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Gonadotropin-releasing hormone (GnRH) vaccines have been successfully used for the inhibition of gonadal development and function, but current GnRH-based vaccines often present variability in the response. Cross-reactive material 197 (CRM197) has been used as carrier molecules to enhance an immune response to associated antigens. So, the synthetic mammalian tandem-repeated GnRH hexamer (GnRH6) gene was integrated into the expression plasmid pET-21a. Recombinant GnRH6-CRM197 protein was subsequently overexpressed in Escherichia coli strain BL21 and purified through Nickel column affinity chromatography and the antigenicity and biological effects of GnRH6-CRM197 were evaluated in rats. Sixteen 4-month-old adult male rats were randomly divided into two groups: the GnRH6-CRM197 group (n = 8) and the control group (n = 8). The GnRH6-CRM197 group rats were subcutaneously immunized with 100 μg of GnRH6-CRM197, administered thrice at 2-week intervals with GnRH6-CRM197.The control group received only a white oil adjuvant. Following the initial immunization, the weights of animals were recorded, and blood samples were collected from the orbital sinus at 4, 4.5, 5, 5.5, 6, 6.5, and 7 months. Serum antibody titers and testosterone concentrations were quantified using ELISA and CLIA, respectively. Additionally, testicular tissues were collected for morphological examination. The results revealed a significant increase in serum GnRH antibody titers (p < 0.05), but a significant decrease in serum testosterone concentrations (p < 0.05), and the weight, length, width, and girth of the testis, and the number of spermatogonia cells, spermatocytes, and sperm cells in the immunized rats. Furthermore, seminiferous tubules revealed significant atrophy and no sperm were observed in the immunized animals. Thus, GnRH6-CRM197 may be an effective antigen and a potential immunocastration vaccine. [ABSTRACT FROM AUTHOR]

Simple Summary: Castration benefits the management of male livestock and improves their meat quality. Immunocastration is more convenient and aligns with animal welfare, compared to surgical castration. In this study, two subunit vaccines were designed by fusing the conserved antigenic epitopes of porcine FSHR and LHR with the T-helper epitope region of the diphtheria toxin (DTT) and validated in male Sprague Dawley rats. The results showed that both vaccines induced antibody production in rats to some extent, inhibited testicular development, and reduced testicular function. Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) play key roles in regulating testosterone secretion and spermatogenesis in male mammals, respectively, and they maintain the fertility of male animals by binding to their corresponding receptors. We designed and prepared a recombinant LH receptor (LHR) subunit vaccine and a recombinant FSH receptor (FSHR) subunit vaccine and used male Sprague Dawley (SD) rats as a model to examine their effects on testicular development, spermatogenesis, and testosterone secretion in prepubertal and pubertal mammals. Both vaccines (LHR-DTT and FSHR-DTT) significantly decreased the serum testosterone level in prepubertal rats (p < 0.05) but had no effect on the testosterone secretion in pubertal rats; both vaccines decreased the number of cell layers in the seminiferous tubules and reduced spermatogenesis in prepubertal and pubertal rats. Subunit vaccine FSHR-DTT decreased the sperm density in the epididymis in both prepubertal and pubertal rats (p < 0.01) and lowered testicular index and sperm motility in pubertal rats (p < 0.05), whereas LHR-DTT only reduced the sperm density in the epididymis in pubertal rats (p < 0.05). These results indicate that the FSHR subunit vaccine may be a promising approach for immunocastration, but it still needs improvements in effectiveness. [ABSTRACT FROM AUTHOR]

