Testosterone-treated Females Research Articles

Developmental Programming: Deficits in Reproductive Hormone Dynamics and Ovulatory Outcomes in Prenatal, Testosterone-Treated Sheep1

Prenatal testosterone excess leads to neuroendocrine, ovarian, and metabolic disruptions, culminating in reproductive phenotypes mimicking that of women with polycystic ovary syndrome (PCOS). The objective of this study was to determine the consequences of prenatal testosterone treatment on periovulatory hormonal dynamics and ovulatory outcomes. To generate prenatal testosterone-treated females, pregnant sheep were injected intramuscularly (days 30-90 of gestation, term=147 days) with 100 mg of testosterone-propionate in cottonseed oil semi-weekly. Female offspring born to untreated control females and prenatal testosterone-treated females were then studied during their first two breeding seasons. Sheep were given two injections of prostaglandin F2alpha 11 days apart, and blood samples were collected at 2-h intervals for 120 h, 10-min intervals for 8 h during the luteal phase (first breeding season only), and daily for an additional 15 days to characterize changes in reproductive hormonal dynamics. During the first breeding season, prenatal testosterone-treated females manifested disruptions in the timing and magnitude of primary gonadotropin surges, luteal defects, and reduced responsiveness to progesterone negative feedback. Disruptions in the periovulatory sequence of events during the second breeding season included: 1) delayed but increased preovulatory estradiol rise, 2) delayed and severely reduced primary gonadotropin surge in prenatal testosterone-treated females having an LH surge, 3) tendency for an amplified secondary FSH surge and a shift in the relative balance of FSH regulatory proteins, and 4) luteal responses that ranged from normal to anovulatory. These outcomes are likely to be of relevance to developmental origin of infertility disorders and suggest that differences in fetal exposure or fetal susceptibility to testosterone may account for the variability in reproductive phenotypes.

XenopusVocalizations Are Controlled by a Sexually Differentiated Hindbrain Central Pattern Generator

Male and female African clawed frogs (Xenopus laevis) produce rhythmic, sexually distinct vocalizations as part of courtship and mating. We found that Xenopus vocal behavior is governed by a sexually dimorphic central pattern generator (CPG) and that fictive vocalizations can be elicited from an in vitro brain preparation by application of serotonin or by electrical stimulation of a premotor nucleus. Male brains produced fictive vocal patterns representing two calls commonly produced by males in vivo (advertisement and amplectant call), as well as one call pattern (release call) that is common for juvenile males and females in vivo but rare for adult males. Female brains also produced fictive release call. The production of male calls is androgen dependent in Xenopus; to test the effects of androgens on the CPG, we examined fictive calling in the brains of testosterone-treated females. Both fictive male advertisement call and release call were produced. This suggests that all Xenopus possess a sexually undifferentiated pattern generator for release call. Androgen exposure leads to a gain-of-function, allowing the production of male-specific call types without prohibiting the production of the undifferentiated call pattern. We also demonstrate that the CPG is located in the brainstem and seems to rely on the same nuclei in both males and females. Finally, we identified endogenous serotonergic inputs to both the premotor and motor nuclei in the brainstem that may regulate vocal activity in vivo.

Androgen-dependent maternal effects on offspring fitness in zebra finches

There is accumulating evidence that maternal hormones may play a role in offspring sex adjustment, but little is known about the costs of such hormone-mediated mechanisms. Recent studies have reported sex-specific effects of hormones on offspring viability. Specifically, we previously found that elevating the plasma androgen level in mothers results in a male-biased offspring primary sex ratio, but it affects the viability of sons negatively and daughters positively in zebra finches (Taeniopygia guttata; Rutkowska and Cichon, Anim Behav, 71:1283–1288, 2006). In this study, we studied further fitness consequences of exposure to elevated yolk androgen levels in zebra finches. We measured growth rate and cellular immune response of nestlings that hatched from eggs laid by females injected with testosterone during egg laying and nestlings of unaffected control females. We found that sons of testosterone-treated females grew slower in comparison to sons of control females. The significant interaction between experimental group and offspring sex indicates that sons of testosterone-treated mothers suffered impaired immune responsiveness while daughters seemed to benefit from elevated androgen level in terms of enhanced immune responsiveness. We found no effects of androgens on offspring performance at adulthood—neither fecundity of females nor attractiveness of males was affected. We conclude that the benefits of biasing sex ratio towards males by increasing androgen level in the yolk may be limited due to negative effects on male offspring performance early in life.

