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The role of the epididymis in descensus testis and the topographical relationship between the testis and epididymis from the sixth month of pregnancy until immediately after birth.

The position of the testis, the relationship between the epididymis and the testis, as well as the development and regression of the gubernaculum were investigated in 18 testicles of children from the 26th week of pregnancy until a few weeks after birth. The most important role in descensus testiculorum is ascribed to the differentiation of the epididymis and the ductus deferens. It is androgen dependent. The testis descends in the processus vaginalis, being attached to its dorsal wall.

Epididymis

Androgen-dependent accumulation of carnitine by rat epididymis after injection of [3H]butyrobetaine in vivo.

After i.m. injection of [3H]butyrobetaine into rats, the accumulation of carnitine into the epididymis, prostate gland, seminal vesicles, testis and heart was studied. The concentration of radiolabeled carnitine into the cauda epididymis increased linearly with time up to 72 h after the injection of the precursor, while its level in the prostate and seminal vesicles decreased rapidly. Very low levels of carnitine were found in the testis. Castration reduced the carnitine accumulation by cauda epididymis to 6% of the control levels while treatment of castrated animals with testosterone propionate (500 mug/day) partly restored the carnitine uptake. Similar treatment with 17beta-oestradiol valerate or 17alpha-hydroxyprogesterone had no effect. Surprisingly, cyproterone acetate (5 mg/day) also significantly stimulated carnitine accumulation by the epididymis to a level above that of the castrated controls. Simultaneous injection of both cyproterone acetate and testosterone propionate to castrated animals caused an additive effect of these steroids. This indicated that cyproterone acetate in this system is working as a weak androgen. Treatment of rats with 17beta-oestradiol valerate also reduced carnitine accumulation by the cauda epididymis. This is due to suppression of pituiatry gonadotrophin secretion, since concommitant treatment with testosterone propionate (500 mug/day) caused a normalization of the carnitine uptake. Treatment of intact rats with cyproterone acetate significantly reduced the epididymal weight, but not the carnitine accumulation. 17alpha-Hydroxyprogesterone treatment had no effect either on the epididymal weight or the accumulation of the carnitine. Unilateral orchiectomy reduced the carnitine accumulation by the cauda epididymis to about 40% of that occurring in the non-operated control side. This indicates that the luminal contact between the testis and epididymis or the luminal content of the epididymis itself is of importance for the androgen-dependent metabolic process occurring in the cauda epididymis. Castration or hormone treatment did not change the conversion of butyrobetaine to carnitine, or the carnitine uptake by heart. Carnitine uptake by the testis after [3H]butyrobetaine injection was rather low and this would exclude the possibility of synthesis of carnitine in the testis as a source of epididymal carnitine. Carnitine only accumulated in the cauda epididymis in vivo 4 to 96 h after injection of [3H]butyrobetaine. The presence of radioactively labeled butyrobetaine or methylcholine was not detected.

Animals

Functional maturation of the epididymis in the rat.

Weight, histological and biochemical changes in the rat epididymis were investigated during prepubertal, pubertal and postpubertal periods. The phase of most rapid growth of the epididymis commenced at 21 days and extended to 60 days of age; this period corresponded closely to the onset of androgen production at 3 weeks and stabilization of the leydig cell number at 60 days. Histological differentiation in the caput epididymis started before sperm entry and was complete in the cauda only several days after the spermatozoa had appeared. The presence of appreciable quantities of glycerophosphorylcholine (GPC) and sialic acid in the epididymis of 21-day-old rats suggested inherent secretory ability of the epididymal epithelium. The concentrations of GPC, sialic acid, phospholipids and glycogen in the epididymis gradually increased with age, but each came under the influence of androgen at a different age. There was no evidence to suggest that the presence of spermatozoa has a stimulatory effect on the epididymis. Maximal secretory activity of the epididymis became established only by 90 days of age.

Animals

Free amino acids in the caput and the cauda epididymis of adult rats.

