Effects of GnRH immunization on the reproductive axis and thymulin

  1. Yunsheng Li
  1. College of Animal Sciences and Technology, Anhui Agricultural University, 130, Changjiang West Road, Hefei, Anhui 230036, People's Republic of China
    1College of Life Science, Fujian Agriculture and Forestry University, Fuzhou 350002, People's Republic of China
    2The Biotechnology Center of Anhui Agriculture University, Hefei, People's Republic of China
  1. Correspondence should be addressed to F Fang; Email: fgfang{at}ahau.edu.cn
  1. Figure 1

    Detection of GnRH antibody titers. Measurements were taken every 2 weeks from the first immunization. Absorbance readings taken at 450 nm showed increased antibody titers from representative sera taken from male rats immunized with GnRH-tandem-OVA compared with Al(OH)3 adjuvant control (n=8). *P<0.01, **P<0.001.

  2. Figure 2

    Levels of several hormones in serum of control and immunized rats (n=8). Measurements were taken every 2 weeks from the first immunization. (A) Testosterone levels in serum. (B) LH levels in serum. (C) FSH levels in serum. (D) Thymulin levels in serum. The levels of detection for testosterone, LH and FSH were all 0.02 ng/ml. The sensitivity of thymulin was 10.0 pg/ml. *P<0.05; **P<0.01; ***P<0.001.

  3. Figure 3

    The levels of thymulin and GnRH in the hypothalami of control and immunized rats (n=8). Measurements were taken every 2 weeks from the final booster immunization. (A) GnRH levels in homogenized hypothalamus tissue of control and immunized rats. (B) Thymulin levels in homogenized hypothalami. The sensitivities of GnRH and thymulin were 5.2 and 10.0 pg/ml respectively. HW, hypothalamus weight. *P<0.05; **P<0.01.

  4. Figure 4

    Protein expression of GnRH and GFAP in hypothalamic nuclei of control and immunized rats (n=5). (A and B) Paraventricular nucleus (Pa) of control and immune rats respectively; (C and D) medial preoptic nucleus (MPN) respectively; (E and F) arcuate nucleus (Arc) respectively; (G and H) median eminence (ME) respectively. (I and J) Average fluorescence intensities (AFIs) of GnRH and GFAP in different areas of the hypothalamus respectively. A dotted line indicated the nuclei of the hypothalamus. Green indicates a positive reaction of GnRH with IgG-FITC, orange a positive reaction of glial fibrillary acidic protein (GFAP) with IgY-Cy5, and their merge image was marked. TV: third ventricle, bar=50 μm. AFIs of GnRH (E) and GFAP (F) in nuclei of rat hypothalami were analyzed. AFI was obtained and analyzed in two 40× object lens microscopic fields of every nucleus per hypothalamus of five rats in two treatments using the FV10.ASW 3.0 software and Image Pro-Plus 6.0 (Media Cybernetics Co., Rockville, MD, USA). Significant differences between groups were determined using an unpaired Student's t-test (n=10; two slices per hypothalamic nuclei). *P<0.05, **P<0.01.

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