Harderianization is another sexual dimorphism of rat exorbital lacrimal gland

The exorbital lacrimal glands (ELG) of rats were examined for both sexes to determine what degree of harderianization occurred as a function of age and after castration, and to investigate its time course and origin in ELG. Light microscopically, very small Harderian foci were seen in the ELG of both sexes at 3 weeks of age. As the male rats became older, the relative volume of the Harderian gland (HG) cells in the ELG increased. At age 6 months, the value was 1.25 +/- 0.31% in males and 0.13 +/- 0.05% in females (p less than 0.05). After castration, a significant decrease (0.21 +/- 0.01%, p less than 0.05) was observed in that of male ELG. In contrast, in female ELG, HG cells were inconspicuous and the relative volume of those did not vary during this experimental period or after castration. It appeared that the HG cells had developed from undifferentiated basal cells of the acini and the intercalated ducts in the ELG at age 2-6 months. Then, at age 22 months, they also probably developed from those of the excretory ducts of the ELG.     

Demonstration and characterization of cytosol androgen receptor in rat exorbital lacrimal gland

The presence of a macromolecule which binds androgen with a high affinity and a low capacity was demonstrated in the cytosol of the lacrimal glands of male and female rats. Evidence was found that this macromolecule was a protein by treatment with protease, trypsin or heat. A specific 8-8.5 S peak was obtained in both sexes by glycerol gradient centrifugation in low salt condition, whereas a specific 5.2 S peak was found in high salt condition. This protein could bind to DNA-cellulose after treatment of androgen-cytosol complexes by warming (25 degrees C 15 min) or exposure under high salt (0.4 M KCl). These results suggested that this protein was an androgen receptor.     

Leucine aminopeptidase (LAP) activity and sexual dimorphism in rat exorbital lacrimal gland

Leucine amonopeptidase (LAP) activity was histochemically studied in the rat exorbital lacrimal gland. The enzyme is present in the secreting cells of both male and female prepuberal rats, in vivo and in monolayer cultures, while in the adult rat it is demonstrable only in the female. Furthermore, LAP is influenced by the sex hormones, disappearing in the female gland after testosterone treatment and appearing in the adult male gland after administration of estradiol or cyproterone acetate. These results show that the behaviour of LAP can be considered as another character of sexual dimorphism of the rat exorbital gland. Furthermore, our findings, showing an inverse relationship between LAP activity and the PAS positivity of the secretion products, suggest the hypothesis that the presence of this exopeptidase could induce qualitative modifications of one or more secreted glycoprotein.     

The effects of vinblastine on acinar cells of the exorbital lacrimal gland of the rat

Summary The effects of vinblastine treatment on acinar cells of the rat exorbital lacrimal gland were studied by electron microscopy. Experimental animals of both sexes were given single intraperitoneal injections of (1) vinblastine (4mg/kg body weight) at 1 to 24 h before sacrifice; (2) pilocarpine (20 mg/kg b.w.) for 1 h; or (3) vinblastine for l h followed by pilocarpine for 1 h.Vinblastine treatment caused a number of changes including autophagocytosis, formation of intracisternal granules, and alteration of secretory granules. These changes varied in extent and onset between male and female rats. In addition, the Golgi apparatus was reduced in size and dispersed throughout the cytoplasm. Mitotic figures were commonly observed. Moreover, vinblastine inhibited the pilocarpine-stimulated degranulation of the acinar cells.In view of the known anti-microtubular action of vinblastine, these results suggest that microtubules are involved in various aspects of the transport, packaging, and secretion of exportable proteins in the lacrimal gland. Additionally, autophagocytosis and alteration of secretory granules may partially result from the interaction of vinblastine with membranes.The authors thank Mr. Steve Coriell and Mr. Steve Floyd for preparing the micrographs. Robert Kelly also thanks Dr. George Chapman for his support during the initial phase of this project.     