The time of puberty onset is crucial for female animal, as it can affect the generation interval, feeding costs and the utilization of animals. However, little is known about the mechanism of hypothalamic lncRNAs (long noncoding RNAs) in regulatory goat puberty onset. Therefore, genome‐wide transcriptome analysis was performed in goats to clarify the roles of hypothalamic lncRNAs and mRNAs in the onset of puberty. In the present study, the co‐expression network of differentially expressed (DE) mRNAs in goat hypothalamus identified FN1 as the hub gene, and ECM‐receptor interaction, Focal adhesion and PI3K‐Akt signalling pathways are involved in puberty. We also observed the crucial hub transcription factors (TCF12, STAT1, STAT2, GATA3 and TEAD4) associated with reproduction and puberty. Then, genetic correlation analysis of DE mRNAs and DE lncRNAs identified the key lncRNAs involved in puberty. This research supplies a resource for transcriptome studies in goat puberty and indicated DE lncRNAs in the ECM‐receptor interaction pathway were novel candidate regulators for genetic studies on female reproduction. [ABSTRACT FROM AUTHOR]

MicroRNA-1 (miR-1) has been shown to play an important role in muscle growth and development, however, it was mainly discovered in model animals. To explore the function and mechanism of miR-1 in goat, we firstly explored the expression profile of miR-1 in goat tissues and cells. Furthermore, the target gene of miR-1 was predicted, and the relationship between miR-1 and one of its target genes, histone deacetylase 4 (HDAC4), was analyzed through double luciferase reporter assay, real-time PCR, and western blot. It was found that the miR-1 is most abundantly expressed in goat heart and skeletal muscle tissue. Meanwhile, the expression of miR-1 showed an increasing tendency from new-born goats to the 7-month-old goats, and then its expression decreases as the goats mature further. In addition, the expression levels of miR-1 decreased in goat skeletal muscle satellite cells with the algebraic increasing of cells. At last, the results showed that HDAC4 is a target gene of miR-1 in goat, and miR-1 can inhibit the post-transcriptional expression of HDAC4, but had no significant influence on the mRNA level of HDAC4. It was hypothesized that miR-1 promotes muscle development by inhibiting the post-transcriptional expression of HDAC4 in goat. [ABSTRACT FROM AUTHOR]

RNA sequencing performed on goat matured oocytes and preimplantation embryos generated in vivo enabled us to define the transcriptome for goat preimplantation embryo development. The largest proportion of changes in gene expression in goat was found at the 16-cell stage, not as previously defined at the 8-cell stage, and is later than in other mammalian species. In all, 6482 genes were identified to be significantly differentially expressed across all consecutive developmental stage comparisons, and the important signalling pathways involved in each development transition were determined. In addition, we identified genes that appear to be transcribed only at a specific stage of development. Using weighted gene coexpression network analysis, we found nine stage-specific modules of coexpressed genes that represent the corresponding stage of development. Furthermore, we identified conserved key members (or hub genes) of the goat transcriptional networks. Their association with other embryo genes suggests that they may have important regulatory roles in embryo development. Our cross-mammalian species transcriptomic comparisons demonstrate both conserved and goat-specific features of preimplantation development. Using the RNA sequencing technologies, we obtained the first complete transcriptome of in vivo goat mature oocytes and preimplantation embryos. Embryonic genome activation occurred at the 16-cell stage, and unique gene regulators of embryo development were identified. Comparison of transcriptomes between mammalian species indicated both conserved and goat-specific features of preimplantation development. [ABSTRACT FROM AUTHOR]

NESFATIN‐1 acts as a neuroendocrine hormone to suppress gonadotropin secretion in the female goldfish and to prevent germinal vesicle breakdown of oocytes in the zebrafish. However, the expression and function of NESFATIN‐1 in meiotic maturation and development of porcine oocytes remains elusive. Genomic structure of porcine NESFATIN‐1 precursor nucleobindin 2 (NUCB2) is first characterized in detail and an evolutionally closer relationship of NESFATIN‐1 between pig and rat is shown by phylogenetic analysis of multiple species. Additionally, immunofluorescence analysis revealed that NESFATIN‐1 is predominantly expressed and localizes on the membrane of both theca cells and granulosa cells, but not expressed in oocytes. Real‐time quantitative polymerase chain reaction showed that the abundance of NESFATIN‐1 transcripts in granulosa cells progressively decreases during the developmental transition from small follicles to large follicles. Correspondingly, NESFATIN‐1 could significantly enhance both the cleavage and blastocyst rate of parthenogenetically activated oocytes from small follicles (p < 0.05), whereas it did not affect meiotic maturation and development of oocytes from large follicles. Interestingly, we found that NESFATIN‐1 significantly improves meiotic maturation of oocytes cultured in chemically defined medium in the absence of pyruvate compared with the control group (p < 0.05), suggesting that the NESFATIN‐1 as a substitute for pyruvate exerts beneficial effects on porcine oocyte maturation. In conclusion, these results demonstrate that NESFATIN‐1 facilitates both meiotic maturation and development of porcine oocytes. [ABSTRACT FROM AUTHOR]