To examine the content and the composition of free amino acids in the intraluminal fluid of rat epididymis, the fluids were obtained by light pressure on the dissected tissues. The amount of the total free amino acids in the pressed fluid from the caput epididymis was significantly higher than those of the cauda epididymis and the testis. Glu and Gln were predominant amino acids in the caput, and their amounts occupied more than half of the total ninhydrin reactive compounds. Such a high concentration of Glu and Gln was not observed either in the cauda or in the testis. Castration decreased Glu and increased Gln in amount. Testosterone treatment to castrated animals did not restore Glu and Gln contents in the pressed fluid from the caput epididymis to the level observed in intact rats completely. Therefore, it was assumed that a large amount of Glu in the caput was due to many factors; secretion and metabolism of epithelial cells of the gland which might be regulated by androgen, inflow of rete testis fluid, and sperm metabolism of amino acids in the epididymis. The results obtained from the caput epididymis to which the efferent duct of the testis was ligated also supported this interpretation.

Amino Acids

Metabolism of testosterone by the epididymis and ventral prostate of rat and its inhibition by cyproterone acetate.

The metabolism of 3H-testosterone by the epididymis and accessory organs of adult male rats exposed continuously to microdoses of cyproterone acetate from subcutaneous capsules were studied. The major metabolite of 3H-testosterone in the epididymis, vas deferens and ventral prostate of control rat was dihydrotestosterone while the formation of androstanediol by these tissues was low. The highest percentage of DHT was formed by the ventral prostate and cauda epididymis. In rats exposed to cyproterone acetate for four months, the conversion of testosterone to DHT was inhibited in all the tissues but maximally in the ventral prostate and cauda epididymis. In these rats, the secretory function of the ventral prostate was normal while that of the epididymis was markedly decreased. These data are discussed based on the differential thresholds of androgens required to regulate the functions of the accessory organs.

Animals

Activity and androgenic control of glycolytic enzymes in the epididymis and epididymal spermatozoa of the rat.

1. Procedures were developed for the extraction and assay of glycolytic enzymes from the epididymis and epididymal spermatozoa of the rat. 2. The epididymis was separated into four segments for analysis. When rendered free of spermatozoa by efferent duct ligation, regional differences in enzyme activity were apparent. Phosphofructokinase, glycerol phosphate dehydrogenase and glucose 6-phosphate dehydrogenase were more active in the proximal regions of the epididymis, whereas hexokinase, lactate dehydrogenase and phosphorylase were more active in the distal segment. These enzymes were less active in the epididymis of castrated animals and less difference was apparent between the proximal and distal segments. However, the corpus epididymidis from castrated rats had lower activities of almost all enzymes compared with other epididymal segments. 3. Spermatozoa required sonication to obtain satisfactory enzyme release. Glycolytic enzymes were more active in spermatozoa than in epididymal tissue, being more than 10 times as active in the case of hexokinase, phosphoglycerate kinase and phosphoglycerate mutase. 4. The specific activities of a number of enzymes in the epididymis were dependent on the androgen status of the animal. These included hexokinase, phosphofructokinase, aldolase, glyceraldehyde phosphate dehydrogenase, phosphoglycerate kinase, pyruvate kinase, glycerol phosphate dehydrogenase, glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and phosphorylase. 5. The caput and cauda epididymidis differed in the extent to which enzyme activities changed in response to an altered androgen status. The most notable examples were hexokinase, phosphofructokinase, aldolase, phosphoglycerate kinase, 6-phosphogluconate dehydrogenase and phosphorylase.

Animals

Postnatal study on the histochemistry of epididymis in buffalo (Bubalus bubalis).

The histochemical localisation and development of carbohydrates, acid mucopolysaccharides, lipids and desoxyribonucleic acid has been determined in the head, body and tail segments of buffalo epididymis at different stages of postnatal development from 3-76 weeks of age. The cytoplasm contains numerous Schiff-positive, diastase-resistant granules which are abundant in the apical region of the epithelial cells at 3-52 weeks. They decrease in number at 72-76 weeks and instead, a diffuse Schiff-positive material is observed in the cytoplasm of the cells, particularly in regions III-VI of the epididymis. In all the stages of development, the basement membrane and the luminal border of the epithelial cells have strong PAS-positive reaction. The intertubular connective tissue is mildly PAS-reactive, with moderate activity on the endothelial lining and smooth muscles of blood vessels. The capsule shows an intense Schiff-positive material in the fibers while the stereocilia are mildly reactive. PAS activity is less in region I and greater in regions V-VI as compared with the other regions of the epididymis. A moderate quantity of lipids is present in the epithelial cells and smooth muscles of the tubules. Sudanophilia is more pronounced in the tail region as compared with the head and body regions of the epididymis. Acid mucopolysaccharides are present, minutely, in the epithelial cell cytoplasm, with moderate activity on the stereocilia of the tubules. The Feulgen reaction is deep in region I and light to moderate in the other regions of the epididymis.