Relationship between the golgi apparatus, gerl, and secretory granules in acinar cells of the rat exorbital lacrimal gland

The method of secretory granuleformation in the acinar cells of the rat exorbital lacrimal gland was studied by electron microscope morphological and cytochemical techniques. Immature secretory granules at the inner face of the Golgi apparatus were frequently attached to a narrow cisternal structure similar to GERL as described in neurons by Novikoff et al. (Novikoff, P. M., A. B. Novikoff, N. Quintana, and J.-J. Hauw. 1971. J. Cell Bio. 50:859-886). In the lacrimal gland. GERL was located adjacent to the inner Golgi saccule, or separated from it by a variable distance. Portions of GERL were often closely paralleled by modified cisternae of rough endoplasmic reticulum (RER), which lacked ribosomes on the surface adjacent to GERL. Diaminobenzidine reaction product of the secretory enzyme peroxidase was localized in the cisternae of the nuclear envelope, RER, peripheral Golgi vesicles, Golgi saccules, and immature and mature secretory granules. GERL was usually free of peroxidase reaction product or contained only a small amount. Thiamine pyrophosphatase reaction product was present in two to four inner Golgi saccules; occasionally, the innermost saccule was dilated and fenestrated, and contained less reaction product than the next adjacent saccule. Acid phosphatase (AcPase) reaction product was present in GERL, immature granules, and, rarely, in the innermost saccule, but not in the rest of the Golgi saccules. Thick sections of AcPase preparations viewed at 100 kV revealed that GERL consisted of cisternal, and fenestrated or tublular portions. The immature granules were attached to GERL by multiple connections to the tublular portions. These results suggest that, in the rat exorbital lacrimal gland, the Golgi saccules participate in the transport of secretory proteins, and that GERL is involved in the formation of secretory granules.     

beta-Adrenergic receptors stimulated peroxidase secretion from rat lacrimal gland

Incubation of rat extraorbital lacrimal gland slices with the beta-agonist isoproterenol caused peroxidase secretion but no K+ release. The peroxidase secretion was inhibited by propranolol. Addition of dibutyryl cyclic AMP or adenosine 3'5'-cyclic phosphorothioate to lacrimal slices produced peroxidase secretion at a higher rate than that obtained with optimal concentration of isoproterenol. Methyl isobutylxanthine is also a strong stimulator of peroxidase secretion. Peroxidase activity was determined by a modified sensitive guaiacol method. Membrane fraction of lacrimal cells was shown to contain an isoproterenol-stimulated adenylate cyclase activity. It is therefore suggested that there is a beta-adrenergic receptor in the rat lacrimal gland and that its stimulation causes activation of an adenylate cyclase which leads to peroxidase secretion.     

Control of estrogen and androgen receptors in the rat pineal gland by catecholamine transmitter

Sucrose gradient studies of rat pineal cytosol incubated with ³H-estradiol (female pineals) or ³H-5 α -dihydrotestosterone (male pineals) revealed a radioactivity peak in the 8 S region which disappeared after superior cervical ganglionectomy or incubation with excess unlabeled hormone. Ganglionectomy decreased significantly estradiol and testosterone uptake by the pineal gland $\text{in vitro}$ as well as high affinity binding to pineal cytoplasmic and nuclear components. Norepinephrine treatment counteracted all the effects of ganglionectomy but was unable to modify hormone uptake and binding by the pineal gland of sham-operated controls. Pre-treatment with actinomycin D or propranolol but not with phentolamine impaired norepinephrine effects; propranolol blockage however was only partial. Administration of isoproterenol, L-dopa or phentolamine increased hormone uptake by denervated pineals. The effects of isoproterenol were also observed $\text{in vitro}$ and were blocked by propranolol. These results indicate that sex steroid receptors in the pinealocytes are controlled by norepinephrine via beta-adrenergic receptors and that depletion of neural norepinephrine enhanced responsiveness of pineal hormone receptors to exogenous catecholamines.     

