Sexual dimorphism in the Harderian gland proteins of the golden hamster

SDS polyacrylamide gel electrophoresis of male and female hamster Harderian gland homogenates has shown a clear-cut sexual dimorphism. At least three major proteins present in the male gland are missing from the female gland. Two of the above are associated with the tubular clusters of the male gland while the third seems to be a structural component.     

Analysis of ear sebum of the Syrian hamster (Mesocricetus auratus) reveals pronounced sexual dimorphism

External ear of male and female Syrian hamsters (Mesocricetus auratus) were extracted with hexane and separated by class on thin-layer chromatography (TLC). Separated lipid classes were eluted and saponified, and non-saponifiable lipids further characterized by TLC, gas-liquid chromatography (GLC), UV and IR spectroscopy and functional group analyses. Many sex differences were observed, most notably the presence of sex-specific sterols of males and females. Mature animals were found to have greater quantities of ear sebum, but the characteristic qualitative lipid profiles of each sex were apparent in immature animals.     

Androgenic control of immunoreactive somatostatin in the Harderian gland of the Syrian hamster

Harderian glands of Syrian hamsters contained measurable levels of immunoreactive somatostatin. After an extraction procedure, serial dilutions of tissue were assayed and showed parallelism in the displacement curve with dilutions of purified somatostatin standard in the radioimmunoassay. Somatostatin concentrations were higher in female hamsters (10.0 +/- 2.1 ng/mg protein) than in males (2.6 +/- 0.4 ng/mg protein). Castrated males had somatostatin values in the range of females (12.4 +/- 2.3 ng/mg protein) at 1 month after gonadectomy. Testosterone implants prevented the rise of Harderian gland somatostatin in castrated males. Gonadectomized males had lower somatostatin content in the gland than did control males (1.0 +/- 0.2 ng/mg protein) at 2 months after castration. Somatostatin values in females were unaffected by gonadectomy, but there were variations during the oestrous cycle, with a nadir detected at dioestrus-1 and maximal values coincident with the day of the ovulation.     

Gender differences and time course of castration-induced changes in porphyrins, indoles, and proteins in the Harderian glands of the Syrian hamster.

Sexual differences and the effects of orchidectomy were determined for porphyrin and melatonin concentrations and for the activities of the enzymes N-acetyltransferase and hydroxyindole-O-methyltransferase, which synthesize melatonin from serotonin, in the Harderian glands of the Syrian hamster. Porphyrin concentrations in intact males were about 1/400th those of intact females. Castration for 1 week increased male Harderian porphyrin concentrations 10-fold; by 3 weeks, castrated male porphyrin levels were 140 times those of control values. N-Acetyltransferase activity in intact male Harderian glands was about 4 times that of females. Castration led to a drop in N-acetyltransferase activity to female levels within 2 weeks. Hydroxyindole-O-methyltransferase activity was 7 times higher in females than in males and castration had no effect on male Harderian hydroxyindole-O-methyltransferase activity. Neither gender nor castration influenced Harderian melatonin concentrations. Soluble proteins in Harderian glands from male and female hamsters and from male hamsters castrated for 1 and 4 weeks were examined by sodium dodecyl sulfate--polyacrylamide gel electrophoresis. The gel profiles revealed several differences among the protein distribution in male and female gland lysates. Orchidectomy led to a female protein pattern within 4 weeks.     

In vivo relationship between monoamine oxidase type B and alcohol dehydrogenase: effects of ethanol and phenylethylamine

