Androgenic control of the harderian gland in the male golden hamster.

In the golden hamster, there are marked sex differences in the Harderian gland. Male glands (which are heavier than female glands) possess two cell forms (Type I and Type II cells); female glands only exhibit the former. Female (but not male) glands contain large amounts of porphyrin, which are readily visible as solid depositions within the lumina. The weight, histology and porphyrin content of the Harderian gland was examined in intact adult male hamsters and in male hamsters castrated for 1,2 or 8 months. Castration resulted in a significant reduction in the weight of the gland, the disappearance of Type II cells, and the presence in the gland of solid porphyrin accretions. The levels of copro- and (especially) protoporphyrin were greatly increased. These changes were more marked with time after castration. When the ability of diverse androgens (testosterone, 5alpha-dihydrotestosterone, androst-4-ene-3,17-dione (androstenedione), dehydroepiandrosterone and androsterone) to prevent these changes was tested, testosterone and androstenedione maintained glandular weight. All the androgens maintained normal frequencies of Type II cells and all except dehydroepiandrosterone prevented deposition of porphyrin. The potencies of the various androgens in maintaining normal Harderian gland morphology and activity are compared with their effects on other somatic variables and sexual behaviour.     

Regulation of the androgen receptors in the Harderian gland of the male Syrian hamster: Influence of photoperiod, castration, and chronic melatonin treatment

Male Syrian hamsters that were exposed for 8 weeks to short photoperiod (LD 10:14) or treated with melatonin in the late afternoon under long photoperiod conditions (LD 14:10) had a significantly higher content of androgen receptors in the Lipidex-purified soluble fractions isolated from the Harderian glands as compared to the long photoperiod (LD 14:10) exposed controls. Simultaneous computer-assisted analyses of all series of saturation and competition experiments revealed that the numerical value of the apparent Kd, as determined by using the synthetic androgen R-1881 (methyltrienolone), was not different between the experimental groups, and ranged from 0.050 to 0.067 nM. Of the principal natural androgens, testosterone (T) was most potent in inhibiting methyltrienolone binding to the receptor (Ki values from 0.33 to 0.55 nM), and 5 alpha-dihydrotestosterone (DHT) and delta 4-androstenedione (AD) were less effective (Ki values between 1 and 1.9 nM). In the hypothalami and pituitaries of the same animals, used in parallel control assays, DHT was twice as potent as T. Short-term castration (24 hr post-orchidectomy) did not result in significant changes in the receptor binding characteristics. Following 8 weeks exposure to a long photoperiod (LD 14:10) the Bmax values demonstrated a four-fold increase in castrated animals (179 fmoles/mg protein vs. 47 fmoles/mg protein) over intact controls. The relative binding affinity of the major androgens under these conditions remained unchanged, with the exception of AD, where a five-fold increase in the numerical Ki values (decrease in the binding affinity) was recorded (Ki = 9.6 nM).(ABSTRACT TRUNCATED AT 250 WORDS)     

Sequence analysis and androgen regulation of MHG07 (Male harderian gland) mRNA in male hamster harderian gland

The hamster Harderian gland (HG), a compound tubuloalveolar gland located in the orbital cavity, displays sex dimorphism. The present study focuses on the sequence analysis of a cDNA clone named MHG07 and on the regulation of its expression by steroid hormones. MHG07 mRNA (5.0 kb) is expressed in male HG only. The MHG07 cDNA (1.74 kb) shows an ORF of 94 amino acids and has no significant homologies with other polypeptides/genes. Castration leads to the disappearance of MHG07 mRNA after 4 days, whereas treatment with testosterone impairs the effect of castration. No MHG07 mRNA has been found in either rat or murine HGs. Androgen (A) administration to female hamsters induces the appearance of MHG07 mRNA. In primary culture of male hamster HG, androgens increase the MHG07 expression and this effect is blocked by both flutamide and cycloheximide. Dose-response experiments show that, at low A concentration (10(-12) M), the MHG07 was higher than that of the control (2-fold). This effect reaches its zenith at 10(-8) M (10-fold). This picture is paralleled by androgen receptor mRNA expression. It is argued that the expression of MHG07 is under androgenic control.     