Androgen receptors in rat and human prostate

In intact adult rats almost all androgen receptor (AR) sites of the rat ventral prostate (RVP) are occupied by endogenous dihydrotestosterone, and about 80% of these sites are nuclear. Nuclear AR disappears rapidly after castration (half-life of 3 h). The amount of cytosolic AR does not change within the initial 36 h, then markedly decreases during the next 2-5 days. An early and specific action of androgen is a remarkable increase of its own receptor. RVP also contains an estradiol receptor (ER) which rapidly disappears after castration and which, contrary to AR, is predominantly localized in the cytosol of stromal elements. The published procedures for steroid receptors grossly underestimate receptors concentrations in normal (NHP) and hyperplastic (BPH) human prostate. We have recently established a reliable method for the measurement of total AR, and we have found no difference in AR concentrations between NHP and BPH. BPH also contains a progesterone receptor and an elusive ER. Finally, we have used specific immunoglobulins in sex hormone binding plasma protein (SBP) for the demonstration of SBP-like immunoreactivity by the indirect immunofluorescence technique. The specific antigenic material was exclusively localized in the cytoplasm of BPH epithelial cells.     

Seasonal expression of androgen receptors, estrogen receptors, and aromatase in the canary brain in relation to circulating androgens and estrogens

Songbirds have a complex neural network for learning and production of song, namely the neural song system. Several nuclei of the song system contain androgen receptors (AR), and the neostriatal nucleus HVc also contains alpha type estrogen receptors (ER). Many songbird species show seasonal changes in both song and the neural song system that are correlated with seasonal variations in the circulating levels of gonadal steroids. However, there is increasing evidence that the sensitivity of the song system to gonadal steroids also changes seasonally. This could involve changes in the expression and activity of steroid receptors and steroid-metabolizing enzymes, such as the estrogen-synthesizing enzyme aromatase (AROM). The seasonal regulation of brain AR, ER, and AROM has not been studied before in the same individual songbirds. In this work, we compared plasma levels of androgens and estrogens, the expression level of AR-, ER-, and AROM-mRNA in the telencephalon, and brain AROM activity in male canaries between autumn (November) and spring (April) periods of high singing activity. Plasma levels of androgens and estrogens were higher in April than in November. The expression level of ER in HVc was higher in November than in April. In contrast, the expression level of AROM in the caudomedial neostriatum was higher in April than in November. However, we found no seasonal differences in the level of expression of AR and the volume of HVc as delimited by AR expression. Thus, AR expression in HVc was not correlated with circulating androgen levels. This study shows that both steroid-dependent and -independent seasonal factors regulate the action of gonadal hormones on the song system. In addition, we report a new site of AROM expression in the songbird brain, the nucleus interfacialis.     

Androgen regulation of secretory component synthesis by lacrimal gland acinar cells in vitro

This study sought to determine whether androgens directly stimulate the production of secretory component (SC) by acinar cells from the rat lacrimal gland. Homogeneous populations of acrinar cells were isolated from lacrimal tissues by serial enzymatic digestion and Ficoll gradient centrifugation and then cultured on reconstituted basement membranes in supplemented, serum-free medium. Acinar cell exposure in vitro to dihydrotestosterone (DHT) resulted in a significant increase in cellular SC output. This hormone action was dose dependent and androgen specific. Testosterone, but not 17 beta-estradiol, progesterone, dexamethasone, or aldosterone, also induced a considerable elevation in acinar cell SC production. The effect of testosterone may not require intracellular enzymatic conversion to DHT. The impact of androgens on SC output was associated with enhanced cellular synthesis and secretion and did not involve variations in acinar cell viability or density. Moreover, the SC response to DHT occurred irrespective of whether lacrimal gland acinar cells were obtained from young adult male or female rats. In contrast, the androgen-related rise in SC production was significantly reduced in acinar cells isolated from tissue of orchiectomized and hypophysectomized rats. In summary, these findings demonstrate that androgens directly increase the synthesis of SC by lacrimal gland acinar cells in vitro. This effect, however, may be significantly altered by prior changes in the endocrine environment of acinar cells in vivo.     

