells to interrogate parent-of-origin effects in human embryogenesis.

Endogenous retroviral insertions drive non-canonical imprinting in extra-embryonic tissues

BackgroundGenomic imprinting is an epigenetic phenomenon that allows a subset of genes to be expressed mono-allelically based on the parent of origin and is typically regulated by differential DNA methylation inherited from gametes. Imprinting is pervasive in murine extra-embryonic lineages, and uniquely, the imprinting of several genes has been found to be conferred non-canonically through maternally inherited repressive histone modification H3K27me3. However, the underlying regulatory mechanisms of non-canonical imprinting in post-implantation development remain unexplored.ResultsWe identify imprinted regions in post-implantation epiblast and extra-embryonic ectoderm (ExE) by assaying allelic histone modifications (H3K4me3, H3K36me3, H3K27me3), gene expression, and DNA methylation in reciprocal C57BL/6 and CAST hybrid embryos. We distinguish loci with DNA methylation-dependent (canonical) and independent (non-canonical) imprinting by assaying hybrid embryos with ablated maternally inherited DNA methylation. We find that non-canonical imprints are localized to endogenous retrovirus-K (ERVK) long terminal repeats (LTRs), which act as imprinted promoters specifically in extra-embryonic lineages. Transcribed ERVK LTRs are CpG-rich and located in close proximity to gene promoters, and imprinting status is determined by their epigenetic patterning in the oocyte. Finally, we show that oocyte-derived H3K27me3 associated with non-canonical imprints is not maintained beyond pre-implantation development at these elements and is replaced by secondary imprinted DNA methylation on the maternal allele in post-implantation ExE, while being completely silenced by bi-allelic DNA methylation in the epiblast.ConclusionsThis study reveals distinct epigenetic mechanisms regulating non-canonical imprinted gene expression between embryonic and extra-embryonic development and identifies an integral role for ERVK LTR repetitive elements.

A method for identifying allele-specific hydroxymethylation

ABSTRACTWe previously identified sequence-dependent allele-specific methylation (sd-ASM) in adult human peripheral blood leukocytes, in which ASM occurs in cis depending on adjacent polymorphic sequences. A number of groups have identified sd-ASM sites in the human and mouse genomes, illustrating the prevalence of sd-ASM in mammalian genomes. In addition, sd-ASM can lead to sequence-dependent allele-specific expression of neighbouring genes. Imprinted genes also often exhibit parent-of-origin–dependent allele-specific methylation (pd-ASM), which causes parent-of-origin–dependent allele-specific expression. However, whether most of the already known sd-ASM and pd-ASM sites are methylated or hydroxymethylated remains unclear due to technical restrictions. Accordingly, a novel method that enables examination of allelic methylation and hydroxymethylation status and also overcomes the drawbacks of conventional methods is needed. Such a method could also be used to elucidate the mechanisms underlying polymorphism-associated inter-individual differences in disease susceptibility and the mechanism of genomic imprinting. Here, we developed a simple method to determine allelic hydroxymethylation status and identified novel sequence- and parent-of-origin–dependent allele-specific hydroxymethylation sites. Correlation analyses of TF binding sequences and methylation or hydroxymethylation between three mouse strains revealed the involvement of Pax5 in strain-specific methylation and hydroxymethylation in exon 7 of Pdgfrb.

Ducklings imprint on chromatic heterogeneity

Avian filial imprinting is a rapid form of learning occurring just after hatching in precocial bird species. The acquired imprint on either or both parents goes on to affect the young bird’s survival and social behaviour later in life (Bateson in Biol Rev 41:177–217, 1966). The imprinting mechanism is specialized but flexible, and causes the hatchling to develop high-fidelity recognition and attraction to any moving stimulus of suitable size seen during a predefined sensitive period. It has been observed (Martinho and Kacelnik in Science 353:286–288, 2016; Versace et al. in Anim Cogn 20:521–529, 2017) that in addition to visual and acoustic sensory inputs, imprinting may incorporate informational rules or abstract concepts. Here we report a study of mallard ducklings (Anas platyrhynchos domesticus) undergoing imprinting on the chromatic heterogeneity of stimuli, with a focus on how this may be transferred to novel objects. Ducklings were exposed to a series of chromatically heterogeneous or homogeneous stimuli and tested for preference between two novel stimuli, one heterogeneous and the other homogeneous. Exposure to heterogeneity significantly enhanced preference for novel heterogeneous stimuli, relative to ducklings exposed to homogeneous stimuli or unexposed controls. These findings support the view that imprinting does not rely solely on exemplars, or snapshot-like representations of visual input, but that instead young precocial animals form complex multidimensional representations of the target object, involving abstract properties, either at the time of learning, or later, through generalization from the learnt exemplars.