Influences of photoperiod on nuclear androgen receptor occupancy in neuroendocrine tissues of the golden hamster

Day length regulates the negative feedback potency of gonadal steroids upon luteinizing hormone (LH) in seasonal breeders such as the golden hamster. We have used an exchange assay employing 3H-R1881 to determine whether nuclear androgen plus receptor levels in the preoptic area, medial basal hypothalamus, or anterior pituitary differ between male hamsters maintained in long or short days. Cell nuclear androgen plus receptor levels in brain and anterior pituitary were significantly lower in intact males maintained in short days; these differences reflected significant decreases in testis size and serum testosterone (T levels upon exposure to inhibitory photoperiods. In castrated males in which serum T levels were 'clamped' by the insertion of T-filled Silastic capsules, exposure to short days was not correlated with an increase in preoptic area, medial basal hypothalamus, or anterior pituitary receptor occupancy even though T's negative feedback actions upon LH were clearly enhanced. In contrast, there were instances in which androgen receptor occupation was elevated in males exposed to long days. Our results suggest that in the male golden hamster, the well-documented increase in the ability of T to suppress LH secretion in short photoperiods cannot be attributed to an increase in receptor-mediated uptake and nuclear accumulation of androgen in target cells in the brain and anterior pituitary gland.     

Androgen receptor in the Harderian gland of Rana esculenta

An androgen receptor has been identified in the cytosolic and nuclear extracts of the Harderian gland of the frog, Rana esculenta. A single class of high-affinity binding sites was found: Kd = 1.9 +/- 1.3 (S.D.) nmol/l (n = 26) for the cytosolic extract and Kd = 0.9 +/- 0.8 nmol/l (n = 15) for the nuclear extract. The presence of binding activity in both nuclear and cytosolic extracts and the low rate of ligand-receptor dissociation are characteristics that distinguish this receptor from a steroid-binding protein. The Kd did not show any sex difference and did not exhibit any secretory activity-related change. Binding in both cytosolic and nuclear extracts was specific for androgens (testosterone = 5 alpha-dihydrotestosterone); oestradiol-17 beta showed a 30% cross-reaction; moreover, specific binding of [3H]oestradiol-17 beta was not detectable. The binding capacity of the Harderian gland increased progressively in both fractions from October to December, reaching a peak in May, and decreased suddenly during July to August. The lack of any morphological sex-related difference in the Harderian gland of the green frog might be accounted for by the high amount of circulating androgens as well as a similar concentration of androgen receptor in both sexes.     

Evidence for melatonin synthesis in rodent Harderian gland: A dynamic in vitro study