Contents: Epigenetics plays an important role in controlling female puberty. Both DNA methylation and long non‐coding RNAs (lncRNA) regulate the initiation of puberty by affecting the expression of genes related to puberty. While recent studies have indicated that DNA methylation of lncRNA represses the expression of lncRNA, its role in regulating puberty remains unclear. To explore the mechanism between DNA methylation and lncRNAs during puberty onset, we performed whole‐genome bisulphite sequencing (WGBS) and RNA‐sequencing (RNA‐seq). We found that DNA methylation was inversely correlated to gene expression levels during puberty. Methylation levels gradually decreased near the transcription initiation site and were present at high levels in the exon, intron and 3′ untranslated regions. In the promoter, lncRNA expression was negatively related to DNA methylation. We reported hypermethylation in the gene body and downstream of the lncRNA compared with upstream regions. In GO and KEGG analyses, we found enriched target genes of lncRNA, XLOC_960044 and XLOC_767346. During puberty, methylation of these genes increased while expression decreased. Our study indicates that DNA methylation of the promoter is negatively correlated with lncRNA during puberty onset, and methylation regulates the initiation of puberty via lncRNA, which provides new insight into the epigenetic mechanism of puberty onset. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNAs) play important roles in almost all biological processes. However, there is little information on the effects of lncRNAs on ovulation and lambing rates. In the present study, we used high-throughput RNA sequencing to identify differentially expressed lncRNAs between the ovaries of multiparous (Mul) and uniparous (Uni) Anhui White goats. Among the 107,255,422 clean reads, 183,754 lncRNAs were significantly differentially expressed between the Uni and Mul. Among them, 455 lncRNAs were co-expressed between the two samples, whereas, 157,523 lncRNAs were uniquely expressed in the Uni, and 25,776 uniquely lncRNAs were expressed in the Mul. Through Cis role analysis, 24 lncRNAs were predicted to overlap with cis-regulatory elements, which involved in Progesterone-mediated oocyte maturation, Steroid biosynthesis, Oocyte meiosis, and gonadotropin-releasing hormone (GnRH) signaling pathway. These 4 pathways were related to ovulation, and the KEGG pathway analysis on target genes of the differentially expressed lncRNAs confirmed this results. In addition, 10 lncRNAs harbored precursors of 40 miRNAs, such as TCONS_00320849 related to a mature miRNA sequence, miR-365a, which was reported to be related to proliferation, were annotated in the precursor analysis of miRNAs. The present expand the understanding of lncRNA biology and contribute to the annotation of the goat genome. The study will provide a resource for lncRNA studies of ovulation and lambing. [ABSTRACT FROM AUTHOR]

Female pubertal development is tightly controlled by complex mechanisms, including neuroendocrine and epigenetic regulatory pathways. Specific gene expression patterns can be influenced by DNA methylation changes in the hypothalamus, which can in turn regulate timing of puberty onset. In order to understand the relationship between DNA methylation changes and gene expression patterns in the hypothalamus of pubertal goats, whole-genome bisulfite sequencing and RNA-sequencing analyses were carried out. There was a decline in DNA methylation levels in the hypothalamus during puberty and 268 differentially methylated regions (DMR) in the genome, with differential patterns in different gene regions. There were 1049 genes identified with distinct expression patterns. High levels of DNA methylation were detected in promoters, introns and 3′-untranslated regions (UTRs). Levels of methylation decreased gradually from promoters to 5′-UTRs and increased from 5′-UTRs to introns. Methylation density analysis demonstrated that methylation level variation was consistent with the density in the promoter, exon, intron, 5′-UTRs and 3′-UTRs. Analyses of CpG island (CGI) sites showed that the enriched gene contents were gene bodies, intergenic regions and introns, and these CGI sites were hypermethylated. Our study demonstrated that DNA methylation changes may influence gene expression profiles in the hypothalamus of goats during the onset of puberty, which may provide new insights into the mechanisms involved in pubertal onset. [ABSTRACT FROM AUTHOR]