Age Factors

Pairing of spermatozoa in the epididymis of the opossum (Didelphis virginiana): a scanning electron microscopic study.

Spermatozoa of the opossum, Didelphis virginiana, were taken from three different regions of the epididymis and examined by scanning electron microscopy, phase-contrast and dark-field microscopy. It has been demonstrated that marked morphological changes occur in spermatozoa during their passage through the epididymis. In the first segment or head of the epididymis, spermatozoa are non-motile and the long axis of their nuclei is perpendicular to that of the tail. The nucleus is V-shaped with a thick and a thin arm. At the junction of the two arms. At the junction of the two arms, where the arms join, the articular fossa receives the capitulum of the connecting piece which attaches the head of the spermatozoon to the tail. Spermatozoa from the central region of the epididymis show a re-orientation of the nuclei which now lie parallel to the long axis of the tail. Both paired and unpaired spermatozoa are found and show an increase in motility. In the caudal region of the epididymis all spermatozoa are paired. Pairing occurs as a result of the close apposition of adjacent cell membranes covering the acrosomes of the larger arms.

Animals

[P. Metastasis of prostatic carcinoma to the epididymis].

When reviewing the bibliography of this rare metastatic location of cancer of the prostate gland, we have only been able to compile the observations published by HUMPHREY (1), BROTHERUS (2) and SMITH (3) wo describe cases with the same location and characteristics. All the observations of metastasis in the epididymis of cancer of the prostate gland constitute unexpected findings during the course of surgical castration operations. The morphology is always the same, small indurated nodules respecting the rest of the epididymis. On the other hand, metastases in the epididymis of extraurinary adenocarcinomas most of which were digestive, were not discovered until the increased size and pain in the testicle attracted the patient's attention and the exploration led to the identification of the tumour in the epidiymis (2, 4, 5). However, primitive malignant tumours of the epididymis constitute an infrequent lesion. In the literature there were found to be more primitive tumours in relation to the metastatic ones and it was difficult to establish the diagnosis of a primitive tumour of the epididymis without having previously ruled out the possibility of other distant neoplasias which are sometimes even asymptomatic.

Adenocarcinoma

The features of sperm maturation in the epididymis of a marsupial, the brushtailed possum Trichosurus vulpecula.

Possum spermatozoa undergo a distinctive process of maturation in the epididymis, as shown by change in the properties of the sperm surface, by modification of their morphology and by their increasing capacity for progressive motility. Modification of the sperm surface over the head and tail is demonstrated by the different affinities of sperm from successive regions of the epididymis for FITC-conjugated wheat germ agglutinin and concanavalin A, and for cationic ferric oxide colloidal particles. Changes in sperm head morphology are caused by (1) a dramatic reshaping and consolidation of the acrosome in which excess plasma membrane overlying it is sloughed as a cluster of vesicles, (2) a reorientation of the nucleus almost parallel to the axis of the tail and (3) distal movement of the droplet from its initial envelopment of the nucleus to an eccentric position on the anterior segment of the midpiece. Spermatozoa released from the testis and caput epididymidis are essentially immotile or exhibit only lazy uncoordinated movements, whereas many from the corpus and most from the more distal regions of the epididymis display an energetic, progressive motility imparted by a rapid and stiff tail beat of narrow arc. This maturation of the capacity for motility is accompanied by an enhanced stability of the dense fibers and sheath, which became more resistant to the disruptive action of SDS and DTT, and by changes in the ultrastructure of the sperm tail. These include modification of the matrix of the mitochondria and also an unusual differentation of the midpiece into two distinct segments. The anterior segment is defined by profuse peri-mitochondrial stacks of membranes which developed as spermatozoa pass through the epididymis. These membranes, although prominent in mature spermatozoa fixed in situ, appear sparse and disorganised in spermatozoa fixed after 15 to 30 minutes of active motility in physiological medium, suggesting their possible utilisation in motile spermatozoa. The posterior segment is characterised by a thick peri-mitochondrial cytoplasmic sleeve, by spirally arranged parallel fibrous bands immediately beneath the plasma membrane and, subsequently, as spermatozoa pass into the lower corpus epididymidis, by rows of flask-shaped surface invaginations which develop between the spiral bands. Despite broad similarities in the features of sperm maturation in this marsupial and in eutherian mammals, there are distinct differencesin the structural organisation of their spermatozoa, particularly in the pserm head. Until more is known of the details of fertilisation in marsupials the significance of these differences will remain unclear.

