Journal: International Journal of Molecular Sciences

Article Title: Gonadotropin-Releasing Hormone in Regulation of Thymic Development in Rats: Profile of Thymic Cytokines

doi: 10.3390/ijms20164033

Figure Lengend Snippet: Ontogenetic pattern of the gonadotropin-releasing hormone (GnRH) receptor expression in the developing thymus. ( A ) mRNA of GnRH receptor revealed by RT-PCR in the rat thymus on embryonic days (ED) 16, 17, 18, 19, 21 and postnatal day 3 (PND3). ( B ) Western blot assay of GnRH receptor in the rat thymus on ED17, ED18, ED19, ED20, ED21, and PND3. The anterior pituitary (PND3) was used as a positive control (+). ( C ) Protein expression of GnRH receptor in thymocytes and thymic stromal elements on ED18. Plots represent the optic density of the corresponding bands. Bars indicate the means ± SEM of three independent experiments; * p < 0.05 using one-way ANOVA test.

Article Snippet: Separated proteins were transferred to a nitrocellulose membrane in transfer buffer (25 mM Tris-HCl, pH 7.5, 192 mM glycine, 20% ethanol) and blots were incubated overnight at 4 °C with antibodies to GnRH receptor (1:1000, Alomone Labs, Jerusalem, Israel) or actin (1:10,000, Sigma, St. Louis, Mo, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Positive Control

Journal: International Journal of Molecular Sciences

Article Title: Gonadotropin-Releasing Hormone in Regulation of Thymic Development in Rats: Profile of Thymic Cytokines

doi: 10.3390/ijms20164033

Figure Lengend Snippet: Schematic representation of GnRH effects on the thymus development in fetal and early postnatal rats. During prenatal development up to ED20–21, before the establishment of the HPG endocrine regulations, hypothalamic GnRH can be released to general circulation and provide a direct effect on the morphogenesis of thymus. Since the development of blood–brain barrier and the establishment of the HPG axis in early postnatal life GnRH is involved in the bidirectional programming of both neuroendocrine and immune functions via gonadotropins and sex steroids. In addition to the neuroendocrine regulation, the effects of GnRH synthesized in thymus can be realized by autocrine/paracrine mechanisms. The developmental pattern of the GnRH receptor expression in the thymus is in favor of this assumption. ED—embryonic day, PND—postnatal day, HP—hypophysis.

Article Snippet: Separated proteins were transferred to a nitrocellulose membrane in transfer buffer (25 mM Tris-HCl, pH 7.5, 192 mM glycine, 20% ethanol) and blots were incubated overnight at 4 °C with antibodies to GnRH receptor (1:1000, Alomone Labs, Jerusalem, Israel) or actin (1:10,000, Sigma, St. Louis, Mo, USA).

Techniques: Synthesized, Expressing

Journal: Heliyon

Article Title: Brucella abortus RB51 Δ leuB expressing Salmonella FliC conjugated gonadotropins reduces mouse fetal numbers: A possible feral swine brucellosis immunocontraceptive vaccine

doi: 10.1016/j.heliyon.2021.e06149

Figure Lengend Snippet: Anti-his-tag Western blot. Fluorescent anti-his-tag antibodies were applied to blotted protein extracts. Lane-1 consists of the BioRad Precision Plus Protein Ladder. Lane-2 consists of protein extract from strain RB51L, showing no significant bands. Lane-3 consists of protein extract from strain RB51LGnRH isolated from the spleens of BABLB/c mice 14 days post inoculation, with a his-tag protein between 37 kDa and 50 kDa. This corresponds with the theoretical size of the his-tagged FliC-GnRH protein (≈40kDa). The original pictograph is represented in Supplemental Figure 4 .

Article Snippet: The presence of GnRH and FSH antibodies was confirmed via absorbance values with the following protocol: Antigens (Provided by Boster and BEI Resources, VA, USA) were diluted in PBS to a concentration of 20 μg/ml.