To explore if the regulation at post-transcriptional level of follicular phase (Fols) to luteal phase (Luts) transition occurs in the ovaries of Anhuai goats, the differentially expressed microRNAs (miRNAs) of ovaries in the Fols and Luts were analyzed using Solexa sequencing in the study. In total, 320 known miRNAs were co-expressed in the two phases, 339 and 353 known miRNAs were expressed in the ovary in the Fols and Luts, respectively. In addition, 45 novel miRNAs were co-expressed in the two phases, 70 and 94 novel miRNAs were expressed in the ovary in the Fols and Luts, respectively. Let-7f was the highest expressed significantly different known miRNA in the two phases, and mir-159 was the highest expressed significantly different novel miRNA in the two phases, which may participate in the follicular-luteal transition of Anhuai goats. GO annotation and KEGG pathway analysis were applied to analyze the target genes of differentially expressed miRNAs detected in the two phases. The results will help to further understand the role of miRNAs in the regulation of follicular to luteal transition in goat ovaries. [ABSTRACT FROM AUTHOR]

The low full-term developmental efficiency of porcine somatic cell nuclear transfer (SCNT) embryos is mainly attributed to imperfect epigenetic reprogramming in the early embryos. However, dynamic expression patterns of histone methylation involved in epigenetic reprogramming progression during porcine SCNT embryo early development remain to be unknown. In this study, we characterized and compared the expression patterns of multiple histone methylation markers including transcriptionally repressive (H3K9me2, H3K9me3, H3K27me2, H3K27me3, H4K20me2 and H4K20me3) and active modifications (H3K4me2, H3K4me3, H3K36me2, H3K36me3, H3K79me2 and H3K79me3) in SCNT early embryos from different developmental stages with that from in vitro fertilization (IVF) counterparts. We found that the expression level of H3K9me2, H3K9me3 and H4K20me3 of SCNT embryos from 1-cell to 4-cell stages was significantly higher than that in the IVF embryos. We also detected a symmetric distribution pattern of H3K9me2 between inner cell mass (ICM) and trophectoderm (TE) in SCNT blastocysts. The expression level of H3K9me2 in both lineages from SCNT expanded blastocyst onwards was significantly higher than that in IVF counterparts. The expression level of H4K20me2 was significantly lower in SCNT embryos from morula to blastocyst stage compared with IVF embryos. However, no aberrant dynamic reprogramming of H3K27me2/3 occurred during early developmental stages of SCNT embryos. The expression of H3K4me3 was higher in SCNT embryos at 4-cell stage than that of IVF embryos. H3K4me2 expression in SCNT embryos from 8-cell stage to blastocyst stage was lower than that in the IVF embryos. Dynamic patterns of other active histone methylation markers were similar between SCNT and IVF embryos. Taken together, histone methylation exhibited developmentally stage-specific abnormal expression patterns in porcine SCNT early embryos. [ABSTRACT FROM AUTHOR]