Testosterone induction of male‐typical sexual behavior is associated with increased preoptic NADPH diaphorase and citrulline production in female whiptail lizards

In rodents, male-typical copulatory behavior is generally dependent on gonadal sex steroids such as testosterone, and it is thought that the mechanism by which the hormone gates the behavior involves the gaseous neurotransmitter nitric oxide. According to one model, testosterone induces an up-regulation of nitric oxide synthase (NOS) in the preoptic area, increasing nitric oxide synthesis following exposure to a sexual stimulus. Nitric oxide in turn, possibly through its effect on catecholamine turnover, influences the way the stimulus is processed and enables the appropriate copulatory behavioral response. In whiptail lizards (genus Cnemidophorus), administration of male-typical levels of testosterone to females induces the display of male-like copulatory responses to receptive females, and we hypothesized that this radical change in behavioral phenotype would be accompanied by a large change in the expression of NOS in the preoptic area. As well as comparing NOS expression using NADPH diaphorase histochemistry between testosterone-treated females and controls, we examined citrulline immunoreactivity (a marker of recent nitric oxide production) in the two groups, following a sexual stimulus and following a nonsexual stimulus. Substantially more NADPH diaphorase-stained cells were observed in the testosterone-treated animals. Citrulline immunoreactivity was greater in testosterone-implanted animals than in blank-implanted animals, but only following exposure to a sexual stimulus. This is the first demonstration that not only is NOS up-regulated by testosterone, but NOS thus up-regulated is activated during male-typical copulatory behavior.

Maternal testosterone affects the primary sex ratio and offspring survival in zebra finches

Female birds have repeatedly been reported to adjust the primary sex ratio of their offspring to environmental, social and physiological cues. However, the mechanism behind sex adjustment remains unknown. It has been suggested that maternal hormones may constitute an important mediator in this mechanism, as androgen levels differ between eggs bearing male and female embryos. To evaluate whether the level of maternal androgens affects the offspring sex ratio, we injected female zebra finches, Taeniopygia guttata, with testosterone during egg laying. The sex ratio of eggs laid after testosterone administration became significantly male biased, compared to eggs laid by control females that received a vehicle injection. However, sons of testosterone-treated females suffered lower hatching success. In contrast, daughters seemed to benefit from elevated androgen level in terms of future survival prospects. The opposite effects on male and female offspring may constitute an important constraint on maternal androgen allocation to the eggs and reduce the benefits of biasing the sex ratio towards males by increasing the testosterone level.

Testosterone enhances early cardiac remodeling after myocardial infarction, causing rupture and degrading cardiac function

Cardiac rupture can be fatal after myocardial infarction (MI). Experiments in animals revealed gender differences in rupture rate; however, patient data are controversial. We found a significantly higher rupture rate in testosterone-treated female mice within 1 wk after MI, whereas castration in males significantly reduced rupture. We hypothesized that testosterone may adversely affect remodeling after MI, exaggerating the inflammatory response and increasing cardiac rupture, whereas estrogen may be cardioprotective, attenuating early remodeling and reducing rupture rate. We studied the effect of gender and hormone manipulation on morphological and histological changes during early remodeling after MI in 4-wk-old male and female C57BL/6J mice and how these events could affect cardiac function. Females were randomly divided into 1) sham ovariectomy + placebo (s-ovx + P), 2) s-ovx + testosterone (T), 3) ovx + P, and 4) ovx + T; males were divided into 1) sham castration + P (s-cas + P), 2) s-cas + 17beta-estradiol (E), 3) cas + P, and 4) cas + E. At 6 wk after gonadectomy and hormone manipulation, MI was induced. Mice were randomly killed 1, 2, 4, 7, and 14 days after MI. The left ventricle was weighed and sectioned for evaluation of MI size, infarct expansion index (IEI), and neutrophil infiltration. Transthoracic echocardiography was performed in conscious mice in the 14-day group before organ harvest. Cardiac rupture rate and IEI were significantly higher in testosterone-treated females and noncastrated males than in controls; these effects were accompanied by enhanced neutrophil infiltration and pronounced deterioration of cardiac function and left ventricular dilatation. Ovariectomy in females and estrogen supplementation in males did not confer significant protection from cardiac rupture, IEI, or neutrophil infiltration. We concluded that, in mice, high testosterone levels enhance acute myocardial inflammation, adversely affecting myocardial healing and early remodeling, as indicated by increased cardiac rupture, and possibly causing deterioration of cardiac function after MI, and, conversely, estrogen seems to have no significant protective effect in the acute phase after MI.