Techniques: Western Blot, Isolation

Journal: Heliyon

Article Title: Brucella abortus RB51 Δ leuB expressing Salmonella FliC conjugated gonadotropins reduces mouse fetal numbers: A possible feral swine brucellosis immunocontraceptive vaccine

doi: 10.1016/j.heliyon.2021.e06149

Figure Lengend Snippet: GnRH and porcineFSHβ IgG antibody ELISAs from male and female BALB/c mice. (a) Fifty microliters of serum collected 81 days post vaccination from both male and female mice were tested in duplicate for detection of anti-GnRH IgG antibodies. All group values were compared via One-way ANOVA, and the statistically significant differences are shown. The OD 450 values from the serum of mice in the strain RB51LGnRH immunized group were significantly higher than those from the PBS control group with a p value of <0.0001. The OD 450 values from the serum of mice in the Improvest® immunized group were significantly higher than those from the control group PBS, with the p value of <0.0001. The OD 450 values from the serum of mice in the strain RB51LGnRH immunized group were significantly higher than those from the RB51L control group with a p value of 0.02. (b) Fifty microliters of serum collected 81 days post vaccination from both male and female mice were tested in duplicate for detection of porcineFSHβ IgG antibodies. All group values were compared via One-way ANOVA, and the statistically significant differences are shown. The OD 450 values from the serum of mice in the strain RB51LF immunized group were significantly higher than those from the PBS control group with a p value of <0.0001 and the control RB51L group with a p value of 0.0011. This shows that there was an immune response amounted to the vaccines, which can be further seen by the other data sets. There were no significant differences between groups without brackets and p values.

Article Snippet: The presence of GnRH and FSH antibodies was confirmed via absorbance values with the following protocol: Antigens (Provided by Boster and BEI Resources, VA, USA) were diluted in PBS to a concentration of 20 μg/ml.

Techniques: Control, Vaccines

Journal: Frontiers in Neuroscience

Article Title: Neuroendocrine regulation of gonadotropin secretion in seasonally breeding birds

doi: 10.3389/fnins.2013.00038

Figure Lengend Snippet: Schematic model of the mechanism of seasonal reproduction in birds. Seasonal breeding birds detect environmental signals, such as light, temperature, water, and food availability, predator pressure, etc. to time reproduction. Their reproductive activity is also shaped by social interactions with the conspecifics, in terms of sound, sight, contact, etc. Hypothalamic neurons in the bran integrate external and internal signals, and regulate reproduction by releasing neurohormones at the median eminence (ME) to regulate the pituitary hormone secretion. The anterior pituitary gland synthesizes and releases gonadotropins (LH, FSH), and gonadotropins act on the gonads to stimulate gametogenesis and sex steroid secretion. Sex steroids induce secondary sexual characteristics of the organism. A hypothalamic neuropeptide, GnRH1, activates reproduction by stimulating gonadotropin synthesis and release via its receptor (GnRH-R) expressed on the gonadotropes. Another hypothalamic neuropeptide, GnIH, may inhibit gonadotropin synthesis and release directly by acting on the pituitary or indirectly by decreasing the activity of GnRH1 neurons via its receptor (GnIH-R) expressed on the gonadotropes and GnRH neurons. GnRH2 neurons may stimulate reproductive behavior, and GnIH may inhibit reproductive behavior by inhibiting GnRH2 neurons. GnRH and GnIH neurons in the hypothalamus and their receptors in the pituitary are also regulated by internal signals, such as testosterone (T), estradiol (E2), and progesterone (P4). It is hypothesized that locally produced triiodothyronine (T 3 ) from thyroxine (T 4 ) by the action of type 2 iodothyronine deiodinase (DIO2) at the mediobasal hypothalamus stimulates the release of GnRH1. The function of GnRH1 neurons is also regulated by transcription of the GnRH gene. Because melatonin, a nocturnal hormone, stimulates the synthesis and release of GnIH in birds, GnIH may regulate daily rhythm of gonadotropin secretion. GnIH may also temporally suppress gonadotropin secretion when environmental or social condition is unfavorable.

Article Snippet: Three GnRH receptor subtypes (type I, II, III) have been identified with distinct distributions and functions in vertebrates (Millar et al., ).

Techniques: Activity Assay, Produced