Changes in nucleolar structure,number and size in cellular activation and inactivation

In a cytophysiological study it was investigated whether in juvenile trout gonadal steroids stimulate the gonadotropic (GTH)-cells directly or indirectly via the brain. Pituitaries of donor animals were transplanted into the caudal musculature of testosterone-treated and non-testosterone-treated host fish. Testosterone treatment caused an increase in GTH-content in the in situ pituitaries and in the grafts. Accordingly, the gonadotrops displayed ultrastructural changes such as the appearance of well-developed Golgi systems and large globules. The stimulation of the morphological development of gonadotrops and of synthesis and storage of GTH in the allografted pituitaries indicates that testosterone affects the GTH-cells directly. In untreated juvenile trout the gonadotropin content of the pituitary and the gonadotropin concentration in the plasma vary with the time of year. This variation and the role of testosterone and gonadotropin-releasing hormone on the release of GTH are discussed.     

Effect of 17α-methyltestosterone,estradiol-17β and synthetic LHRH on production of gonadotropic hormone in pituitaries of rainbow trout (organ culture)

Pituitary glands from 6-month-old sexually immature female rainbow trout, Salmo gairdneri, were kept in organ culture for 48 or 72 h. Certain groups of pituitaries were cultivated for 48 h on either control medium or medium with 17 alpha-methyltestosterone (MT), or with estradiol-17 beta (E2) in concentrations of 8.5 X 10(-7) M. Other groups of pituitaries were cultivated for 72 h on control medium, or for 48 h on either control medium or MT-medium or E2-medium, and subsequently for 24 h on medium with synthetic LHRH in concentrations of 8.5 X 10(-7) M and 8.5 X 10(-10) M. Gonadotropic (GTH) cells are identified by Alcian Blue-Periodic Acid Schiff-Orange G staining and the double-antibody immunoenzyme-cytochemical technique using anti-carp beta GTH as the first antibody. A quantitative histological procedure was used to study the nuclear size of the GTH cells in response to the different hormones. Secretory activity was estimated by measuring the gonadotropin (GTH) content in extracts of pituitaries, plasma, and the culture media every 24 h by radioimmunoassay. Cultivation on MT- or E2-enriched medium results in an increase of the total amount of GTH in the pituitary and medium, an accumulation of GTH in GTH-cells (approximately 20 percentage points) and an increase in their nuclear size, indicating a stimulation of GTH synthesis. However, autonomous GTH-release is not affected by these steroids. Subsequent cultivation of the pituitaries for 24 h with LHRH causes stimulation of GTH synthesis (approximately 20 percentage points). Preincubation with steroids increases the GTH synthesis capacity of LHRH only when used in a concentration of 8.5 X 10(-10) M. Moreover, 8.5 X 10(-7) M LHRH causes a stimulation of GTH-release. Preincubation of the pituitaries with steroids increases the responsiveness of GTH-cells to LHRH. It is concluded that GTH-production in pituitaries of immature female rainbow trout can be directly influenced by gonadal steroids and by a hypophysiotropic substance.     

Effects of Castration and Androgen-treatment on Pituitary and Testes of the Three-spined Stickleback,Gasterosteus aculeatus L., in the Breeding Season

Methyltestosterone (MT) (35 μg/l aquarium water) suppresses steroid production in the testes of sham-operated fish, as studied enzyme histochemically. This treatment also diminishes the volume of the proximal pars distalis, as well as the area of the gonadotropic (GTH) cell nuclei. Such effects were not found in MT-treated castrated fish. Castration alone led to a partial degranulation of the GTH-cells as compared with sham-operated controls. Extensively dilated granulated endoplasmic reticulum was found in the GTH-cells of some of the castrated animals and also in GTH-cells of some castrated fish that had been treated with MT. The possibility of a non-steroidal feedback mechanism from testes to pituitary is discussed. No effects of the treatments were observed on prolactin cells and on the second basophilic cell type (GTH II?, TSH?).     