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Neonatal sex steroids affect responses to Seoul virus infection in male but not female Norway rats

Previous studies illustrate that after inoculation with Seoul virus (i.e., the naturally occurring hantavirus found in Norway rats), adult male rats produce higher antibody responses, exhibit higher Th1 responses (i.e., IgG2a, IL-2, and IFNγ), and shed virus longer than females, but these difference are not altered by manipulation of sex steroids in adulthood. To determine whether sex steroid hormones organize adult responses to hantavirus infection, at 2–4 days of age, male rats were gonadectomized and female rats were injected with testosterone. As adults, animals were inoculated with 10 4pfu of Seoul virus. Neonatally gonadectomized males (NGM), control females (CF), and neonatal testosterone-treated females (NTF) had lower anti-Seoul virus IgG and IgG2a responses than control males (CM) 20, 30, and 40 days post-inoculation. Neonatal testosterone treatment had no effect on female antibody responses to infection. NGM, CF, and NTF shed virus in saliva and feces for a shorter duration than CM. There was no effect of neonatal hormone manipulation either on the percentage of animals with detectable virus or on the number of virus copies within each target organ. Genetic males, regardless of hormone manipulation, had higher virus replication in lung tissue than did genetic females. Neonatal sex steroids influence the sexual dimorphism in host immune function but do not modify virus replication in target tissues.

GABA mediates steroid-induced astrocyte differentiation in the neonatal rat hypothalamus.

Our previous work has demonstrated that astrocytes in the developing arcuate nucleus of the rat hypothalamus are sexually dimorphic as a result of differential exposure to oestradiol. Moreover, our experiments in neonatal rats suggest an absence of oestrogen receptors (ER) in arcuate nucleus astrocytes in vivo. This, along with the conspicuous lack of evidence in the literature confirming the presence of ER in arcuate nucleus astrocytes of the neonatal rat brain, led us to question the mechanism by which oestrogen induces changes in arcuate nucleus astrocyte morphology. Based on our previous findings that oestradiol increases gamma-aminobutyric acid (GABA) levels in the neonatal rat arcuate, we hypothesize that GABA is released from neighbouring oestrogen-sensitive neurones and alters arcuate nucleus astrocyte morphology. Here, we report that in vivo reduction of GABA synthesis in the neonatal rat brain by antisense oligodeoxynucleotides to glutamic acid decarboxylase prevented gonadal steroid-induced astrocyte differentiation in males and testosterone-treated females. Conversely, activation of GABAA receptors with the agonist muscimol increased astrocyte differentiation in females in the absence of gonadal steroids. Given that GABA is made only in neurones and that its synthesis is increased by oestradiol, we conclude that it acts as a diffusible factor inducing the differentiation of neighbouring astrocytes.

Sex difference in the phosphorylation of cAMP response element binding protein (CREB) in neonatal rat brain

On the day of birth, a surge in testicular testosterone release in male rats is critical for sexual differentiation of the brain. Steroid hormones function by binding to intracellular steroid receptors and altering gene expression; however, little is known about the signal transduction pathways altered as a consequence of steroid hormone action in developing brain. We investigated whether the increase in testosterone at birth alters the phosphorylation of CREB, a major signal transduction protein. Adjacent brain sections from male and female pups were immunocytochemically stained for serine 133 phosphorylated CREB (pCREB) or total CREB on the day of birth. Males had more pCREB-immunoreactive positive cells than females in the medial preoptic area, ventromedial nucleus of the hypothalamus, the arcuate nucleus, and the CA1 region of the hippocampus, but not in two thalamic nuclei. There was no sex difference in total CREB immunoreactive cell number. To determine if the sex difference in pCREB persisted 24 h after birth and whether the difference was due to testosterone, newborn female pups were injected with 100 μg of testosterone propionate, and male and control female pups were injected with vehicle. Twenty-four hours later, adjacent brain sections were immunocytochemically stained for either pCREB or CREB. We found that males and testosterone-treated females had more pCREB in the ventromedial nucleus of the hypothalamus contrasted to control females. There were no group differences in pCREB or CREB in any other area examined. These results indicate that some of the effects of testosterone in developing brain occur via pathways associated with the phosphorylation of CREB.