RNA insertion in DNA as the imprint moiety: the fission yeast paradigm.

This review elaborates on the findings of a new report which possibly resolves the biochemical nature of a novel type of DNA imprint as ribonucleotide and the mechanism of its formation during cell differentiation in fission yeast. The process of mating-type switching in fission yeast, Schizosaccharomyces pombe, displays characteristics of a typical mammalian stem cell lineage, wherein a cell divides to produce an identical cell and a differentiated cell after every two cell divisions. This developmental asymmetry has been ascribed to play a role in generation of a DNA strand-specific imprint at the mat1 locus during lagging strand synthesis and its segregation to one of the two daughter cells by the process of asymmetric, semi-conservative DNA replication. The nature of this imprint and mechanisms of its generation have been a subject of research and debate. A recent report by Singh et al. (Nucleic Acids Res 47:3422-3433. https://doi.org/10.1093/nar/gkz092 , 2019) provides compelling evidence in support of a ribonucleotide as the imprint moiety within the mat1 DNA and demonstrates the role of Mcm10/Cdc23, an important, evolutionarily conserved component of DNA replication machinery in eukaryotes, in installing the imprint through a non-canonical primase activity and interaction with DNA Polα and Swi1. The high degree of conservation of DNA replication machinery, especially the presence of the T7 gene 4 helicase/primase domain in the mammalian orthologs of Mcm10 suggests that similar mechanisms of DNA imprinting may play a role during cell differentiation in metazoans.

The mouse Necdin gene is expressed from the paternal allele only and lies in the 7C region of the mouse chromosome 7, a region of conserved synteny to the human Prader-Willi syndrome region.

Prader-Willi syndrome (PWS) is a neurogenetic disorder resulting from the loss of paternal expression of gene(s) localized in the 15q11-q12 region. A new human gene encoding a putative protein with high homology to the mouse NECDIN protein has recently been characterized and mapped to chromosome 15q11-q12. It is expressed from the paternal allele only, suggesting its potential involvement in PWS. We now report the localization of the mouse Necdin gene in a region of conserved synteny to the human PWS region. We demonstrate the paternal specific expression of Necdin in the mouse central nervous system, and show that parental alleles display a differential methylation profile in the coding region. Finally, fluorescence in situ hybridization analysis reveals an asynchronous pattern of replication at the Necdin locus. These results clearly demonstrate imprinting of the mouse Necdin gene. Mouse models will be powerful tools in the study of human PWS phenotype and imprinting mechanisms.

Evolution of viviparity in mammals: what genomic imprinting tells us about mammalian placental evolution.

Genomic imprinting is an epigenetic mechanism of regulating parent-of-origin-specific monoallelic expression of imprinted genes in viviparous therian mammals such as eutherians and marsupials. In this review we discuss several issues concerning the relationship between mammalian viviparity and genomic imprinting, as well as the domestication of essential placental genes: why has the genomic imprinting mechanism been so widely conserved despite the evident developmental disadvantages originating from monoallelic expression? How have genomic imprinted regions been established in the course of mammalian evolution? What drove the evolution of mammalian viviparity and how have genomic imprinting and domesticated genes contributed to this process? In considering the regulatory mechanism of imprinted genes, reciprocal expression of paternally and maternally expressed genes (PEGs and MEGs respectively) and the presence of several essential imprinted genes for placental formation and maintenance, it is likely that complementary, thereby monoallelic, expression of PEGs and MEGs is an evolutionary trade-off for survival. The innovation in novel imprinted regions was associated with the emergence of imprinting control regions, suggesting that genomic imprinting arose as a genome defence mechanism against the insertion of exogenous DNA. Mammalian viviparity emerged in the period when the atmospheric oxygen concentration was the lowest (~12%) during the last 550 million years (the Phanerozoic eon), implying this low oxygen concentration was a key factor in promoting mammalian viviparity as a response to a major evolutionary pressure. Because genomic imprinting and gene domestication from retrotransposons or retroviruses are effective measures of changing genomic function in therian mammals, they are likely to play critical roles in the emergence of viviparity for longer gestation periods.