Effects of gonadal steroids during pubertal development on androgen and estrogen receptor‐α immunoreactivity in the hypothalamus and amygdala

Perinatal development is often viewed as the major window of time for organization of steroid‐sensitive neural circuits by steroid hormones. Behavioral and neuroendocrine responses to steroids are dramatically different before and after puberty, suggesting that puberty is another window of time during which gonadal steroids affect neural development. In the present study, we investigated whether the presence of gonadal hormones during pubertal development affects the number of androgen receptor and estrogen receptor α‐immunoreactive (AR‐ir and ERα‐ir, respectively) cells in limbic regions. Male Syrian hamsters were castrated either before or after pubertal development, and 4 weeks later they received a single injection of testosterone or oil vehicle 4 h prior to tissue collection. Immunocytochemistry for AR and ERα was performed on brain sections from testosterone‐treated and oil‐treated males, respectively. Adult males that had been castrated before puberty had a greater number of AR‐ir cells in the medial preoptic nucleus than adult males that had been castrated after puberty. There were no significant differences in ERα‐ir cell number in any of the brain regions examined. The demonstration that exposure to gonadal hormones during pubertal development is associated with reduced AR‐ir in the medial preoptic nucleus indicates that puberty is a period of neural development during which hormones shape steroid‐sensitive neural circuits. © 2000 John Wiley & Sons, Inc. J Neurobiol 44: 361–368, 2000     

Effects of gonadal steroids during pubertal development on androgen and estrogen receptor-alpha immunoreactivity in the hypothalamus and amygdala

Perinatal development is often viewed as the major window of time for organization of steroid-sensitive neural circuits by steroid hormones. Behavioral and neuroendocrine responses to steroids are dramatically different before and after puberty, suggesting that puberty is another window of time during which gonadal steroids affect neural development. In the present study, we investigated whether the presence of gonadal hormones during pubertal development affects the number of androgen receptor and estrogen receptor alpha-immunoreactive (AR-ir and ER alpha-ir, respectively) cells in limbic regions. Male Syrian hamsters were castrated either before or after pubertal development, and 4 weeks later they received a single injection of testosterone or oil vehicle 4 h prior to tissue collection. Immunocytochemistry for AR and ER alpha was performed on brain sections from testosterone-treated and oil-treated males, respectively. Adult males that had been castrated before puberty had a greater number of AR-ir cells in the medial preoptic nucleus than adult males that had been castrated after puberty. There were no significant differences in ER alpha-ir cell number in any of the brain regions examined. The demonstration that exposure to gonadal hormones during pubertal development is associated with reduced AR-ir in the medial preoptic nucleus indicates that puberty is a period of neural development during which hormones shape steroid-sensitive neural circuits.     

Gonadal hormones modulate the display of conditioned defeat in male Syrian hamsters

It has been widely reported that gonadal hormones influence the display of aggression in Syrian hamsters; conversely, much less is known about whether gonadal hormones modulate submissive/defensive behaviors in these animals. Following social defeat, male hamsters no longer display normal territorial aggression but instead display submissive/defensive behavior in the presence of a smaller opponent, a phenomenon we have termed conditioned defeat (CD). The purpose of the present study was to examine the effect of gonadal hormones on the display of CD in male hamsters. In Experiment 1, males were castrated or sham-operated. The castrated males were significantly more submissive following social defeat relative to their intact counterparts. The increased submissive behavior in the castrated males during CD testing was particularly surprising, given the fact that they were attacked significantly less during CD training. In Experiment 2a, males were castrated and given hormone replacement. Castrated males treated with testosterone or dihydrotestosterone displayed significantly less submissive behavior following social defeat than did those treated with cholesterol or estradiol. Finally, in Experiment 2b, there was no effect of hormone replacement on aggressive behavior in non-defeated hamsters suggesting that the decrease in submissive behavior in males treated with dihydrotestosterone or testosterone is specific to being previously defeated. Taken together the data indicate that the presence of androgens reduces the display of submission in defeated male hamsters. More importantly, these findings suggest that androgens may have a protective effect against the development of depression-like or anxiety-like behaviors following exposure to an ethologically relevant stressor.     

