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Effects of castration and of testosterone replacement on alpha(1)-adrenoceptor subtypes in the rat vas deferens.

The contractions of the rat vas deferens in response to noradrenaline are mediated through alpha(1A)-adrenoceptors. We observed participation of alpha(1B)-adrenoceptors in these contractions after castration. We now investigated the time course of this plasticity and the effects of testosterone by determining the actions of competitive antagonists on noradrenaline-induced contractions after 7, 14, 21 and 30 days of castration. BMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4.5]decane-7,9-dione dihydrochloride) antagonised noradrenaline-induced contractions in control and castrated rats with low pA(2) values (approximately = 6.8). In control vas deferens, WB 4101 (2-(2,6-dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane hydrochloride) had a slope in the Schild plot no different from 1.0, while slopes lower than 1.0 (approximately 0.6) were observed for vas deferens from castrated rats. Chloroethylclonidine was ineffective in the control vas while it inhibited noradrenaline-induced contractions in vasa from castrated rats and converted the complex antagonism by WB 4101 into simple competitive antagonism. Treatment of castrated rats with testosterone prevented the effects of castration. The results suggest that alpha(1B)-adrenoceptors are detectable in vas deferens from at least the 7th through the 30th day after castration and that testosterone prevents this plasticity.

Animals↗

The enhancing effects of amastatin, phosphoramidon and captopril on the potency of [Met5]-enkephalin in rat vas deferens.

The enkephalin-inactivating enzymes in rat vas deferens were studied by using the relatively specific inhibitor of each enzyme. The results showed that the rat vas deferens, like the other three preparations, guinea-pig ileum, mouse vas deferens and striatal membranes of guinea-pig brain, which had been investigated previously, contained three distinct enkephalin-hydrolyzing peptidases. Additionally, the enkephalin-hydrolyzing aminopeptidase, endopeptidase-24.11 and peptidyl dipeptidase A in rat vas deferens were found to be inhibited maximally with 1 microM of amastatin, 1 microM of phosphoramidon and 1 microM of captopril, respectively. In contrast to these three enzymes, both L-tyrosyl-L-tyrosine-sensitive dipeptidyl aminopeptidase and D-phenylalanine-sensitive carboxypeptidase were suggested not to be involved significantly in the inactivation of exogenously given enkephalin in rat vas deferens. The characteristics of the enkephalin-degradative enzymes in rat vas deferens were discussed in terms of their similarities to and differences from those in the other preparations.

Animals↗

Failure of non-adrenergic transmission in the rat vas deferens.

Repetitive field stimulation of the vas deferens of the rat with long trains of pulses (90 pulses, 10 Hz, 1 msec) at intervals of 30 or 60 sec produced a decline in the heights of contractile responses. This decline in responses was much more pronounced in phentolamine (5 microM)-treated than in untreated preparations and in the prostatic rather than in the epididymal end of the vas deferens. The decline in the responses was dependent on the number of pulses/train, the frequency of pulses in the train and the interval between successive trains. Potassium chloride (KCl)-induced and adenosine triphosphate (ATP)-induced contractions were largely unaltered after 30 min of stimulation. The decline in the responses was significantly reduced in low but not in high extracellular calcium. 4-Aminopyridine (4-AP) caused an initial potentiation of contractile responses, followed by a faster decline of responses. The decline in responses was more intense in vas deferens from reserpine-treated animals. The results support the view that electrical stimulation-induced decay of motor transmission in the vas deferens of the rat is caused by a specific failure of the non-adrenergic transmission.

4-Aminopyridine↗

5'-flanking and intragenic sequences confer androgenic and developmental regulation of mouse aldose reductase-like gene in vas deferens and adrenal in transgenic mice.