Effects of testosterone propionate and nandrolone decanoate on body composition and lipoprotein concentrations in the rat

AbstractAnabolic–androgenic steroids (AAS) are used illicitly by athletes, bodybuilders and polydrug users. Cardiovascular complications are associated with their use and adverse lipoprotein alterations have been implicated as a contributing factor. In this experiment male and female rats were administered testosterone propionate, nandrolone decanoate or vehicle (peanut oil), 1 mg/kg/day s.c. for 14 days. Only the testosterone‐treated females experienced a significant increase in total body weight gain compared to peanut oil controls. No changes in food and water intake or excretion were noted between these two groups. Total body water increased in the testosterone‐treated female rats. Changes in lipoprotein profiles in female testosterone rats over the treatment period indicated a significant decrease in high‐density lipoproteins (HDL) compared to peanut oil‐treated animals.This study indicates that the treatment protocol used may serve as a model to further investigate the mechanisms underlying the effects of AAS.

CAN TESTOSTERONE INDUCE THE MALE WHISTLE-SHAKE CALL IN FEMALE SHELDUCKS TADORNA TADORNA?

ABSTRACT We investigated by application of testosterone-proprionate whether females of the dimorphic Shelduck Tadorna tadorna are capable of producing vocalisations typical of the male sex. We found that hormone-treated females generated the whistle-shake call, an epigamie vocalisation pattern of male Shelducks. Although the female whistle- shake call was male-like in its syntactic form, we observed significant differences in both the phonetic and temporal characteristics of male and female whistle-shake calls. None of the testosterone-treated females was found to produce real male-typical elements. All whistle-shake calls of these females consisted of elements which are also used in calls of untreated females, e.g. the agagak-call. The present findings indicate that the sexual dimorphism in Shelduck vocalisations is not due to early organising actions of sex hormones influencing the differentiation of the brain. It results mainly from adult sex differences in the endogenous testosterone production. Only males produce testosterone in considerable amounts. The inability of testosterone-treated female Shelducks to generate male-typical elements might be caused by limitations in the sound-producing organ, the syrinx. Male Shelducks have associated with the syrinx a large resonating chamber, the bulla ossea, which is lacking in females.

Effects of testosterone on cell-mediated and humoral immunity in non-breeding adult European starlings

One of the primary assumptions of the immunocompetence hypothesis is that testosterone is immunosuppressive. Although many studies in birds and mammals have supported this assumption, conflicting results have been reported in a variety of species. We investigated the effects of testosterone manipulation on both cell-mediated and humoral immunity in adult songbirds, European starlings (Sturnus vulgaris). Male and female starlings were wild-caught, housed in the laboratory, and implanted with either empty silastic capsules or capsules containing testosterone. Six weeks after implantation, humoral immunity was assessed by injecting the birds with a novel antigen, keyhole limpet hemocyanin, and measuring specific antibody responses 10 and 15 days later via an enzyme-linked immunosorbant assay. Cell-mediated immunity was assessed 7 weeks after implantation via intradermal injection of the T-cell mitogen phytohemagglutinin into the wing web and measuring the degree of swelling 24 h later. Antibody responses to antigenic challenge were significantly suppressed in testosterone-treated females 10 days post-injection and in both sexes 15 days post-injection. Furthermore, there was a significant inverse relationship between individual variability in antibody responsiveness and plasma testosterone concentrations. Cell-mediated responses to phytohemagglutinin stimulation were also significantly suppressed in testosterone-treated males compared to same-sex controls. Testosterone treatment significantly increased plasma corticosterone concentrations compared to controls, and the possibility of this effect mediating the immunosuppressive effects of testosterone is discussed. The present study is among the first to demonstrate testosterone-induced suppression of both cell-mediated and humoral immunity in a species of songbird.

Effect of testosterone on the distribution of vasotocin immunoreactivity in the brain of the zebra finch, Taeniopygia guttata castanotis

The distribution of vasopressin or vasotocin immunoreactive cells and fibers in the lateral septum and the bed nucleus of the stria terminal is are sexually dimorphic in many vertebrates including several species of birds examined to date. We examined the vasotocin-like immunoreactivity in the zebra finch brain. Male birds had a higher level of immunoreactive staining in some telencephalic and diencephalic regions. The density of immunostaining increased in the testosterone-treated females to levels typically seen in males. The sexual dimorphism and testosterone dependence of the vasotocin-like immunoreactivity are similar to that found in the canary. Thus this pattern of vasotocin localization and testosterone dependence may be a general feature in brains of passerine songbirds.