Short-day increases in aggression are inversely related to circulating testosterone concentrations in male Siberian hamsters (Phodopus sungorus)

Many nontropical rodent species display seasonal changes in both physiology and behavior that occur primarily in response to changes in photoperiod. Short-day reductions in reproduction are due, in part, to reductions in gonadal steroid hormones. In addition, gonadal steroids, primarily testosterone (T), have been implicated in aggression in many mammalian species. Some species, however, display increased aggression in short days despite basal circulating concentrations of T. The goal of the present studies was to test the effects of photoperiod on aggression in male Siberian hamsters (Phodopus sungorus) and to determine the role of T in mediating photoperiodic changes in aggression. In Experiment 1, hamsters were housed in long and short days for either 10 or 20 weeks and aggression was determined using a resident-intruder model. Hamsters housed in short days for 10 weeks underwent gonadal regression and displayed increased aggression compared to long-day-housed animals. Prolonged maintenance in short days (i.e., 20 weeks), however, led to gonadal recrudescence and reduced aggression. In Experiment 2, hamsters were housed in long and short days for 10 weeks. Half of the short-day-housed animals were implanted with capsules containing T whereas the remaining animals received empty capsules. In addition, half of the long-day-housed animals were castrated whereas the remaining animals received sham surgeries. Short-day control hamsters displayed increased aggression compared to either castrated or intact long-day-housed animals. Short-day-housed T treated hamsters, however, did not differ in aggression from long-day-housed animals. Collectively, these results confirm previous findings of increased aggression in short-day-housed hamsters and suggest that short-day-induced increases in aggression are inversely related to gonadal steroid hormones.     

Hormones and social behavior in the lizard, Anolis carolinensis

The purpose of this experiment was to study the effects of homologous and heterologous gonadal hormones on sexual and aggressive behavior in a reptilian species. Thirty adult male and thirty adult female lizards (Anolis carolinensis) were divided into 10 groups of six each (five groups per sex) and each group was given one of five treatments: either left intact, sham-castrated and injected with the hormone vehicle, castrated and injected with the hormone vehicle, castrated and injected with estradiol benzoate, or castrated and injected with testosterone propionate. After a week of visual isolation and daily hormone injection, animals were tested four times, twice with a stimulus animal of each sex. Females treated with estrogen were receptive, but did not court. Females treated with androgen were receptive and also courted and pursued stimulus females as frequently as males given androgen. No males in any group were receptive, and thus the female appears to be more capable of heterotypical sexual behavior than the male. Castrated males failed to court. Courtship and pursuit of stimulus females was readily stimulated in males with testosterone, and weakly stimulated by estrogen. Intact males were very aggressive, but lower levels of aggression were independent of gonadal hormones, as was subordination (head-nodding). The results for aggression and subordination are interpreted with reference to naturally-occurring Anolis behavior, and the results for sexual behavior are compared with similar experiments with mammals and birds.     

The effects of aromatizable androgens,non-aromatizable androgens,and estrogens on gonadotropin release in castrated African catfish,Clarias gariepinus (Burchell)

Summary To study the feedback mechanism of gonadal hormones on GTH secretion in male African catfish, the effects of castration and steroid replacement on GTH release, pituitary GTH content, and ultrastructural appearance of gonadotropes were investigated.Castration resulted in an increase in plasma GTH levels, a decrease in pituitary GTH content, and a degranulation of many gonadotropes. The aromatizable androgens testosterone and androstenedione were able to abolish the castration-induced increase in plasma GTH. This was accompanied with a restoration of pituitary GTH content and a regranulation of gonadotropes. The non-aromatizable androgens 5-dihydrotestosterone and 11-hydroxyandros tenedione did not have these effects. Replacement with estrone or estradiol resulted in an increase in pituitary GTH, however, without abolishing the elevated plasma GTH levels; ultrastructurally, many gonadotropes showed a welldeveloped granular endoplasmic reticulum together with a regranulation.The results of the present study indicate the significance of androgen aromatization in the feedback mechanism of gonadal steroids on the brain-pituitary axis.     

Testosterone regulates the density of dendritic spines in the male preoptic area

Male-typical behavior is dependent on testosterone. Castrated males gradually stop mating and engaging in sexual behaviors. Castrates treated with testosterone regain motivation and sex behaviors over time. Although this effect is well characterized, the specific mechanisms by which testosterone treatment recovers sexual behaviors remain unknown. The medial preoptic area (MPOA) is a likely site for testosterone's action on copulation. The integrity of the area is essential for the expression of male sex behavior; and the MPOA is densely populated with receptors for gonadal steroids. Moreover testosterone appears to regulate synaptic efficacy in the MPOA. Exposure to sexually relevant stimuli stimulates the MPOA but only in the presence of circulating testosterone. Sites afferent to the area respond to similar exposure independent of the hormonal milieu suggesting that testosterone mediates communication between the MPOA and its afferents. The protracted time course suggests that the effects of steroidal manipulation are mediated by structural changes. The present experiment evaluated this hypothesis by comparing dendritic spine density among Syrian hamsters that were castrated, castrated and treated with testosterone, or were left gonadally intact. Brains were sectioned and stained using the rapid Golgi stain protocol (FD Neurotechnologies, Baltimore), and the spine density, dendrite length, and the number of branches were compared among groups. Intact and testosterone replaced animals had more spines and greater spine density but did not differ in dendrite length and branching from castrated animals. These results suggest that existing dendrites increase the number of spines available for synapse formation but do not extend their dendrites in response to testosterone treatment.     