The MVDP (mouse vas deferens protein) gene, which encodes an aldose reductase-like enzyme, is mainly expressed in vas deferens epithelium and adrenal cortex. Vas deferens MVDP gene transcription was known to be under androgenic control, we now have evidence for androgen and probable ACTH responsiveness of the MVDP gene in the adrenal. To analyze the role of potential regulatory regions in hormonal, developmental, and tissue-specific aspects of MVDP regulation, we generated transgenic mice harboring MVDP-CAT fusion genes. The constructs carried either -1.8 or -0.5 kb 5'-flanking sequence attached to the chloramphenicol acetyltransferase gene in presence or absence of a 3.5-kb intragenic fragment in a downstream position. We show that at least two regions ensure proper gene regulation in vivo. The first, located within the 1.8-kb promoter fragment, directs tissue specificity; positive elements necessary for vas deferens and adrenal expression lay within positions -1804 to -510 and -510 to +41, respectively. The second, located within the 3.5-kb intragenic fragment spanning intron 1 to intron 2, increases percentage of expressing lines and behaves as a vas deferens-specific enhancer. Hormonal and developmental control of transgenes closely parallel endogenous gene regulation. Androgen and ACTH responsiveness in adrenals is conferred by 0.5-kb promoter, whereas in vas deferens, full androgenic response of the 1.8-kb promoter required the 3.5-kb intragenic fragment. Thus, vas deferens and adrenals use distinct cis-acting elements to direct and regulate the expression of the MVDP gene.

Adrenal Glands↗

On the transportation of spermatozoa in the vas deferens.

In this report the intraluminal pressure of the vas deferens was measured in vivo in male cross-bred dogs. The measurement of the pressure elucidated that the vas deferens repeated autonomic contraction. This contraction was not the so-called peristalsis but the entire portion of the vas deferens contracted simultaneously. Stimulation of the hypogastric nerves resulted in a prominent elevation of the pressure in the proximal portion of the vas deferens, i.e., the portion approximate to the epididymis, followed 5-10 seconds later by a rise in the distal portion, i.e., the portion approximate to the prostate. These results suggest that the spermatozoa present near the proximal end of the vas deferens are transported littly by little by the autonomic movements of the vas deferens and that on ejaculation the spermatozoa are pressed forward from the proximal end of the vas deferens to its peripheral end by the stimuli through the hypogastric nerves.

Animals↗

Activation of alpha 1-adrenoceptors increases [3H]inositol metabolism in rat vas deferens and caudal artery.

Rings of rat vas deferens and caudal artery were incubated with [3H]inositol in Krebs-Ringer bicarbonate buffer containing 10 mM lithium chloride, and the production of water-soluble [3H]inositol phosphates was monitored. Norepinephrine increased [3H]inositol phosphate accumulation 7-fold in rings from vas deferens and 3-fold in rings from caudal artery. Epinephrine, phenylephrine and methoxamine were as effective as norepinephrine, suggesting that these drugs are full agonists in causing this response. Prazosin, phentolamine and yohimbine completely blocked the stimulation by norepinephrine in both tissues with potencies typical of blockade of alpha 1-adrenoceptors. Despite a substantial receptor reserve for alpha 1-adrenoceptor mediated contractile responses, clonidine, p-amino-clonidine, phenylpropanolamine and ephedrine can only cause a partial contractile response in rat vas deferens. However, all of these partial agonists were either as effective or more effective in increasing [3H]inositol phosphate accumulation in rat vas deferens as they were in activating a contractile response. These data suggest that alpha 1-adrenoceptors increase phosphatidylinositol turnover in rat vas deferens and caudal artery, and that there may be a receptor reserve for alpha 1-adrenoceptor mediated increases in [3H]inositol phosphate accumulation in these smooth muscles.

Adrenergic alpha-Agonists↗

Inhibitory histamine receptors in the rat vas deferens.

The effect of histamine on the field stimulated rat vas deferens preparation was studied and compared to mouse vas deferens. Histamine produced a concentration dependent inhibition of rat vas deferens (ID50 14 microM) similar to that seen with mouse vas deferens (ID50 2.4 microM). Unlike mouse however, where only H2 receptors are observed, the histamine inhibition of rat vas deferens was attenuated by both H1, (diphenhydramine) and H2 (cimetidine) antagonists. It is concluded that rat vas deferens contains both H1 and H2 receptor types.

