Differential mechanisms for regulation of the stress response across latitudinal gradients.

We examined plasticity of the stress response among three populations of the white-crowned sparrow (Zonotrichia leucophrys). These populations breed at different elevations and latitudes and thus have breeding seasons that differ markedly in length. We hypothesize that in populations where birds raise only one or rarely two broods in a season, the fitness costs of abandoning a nest are substantially larger than in closely related populations that raise up to three broods per season. Thus individuals with short breeding seasons should be less responsive to stressors and therefore less likely to abandon their young. In our study, baseline and handling-induced corticosterone levels were similar among populations, but corticosteroid-binding globulins differed, leading to a direct relationship between stress-induced free corticosteroid levels and length of breeding season. There were also population-specific differences in intracellular low-affinity (glucocorticoid-like) receptors in both liver and brain tissue. Although investigations of population-based differences in glucocorticoid secretion are common, this is the first study to demonstrate population-level differences in binding globulins. These differences could lead to dramatically different physiological and behavioral responses to stress.

Effects of acute treatment with 8-OH-DPAT and fluoxetine on aggressive behaviour in male song sparrows (Melospiza melodia morphna).

The role of serotonin in modulating male aggressive behaviour was investigated in male song sparrows, Melospiza melodia morphna, using two different serotonergic drugs, fluoxetine and 8-OH-DPAT. Fluoxetine is a selective serotonin reuptake inhibitor of the neuronal reuptake pump increasing synaptic concentrations of serotonin, and 8-OH-DPAT is a specific serotonin (5-HT1A) receptor agonist. The serotonergic control of aggression in passerines has not been previously investigated. We examined these behaviours within a controlled setting using a laboratory simulated territorial intrusion, with a hierarchical scale to quantify male-male aggressive behaviour. Utilizing this scale, we quantified the extent of male aggressive behaviour in two experiments. In experiment 1, song sparrows were given 100 micro l, s.c. injections of either fluoxetine (10 mg/kg) or 8-OH-DPAT (1 mg/kg). Experiment 2 was a dose-response study using three doses of 8-OH-DPAT (0.1, 1 and 10 mg/kg). In both studies, aggressive behaviour was measured 1 h after injection for 10 min in response to the presence of a novel male decoy combined with playback of conspecific song. Both drugs significantly reduced male aggressive behaviour, and 8-OH-DPAT did so in a dose-dependent manner. The effect of the two drugs upon general activity was also measured using infra-red perch hop detectors. Activity levels were not effected by either fluoxetine or 8-OH-DPAT at all of the respective doses, indicating that the reduction in aggressive behaviour was specific. These results demonstrate that, in a passerine species, the serotonergic system negatively regulates male-male aggressive behaviour. These results further demonstrate that aggression can be effectively studied in a laboratory setting and natural aggressive responses can be elicited using this method.

Androgen binding profiles of two distinct nuclear androgen receptors in Atlantic croaker (Micropogonias undulatus).

In the present study, the binding affinities of 28 androgens for two nuclear androgen receptors (AR), termed AR1 and AR2, in Atlantic croaker (Micropogonias undulatus) brain and ovarian tissues, respectively, were determined using competitive binding assays. The 5alpha-reduction of steroids, in general, increased the metabolite's binding affinity for AR2 while decreasing it for AR1. In addition, few androgens bound to AR1 with high affinity and modifications to the basic 3-ketone,4-ene,17beta-hydroxy structure of testosterone usually reduced its binding affinity for AR1. However, androgens with ketone groups at the 3- and 17-position bound with high affinity to AR1 provided that the androgen had either a 5alpha-reduced A-ring or a third ketone group at the 11-position. This suggests that there may be several high affinity conformations that AR1 can occupy depending upon whether an androgen possesses a ketone or a hydroxyl group at the 17-position. The binding of androgens to AR2 showed a more predictable pattern, 5alpha-reduced steroids bound better than 4-ene steroids and any changes to the basic 3-keto,17-hydroxy motif of 5alpha-dihydrotestosterone lowered the binding affinity of a steroid. However, these structural changes often caused only minor decreases in binding affinity, such that AR2 has a broader affinity for androgens and a greater affinity than AR1 for structurally diverse androgens. Widely different androgen binding affinities of AR1 and AR2 suggest that these two nuclear androgen receptors may mediate the physiological actions of different androgens in teleosts.