Carbonic anhydrase isozyme VI in rat lacrimal gland

Using monoclonal antibody specific to rat carbonic anhydrase isozyme VI (CA VI), the isozyme was localized in the lacrimal gland. A minority of acini (less than 10% of the total) contained a few immunoreactive acinar cells. Enzyme histochemistry indicated that the CA VI-positive cells were the only cells possessing CA in the lacrimal acini. In the acinar cells, the reaction product for CA VI was distributed in the secretory granules and cytosol between secretory granules. Except for mitochondrial enzyme (CA V) activity, the intracellular distribution of enzyme activity was similar to that of CA VI immunoreactivity, suggesting that rat lacrimal acinar cells contain only CA VI and CA V. CA VI in the secretory granules was discharged into the acinar lumen and is considered to carry out its function on the surface of the conjunctiva and cornea. The cytosolic CA VI may function in situ and be involved in electrolyte and water secretion by the acinar cells. Polyclonal antibody to rat erythrocyte CA (CA I and CA II) stained only the interlobular ducts. In contrast, all the ductal elements exhibited CA enzyme activity. This discrepancy between immunohistochemistry and enzyme histochemistry suggests the presence of CA isozyme(s) other than CA I, CA II and CA VI in the lacrimal duct.     

Androgen and estrogen receptors: potential of crystallography in the fight against cancer

Androgen (AR) and estrogen (ERalpha and ERbeta) receptors are primary targets in the treatment of hormone-sensitive tumors such as prostate or breast cancers. Because of their diverse and important roles in normal and pathologic physiology, these nuclear receptors have prompted intense research. Here, we review how structural studies conducted over the past several years on AR and ERs have provided significant advances in our comprehension of androgen and estrogen signaling and how this information can be used in the fight against cancer.     

Androgen receptors in the rat epididymis and their hormonal control.

The concentrations of cytoplasmic receptor sites for androgens in the caput, corpus and cauda epididymidis, and the effect of ligation of the efferent ducts and testosterone treatment after bilateral castration on the concentration of receptors in the caput have been measured. Androgen receptors in the ventral prostate have been measured in the same animals for comparison. The caput has the highest concentration of receptor sites, the corpus the lowest. The ligation of the efferent ducts has no effect on this concentration which is dependent on testicular androgens. The present data do not yet allow explanation of the differential response of the different regions of the epididymis and of the other accessory glands to the administration of androgens.     

Persistent changes in gene expression induced by estrogen and progesterone in the rat mammary gland.

Epidemiological studies have consistently shown that an early full-term pregnancy is protective against breast cancer. We hypothesize that the hormonal milieu that is present during pregnancy results in persistent changes in the pattern of gene expression in the mammary gland, leading to permanent changes in cell fate that determine the subsequent proliferative response of the gland. To investigate this hypothesis, we have used suppression subtractive hybridization to identify genes that are persistently up-regulated in the glands of E- and progesterone (P)-treated Wistar-Furth rats 28 d after steroid hormone treatment compared with age-matched virgins. Using this approach, a number of genes displaying persistent altered expression in response to previous treatment with E and P were identified. Two markers have been characterized in greater detail: RbAp46 and a novel gene that specifies a noncoding RNA (designated G.B7). Both were persistently up-regulated in the lobules of the regressed gland and required previous treatment with both E and P for maximal persistent expression. RbAp46 has been implicated in a number of complexes involving chromatin remodeling, suggesting a mechanism whereby epigenetic factors responsible for persistent changes in gene expression may be related to the determination of cell fate. These results provide the first support at the molecular level for the hypothesis that hormone-induced persistent changes in gene expression are present in the involuted mammary gland.