ABSTRACT: Melatonin content and release from Harderian glands (HGs) has been measured by an in vitro perifusion technique in three rodent species: Wistar rat, Syrian hamster, and Siberian hamster. Melatonin immunoreactive concentrations in HGs of animals killed at 10.00 hr were 0.31 ± 0.031 pg/mg gland in male Wistar rat, 0.54 ± 0.026 pg/mg gland in male Siberian hamster, 0.17 ± 0.070 and 0.20 ± 0.059 pg/mg gland in male and female Syrian hamster, respectively. In all species examined, isolated HGs perifused for 9–15 hr released melatonin but did not stabilize their melatonin release rate. No sex-related difference could be noted in the HG melatonin release rate. The total amount of melatonin released over a 15 hr long perifusion was about 0.075 ± 0.004 ng/15 h/mg gland and 0.063 ± 0.010 ng/15 hr/mg gland in male and female Wistar rat, respectively; 0.155 ± 0.019 ng/15 hr/mg gland and 0.141 ± 0.006 ng/15 hr/mg gland in male and female Siberian hamster, respectively; 0.035 ± 0.003 ng/15 hr/mg gland and 0.045 ± 0.004 ng/15 hr/mg gland in male and female Syrian hamster, respectively. This amount, which is higher than the tissue levels, demonstrates the de novo melatonin synthesis. This is confirmed by the fact that infusion of the indoleamine precursor, tryptophan (TRP), stimulated melatonin secretion from HGs. The melatonin release is increased by 2.5-fold in male and female Wistar rat, 1.5-fold in male and female Siberian hamster, and 2.0- and 3.0-fold in male and female Syrian hamster, respectively. Treatment with a TRP hydroxylase inhibitor, para-chlorophenylalanine, reduced basal melatonin release and inhibited the TRP-induced melatonin stimulation. Kinetics and amounts of melatonin released were not affected by pinealectomy, ruling out a possible plasmatic origin of the HG melatonin. Isoproterenol, a β-adrenergic agonist, and dibutyryl cyclic AMP, a cyclic AMP analogue, failed to stimulate HG melatonin secretion. In conclusion, these results confirm the presence of melatonin in the HGs and demonstrate that melatonin is synthesized in and released from isolated rodent HGs.     

Harderian gland N-acetyltransferase activity in the male Syrian hamster: effects of gonadectomy, short photoperiod exposure, or subcutaneous melatonin implants

The activities of NAT and HIOMT and the melatonin concentration in the Harderian glands of intact, gonadectomized, and gonadally-regressed male Syrian hamsters were studied. To produce gonadal regression, hamsters were exposed to either artificial or naturally short photoperiods. NAT activity of castrated and gonadally-regressed hamsters was always less in comparison to that of animals with intact gonads. Castrated hamsters exposed to long days showed higher NAT activity than that of castrated animals exposed to short photoperiods indicating that light may have some influence on Harderian NAT independent of the gonadal status. Also, gonadally-regressed hamsters exposed to long photoperiods exhibited higher NAT activity in comparison to gonadally-regressed animals exposed to short days. The HIOMT activity and melatonin content of Harderian glands in all these groups of male Syrian hamster were very low.     

Chronic administration of melatonin induces changes in porphyrins and in the histology of male and female hamster Harderian gland: Interrelation with the gonadal status

In this paper, we have investigated the influence of melatonin on the histology and porphyrin content of the Syrian hamster Harderian glands. Daily afternoon injections of 25 micrograms of melatonin to female hamsters for 12 weeks resulted in the discontinuity of estrous cyclicity, a marked decrease in the Harderian gland intraluminal area occupied by porphyrins, and in a significant rise in the number of Type II cells. A similar decrease in porphyrins was observed after 8 weeks of ovariectomy. However, if the melatonin injections were given for only 8 weeks (without inducing gonadal atrophy), no changes were observed in the area occupied by intraluminal porphyrins, suggesting that the effects of melatonin in female Syrian hamsters might be associated with the subsequent gonadal atrophy. Castration of male hamsters induced a significant increase in porphyrins and a clear drop in the number of Type II cells. These changes were totally prevented when melatonin was administered daily from the day of castration. Our results suggest that melatonin, at least in male Syrian hamsters, plays a role in Harderian metabolism, acting directly on the Harderian secretory cells or indirectly through pituitary hormones.     

Influence of age on sperm concentration in the male golden hamster

Male golden hamsters were separated into three age groups: adolescent, 2-4 months; adult, 12-13 months; aged 20-24 months, and maintained on a reversed light cycle. Body, testis and epididymal weights were recorded at the time of sacrifice. Stock suspensions of sperm were obtained from the cauda epididymidis and serially diluted in saline. Optical density (O.D.) readings and sperm counts were taken for each dilution sample. Data points, O.D. versus sperm counts, were computer fit. Comparison of the regression lines demonstrated a significant difference between the slopes of curves for adolescent males and for adult and aged males (99% confidence level). Mean sperm counts for the adult and aged males were significantly higher than for the young males. The increased counts were accompanied by decreases in O.D. with advancing age. Trypsin treatment of the sperm revealed no effect on O.D. indicating that separation of heads and tails was not responsible for the change in O.D. values of sperm samples. Likewise, comparison of washed and unwashed sperm indicated that epididymal fluid plays no role in determining O.D. values. There were no significant age-related differences in body, testis or epididymal weights.     