Photoperiodic and gonadal steroid regulation of pineal hydroxyindole-O-methyl transferase and N-acetyl transferase in Coturnix quail.

The activities of the melatonin-synthesizing enzymes were determined in pineals of Coturnix quail in response to photoperiodicity and gonadal hormones. Both hydroxyindole-O-methyl transferase (HIOMT) and N-acetyl transferase (NAT) were two-fold higher during exposure to darkness in female and male Coturnix maintained in a gonad-stimulating photoperiod (16L:8D). Castration decreased HIOMT activity in both female and male Coturnix. Administration of diethylstilbestrol, estradiol benzoate and progesterone into castrated females, and testosterone propionate and androstenedione into castrated males, restored HIOMT activity similar to that of intact controls. NAT was not affected by castration or gonadal steroids. These results suggest that the activity of pineal NAT is regulated primarily by photoperiodicity, while HIOMT activity is a consequence of photoperiodic and gonadal steroid regulation.     

Gonadal stage-dependent effects of gonadal steroids on gonadotropin II secretion in the Atlantic croaker (Micropogonias undulatus).

Involvement of gonadal steroids in the control of gonadotropin II (GTH II) (homologous to LH) secretion was investigated in the Atlantic croaker (Micropogonias undulatus) using gonadectomy (Gx) and steroid replacement paradigms. Gonadectomy in males and females during the late gonadal recrudescence phase elicited significant increases in the gonadotropin response to stimulation by an LHRH analog (LHRHa), without altering basal GTH II secretion. Slow-release silicone elastomer implants of testosterone or estradiol significantly inhibited LHRHa-induced GTH II secretion in gonad-intact and Gx males, and in Gx females, whereas 5alpha-dihydrotestosterone, a nonaromatizable androgen, was ineffective. Pretreatment of fish with an aromatase inhibitor, 1,4, 6-androstatrien-3,17-dione, 2 days before the administration of testosterone implants, completely blocked the negative effect of testosterone on LHRHa-induced GTH II secretion in males, but only partially restored it in females. This suggests that the negative feedback of testosterone in males is primarily mediated by its conversion to estradiol at the level of the hypothalamus and/or pituitary gland, while in females the androgen may also exert a direct inhibitory effect on GTH II secretion, probably mediated via an androgen receptor. In addition, estradiol and testosterone exerted positive effects on basal and LHRHa-induced GTH II secretion during the early-recrudescence phase of the gonadal cycle. The steroids switched to a negative effect on LHRHa-induced GTH II secretion once the fish had fully developed gonads, possibly as a mechanism that prevents a precocious surge in GTH II secretion and final gamete maturation until gametogenesis is complete and the environmental conditions are appropriate for spawning.     

Hypertrophy of gonadotropin releasing hormone-containing neurons after castration in the teleost,Haplochromis burtoni

In the African cichlid fish, Haplochromis burtoni, males are either territorial or nonterritorial. Territorial males suppress reproductive function in the nonterritorial males, and have larger gonads and larger gonadotropin-releasing hormone- (GnRH) containing neurons in the preoptic area (POA). We describe an experiment designed to establish the causal relationship between large GnRH neurons and large testes in these males by determining the feedback effects of gonadal sex steroids on the GnRH neurons. Territorial males were either castrated or sham-operated, 4 weeks after which they were sacrificed. Circulating steroid levels were measured, and the GnRH-containing neurons were visualized by staining sagittal sections of the brains with an antibody to salmon GnRH. The soma areas of antibody-stained neurons were measured with a computer-aided imaging system. Completely castrated males had markedly reduced levels of circulating sex steroids [11-ketotestosterone (11 KT) and testosterone (T)], as well as 17β-estradiol (E₂). POA GnRH neurons in castrates showed a significant increase in mean soma size relative to the intact territorial males. Hence, in mature animals, gonadal steroids act as a brake on the growth of GnRH-containing neurons, and gonadal products are not responsible for the large GnRH neurons characteristic of territorial males. © 1992 John Wiley & Sons, Inc.     