Identification of two nuclear androgen receptors in kelp bass (Paralabrax clathratus) and their binding affinities for xenobiotics: comparison with Atlantic croaker (Micropogonias undulatus) androgen receptors.

Two distinct nuclear androgen receptors (ARs) were identified in brain and ovarian tissues of kelp bass, Paralabrax clathratus, termed kbAR1 and kbAR2, which correspond to the two nuclear ARs we have previously characterized in Atlantic croaker, Micropogonias undulatus, termed acAR1 and acAR2. Scatchard analysis of nuclear fractions of whole brain tissue demonstrated that kbAR1 had a single class of high-affinity binding sites for testosterone (T; K(d) of 1. 8 nM and B(max) of 1.0 pmol/g tissue), whereas cytosolic fractions of kbAR2 ovarian tissue had a single class of high-affinity binding sites for dihydrotestosterone (DHT; K(d) of 0.1 nM and B(max) of 0.5 pmol/g tissue). Competition studies showed that both kbAR1 and kbAR2 were specific for androgens. However, kbAR1 bound only T with high affinity, whereas kbAR2 bound DHT, mibolerone, 17alpha-methyl-testosterone, T, and 11-ketotestosterone with high affinity. In addition, we examined the binding affinities of dichlorodiphenyltrichloroethane and its derivatives, several hydroxylated polychlorinated biphenyl (PCB) congeners, PCB mixtures, and the fungicide vinclozolin and its two metabolites M1 and M2 for the two ARs in Atlantic croaker ovarian, testicular, and brain tissues and in kelp bass ovarian and brain tissues. Only 4, 4'-PCB-3-OH and 2',5'-PCB-3-OH demonstrated greater than 50% displacement of [(3)H]testosterone from either acAR1 or kbAR1. In contrast, with the exception of vinclozolin, all of the xenobiotics examined demonstrated binding to acAR2 in testicular and ovarian tissues. The binding affinities were highest in the testicular tissue with M2, 2,2'5'-PCB-4-OH, and o,p'-DDD all binding with EC(50)s less than 10 microM. The binding affinities of xenobiotics to kbAR2 in ovarian tissue were similar to their binding affinities for ovarian acAR2. The finding that AR1 and AR2 possess different binding affinities for natural androgens and synthetic steroids, as well as for xenobiotics, suggests that the activities of androgens and of certain xenobiotics will depend upon the type of AR present within the target tissue.

Characterization of two nuclear androgen receptors in Atlantic croaker: comparison of their biochemical properties and binding specificities.

Two distinct androgen receptors (ARs) with different characteristics were identified in the brain and ovary of Atlantic croaker, Micropogonias undulatus. A nuclear AR, AR1, was identified in the brain that had high affinity binding sites for testosterone (T; Kd, 1.1 +/- 0.15 nM; binding capacity, 1.4 +/- 0.14 pmol/g tissue; n = 16). A second nuclear AR, AR2, was found in the ovary that had high affinity binding sites for 5alpha-dihydrotestosterone (DHT; Kd, 0.62 +/- 0.1 nM; binding capacity, 0.38 +/- 0.06 pmol/g tissue; n = 14). AR2 has physiochemical properties similar to those of other vertebrate ARs. AR2 has high affinity binding for a broad spectrum of natural and synthetic androgens, including 17alpha-methyl-5alpha-dihydrotestosterone, which has a relative binding affinity of DHT = 100% > T > mibolerone > 11-ketotestosterone = 16%, a rapid association (t1/2, 44 min) and a slow dissociation (t1/2, 45 h) rate, as well as specific binding to purified DNA. The cytosolic AR2 interacts with heat shock proteins in a manner similar to other steroid receptors, as sodium molybdate stabilizes the receptor, and it has a 7.4-7.8S sedimentation coefficient in a 5-20% sucrose gradient. In contrast, AR1 is highly specific for only a few androgens, with T = 100% relative binding affinity >> DHT >> 11-ketotestosterone > mibolerone > 17alpha-methyl-5alpha-dihydrotestosterone = 0, has rapid association (t1/2, 15 min) and dissociation (t1/2, 2.6 +/- 0.7 h) rates, and has specific binding to purified DNA upon heat activation. The cytosolic binding component sediments at 5.6-5.7S in a 5-20% sucrose gradient and is not affected by sodium molybdate, which suggests that AR1 does not interact with heat shock proteins in the usual manner. This is the first report of the presence of two different nuclear ARs displaying markedly different steroid binding specificities within a single vertebrate species.