This study was designed to explore the effect of active immunization against maltose binding protein-gonadotropin releasing hormone I hexamer ( MBP- GnRH-I6) on the reproductive function in cats. Each immunized cat was administered twice intramuscularly in the neck at 16 and 20 weeks old. The concentrations of the testosterone and estradiol and the level of anti- GnRH- I antibody in the serum were measured by radioimmunoassay and ELISA, respectively. The results showed that the weight and size of testicles and ovaries, and the concentrations of serum testosterone and estradiol in the immunized animals were lower than those of the control cats ( P < 0.05), but that the levels of anti- GnRH- I antibody were significant higher compared to control animals ( P < 0.05). Testicular tissues from the immunized male cats showed that seminiferous tubules were depauperate with the lumen relatively empty and that the differentiation of spermatogonia was not obvious. Tissues from the immunized female cats showed that the ovaries had many primordial follicles and primary follicles, but no secondary follicle was observed. These results showed active immunization against MBP- GnRH-I6 could make the gonads atrophy and reduce the concentrations of gonadal hormones, which suggested that MBP- GnRH-I6 was a very effective immunogen in the cat. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the efficacy of active immunisation against recombinant maltose binding protein-gonadotropin releasing hormone I (MBP-GnRH-I6) in preventing boar taint. The concentrations of testosterone and anti-GnRH-I antibodies in serum were determined by radioimmunoassay and enzyme linked immunosorbent assay, respectively. The mRNA levels of 3β-hydroxysteroid dehydrogenase (3β-HSD) and 17β-hydroxysteroid dehydrogenase (17β-HSD) in testes were analysed by real time quantitative PCR. The concentrations of androstenone and 3-methylindole in backfat were assayed by HPLC. Active immunisation against MBP-GnRH-I6 increased the serum concentration of anti-GnRH-I antibodies (P<0.05) and the levels of 3β-HSD and 17β-HSD mRNA in testis. However, it reduced the serum concentrations of testosterone (P<0.05) and androstenone. The concentration of 3-methylindole in backfat (P>0.05) was not changed. Thus, we conclude that vaccination against MBPGnRH- I6 is a practical and effective method to suppress the synthesis of androstenone. [ABSTRACT FROM AUTHOR]

All- trans retinoic acid ( t- RA) is a natural component and representative physiologically active metabolite of vitamin A, having multiple physiologic functions. The objective of this study was to evaluate the effect of t- RA on goat oocyte maturation and cumulus cell apoptosis during in vitro maturation ( IVM). Immature goat cumulus-oocyte complexes ( COCs) were matured in vitro in the absence or presence of t- RA at concentrations of 10 nmol/L, 100 nmol/L and 1000 nmol/L. Oocyte maturation and embryo development were assessed by polar body formation and parthenogenetic activation, respectively. Oocyte survival was checked by Trypan blue staining. Apoptosis of cumulus cells was analyzed by terminal deoxynucleotidyl transferase nick end labeling staining and quantitative real-time PCR. In comparison with the control group, 100 nmol/L and 10 nmol/L t- RA significantly improved goat nuclear oocyte maturation and survival ( P < 0.05). Addition of 1000 nmol/L t- RA improved nuclear maturation ( P < 0.05), but had no effect on survival of goat oocytes. t- RA had no positive effect on goat parthenogenetic embryonic cleavage, blastocyst formation or total cell numbers. However, t- RA inhibits the apoptosis of cumulus cells ( P < 0.01). t- RA treatment up-regulated the expression of B-cell lymphoma 2 ( BCL-2), catalase ( CAT) ( P < 0.05) and down-regulated the expression of Caspase-8 ( P < 0.05). In conclusion, t- RA has positive effects on goat oocyte nuclear maturation and reduces apoptotic cumulus cells during IVM. [ABSTRACT FROM AUTHOR]

Abstract: To determine whether hormone-receptor signaling pathways in the thymus are altered by active immunization against gonadotrophin-releasing hormone I (GnRH), 3-week-old Sprague–Dawley male rats received GnRH-tandem-OVA peptides (200μg/ml), and the effects were compared to a control group. Serum testosterone, LH and FSH concentrations were markedly reduced, with severe testicular atrophy, compared to controls, demonstrating effective blockade of the pituitary–gonadal axis. The reduction in LH and FSH concentrations in the thymus of immunized animals was lower than that observed in the serum, where a significant difference (P <0.001) in concentration was observed between both groups. Concentrations of GnRH were increased in the thymus of immunized rats. In thymic tissue, GnRHR, FSHR and LHR demonstrated stronger immunostaining, and AR weaker staining, in the immunized group compared to controls. Reproductive hormone receptor mRNA expression was consistent with protein variations in the immunized thymus. Compared to controls, GnRHR gene levels were significantly increased (P <0.05), however, AR mRNA expression were greatly decreased with immune week-age (P <0.05). Both FSHR and LHR mRNA expression levels were significantly higher in the treated group than in controls in the first three samples (P <0.05). When GnRHR was blocked by an antagonist in thymocytes, all reproductive hormone receptor gene expressions were significantly increased (P <0.001). In summary, these findings suggest that active immunization against GnRH can up-regulate GnRH receptor and gonadotropin receptor signaling, by stimulating thymic autocrine and paracrine function, whereas the androgen receptor is down-regulated due to a lack of testosterone secretion in the thymus. [Copyright &y& Elsevier]