The effect of rapid and depot testosterone and estradiol on spatial performance in water maze

Men and women differ in some cognitive functions including spatial abilities. These differences seem to be affected by sex steroids, but the results are controversial. The aim of this work is to describe the effects of rapid or depot testosterone and estradiol on spatial memory in rats. Thirty-two adult male Wistar rats were divided into 6 groups. Five groups were gonadectomized, and one group was left as control. Castrated groups received sterile oil, testosterone isobutyras, testosterone propionate, estradiol dipropionate or estradiol benzoate. We evaluated spatial performance (escape latency, overall improvement, and time in the quadrant after platform removal) of the rats in a spatial water maze. Animals receiving exogenous sex steroids showed higher plasma concentrations of the particular hormones. Experimental groups improved during the acquisition spatial trials in the water maze. No significant differences between the groups during probe trial were found. In overall improvement, the testosterone depot and estradiol depot groups showed less improvement in comparison to the control groups (P<0.05). No differences in respect to administered hormones were found in corresponding receptor gene expression in hippocampus. In conclusion, exogenous testosterone affects spatial memory of adult castrated males.     

Hormonal Induction of Male Courtship Behavior in the Japanese Newt, Cynops pyrrhogaster

In the breeding season, the sexually mature male newt, Cynops pyrrhogaster, vibrates the tail in front of the female at an early stage of courtship. Effects of prolactin (PRL), gonadotropin (GTH), and sex steroids on this behavior were investigated in the male paired with the female receiving PRL and GTH. The behavior was elicited in the sexually inert male by injections of PRL of bovine, ovine, or bullfrog origin and human chorionic gonadotropin or bullfrog LH and FSH in combination. The effect of PRL or GTH alone was less marked than that of PRL plus GTH, especially in terms of frequency of the behavior. In the hypophysectomized male, combination of PRL and GTH significantly increased both the incidence and frequency of the behavior. However, PRL alone was not effective, and the effect of GTH alone was less pronounced than that in the intact animal receiving GTH injections. The effect of GTH was nullified by castration. In the PRL-treated castrated animal, testosterone or dihydrotestosterone, but not estradiol, was effective in inducing the behavior.     

Sex differences in the activity of mice: modulation by postnatal gonadal hormones

A series of six experiments was performed to examine the influence of postnatal-gonadal-hormone exposure on home-cage activity in Rockland-Swiss albino mice. Intact females were more active than their male counterparts and gonadectomy in adulthood, while reducing levels of the behavior in both sexes, did not eliminate the gender difference. Males that were castrated on the day of birth were more active than animals castrated 5, 10, or 25 days later. Also, females treated with testosterone propionate on the day of birth were less active than oil-treated controls and females exposed to the steroid 10 days after birth. Thus, perinatal exposure to gonadal hormones suppresses adult levels of home-cage activity in mice.     

Effects of photoperiod on pituitary gonadotropin levels in masu salmon

We examined the effects of photoperiod on pituitary levels of two types of gonadotropin (GTH), GTH I and GTH II, in masu salmon Oncorhynchus masou to study their mechanism of synthesis. In Experiment 1, the effects of long or short photoperiod combined with castration were examined using 8-month-old precocious males. Castration was carried out in early August and then the fish were reared under a short (8L16D) or long (16L8D) photoperiod for 60 days. In Experiment 2, the effects of photoperiod combined with testosterone treatment were examined using 12-month-old immature females. Silastic tubes containing testosterone (500 microg /fish) or vehicle were implanted intra-peritoneally in early October. Fish were reared under 16L8D for 60 days, and then half of the fish were transferred to 8L16D, while the remaining fish were kept under 16L8D until Day 90. In Experiment 1, GTH I contents were higher under 16L8D than under 8L16D in the castrated group on Day 30. Moreover, GTH I contents were higher in the castrated group than the control group under 16L8D on Day 30. GTH II contents increased with testicular maturation in the control groups, whereas they remained at low levels in the castrated groups regardless of photoperiodic treatment. In Experiment 2, GTH I contents did not change remarkably in all the groups, while GTH II contents were remarkably increased by testosterone treatment regardless of photoperiodic treatment. These results indicate that the synthesis of GTH I and GTH II are differently regulated by photoperiod and testosterone in masu salmon.     

