Testosterone-dependent plasticity of avian forebrain neurons is not restricted to the song control system.

Testosterone is acting on brain areas involved in the control of sexual behaviour, for example the preoptic area and the song system. We now demonstrate that it also affects other avian brain areas, as exemplified here by measurement of spine densities. Depletion of testosterone by castration or application of cyproterone acetate leads to a decrease in spine density in secondary sensory areas like lateral neo- and hyperstriatum and hyperstriatum accessorium and dorsale, or in associative areas such as the caudal archi- and neostriatum. We conclude that testosterone is acting directly on the spines, and suggest that the mechanism of spine density control by hormones may have arisen because of energy demands.

Effects of castration and testosterone treatment on the activity of testosterone-metabolizing enzymes in the brain of male and female zebra finches.

Recently, we described the distribution of testosterone-metabolizing enzymes (i.e., aromatase, 5 alpha- and 5 beta-reductases) in the zebra finch (Taeniopygia guttata) brain using a sensitive radioenzyme assay combined to the Palkovits punch method. A number of sex-differences in the activity of these enzymes were observed especially in nuclei of the song-control system. The hormonal controls of these differences have now been analyzed by gonadectomizing birds of both sexes and by giving them a replacement therapy with silastic implants of testosterone (T). Five nuclei of the song system (Area X [X], nucleus magnocellularis of the anterior neostriatum [MAN], nucleus robustus archistriatalis [RA], nucleus intercollicularis [ICo], hyperstriatum ventrale, pars caudalis [HVc]) and three preoptic-hypothalamic areas (preoptic anterior [POA], periventricular magnocellular nucleus [PVM], and posterior medial hypothalamic nucleus [PMH]) were studied as well as other limbic and control non-steroid-sensitive areas. The activity of the 5 alpha-reductase was higher in males than in females for the five song-control nuclei and was not affected by the hormonal treatments. The overall activity of this enzyme was not sexually dimorphic in POA and PVM. It was higher in males than in females in intact birds only, and was reduced by gonadectomy and enhanced by T. The activity of the 5 beta-reductase was higher in females than in males in all nuclei of the song system and in POA, but was not influenced by the changes in T level. Both sex and treatment effects were observed in the control of aromatase. The production of estrogens was dimorphic (females greater than males) in RA and PMH. It was increased by T in POA, PVM, and PMH, and also in RA. These data show that some of the sex differences in T-metabolizing enzymes result from the exposure to different levels of T in adulthood (e.g., 5 alpha-reductase in POA and PVM or aromatase in PVM), whereas others persist even if birds are exposed to the same hormonal conditions. These are presumably the result of organizational effects of steroids. The steroid modulation of the aromatase might be related directly to the activation of sexual, aggressive, and nest-building behaviors, whereas the stable dimorphism in 5 alpha- and 5 beta-reductase observed in the nuclei of the song system might be one of the neurochemical bases of the sex differences in the vocal behavior of the zebra finch.

Sex- and age-related differences in the activity of testosterone-metabolizing enzymes in microdissected nuclei of the zebra finch brain.

Many effects of testosterone (T) in the zebra finch (Taeniopygia guttata) can be mimicked by T-metabolites, mainly estradiol and 5 alpha-dihydrotestosterone. We have therefore studied the neuroanatomical distribution of testosterone-metabolizing enzymes by means of the Palkovits punch technique combined with radioenzyme assay in the brain of adult and young male and female zebra finches. The activity of these enzymes was studied by a one-point assay in 5 nuclei of the song system (X, MAN, HVc, RA, ICo), 2 nuclei of the visual system (ectostriatum, nucleus rotundus) and in limbic and hypothalamic areas. Very noticeable was the presence of a very high aromatase activity in the hippocampal and parahippocampal region and in the nucleus taeniae and the absence of this enzyme in ICo. We found a higher aromatase activity in female than male HVc and RA and a higher 5 alpha-reductase activity in MAN, HVc, RA and ICo of males compared to females. The 5 alpha-reductase was more active in the preoptic area of females. A few sex-related differences in the activity of the 5 beta-reductase were also observed (higher activity in females than in males for area X and RA, but difference in the opposite direction for the ectostriatum). The statistical significance of these differences depended, to some extent, on the statistical technique used to demonstrate them, with the sex differences in RA being by far the most robust ones. Many age-related metabolic differences were also detected but these do not have a clear interpretation since the Km of these enzymes also changes with age. Extremely low levels of 5 beta-reductase activity were found in the nuclei of the visual system in adult birds while this enzymatic activity was very high in young birds. The biological significance of this change with age remains obscure. Correlations are thus observed between the neuroanatomical distribution of T-metabolizing enzymes and of androgen and estrogen receptors with the important exception of ICo which has no aromatase but contains high concentrations of estrogen receptors. Testosterone-metabolizing enzymes are however also present in areas which are not known as steroid targets.