Agglutinins

The structure of the epididymis, efferent ductules and ductus deferens of the guinea pig: a light microscope study.

In the guinea pig, the narrow part of the epididymis that traverses the upper pole of the testis and passes downward over the entire length of the gonad is composed exclusively of efferent ductules and the initial segment (zone I) of the epididymal duct. At the beginning of zone II, the narrow contour of zone I expands into a large globular region which lies adjacent to the caudal pole of the testis. The globular region of the guinea pig epididymis is commonly referred to as the cauda epididymidis but in the present study, examination with the light microscope reveals that it is composed of five histologically distinct zones (zones III through VII). A detailed histological analysis of the characteristics of the epithelium in the seven zones of the guinea pig epididymis and in the efferent ductules and ductus deferens was udertaken to obtain a better understanding of structure-function relationships in the epididymis of the guinea pig. It was found that each of the zones could be readily distinguished on the basis of its histological features and primarily on the basis of the appearance of the principal cells.

Animals

Tumors of the epididymis.

Tumors of the epididymis, both primary and secondary, whether benign or malignant are very rare. The elements of the spermatic duct system, whether inside the testis or outside it in the epididymis and spermatic cord, exhibit only 1/20th of that of the germinal epithelium. In accordance with the mesodermal, or mesothelial origin of the whole spermatic ducr system, tumors of the epididymis and spermatic cord are - with few exceptions - mesenchymal tumors, the most frequent of these being the so-called adenomatoid tumor, a benign slowly growing tumor. With regard to the relative frequency of the various extratesticular tumors, a British panel has shown that one quarter of its cases are adenomatoid tumors, a second quarter sarcomas of children and juveniles (rhabdomyosarcomas and embryonic sarcomas), a third quarter sarcomas of adults and aged, while the last quarter includes simple benign mesenchymal tumors, mixed tumors, metastatic tumors, and others. There is only one extremely rare epithelial tumor of the epididymis, the papillary cystadenoma, which seems to be a partial menifestation or forme fuste of Hippel-Lindau's disease. So far only 40 cases have been described in the world literature. Therapy of intrascrotal but extratesticular tumors always ought to be surgical: if the tumor proves to be benign, simple excision suffices. In cases of malignancy, immediate inguinal lymphadenectomy and - as with testicular tumors - lymphography followed by retroperitoneal lymphadenectomy. Whether one should, in principle, then continue treatment either chemotherapeutically or radiotherapeutically, or do so only when there is evidence of metastasis, is a question whiich still remains open.

Adult

Cellular proliferation and renewal in the various zones of the hamster epididymis after colchicine administration.

Cellular proliferation was examined in the various regions of the excurrent ducts of the reproductive system of the male golden hamster after colchicine administration. Each of the two zones of the ductuli efferentes displayed only infrequent mitotic figures. Similarly, the basal cell population of the epididymis as well as the epithelium of the vas deferens exhibited too few mitotic figures to allow meaningful evaluation of cellular proliferation. The following observations were reported. 1. The principal cells of the entire epididymis are characterized by a singular diurnal cycle of proliferative activity reaching its maximum about 3 P.M. and its nadir about 9 A.M. 2. A circadian rhythm of cell division is also found in the principal cells of each of the six zones of the epididymis, although the time of maximal activity may vary from zone to zone. 3. The most actively proliferating region of the excurrent duct system is zone 3 of the epididymis, whereas the least active region is the ductuli efferentes. 4. The renewal rates of the principal cells vary considerably at various points along the excurrent duct system, ranging from cells which renew themselves as many as 10 times to cells which fail to renew themselves at all during the normal life-span of the animal.