Animals↗

Effects of the antidepressant St. John's wort (Hypericum perforatum) on rat and human vas deferens contractility.

PURPOSE: Since sexual dysfunction related to vas deferens smooth muscle contractility is a possible side effect of St. John's wort (SJW) (Hypericum perforatum) we evaluated the effect of this herbal antidepressant on rat and human vas deferens contractility. MATERIALS AND METHODS: The effect of SJW was evaluated on contractions induced by electrical field stimulation or exogenous agonists (alpha,beta-methylene adenosine triphosphate and phenylephrine) in isolated rat and human vas deferens. RESULTS: SJW (1 to 300 microM) decreased in a concentration dependent manner the amplitude of electrical field stimulation and agonist induced contractions with the same potency, suggesting direct inhibition of rat vas deferens smooth muscle. Of the chemical constituents of SJW tested hyperforin but not hypericin or the flavonoids quercitrin, rutin and kaempferol inhibited phenylephrine induced contractions. SJW and hyperforin also inhibited phenylephrine induced contractions in human vas deferens CONCLUSIONS: The results of our study demonstrate that SJW directly inhibits rat and human vas deferens contractility. If confirmed in vivo, these results suggest that SJW might affect sexual function in humans. These results might explain delayed ejaculation described in patients receiving SJW.

Animals↗

Sympathetic denervation does not alter the density or properties of alpha-1 adrenergic receptors in rat vas deferens.

Alpha-1 adrenergic receptors in surgically denervated rat vas deferens were studied using radioligand binding assays of [125I] BE 2254 ([125I]BE) and contraction measurements. Scatchard analysis of saturation isotherms of specific [125I]BE binding showed no change in the affinity or density of binding sites 4, 7 or 14 days after denervation of rat vas deferens. The potency of norepinephrine in inhibiting specific [125I]BE binding was also unchanged 7 days after denervation of vas deferens. The potency of phenylephrine in causing contraction in vitro did not change 4, 7 or 14 days after denervation of vas deferens; however, there was a significant increase in the maximum contractile response to phenylephrine at all time points. After partial inactivation of alpha-1 adrenergic receptors in vitro with phenoxybenzamine, there was an equivalent reduction in the number of [125I]BE binding sites in the control and 14-day denervated vas deferens. The equilibrium dissociation constants calculated from contractile measurements for norepinephrine were the same in the control and denervated tissues. However, there was a 2.2-fold increase in contractile sensitivity to norepinephrine 14 days after denervation and a 3.6-fold increase in contractile sensitivity to methacholine 7 days after denervation. These results show that surgical denervation of the rat vas deferens results in an increase in contractile sensitivity to norepinephrine and methacholine and an increase in maximum contraction. However, there is no change in alpha-1 adrenergic receptor density or properties at any time after denervation. Thus, alterations in alpha-1 adrenergic receptors do not contribute to contractile supersensitivity of denervated rat vas deferens.

Animals↗

Androgen regulation of the mRNA encoding a major protein of the mouse vas deferens.

A cDNA encoding the major mouse vas deferens protein (MVDP) has been cloned and characterized. Using in situ hybridization we have identified the epithelial cells of the vas deferens as the site of synthesis of MVDP mRNA. Northern blot analysis suggests that a high level of an mRNA corresponding to the MVDP gene is present in the mouse vas deferens whereas the amount of MVDP mRNA in vas deferens of other species studied, or in other mouse tissues, even if present, is undetectable. Steady-state levels of MVDP mRNA are decreased by approximately 42% 3 days after castration but a significant hybridization signal is still observed even 50 days after castration. Testosterone treatment for 2 weeks is necessary to completely reverse the effect of castration. In vitro transcription assays on isolated nuclei showed that the hormonal induction of the MVDP gene is achieved mainly at transcriptional level.