Abstract: Ghrelin, the endogenous ligand for the GH secretagogue receptor (GHS-R), has various functions. The expression of ghrelin and growth hormone secretagogue receptor type 1a (GHS-R1a) has been demonstrated in rat and human testis, and ghrelin also affects testosterone (T) secretion in vitro, suggesting a role for this molecule in the direct control of testis function. However, whether this signaling system is present in pig testis, and the association with serum T remains largely unexplored. In this study we investigated the relationship between serum T levels and ghrelin and GHS-R1a gene expression in the testes of intact and immunocastrated boars. Testicular tissue and serum samples were collected from seven intact boars and seven recombinant GnRH-I immunocastrated boars. GHS-R1a gene expression in the testis was higher in immunocastrated than intact boars (2.22±0.71 vs. 0.53±0.23; P <0.01). Mean serum T levels were markedly lower in immunocastrated than intact boars (0.40±0.04 vs. 2.10±0.94ng/mL, P <0.01). Ghrelin gene expression in testis did not differ between both groups of boars. GHS-R1a gene expression was positively correlated with ghrelin gene expression (r =0.68, P <0.001), but negatively correlated with serum T concentrations (r =−0.83, P <0.001). Overall, the existence of ghrelin and GHS-R1a gene expression in pig testis, and the increase in GHS-R1a gene expression in the testes and the inverse correlation with serum T in immunocastrated boars are highly suggestive of a role for ghrelin in the regulation of mammalian testicular function. [Copyright &y& Elsevier]

The objective of the present study was to evaluate the effects of ghrelin on the concentrations of estrogen (E2) and progesterone (P4) in serum and the mRNA expression of estrogen receptor beta (ERβ) and progesterone receptor (PRA++B) in ovary in rats during estrous cycle. Adult female Sprague Dawley rats were intracerebroventricularly (i.c.v.) injected with 3 nmol ghrelin during the estrous cycle, and sacrificed 15 min later. Blood samples and ovaries were collected. The concentrations of serum E2 and P4 were measured by radioimmunoassay, while the amount of ERβ and PRA++B mRNA was assessed by real-time quantitative PCR. Our studies showed that ghrelin could significantly reduce the serum concentration of E2 throughout the estrous cycle ( P < 0.05), the serum level of P4 ( P < 0.05), and the amount of ERβ mRNA during metestrus ( P < 0.05). Meanwhile, the amount of PRA++B mRNA was only reduced during diestrus ( P < 0.05). Overall, our present findings provide the first evidence that i.c.v. injection of ghrelin could reduce the serum concentration of E2 and P4 and the level of ERβ and PRA++B mRNA expression, supporting the role of ghrelin in reproduction. [ABSTRACT FROM AUTHOR]

The present study was designed to determine the effects of ghrelin on in vivo and in vitro secretion of testosterone (T) and the expression of androgen receptor (AR) mRNA in the adult rat testis. The distribution of growth hormone secretagogue receptors (GHS-R1a) in the testis was also investigated. GHS-R1a immunoreactivity presented mainly in Sertoli and Leydig cells, primary spermatocytes, and secondary spermatocytes. Adult rats that were intracerebroventricularly (i.c.v.) administrated different dosages (1 nmol and 3 nmol) of ghrelin could significantly inhibit the secretion of T. The experession of AR mRNA in the testis was also notably reduced with 3 nmol ghrelin. Additionaly, in vitro exposure of the Leydig cells to increasing concentrations of ghrelin resulted in no obvious changes of T secretion in the culture media and AR mRNA expression of Leydig cells. Overall, our data demonstrate that the i.c.v. injection of ghrelin plays a physiological role in T secretion and AR mRNA expression in the testis, further confirming the reproductive role of ghrelin. [ABSTRACT FROM AUTHOR]