Animals

Effect of testosterone propionate on levels of carnitine and testicular androgen binding protein (ABP) in rat epididymis.

Twenty-one day old rats were treated for 10 days with various doses of testosterone propionate (TP) (10 micrograms to 10 mg/day) and the levels of L-carnitine and testicular androgen binding protein (ABP) were measured in the 105,000 x g supernatant fractions of epididymis. Treatment with TP in increasing doses had a biphasic effect on the level of ABP in the epididymis; thus, with doses of TP of 10-100 micrograms/day, the ABP level was reduced in a dose-dependent way, whereas with higher doses of TP (0.2 to 1 mg/day) the extent of reduction of ABP levels was less as the dose of TP increased. Treatment with high doses (5 mg or 10 mg/day) did not change the ABP level compared with non-treated control rats. The concentration of carnitine increased linearly (log dose-response) with increasing doses of TP (10-200 micrograms/day) and there was no further increase after treatment with higher doses of TP. In adult rats TP (175 micrograms and 17.5 mg/day) reduced the level of ABP but not the level of carnitine in the epididymis. These studies suggest, therefore, that ABP is of minor importance for the supply of androgens to the carnitine-concentrating cells in the corpus and cauda epididymis.

Androgen-Binding Protein

The effect of castration and testosterone replacement on specific proteins and androgen levels of the rat epididymis.

The normal weight increase of the epididymis during sexual maturation and its maintenance through adulthood were found to be dependent on the provision of androgens. Binding of [3H]dihydrotestosterone (DHT) to the epididymal 8S cytoplasmic receptor gradually decreased after castration to become undetectable after 25 days. Binding to the androgen binding protein (ABP) was absent 4 days after castration and was not reinduced by 3 weeks of testosterone (T) administration. Unilateral castration for periods of up to 27 days showed the disappearance of ABP with preservation of the 8S receptor on the castrated side, indicating a testicular source for ABP and the epididymal origin of the 8S receptor. The tissue concentrations of T and DHT in the epididymis became undetectable 30 days after castration and were restored to normal values by administration of testosterone in large doses (1.5 mg/100 g BW). Similar results were obtained in rats castrated at 10 days of age and injected with testosterone until 60 days old. The ratio DHT/T was depressed in the castrate and increased with testosterone treatment. The protein content of the epididymis (mg of protein/g wet weight) was also found to be influenced by androgens. Our results show evidence of some mechanisms involved in the trophic effect of androgens upon the epididymis and suggest the possible androgenic control of epididymal 5alpha-reductase activity. They also indicate that a testicular factor is required for the maintenance of the 8S cytoplasmic androgen receptor. It is not known whether this factor is testosterone or some other testicular secretion.

Androgens

Steroids in fluids and sperm entering and leaving the bovine epididymis, epididymal tissue, and accessory sex gland secretions.