Apoptosis during sexual differentiation of the bed nucleus of the stria terminalis in the rat brain

The bed nucleus of the stria terminalis (BST) in the rat forebrain differs between males and females. To test whether apoptosis may contribute to the development of sex differences in the BST, the incidence of apoptosis was determined in sham-treated males and sham-treated females sacrificed on postnatal days (PN) 2, 4, 6, 8, 10, and 12 (PN 1 being day of birth). More apoptotic nuclei were found in the principal nucleus of the BST (BSTpr) in females than in males, whereas the reverse was true for the lateral division of the BST (BSTl). Moreover, the volume of the BSTpr was larger in males than in females, whereas there was no sex difference in the volume of the BSTl. Our results also confirmed earlier reports indicating that the incidence of apoptosis in the central part of the medial preoptic nucleus (MPNc) is higher in females than in males. No sex difference in apoptosis was found in the ventromedial hypothalamus (VMH) and paraventricular nucleus (PVN). The volume of the MPNc and VMH was larger in males than in females, whereas the PVN volume did not differ between males and females. To test whether sex differences in neonatal levels of gonadal steroids may cause sex differences in the incidence of apoptosis in the BSTpr, the incidence of apoptosis was compared between castrated males and females that were treated with testosterone propionate or vehicle on the day of birth. In the BSTpr of gonadal steroid-treated animals, the incidence of apoptosis was lower when compared to animals treated with vehicle, which was also true for the MPNc. These results are consistent with the hypothesis that gonadal steroids contribute to the sexually dimorphic differentiation of the BST by controlling the incidence of apoptosis.     

Effects of exogenous estradiol-17β on early growth and gonadal development of diploid and triploid female rainbow trout (Oncorhyncus mykiss)

Triploidy is a viable condition in teleosts. However, in many salmonids, the triploid condition in the female results in sterility as gametogenesis appears to be disrupted. Although the underlying mechanisms regulating the gonadal development of teleosts have not been clearly elucidated, the reversal of phenotypic sex by the administration of the appropriate exogenous steroid during early development supports the argument that gonadal steroids play a pivotal role in sexual differentiation and subsequent gonad development in these fish. To determine whether the failure of normal ovarian development in triploid female rainbow trout (Oncorhynchus mykiss) is due to an absence or reduction of endogenous sex steroids, ovarian morphology was compared between diploid and triploid juvenile rainbow trout treated with exogenous estradiol-17β (E₂). The ovaries of both untreated and E₂ treated diploid fish, at 145 days post-fertilization, contained synchronously developing oocytes in the perinucleolar stage, whereas ovaries from untreated and estradiol-treated triploid fish of the same age were considerably smaller and devoid of developing oocytes. No differences in the ovaries of triploid untreated fish and triploid fish treated with E₂ were observed. It is reported that exposure to exogenous E₂ during the period of gonadal differentiation is not sufficient to induce oocyte development in triploid rainbow trout. © 1996 Wiley-Liss, Inc.     

Hormonal regulation of adult sympathetic neurons: the effects of castration on neuropeptide Y, norepinephrine, and tyrosine hydroxylase activity

Previous studies utilizing the hypogastric ganglia (HG) have indicated that gonadal steroids exert organizational and activational effects on noradrenergic biochemistry. Bilateral castration of male rodents at birth prevents the normal maturation of tyrosine hydroxylase (T-OH) activity in the HG; castration during adulthood results in a progressive decline in T-OH activity. Testosterone replacement corrects both the ontogenetic and adult functional alterations in the neurotransmitter-synthesizing enzyme. The present studies in adult male rats extend these previous observations and asked the question whether gonadal steroids regulate the neurotransmitters neuropeptide Y (NPY) and norepinephrine (NE) in the HG. Adult rodents were castrated and ganglia T-OH, NPY, and NE were examined at various time points after surgery. All three indices of sympathetic neuron biochemistry declined following castration, but they exhibited different profiles. It appears that hormones may affect enzyme activity and neurotransmitter pools differently within neurons. Testosterone replacement therapy fully restored T-OH activity, and NPY and NE levels in the HG. These studies extend the activational role of testosterone in regulating sympathetic neuron neurotransmitter and neuropeptide levels as well as neurotransmitter-synthesizing enzymes.