A recombinant fusion protein, maltose-binding protein (MBP) - gonadotropin releasing hormone I (GnRH-I), was produced by cloning a gene fragment encoding a tandem repeated GnRH-I hexmer peptide (GnRH-I6) into the pMAL-c4x vector which was subsequently expressed in E. coli TB1. MBP-GnRH-I6 was affinity purified. MBP-GnRH-I6 was verified by SDS-PAGE and Western blot and immunogenicity tested in boars. Injection of the recombinant fusion protein into boars yielded a high-titer antibody specific for GnRH-I. This was followed by serum testosterone and the degeneration of spermatogenesis. These results showed that MBP-GnRH-I6 acted as a strong immunogen and could be a candidate for an immune antifertility vaccine. [ABSTRACT FROM AUTHOR]

Abstract: Immunization using recombinant maltose binding protein–gonadotropin releasing hormone (MBP–GnRH6) altered both testicular development and transcription of the pituitary GnRH receptor (GnRHR) gene in boars. Scrotal measurement and blood samples were taken at 4-week interval after immunization at 9 weeks of age. The concentrations of testosterone and anti-GnRH antibodies in serum were determined by radioimmunoassay and enzyme-linked immunosorbent assay, respectively. The results showed that active immunization with MBP–GnRH6 increased the serum concentration of anti-GnRH antibodies (P <0.05) and reduced the serum concentration of testosterone (P <0.05) as compared with MBP controls. At 25 weeks of age, boars were sacrificed and testes were evaluated histologically. Testicular development was suppressed in the MBP–GnRH6 immunized animals as compared with MBP immunized boars. MBP–GnRH6 immunized pigs exhibited mounting behavior 4 weeks later than MBP immunized boars. No mature spermatozoa were observed from MBP–GnRH6 immunized animals. By real-time quantitative PCR analysis, the amount of GnRHR mRNA in the pituitary tissue was found to be significantly lower in MBP–GnRH6 immunized animals than in controls (P <0.05). These data demonstrate that recombinant MBP–GnRH6 was effective in immunological castration in boars. [Copyright &y& Elsevier]

Simple Summary: This study evaluated the effect of γ-aminobutyric acid transaminase (GABA-T) on reproduction in female rats. Our results showed that GABA-T expressed in the reproductive axis of female rats at different developmental stages, and inhibiting GABA-T affected Rfrp-3, Gnrh and Kiss1 mRNA transcription levels in hypothalamic and reduced the concentration of GABA-T, LH, and P4 in serum. At the same time, the initiation of the puberty was delayed, and led to disordered estrous cycle and reduced breastfeeding performance in adult female rats. This study showed that GABA-T affects the reproductive function of female rats by regulating the levels of Gnrh, Kiss1 and Rfrp-3 in the hypothalamus as well as the concentrations of LH and P4. This study explored the role of γ-aminobutyric acid transaminase (GABA-T) in the puberty and reproductive performance of female rats. Immunofluorescence technique, quantitative real-time PCR (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the distribution of GABA-T and the expression of genes and hormones in female rats, respectively. The results showed that GABA-T was mainly distributed in the arcuate nucleus (ARC), paraventricular nucleus (PVN) and periventricular nucleus (PeN) of the hypothalamus, and in the adenohypophysis, ovarian granulosa cells and oocytes. Abat mRNA level at 28 d was lowest in the hypothalamus and the pituitary; at puberty, it was lowest in the ovary. Abat mRNA level was highest in adults in the hypothalamus; at infancy and puberty, it was highest in the pituitary; and at 21 d it was highest in the ovary. After vigabatrin (GABA-T irreversible inhibitor) was added to hypothalamus cells, the levels of Abat mRNA and Rfrp-3 mRNA were significantly reduced, but Gnrh mRNA increased at the dose of 25 and 50 μg/mL; Kiss1 mRNA was significantly increased but Gabbr1 mRNA was reduced at the 50 μg/mL dose. In prepubertal rats injected with vigabatrin, puberty onset was delayed. Abat mRNA, Kiss1 mRNA and Gnrh mRNA levels were significantly reduced, but Rfrp-3 mRNA level increased in the hypothalamus. Vigabatrin reduced the concentrations of GABA-T, luteinizing hormone (LH) and progesterone (P4), and the ovarian index. Lactation performance was reduced in adult rats with vigabatrin treatment. Four hours after vigabatrin injection, the concentrations of GABA-T and LH were significantly reduced in adult and 25 d rats, but follicle-stimulating hormone (FSH) increased in 25 d rats. In conclusion, GABA-T affects the reproductive function of female rats by regulating the levels of Gnrh, Kiss1 and Rfrp-3 in the hypothalamus as well as the concentrations of LH and P4. [ABSTRACT FROM AUTHOR]