The role of acute ethanol (2.5 g/kg i.p.) and phenylethylamine (100 mg/kg i.p.) on the brain and platelet monoamine oxidase activities, hepatic cytosolic alcohol dehydrogenase, redox state and motor behaviour were studied in male rats. Ethanol on its own decreased the redox couple ratio, as well as, alcohol dehydrogenase activity in the liver whilst at the same time it increased brain and platelet monoamine oxidase activity due to lower Km with no change in Vmax. The elevation in both brain and platelet MAO activity was associated with ethanol-induced hypomotility in the rats. Co-administration of phenylethylamine and ethanol to the animals, caused antagonism of the ethanol-induced effects described above. The effects of phenylethylamine alone, on the above mentioned biochemical and behavioural indices, are more complex. Phenylethylamine on its own, like ethanol, caused reduction of the cytosolic redox ratio and elevation of monoamine oxidase activity in the brain and platelets. However, in contrast to ethanol, this monoamine produced hypermotility and activation of the hepatic cytosolic alcohol dehydrogenase activity in the animals. The results suggest that some of the toxic actions of ethanol in rats may be mediated through the activation of monoamine oxidase type B, with the consequent depletion of the endogenous levels of phenylethylamine. The data appear to support the concept of phenylethylamine involvement in affective disorders.     

Histochemical study of monoamine oxidase latency in the liver of the rat

Summary In a histochemical test system with adrenaline as substrate and nitroblue tetrazolium (NBT) as electron acceptor, an increase of NBT reduction in rat liver sections was found microspectrophotometrically following short hypotonic treatment. Investigations with iproniazide, a monoamine oxidase inhibitor, and non-enzymatic NBT reduction showed that the increased formazan formation was related to the presence of monoamine oxidase. It is suggested that the reason for the observed increase of formazan formation is due to increased permeability of the inner mitochondrial membrane to NBT. Consequently, the increase of monoamine oxidase observed in the histochemical test system does not represent mobilization of a latent activity, but rather complete assessment of activity that is normally present.     

N-acetyltransferase activity and indole contents of the male Syrian hamster Harderian gland: changes during the light:dark cycle

The activities of N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT) and the indole contents of the Harderian glands of male Syrian hamsters were studied throughout a 24-h period. NAT activity exhibited a sharp rise 1 h after lights on, decreasing to basal levels 1 h later. Neither a HIOMT activity nor a melatonin concentration rhythm was detected throughout the 24 h. The 5-hydroxytryptamine (serotonin) concentration was highest during the dark phase reaching a peak at 0300 h; with light onset serotonin levels exhibited a rapid short-term drop. The 5-hydroxytryptophol concentration was highest during the mid- to late photophase; the lowest values to this constituent were measured late in the dark phase and at 1 h after lights on. The 5-hydroxyindole acetic acid concentration of the Harderian glands was rather stable throughout the 24-h period but levels did show a short-lived drop 1 h after light onset. Only a few animals contained detectable amounts of N-acetyl-5-hydroxytryptamine (N-acetylserotonin) in their Harderian glands. In agreement with previous work on the Harderian glands of female Syrian hamsters, the present results in males suggest that light onset is associated with marked changes in Harderian indoleamine metabolism.     

Histochemical localization of monoamine oxidase types A and B in the adrenal gland

Summary The distribution of monoamine oxidase types A and B within the adrenal galdn was studied in several mammals by histochemical methods. Controls showed that the methods were valid. The bovine adrenal medulla contained mostly the B type enzyme, distributed heterogeneously, with some A type associated with endothelium, nerves, and cells surrounding the nerves. The bovine adrenal cortex showed a marked zonation of the two types of monoamine oxidase. The zona glomerulosa contained the B type enzyme and the zona fasciculata and zona reticularis contained the type A enzyme. The adrenal medulla of the dog, cat, and rat demonstrated relatively little enzyme activity and it appeared to be both type A and B. The adrenal cortex of these animals appeared to contain mostly the B type enzyme, except the canine zona reticularis, which contained some A type monoamine oxidase as well.     

Histochemical study of monoamine oxidase latency in the liver of the rat.