Investigation of the Role of Postnatal Testosterone in the Expression of Sex Differences in Behavior in Infant Rhesus Macaques (Macaca mulatta)

In several primate species, males have been shown to exhibit a surge in circulating testosterone during the early postnatal period. This surge has been postulated to play a role in the development of sex differences in behavior. In this study, the role of postnatal testosterone in infant behavioral development was investigated in socially living rhesus macaques. Seven male infants were treated with a GnRH agonist, avorelin, from the first week of life onwards. Ten female infants were exposed to testosterone by implantation of capsules containing testosterone. The behavioral development of these and control infants was recorded from birth to 6 months of age. The sexually dimorphic patterns of play and mounting were not affected by manipulation of postnatal testosterone in either male or female infants. Similarly, most mother–infant interactions were not affected by the hormonal manipulation of infants. Mothers of testosterone-treated females were found to take more responsibility for moving into and out of arm's reach of their infants than mothers of some other groups of infants; however, this measure did not normally differ between mothers of male and female infants. Manipulation of the postnatal testosterone surge does significantly affect penile growth and development, but does not affect the expression of infant sex differences in behavior nor greatly affect the development of the mother–infant relationship in rhesus macaques.

Manipulation of postnatal testosterone levels affects phallic and clitoral development in infant rhesus monkeys.

Male primates exhibit marked elevation of circulating testosterone levels during the early postnatal period. The aim of this project was to test whether experimental manipulation of circulating testosterone levels in male and female infant rhesus monkeys affected development of the external genitalia during the first six months of life. Four groups of infants were studied. Seven control male infants exhibited high circulating testosterone levels during the first three months of life. Seven males were treated with a GnRH agonist (avorelin) from the first week of life onwards, which suppressed the postnatal testosterone surge. Ten control females exhibited low circulating testosterone levels during the early postnatal period. Administration of testosterone to 10 females resulted in high circulating levels in these infants. Fortnightly blood samples and genital measurements were taken from all infants during the first six months of life. Growth of the penis of avorelin-treated males was significantly retarded when compared to control males. Average length of the penis at six months of age was significantly (p = 0.012) smaller for avorelin-treated males (25.2 +/- 2.8 mm) than for control males (37.3 +/- 3.0 mm). Avorelin-treated males attained only around 50% detachment of the prepuce from the glans of the penis, while control males averaged 90% detachment. Treatment of females with testosterone resulted in significant growth of the clitoris in comparison to control females. The growth rates of the penis of control males and clitorides of testosterone-treated females were similar and greatest during the first two months of life. Gain in body weight was not affected by either hormonal manipulation. It is concluded that manipulation of circulating testosterone levels during the early postnatal period affects penile and clitoral development of infant rhesus monkeys. This postnatal period may therefore represent an important stage in penile development in primates.

Male-specific transcription initiation of the C4-Slp gene in mouse liver follows activation of STAT5.

The mouse genes encoding the constitutively expressed complement component C4 and its closely related isoform C4-Slp (sex-limited protein), which is expressed only in male animals of several strains, provide a unique model to study sequence elements and trans-acting factors responsible for androgen responsiveness. Our previous studies have shown that hormonal induction of C4-Slp is mediated by a sex-specific pattern of growth hormone secretion. Promoter analyses in vitro have led to contradictory conclusions concerning the significance of C4-Slp-specific sequences in the 5' flanking region. Mutant mice carrying the H-2(aw18) haplotype, which is characterized by a large deletion in the S region covering the C4 and 21-OHase A genes, permit the direct in vivo analysis of C4-Slp transcription, unhindered by the presence of C4. Run-on analysis of transcription in liver nuclei of males and females of this strain demonstrated a 100-fold higher transcriptional activity in males, essentially determined at the transcription initiation level. The androgen dependence of transcription initiation was confirmed by run-on analysis of testosterone-treated females, where transcriptional activity started after 6 days of androgen treatment and reached male levels after 20 days. Conversely, the growth hormone-regulated activity of transcription factor STAT5 was already detected in liver nuclei after 48 hr of androgen treatment. Furthermore, we demonstrate that activated STAT5 recognizes in vitro two upstream gamma interferon-activated sequence (GAS) elements of the C4-Slp gene, centered at positions -1969 and -1831. We postulate that binding of STAT5 to these C4-Slp-specific GAS elements plays a crucial role in the chromatin remodelings that lead to transcriptional competence of the C4-Slp gene in the liver.

A role for ovarian hormones in sexual differentiation of the brain.

