Specific Activation of Estrogen Receptor Alpha and Beta Enhances Male Sexual Behavior and Neuroplasticity in Male Japanese Quail

Two subtypes of estrogen receptors (ER), ERα and ERβ, have been identified in humans and numerous vertebrates, including the Japanese quail. We investigated in this species the specific role(s) of each receptor in the activation of male sexual behavior and the underlying estrogen-dependent neural plasticity. Castrated male Japanese quail received empty (CX) or testosterone-filled (T) implants or were daily injected with the ER general agonist diethylstilbestrol (DES), the ERα-specific agonist PPT, the ERβ-specific agonist DPN or the vehicle, propylene glycol. Three days after receiving the first treatment, subjects were alternatively tested for appetitive (rhythmic cloacal sphincter movements, RCSM) and consummatory aspects (copulatory behavior) of male sexual behavior. 24 hours after the last behavioral testing, brains were collected and analyzed for aromatase expression and vasotocinergic innervation in the medial preoptic nucleus. The expression of RCSM was activated by T and to a lesser extent by DES and PPT but not by the ERβagonist DPN. In parallel, T fully restored the complete sequence of copulation, DES was partially active and the specific activation of ERα or ERβ only resulted in a very low frequency of mount attempts in few subjects. T increased the volume of the medial preoptic nucleus as measured by the dense cluster of aromatase-immunoreactive cells and the density of the vasotocinergic innervation within this nucleus. DES had only a weak action on vasotocinergic fibers and the two specific ER agonists did not affect these neural responses. Simultaneous activation of both receptors or treatments with higher doses may be required to fully activate sexual behavior and the associated neurochemical events

Percentage of birds that displayed at least one mount attempt (A) or one cloacal contact movement (B) and cumulative frequencies of these two behaviors in the 5 experimental groups (C–D).

<p>Data were analyzed by appropriate analyses of variance or χ2 tests that were followed by post-hoc tests specifically comparing the 4 experimental groups to the controls (see text). Results of these post-hoc comparisons are shown at the top of the bars as follows: * =  p<0.05, ** =  p<0.01 and *** =  p<0.001.</p

Cloacal gland area (in mm²) at the end of the experiment (A), percentage of birds that displayed at least one female-induced rhythmic cloacal sphincter movement (RCSM)(B) and cumulative frequencies of these RCSM in the 5 experimental groups (C).

<p>Data were analyzed by appropriate analyses of variance or c2 tests that were followed by post-hoc tests specifically comparing the 4 experimental groups to the controls (see text). Results of these post-hoc comparisons are shown at the top of the bars as follows: * =  p<0.05, ** =  p<0.01 and *** =  p<0.001.</p

Photomicrographs illustrating the aromatase-immunoreactive perikarya (A) and the vasotocin-immunoreactive fibers (B, C) present within the medial preoptic nucleus (POM) that were quantified in the present study.

<p>Panel A illustrates the dense group of aromatase-immunoreactive neurons that outline the entire POM. The dotted line marks the limits of the POM as they were defined for quantification. Panel B shows the accumulation of vasotocin-immunoreactive fibers in the POM at the level of the anterior commissure. The rectangle drawn with a solid line indicates the area where quantification was performed that is illustrated at higher magnification in panel C. The dotted rectangle indicates how the camera field was originally placed before being moved to its final location (see text). Note that quantification of fibers concerned the steroid-sensitive network located in the POM, not the denser network located more ventrally that originates from the magnocellular neurons. CA: commissural anterior, LFB: latera forebrain bundle. Magnification bar =  500 µm in A–B, 100 µm in C.</p

Modulation of testosterone-dependent male sexual behavior and the associated neuroplasticity.

Steroids modulate the transcription of a multitude of genes and ultimately influence numerous aspects of reproductive behaviors. Our research investigates how one single steroid, testosterone, is able to trigger this vast number of physiological and behavioral responses. Testosterone potency can be changed locally via aromatization into 17b-estradiol which then activates estrogen receptors of the alpha and beta subtypes. We demonstrated that the independent activation of either receptor activates different aspects of male sexual behavior in Japanese quail. In addition, several studies suggest that the specificity of testosterone action on target genes transcription is related to the recruitment of specific steroid receptor coactivators. We demonstrated that the specific down-regulation of the coactivators SRC-1 or SRC-2 in the medial preoptic nucleus by antisense techniques significantly inhibits steroid-dependent male-typical copulatory behavior and the underlying neuroplasticity. In conclusion, our results demonstrate that the interaction between several steroid metabolizing enzymes, steroid receptors and their coactivators plays a key role in the control of steroid-dependent male sexual behavior and the associated neuroplasticity in quail