Effect of androgen on Kiss1 expression and luteinizing hormone release in female rats

1. View larger version:

   • In this window
   • In a new window

   • Download as PowerPoint Slide

   Figure 1

   Morphology of ovaries and expression of Kiss1 mRNA in the anteroventral periventricular nucleus (AVPV) and arcuate nucleus (ARC) of ovary-intact rats. (A) Representative ovaries from non-DHT and DHT rats. Bars = 500 µm. CL, corpus luteum. (B) Kiss1 mRNA expression by in situ hybridization in the AVPV and ARC of non-DHT and DHT rats. Non-DHT, a normal control rat showing diestrus stage. DHT rats were ovary-intact animals. Bars = 100 µm. (C) Histogram indicating number of cells expressing Kiss1 mRNA in the AVPV and ARC. Statistical differences were determined by unpaired Student’s t-test. *P < 0.05 vs non-DHT rats. Numbers in each column represent the number of animals examined. Values are expressed as mean ± s.e.m.
2. View larger version:

   • In this window
   • In a new window

   • Download as PowerPoint Slide

   Figure 2

   Estradiol-17β (E2)-induced luteinizing hormone (LH) surge and expression of Kiss1 mRNA in the AVPV of non-DHT and DHT rats. All animals were ovariectomized and implanted with tubes containing a high concentration of E2. (A) Changes in plasma LH levels were assessed in freely moving conscious rats from 13:00 to 20:00. Values are expressed mean ± s.e.m. (each group, n = 9). Statistical differences were determined by two-way repeated-measures ANOVA (clock time and group), followed by Bonferroni correction. *P < 0.05, vs 13:00. ^#P < 0.05, vs DHT rats. (B) Expression of Kiss1 mRNA by in situ hybridization in the AVPV of representative rats. Bars = 100 µm. (C) Number of Kiss1 mRNA-expressing cells in the AVPV. No significant difference was found between the two groups. Numbers in each column represent the number of animals examined. Values are expressed as mean ± s.e.m. (D) Z-stack images of double staining for Kiss1 and c-Fos in the AVPV of representative rats. Kiss1 mRNA-expressing cells, red; c-Fos-ir cells, green. Bars = 50 μm. After blood sampling, animals were perfused from 17:00 to 18:00. (E) Percentage of c-Fos-ir cells to Kiss1 mRNA-expressing cells. Statistical differences were determined by unpaired Student’s t-test. *P < 0.05 vs non-DHT rats. Numbers in each column represent the number of animals examined. (F) Plasma LH levels of rats utilized for double staining of Kiss1 and c-Fos. Statistical analysis was performed as previously described for (A) (non-DHT, n = 4; DHT, n = 5).
3. View larger version:

   • In this window
   • In a new window

   • Download as PowerPoint Slide

   Figure 3

   GnRH immunoreactivity in the preoptic area (POA) (A) and median eminence (ME) (B) of representative non-DHT and DHT rats. All animals were ovariectomized and implanted with a tube containing a high concentration of E2. Bars of upper panels = 200 µm. Lower panels are a higher magnification of upper panels. Bars = 100 µm (A), 200 µm (B). (C) Number of GnRH-immunoreactive (ir) cells in POA and area of GnRH-ir fibres in ME. No significant difference was found between the two groups. Numbers in each column represent the number of animals examined. Values are expressed as mean ± s.e.m.
4. View larger version:

   • In this window
   • In a new window

   • Download as PowerPoint Slide

   Figure 4

   Expression of Kiss1 mRNA and NKB-ir cells in the ARC and pulsatile LH secretion of non-DHT and DHT rats. All animals were ovariectomized. (A) Kiss1 mRNA-expressing cells (upper panels) and NKB-ir cells (lower panels) in the ARC. Bars = 100 µm. (B) Number of Kiss1 mRNA or NKB-ir cells in the ARC. Statistical differences were determined by unpaired Student’s t-test. *P < 0.05, vs non-DHT rats. Numbers in each column represent the number of animals examined. Values are expressed as mean ± s.e.m. (C) Plasma LH profiles of representative non-DHT OVX and DHT OVX rats. Arrowheads indicate peaks of LH pulses identified by the PULSAR computer program.
5. View larger version:

   • In this window
   • In a new window

   • Download as PowerPoint Slide

   Figure 5

   Effect of rat kisspeptin 10 (kp-10) and GnRH agonist on LH release in non-DHT and DHT rats. Kp-10 was injected (arrow) into the third ventricle (3V) 1 h after commencing blood sampling. Buserelin, a GnRH agonist, was intravenously injected into rats 1 h after commencing blood sampling. Normal diestrus rats were used as a control group (non-DHT). DHT rats were ovary-intact animals. Statistical differences were determined by two-way repeated measures ANOVA (time and group), followed by Bonferroni correction. *P < 0.05, vs blood sample just before kp-10 or buserelin injection (time 0). ^#P < 0.05, vs DHT rats. Each group, n = 6 (A). Non-DHT, n = 4; DHT, n = 6 (B).
6. View larger version:

   • In this window
   • In a new window

   • Download as PowerPoint Slide

   Figure 6

   Expression of androgen receptor (AR) on kisspeptin neurons in the AVPV and ARC of female rats. Double staining for Kiss1 and AR in the AVPV (A) and ARC (B) of normal adult female rats. Lower panels show high magnification of each upper panel. Kiss1 mRNA-expressing cells, red; AR-ir cells, green. Arrowheads indicate coexpression of Kiss1 and AR. Bars = 50 μm.

• © 2017 Society for Endocrinology