Testosterone regulation of androgen receptor levels in the uropygial gland of quails (Coturnix coturnix): a further proof for the androgen dependency of the uropygial gland

In castrated quails supplemented with testosterone by a chronically implanted silastic capsule, removal of the implant resulted in the gradual disappearance of the androgen receptors in both cloacal and uropygial glands. However, after a 7-day time span, the receptors reappeared in both glands but did not reach the original level of implanted birds. On the other hand, the photostimulation of intact birds induced an increase of the receptor content of their uropygial and cloacal glands when compared to sexually quiescent birds. These results show that, the concentration of the androgen receptors of the uropygial and cloacal glands of adult male quails is partly controlled by testosterone. By comparison with the mechanism of the action of testosterone in mammal target organs, our results add weight to the androgen dependency of quails uropygial glands (a sebaceous like organ).     

Coexpression of MT1 and RORalpha1 melatonin receptors in the Syrian hamster Harderian gland

Melatonin acts through several specific receptors, including membrane receptors (MT(1) and MT(2)) and members of the RZR/ROR nuclear receptors family, which have been identified in a large variety of mammalian and nonmammalian cells types. Both membrane and nuclear melatonin receptors have been partially characterized in Harderian gland of the Syrian hamster. Nevertheless, the identities of these receptors were unknown until this study, where the coexistence of MT(1) and RORalpha(1) in this gland was determined by nested RT-PCR followed by amplicon sequencing and Western-blot. Furthermore, the cellular localization of both receptors was determined by immunohistochemistry. Thus, MT(1) receptor was localized exclusively at the basal side of the cell acini, supporting the hypothesis that this receptor is activated by the pineal-synthesized melatonin. On the contrary, although a RORalpha(1)-immunoreactivity was observed in nuclei of epithelial cells of both sexes, an extranuclear specific staining, which was more frequently among those cells of males, was also seen. The implication of this possible nuclear exclusion of RORalpha(1) on the role of this indoleamine against oxidative stress is discussed.     

Steroid hormone binding in the Harderian gland of birds: characteristics of the androgen, estrogen, and progestin receptors of Anas platyrhynchos and Gallus domesticus

A series of experiments was carried out in immature female chicks and ducks to establish whether the avian Harderian gland contains specific receptors for sex steroids. Cytosol preparations of Harderian glands were submitted to hormone saturation analysis using radiolabeled estradiol, ORG-2058, and dimethylnortestosterone as ligands. In addition, the sedimentation characteristics of the hormone-receptor complexes were studied by ultracentrifugation of linear sucrose gradients. The presence of high affinity binding sites for estrogens (Kd = 2.4 and 1.6 nM), progestins (0.8 and 1.0 nM), and androgens (1.0 and 1.0 nM) was indicated in the chick and duck glands, respectively. The sedimentation coefficients were 7-7.5 S, 7-8 S, and 8 S for estrogen, progestin, and androgen receptor-ligand complexes, respectively. The concentration of the androgen receptor was significantly higher in chick than in duck Harderian glands whereas the estrogen and progestin receptor concentrations were similar in both species. A striking finding was the presence of progestin receptors, which apparently do not exist in the glands of many mammals. Priming with estrogens did not modify the concentration of ORG-2058 binding sites in either species studied, indicating that gland progestin receptor is not estrogen-regulated. Overall the data suggest intracellular mechanisms whereby circulating gonadal hormones regulate avian Harderian gland function.