A series of complex processes regulate muscle development, and lncRNAs play essential roles in the regulation of skeletal myogenesis. Using RNA sequencing, we profiled the lncRNA expression during goat (Capra hircus) skeletal muscle development, which included seven stages across fetal 45 (F45), 65 (F65), 90 (F90), 120 (F120), 135 (F135) days, born for 24 h (B1) and 90 (B90) days. A total of 15,079 lncRNAs were identified in the seven stages, and they were less conservative with other species (human, cow, and mouse). Among them, 547 were differentially expressed, and they divided the seven stages into three functional transition periods. Following weighted gene co-expression network analysis (WGCNA), five lncRNA modules specific for developmental stages were defined as three types: 'Early modules', 'late modules', and 'individual-stage-specific modules'. The enrichment content showed that 'early modules' were related to muscle structure formation, 'late modules' participated in the 'p53 signaling pathway' and other pathways, the F90-highly related module was involved in the 'MAPK signaling pathway', and other pathways. Furthermore, we identified hub-lncRNA in three types of modules, and LNC_011371, LNC_ 007561, and LNC_001728 may play important roles in goat skeletal muscle. These data will facilitate further exploration of skeletal muscle lncRNA functions at different developmental stages in goats. [ABSTRACT FROM AUTHOR]

To investigate the effect of gonadotropin-releasing hormone 2-multiple antigen peptide (GnRH2-MAP) on reproductive function. In our study, 20-day-old male rats (n = 90) were randomly allocated to one of three treatment groups: GnRH2-MAP immunization, GnRH2 immunization, and non-immunized control groups. The immunized animals were administered three doses of GnRH2-MAP or GnRH2 vaccines from 0 to 6 weeks at 2-week intervals. The control group only received oil adjuvant. Blood and right testis samples were collected, and the left testis was weighed and its volume was measured at 0, 2, 4, 6, 8, 10 and 12 weeks after the first immunization. The serum antibody titer and testosterone concentration were determined by ELISA, and the right testis samples were collected for histological analysis. The results revealed that the serum of vaccinated rats elicited a significantly higher antibody titer and a lower T concentration compared with the control group two weeks after the first immunization ( P  < 0.05), but the highest antibody titer and lowest T concentration were found in animals treated with GnRH2-MAP ( P  < 0.05). The second immunization resulted in a significant decrease in testicular weight and volume ( P  < 0.05) in both immunized groups compared to the control, but these values were significantly lower in the GnRH2-MAP group than in the GnRH2 group. Furthermore, seminiferous tubules revealed more significant atrophy in the GnRH2-MAP group than in the GnRH2 group, and no sperm were observed in rats of the GnRH2-MAP group. Thus, GnRH2-MAP may be an effective antigen and a potential immunocastration vaccine with higher effectiveness. [ABSTRACT FROM AUTHOR]