M28 - MATERNAL AGE AT BIRTH IS ASSOCIATED WITH OFFSPRING LEUKOCYTE TELOMERE LENGTH IN FAMILIES WITH BIPOLAR DISORDER

Background There is already a number of reports correlating the effect of parents’ ages with the risk of schizophrenia, but studies looking for an association between parental age and Bipolar Disorder (BD) are scarce and with inconsistent findings. BD has recently been related to a process of accelerated aging, with studies showing association with Leukocyte Telomere Length (LTL) in this population. However, little is known regarding the inheritance of telomere length, with some studies estimating the heritability between 36–86%. About the effect of a parent' age determining LTL, just a few studies have noted longer telomere lengths among offspring of older mothers, and none have shown an interaction or association with a psychiatric disorder. The present investigation assessed the impact of maternal and paternal age at birth as a determinant for the offspring's LTL with and without BD diagnosis within families with several members affected by this disease. Methods Telomere length (T) was estimated in a sample of 144 individuals, including 60 BD patients from 18 Brazilian families, which was measured in relation to the single copy gene (S) – β-globin gene (HBB) – using a singleplex real time PCR, providing a ratio of number of copies of T by S (T/S). The analysis were obtained by a linear mixed model (LMM). Results We found a positive association between maternal age at childbirth and offspring early life telomere length (β=0.012; p=0.015), whereas there was no association of paternal age on offspring telomere length (β=0.008; p=0.232). The LMM analysis also showed a significant positive relationship on interaction between bipolarity and maternal age demonstrates a strong influence on telomere length (β=0.022; p=0.024). The same analysis showed no association on interaction between bipolarity and paternal age (β=0.009; p=0.223). Discussion An association between maternal age and offspring LTL and the interaction with BD was observed. It could be speculated that an influence of oxidative stress during first pregnancy, changes in mitochondrial DNA or even other parent-specific imprinting mechanism, such as DNA methylation, relevant to the neurodevelopment of the brain. To our knowledge, this is the first study evaluating association of maternal age and telomere length in the offspring of families with several members affected by BD. Additional studies are needed to confirm these preliminary findings.

Enhancer-driven alternative promoters of imprinted genes.

In the current study, we characterized the expression and histone modification profiles of the alternative promoters found within imprinted Igf2r, Mest, Zac1, Peg3, Snrpn and non-imprinted Myc loci. In terms of expression pattern, the alternative promoters are highly tissue-specific, which is in a stark contrast to the ubiquitous expression of the corresponding main promoters. The alternative promoters are associated with the histone modification mark H3K4me1, but not with H3K4me3, which is frequently associated with the main promoters. Phylogenetic analyses also indicated that the majority of the alternative promoters are unique to the mammalian lineage, further suggesting the recent formation of these promoters during mammalian evolution. Overall, this study suggests that the alternative promoters of imprinted loci may have been derived from enhancers in recent evolutionary times and co-evolved with the genomic imprinting mechanism.

Sensory mechanisms of natal stream imprinting and homing in Oncorhynchus spp.

Juvenile Oncorhynchus spp. can memorise their natal stream during downstream migration; juveniles migrate to feed during their growth phase and then they migrate long distances from their feeding habitat to their natal stream to reproduce as adults. Two different sensory mechanisms, olfaction and navigation, are involved in the imprinting and homing processes during short-distance migration within the natal stream and long-distance migration in open water, respectively. Here, olfactory functions are reviewed from both neurophysiological studies on the olfactory discrimination ability of natal stream odours and neuroendocrinological studies on the hormonal controlling mechanisms of olfactory memory formation and retrieval in the brain. These studies revealed that the long-term stability of dissolved free amino-acid composition in the natal stream is crucial for olfactory imprinting and homing. Additionally, the brain-pituitary-thyroid and brain-pituitary-gonadal hormones play important roles in olfactory memory formation and retrieval, respectively. Navigation functions were reviewed from physiological biotelemetry techniques with sensory interference experiments during the homing migration of anadromous and lacustrine Oncorhynchus spp. The experiments demonstrated that Oncorhynchus spp. used compass navigation mechanisms in the open water. These findings are discussed in relation to the sensory mechanisms involved in natal stream imprinting and homing in Oncorhynchus spp.