In a histochemical test system with adrenaline as substrate and nitroblue tetrazolium (NBT) as electron acceptor, an increase of NBT reduction in rat liver sections was found microspectrophotometrically following short hypotonic treatment. Investigations with iproniazide, a monoamine oxidase inhibitor, and non-enzymatic NBT reduction showed that the increased formazan formation was related to the presence of monoamine oxidase. It is suggested that the reason for the observed increase of formazan formation is due to increased permeability of the inner mitochondrial membrane to NBT. Consequently, the increase of monoamine oxidase observed in the histochemical test system does not represent mobilization of a latent activity, but rather complete assessment of activity that is normally present.     

N-acetyltransferase activity, hydroxyindole-O-methyltransferase activity, and melatonin levels in the Harderian glands of the female Syrian hamster: changes during the light:dark cycle and the effect of 6-parachlorophenylalanine administration

The activities of NAT and HIOMT and the melatonin content of the Harderian glands of female Syrian hamsters were studied. When hamsters were kept under a light:dark cycle of 14:10 (lights on at 06.00 h), NAT activity exhibited a sharp, short term rise at one hour after lights on. Simultaneously, the activity of HIOMT, which forms melatonin, exhibited a rapid decline. Melatonin levels, like HIOMT activity, also showed a precipitous drop at one hour after light onset. After the respective changes, both NAT and HIOMT activity reverted back to night time levels. Melatonin levels remained depressed for several hours but by 1400 h (8 hours after lights on), nighttime melatonin values were re-established. Treatment of female hamsters with PCPA, a trytophan hydroxylase inhibitor, led to depressed levels of Harderian melatonin without affecting the activities of either NAT or HIOMT.     

Ultrastructure of the blood vessels in the Harderian gland of the hamster (Mesocricetus auratus): existence of sinusoids

The Harderian gland blood supply of female and male hamsters was studied using light and electron microscopy. A profuse vascularization surrounding secretory acini was observed. Among the blood vessels, the existence of large and irregular sinusoidal capillaries was apparent. These sinusoids appeared in close association to the basal aspect of the secretory cells. Typical, small, fenestrated capillaries were also observed within the connective tissue. The existence of this particular vascularization together with other morphological features of the secretory cell basal pole suggest a possible endocrine function of these orbital glands.     

Sexual dimorphism in N-acetyltransferase activity, hydroxyindole-O-methyltransferase activity, and melatonin content in the Harderian gland of Syrian hamsters: changes following gonadectomy

The activities of N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT) and the melatonin content of the Harderian glands of intact and gonadectomized male and female Syrian hamsters were studied. NAT activity in intact male Harderian glands was twice that of the female. Prepubertal or adult castrated males exhibited a decrease in NAT activity to a level comparable to that seen in the female. Testosterone implants in the castrated males led to a recovery of the original male NAT levels. Intact male hamsters had very low levels of Harderian HIOMT activity and melatonin content in comparison with the glands of the females. Prepubertal gonadectomy but not castration of adult males raised the levels of HIOMT activity and the melatonin content to those of the females. Bilateral ovariectomy had no effect on melatonin content, NAT activity, or HIOMT activity in the female hamster Harderian gland.     

Marked sexual dimorphism of lacrimal gland peroxidase in hamster: repression by androgens and estrogens

Peroxidase secreted in tears by the lacrimal glands is a marker of secretory activity of these glands and is believed to have an antimicrobial function. We report for the first time a marked sex difference in lacrimal gland (LG) peroxidase in hamsters ( approximately 3.4-fold higher activity in females), which is due to an unusual repression by physiological levels of androgens in males. LG peroxidase activity was markedly induced in a time-dependent manner after gonadectomy in males and also females ( approximately 8- and 2-fold, respectively) and was strongly repressed by androgen treatment in a dose- and time-dependent manner. Estrogen treatment of gonadectomized hamsters could also repress LG peroxidase but not below female levels. These repressions by androgens and estrogens were significantly prevented upon co-treatment with their respective receptor antagonists. Western blotting showed that differences in LG peroxidase specific activity, in different sex hormonal states and treatments were due to changes in the levels of peroxidase protein in LG. A tear peroxidase with a clear sex difference suggests that it might also have other novel function(s) in hamster tears.