Historically, studies of the role of endogenous hormones in developmental differentiation of the sexes have suggested that mammalian sexual differentiation is mediated primarily by testicular androgens, and that exposure to androgens in early life leads to a male brain as defined by neuroanatomy and behavior. The female brain has been assumed to develop via a hormonal default mechanism, in the absence of androgen or other hormones. Ovarian hormones have significant effects on the development of a sexually dimorphic cortical structure, the corpus callosum, which is larger in male than in female rats. In the females, removal of the ovaries as late as Day 16 increases the cross-sectional area of the adult corpus callosum. Treatment with low-dose estradiol starting on Day 25 inhibits this effect. Female callosa are also enlarged by a combination of daily postnatal handling and exogenous testosterone administered prior to Day 8. The effects of androgen treatment are expressed early in development, with males and testosterone-treated females having larger callosa than control females as early as Day 30. The effects of ovariectomy do not appear until after Day 55. These findings are more consistent with other evidence of a later sensitive period for ovarian feminization as compared to androgenic masculinization.

Serum testosterone concentration, efficiency of estrus detection and libido expression in androgenized beef cows

Twenty multiparous, cyclic, nonlactating beef cows were blocked by dominance rank and randomly and equally allotted to 1 of 4 treatment groups: an untreated control group, a synovextreated group which received 8 Synovex-H® implants with no additional hormones, a testosterone-treated group which received 500 mg, im and 1500 mg, sc testosterone enanthate on Day 1 with additional 1000 mg, sc doses of testosterone enanthate every 14 d, and a synovex+testosterone-treated group which received 8 Synovex-H® implants with 500 mg, im and 1500 mg, sc testosterone enanthate on Day 1 only. Blood samples were collected via jugular venipuncture once a week beginning 3 wk prior to start of treatment. In addition, samples were collected just prior to treatment; once a day for 1 wk after initiation of treatment; and then twice a week until 225 d after treatment. Efficiency of estrus detection was assessed 22 d prior to start of treatment and every 14 d thereafter for 98 d, using estrus detection trials with synchronized females or modified libido tests. Scores for estrus detection trials included total mounts in 1 h and the percentage of estrous cows detected. Libido was scored on a scale of 0 through 6. All testosterone treatments raised plasma testosterone concentrations above control and pretreatment levels (testosterone and synovex+testosterone>synovex>control; all P<0.05). Synovex-, testosterone- and synovex+testosterone-treated females performed more mounts in 1 h than the controls (18, 9, 6 and 1, respectively; all P<0.05). All testosterone-treated cows mounted a higher number of estrous females than the controls (P<0.05). Only synovex+testosterone- and testosterone-treated cows received libido scores above pretreatment and control values. However, libido of testosterone-treated cows decreased over time, while that of synovex+testosterone-treated females remained high until Day 98. Libido scores correlated positively with the number of mounts in 1 h and the percentage of estrous females detected (0.70 and 0.44, respectively), and the correlation coefficient for these two factors was 0.63. In conclusion, the synovex+testosterone treatment was most effective for producing estrus detector females and libido testing was useful for evaluating sexual activity in androgenized females.

The Kidney Androgen-regulated Protein Promoter Confers Renal Proximal Tubule Cell-specific and Highly Androgen-responsive Expression on the Human Angiotensinogen Gene in Transgenic Mice

Transgenic mice were generated containing a 1542-base pair fragment of the kidney androgen-regulated protein (KAP) promoter fused to the human angiotensinogen (HAGT) gene with the goal of specifically targeting inducible expression of renin-angiotensin system components to the kidney. High level expression of both KAP-HAGT and endogenous KAP mRNA was evident in the kidney of male mice from two independent transgenic lines. Renal expression of the transgene in female mice was undetectable under basal conditions but could be strongly induced by administration of testosterone. Testosterone treatment did not cause a transcriptional induction in any other tissues examined. However, an analysis of six androgen target tissues in males revealed that the transgene was expressed in epididymis. No other extra-renal expression of the transgene was detected. In situ hybridization demonstrated that expression of HAGT (and KAP) mRNA in males and testosterone-treated females was restricted to proximal tubule epithelial cells in the renal cortex. Although there was no detectable human angiotensinogen protein in plasma, it was evident in the urine, consistent with a pathway of synthesis in proximal tubule cells and release into the tubular lumen. These results demonstrate that 1542 base pairs of the KAP promoter is sufficient to drive expression of a heterologous reporter gene in a tissue-specific, cell-specific, and androgen-regulated fashion in transgenic mice.