Maternal Eed knockout causes loss of H3K27me3 imprinting and random X inactivation in the extraembryonic cells.

Genomic imprinting is essential for mammalian development. Recent studies have revealed that maternal histone H3 Lys27 trimethylation (H3K27me3) can mediate DNA methylation-independent genomic imprinting. However, the regulatory mechanisms and functions of this new imprinting mechanism are largely unknown. Here we demonstrate that maternal Eed, an essential component of the Polycomb group complex 2 (PRC2), is required for establishing H3K27me3 imprinting. We found that all H3K27me3-imprinted genes, including Xist, lose their imprinted expression in Eed maternal knockout (matKO) embryos, resulting in male-biased lethality. Surprisingly, although maternal X-chromosome inactivation (XmCI) occurs in Eed matKO embryos at preimplantation due to loss of Xist imprinting, it is resolved at peri-implantation. Ultimately, both X chromosomes are reactivated in the embryonic cell lineage prior to random XCI, and only a single X chromosome undergoes random XCI in the extraembryonic cell lineage. Thus, our study not only demonstrates an essential role of Eed in H3K27me3 imprinting establishment but also reveals a unique XCI dynamic in the absence of Xist imprinting.

Maternal Protein Restriction Induces Alterations in Hepatic Unfolded Protein Response-Related Molecules in Adult Rat Offspring.

Intrauterine growth restriction (IUGR) leads to the development of metabolic syndrome in adulthood. To explore the potential mechanisms of metabolic imprinting, we investigated the effect of malnutrition in utero on hepatic unfolded protein response (UPR)-related genes in IUGR offspring. An IUGR rat model was developed by feeding a low-protein diet to pregnant rats. The expression levels and activity of hepatic UPR genes were analysed by quantitative PCR (qPCR) arrays and western blotting. The hepatic UPR molecules heat-shock 70-kDa protein 4l (Hspa4l), mitogen-activated protein kinase 10 (Mapk10), and endoplasmic reticulum to nucleus signalling 2 (Ern2) were markedly downregulated in IUGR foetuses, but the expression of Mapk10 and Ern2 returned to normal levels at 3 weeks postnatal. In contrast, cAMP responsive element binding protein 3-like 3 (Creb3l3) was upregulated in hepatic tissues at embryo 20(E20), then restored to normal in adulthood (12 weeks). The protein levels of activating transcription factor 2 (Atf2) and Atf6, two key factors of the UPR pathway, were upregulated in the livers of IUGR foetuses, and the latter remained upregulated until 12 weeks. Combined with our previous findings showing an increase in hepatic gluconeogenesis enzymes in IUGR offspring, we speculated that aberrant intrauterine milieu impaired UPR signalling in hepatic tissues; these alterations early in life might contribute to the predisposition of IUGR foetuses to adult metabolic disorders.

Fabrication of large-area V-groove microstructures using gasbag-pressuring edge-irradiating UV imprinting

UV imprinting has been one of the most productive replication techniques due to its low imprinting pressure and temperature. The conventional UV resin is cured by direct irradiation from the bottom. Space management and energy consumption become problems as the areas are getting larger and larger. Another problem is the uniformity of imprinting pressure, which is higher near the center with conventional platen-pressuring imprinting. In this study, we develop an innovative side-emitting UV-curing gasbag-pressuring imprinting mechanism for large area microstructure replication. An imprinting system based on such integration has been designed and implemented. V-groove microstructures has been imprinted onto a PMMA plate with dimensions of 380 mm × 230 mm × 3 mm for light enhancement. The optical performance and the profile of microstructures have been verified.

The 1918 Influenza Pandemic: Looking Back, Looking Forward.

In commemoration of the centennial of the 1918 influenza pandemic, the American Journal of Epidemiology has convened a collection of 12 articles that further illuminate the epidemiology of that pandemic and consider whether we would be more prepared if an equally deadly influenza virus were to emerge again. In the present commentary, we place these 12 articles in the context of a growing body of work on the archeo-epidemiology of past pandemics, the socioeconomic and geographic drivers of influenza mortality and natality impact, and renewed interest in immune imprinting mechanisms and the development of novel influenza vaccines. We also highlight persisting mysteries in the origins and severity of the 1918 pandemic and the need to preserve rapidly decaying information that may provide treasure troves for future generations.