Ten steroids which may have a role in the process of sperm maturation within the epididymis were quantified by competitive protein binding or radioimmunoassay. Rete testis fluid (RTF) carrying testicular sperm into the epididymis was rich in dehydroepiandrosterone and testosterone (21 +/- 2 and 33 +/- 3 ng/ml) while cauda eipididymal plasma (CEP) around sperm which have completed maturation had high levels of progesterone, dihydrotestosterone, 3beta-androstanediol, dehydroepiandrosterone and testosterone (7.4 +/- 0.8, 20.3 +/- 1.1, 6.5 +/- 0.4, 8.0 +/- 0.7 and 11.5 +/- 0.7 ng/ml). About 4 mug of steroids enter the epidymis daily in RTF, but less than 1% was found in CEP; the balance presumably was absorbed by the epithelium in the proximal caput epididymidis. Nevertheless, tissue levels of total 17beta-OH androgens were lower in the proximal caput than in the distal caput or corpus epididymidis. In all zones of the epididymis, dihydrotestosterone accounted fro about 70% of the total 17beta-OH androgens found in the nuclear fraction. In the cytoplasmic fraction, however, dihydrotestosterone predominated only in the distal caput and corpus epididymidis. In the cauda epididymidis, CEP and sperm probably accounted for less than 35% of the total 17beta-OH androgens and less than 25% of the dihydrotesterone. The progesterone concentration of the cauda than in the caput epidymidis. Twice washed testicular sperm contained more testosterone than cauda epididymal or ejaculated sperm (16.6 +/- 1.9, 1.6 +/- 0.2 and 1.5 +/- 0.3 ng/10(9) sperm, respectively), but less progesterone (0.5 +/- 0.1, 1.3 +/- 0.2 and 1.0 +/- 0.4 ng/10(9) sperm, respectively). As a consequence of mixture with estrogen-rich prostatic fluid (150 +/- 9 pg/ml), ejaculated sperm contained a relatively high amount of estrogens (112 +/- 15 pg/10(9) sperm). These studies revealed marked differences in steroid profiles of fluids entering and leaving the epididymis and of infertile testicular and fertile cauda epididymal sperm.

Androstane-3,17-diol

Regulation by testosterone and serum protein of DNA synthesis in the developing epididymis of the rat.

Rates of DNA synthesis were measured as an index of cellular proliferation during the pubertal development of the rat epididymis. A highly reproducible pattern of DNA synthesis was defined by (1) a prepubertal, testosterone-insensitive peak of DNA synthesis at 25 days; (2) a dramatic decrease in DNA synthesis with the onset of puberty; (3) a major burst of testosterone-dependent synthesis peaking at 40 days in the head of the epididymis and at 40-60 days in the tail; (4) a fall to low levels as the adult organ weight was attained. An organ culture system was defined and utilized to analyse further the hormonal dependence of DNA synthesis in the epididymis. Testosterone and dihydrotestosterone failed to activate DNA synthesis at any stage of development in vitro. DNA synthesis was stimulated 100-300% by insulin at supra-physiological concentrations and by protein serum factor(s) at physiological concentrations. The serum activity was stable to heat treatment at 60 degrees C, destroyed by heating at 70 degrees C, and was present in the sera of hypophysectomized animals. These results indicate a primarily 'permissive' role for the action of testerone on DNA synthesis in the epididymis: testosterone acts to permit the expression of a developmental 'programme' of cell proliferation which is activated by specific protein(s) in serum.

Age Factors

Influence of androgens on the weights of the male accessory reproductive organs and on the activities of mitochondrial enzymes in the epididymis of the rat.

The influence of androgens on the male accessory glands of the rat was assessed in terms of changes in weight and of the specific activity of the mitochondrial enzymes, succinate dehydrogenase, glycerolphosphate dehydrogenase and pyruvate carboxylase, in the epididymis. In some instances, the activity of the cytoplasmic enzymes, hexokinase and phosphofructokinase, was also measured and the influence of androgens on these enzymes was found to be similar to that on the mitochondrial enzymes. After the administration of androgen to castrated rats the specific activity of enzymes reached a new steady state sooner than did epididymal weight. The time taken for the specific activity of the enzymes to reach a new steady state after the removal of androgen was variable, depending on the enzyme and the region of the epididymis. This time was generally longer, however, than the time taken for induction, and in the case of glycerolphosphate dehydrogenase, the decline of activity was slower in the cauda than in the caput. In castrated animals, about 100 times as much androgen was required to attain maximum tissue weight as was required to attain maximum enzyme activity. The epididymis, prostate and seminal vesicles responded similarly to androgen in terms of the dose-response pattern and the time taken for tissue weight to attain a new steady-state value, although the gain in weight of the epididymis relative to its weight in unstimulated control animals was less than the relative gain of the other accessory glands. Enzymes in the cauda epididymidis required lower amounts of androgen to elicit maximum activity than were required by those in the caput. The rate of change in the accessory glands in attaining new steady-state levels of tissue weight and enzyme activity was independent of the dose of androgen except during the first few days of hormone administration. Androgens were the most effective steroids in stimulating an increase of tissue weight and enzyme activity, although some changes were induced by oestradiol-3-benzoate and progesterone.

Animals