Changes in the activity of testosterone-metabolizing enzymes in the brain of male and female zebra finches during the post-hatching period.

The kinetic properties of the testosterone-metabolizing enzymes were studied in the hypothalamus of adult and young zebra finches of both sexes. Estradiol, 5 alpha-dihydrotestosterone, 5 beta-dihydrotestosterone and 5 beta-androstane-3 alpha,17 beta-diol were identified as metabolites of testosterone in males and females at different ages between 5 days post-hatch and adulthood. During maturation, the maximum velocity (Vmax) of the aromatase and 5 beta-reductase decreased in males and females while the affinity of these enzymes for the substrate increased (decrease in km). These changes were more pronounced for 5 beta-reductase than for aromatase. The affinity change of the 5 beta-reductase occurred progressively during the post-hatching development between 5 and 30 days and is thus probably a true developmental process. In the case of the aromatase, the change in affinity occurred much later (after 30-40 days) and is thus probably related to the sexual maturation. These kinetic changes during development are directly related to the roles played by testosterone and its metabolites, in particular estradiol, in the differentiation and activation of reproductive behavior in the zebra finch. In particular, the dramatic decrease in 5 beta-reductase activity during sexual maturation should correspond to a potentiation of testosterone action in the brain.

Urinary oestrogen and progesterone excretion before and during pregnancy in a pied bare-face tamarin (Saguinus bicolor bicolor).

Oestrone, oestradiol-17 beta and progesterone were measured by radioimmunoassay in daily urine samples after pairing and during subsequent pregnancy in a pied bare-face tamarin. On the basis of excretion profiles an ovarian cycle length of about 3 weeks and a gestation length of about 160 days are suggested. Oestrone was the predominant urinary oestrogen excreted by the non-pregnant and pregnant pied bare-face tamarin, the oestrone/oestradiol ratio being greater than 100:1. The results suggest that steroid monitoring can provide useful information about reproductive physiology in this species of tamarin.

Brain testosterone metabolism during ontogeny in the zebra finch.

The testosterone metabolism in different parts of the brain and in the syrinx was studied by an in vitro radioenzyme assay in male and female zebra finch (Taeniopygia guttata castanotis) at various ages between 1 day post-hatch and adulthood. The 5 beta-reductase appeared in all experiments as the main enzyme involved in testosterone metabolism. Two experiments allowed to determine the kinetic parameters of the 5 beta-reductase. It was shown that the Vmax of the 5 beta-reduction is much higher in very young animals than in adults but that the Km of the enzyme is smaller (higher affinity) in the latter than in the former. A major decrease of 5 beta-reductase activity is observed during the ontogeny of both males and females in all the tissue samples which were studied including the hypothalamus and brain regions containing nuclei of the song system. This high reductase activity which produces behaviorally inactive compounds is likely to play a protective role during the sexual differentiation of the birds and in this way prevent the masculinization in females of the song system by the relatively high levels of circulating androgens.

[Effect of testosterone on the beginning of singing in the zebra finch (Taeniopygia guttata castanotis)]. / Der Einfluss von Testosteron auf den Gesangsbeginn beim Zebrafinken (Taeniopygia guttata castanotis)

In young male Zebra finches, singing was advanced in time by injections of testosterone-propionate. The juvenile song, which normally occurs on day 34 to 37, could be shifted to day 18 to 19, when injection took place at the 16th day; either earlier or later injections delayed singing (at 25 to 35 days). The advanced juvenile song differs from the normal one, mainly the rhythm is less mature. There seems to be a point prior to which juvenile song cannot be induced, depending possible on the maturation of central nervous motor centers.