Identification of a novel antisense noncoding RNA, ALID, transcribed from the putative imprinting control region of marsupial IGF2R

BackgroundGenomic imprinting leads to maternal expression of IGF2R in both mouse and opossum. In mouse, the antisense long noncoding (lnc) RNA Airn, which is paternally expressed from the differentially methylated region (DMR) in the second intron of Igf2r, is required to silence the paternal Igf2r. In opossum, however, intriguingly, the DMR was reported to be in a different downstream intron (intron 11) and there was no antisense lncRNA detected in previous analyses. Therefore, clarifying the imprinting mechanism of marsupial IGF2R is of great relevance for understanding the origin and evolution of genomic imprinting in the IGF2R locus. Thus, the antisense lncRNA associated with the marsupial DMR can be considered as the ‘missing link’. In this study, we identified a novel antisense lncRNA, ALID, after detailed analysis of the IGF2R locus in an Australian marsupial, the tammar wallaby, Macropus eugenii, and compared it to that of the grey short-tailed opossum, Monodelphis domestica.ResultsTammar IGF2R showed maternal expression and had a maternally methylated CpG island (CGI) in intron 12 as well as a promoter CGI without differential methylation, but none in the second intron. Re-analysis of the IGF2R of opossum detected the CGI in intron 12, not intron 11, as previously reported, confirming that the DMR in intron 12 is conserved between these marsupials and so is the putative imprinting control region of marsupial IGF2R. ALID is paternally expressed from the middle of the DMR and is approximately 650 bp long with a single exon structure that is extremely short compared to Airn. Hence, the lncRNA transcriptional overlap of the IGF2R promoter, which is essential for the Igf2r silencing in the mouse, is likely absent in tammar. This suggests that fundamental differences in the lncRNA-based silencing mechanisms evolved in eutherian and marsupial IGF2R and may reflect the lack of differential methylation in the promoter CGI of marsupial IGF2R.ConclusionsOur study thus provides the best candidate factor for establishing paternal silencing of marsupial IGF2R without transcriptional overlap, which is distinct from the Igf2r silencing mechanism of Airn, but which may be analogous to the mode of action for the flanking Slc22a2 and Slc22a3 gene silencing in the mouse placenta.

Molecular characterization and gene expression of syntaxin-1 and VAMP2 in the olfactory organ and brain during both seaward and homeward migrations of chum salmon, Oncorhynchus keta

Anadromous Pacific salmon (Genus Oncorhynchus) imprint odorants from their natal streams during their seaward migration, and adult salmon use olfaction to identify their natal streams during their homeward migration. However, little is known about the molecular mechanisms of olfactory imprinting in the salmon nervous system. Our previous study suggested that the snap25s gene (encoding a soluble N-ethylmaleimide-sensitive factor attachment protein receptor [SNARE] protein) is involved in pre-synaptic functions for olfactory imprinting and/or olfactory memory retrieval in chum salmon (O. keta). In this study, the expression of other SNARE proteins was analyzed in chum salmon brains. Three cDNAs, encoding salmon SNARE proteins (STX-1a, STX-1b, and VAMP2), were isolated and sequenced, which are well-conserved among vertebrates. Quantitative PCR detected the expression of stx1s and vamp2 in all regions of the brain, and especially highly in the olfactory bulb (OB) and telencephalon. The expression levels of snares in the olfactory rosette (OR) were higher during seaward migration than in adult life stages, subsequently vamp2 in the OB and telencephalon increased during seaward migration, corresponding well with development of the olfactory nervous system. Both stx1s in the OB and stx1b in the telencephalon were elevated in the seaward period, whereas stx1a in the telencephalon increased continuously until the feeding period. Both stx1s in the telencephalon increased in the last phase of upriver migration, possibly related to the retrieval of imprinted memory. Our results indicated the involvement and distinct roles of upregulated snares in synaptic plasticity for olfactory imprinting and/or olfactory memory retrieval in Pacific salmon.