Aldehyde Reductase↗

Cell- and region-specific localization of lysosomal and secretory proteins and endocytic receptors in epithelial cells of the cauda epididymidis and vas deferens of the adult rat.

The epithelial cells lining the cauda epididymidis and vas deferens are active in endocytosis and have an abundance of lysosomes and a well-characterized secretory apparatus. However, little is known about the nature of lysosomal proteins contained within lysosomes, the types of receptors on the cell surface, and the types of proteins secreted by these cells. In the present study, cathepsins A, D, B, and sulfated glycoprotein (SGP)-1, well-characterized lysosomal proteins, as well as SGP-2, a secretory protein and low-density lipoprotein receptor-related protein-2 (LRP-2), an endocytic receptor, were immunolocalized at the light-microscopic level within epithelial cells of the cauda epididymidis and vas deferens. Principal cells showed numerous intensely reactive lysosomes for cathepsins A, D, and SGP-1 in all regions of the cauda and vas deferens and for cathepsin B only in the cauda epididymidis. Basal cells were intensely reactive for cathepsin A, unreactive for cathepsins D and B, and weakly reactive for SGP-1 in the cauda region. In the vas deferens, these cells were intensely reactive for cathepsin A and SGP-1 and unreactive for cathepsin B; in the case of cathepsin D, basal cells were weakly reactive in the proximal vas deferens but intensely reactive in the middle and distal vas deferens. Clear cells, present in the cauda region and proximal vas deferens, were intensely reactive for cathepsin A, weakly reactive for SGP-1, and unreactive for cathepsins D and B, while narrow cells found mainly in the proximal vas deferens were intensely reactive for cathepsins A, D, and SGP-1 and unreactive for cathepsin B. Thus, the expression of different lysosomal enzymes in the cauda epididymidis and vas deferens is not only cell- but also region-specific, suggesting differences in the type of substrates internalized by these cells. SGP-2, a secretory protein, showed a checkerboardlike staining pattern in the cytoplasm of principal cells of the cauda epididymidis, while the cytoplasm of all principal cells were intensely reactive in the vas deferens. This type of reaction, as well as staining of sperm, suggests that SGP-2 is secreted into the lumen, where it functions in relation to sperm. The endocytic receptor LRP-2 was noted only on the apical surface of principal cells of the cauda and vas deferens and in spherical structures indicative of endosomes suggestive of their role in the uptake of various ligands, including SGP-2, for which it has a high binding affinity. Thus SGP-2 in the cauda and vas deferens is not only secreted but endocytosed by principal cells, suggestive of an active turnover in the lumen. In summary, the epithelial cells of the cauda and vas deferens show marked differences in expression of lysosomal proteins, SGP-2, and LRP-2 suggestive of differences in their functional activity while sperm are stored and protected in these regions.

Animals↗

Effects of Carica papaya seed extracts on the physiology of the vas deferens of albino rats.

The physiology of vas deferens after administrating papaya seed extract (1 mg/0.2 ml/day/rat) for 7 and 15 days was studied with reference to organ weights, fertility rate, some biochemical parameters and contractile pattern of isolated right and left vas deferens. Studies on combined extract + ascorbic acid feeding and discontinuation of treatment for 1 and 2 1/2 months were also carried out. The results revealed that the treatment altered the biochemical parameters except cholesterol levels and the contractile pattern was reduced in comparison to control. However, significant recovery occurred by withdrawal of treatment in all parameters studied. AA feeding was efficaceous in restoring the androgenicity of vas deferens under treated conditions. On the whole, the distal vas deferens was affected more by the seed extract treatment than proximal vas and recovery was slower probably due to its higher threshold requirement for androgen. The results are discussed in the light of recent work.

Animals↗

Ectopia of the vas deferens.

Ectopic location of the vas deferens is a rare congenital abnormality that has a spectrum of presentations. Such cases have been associated with anorectal anomalies. Knowledge of distal Wolffian duct embryology may help to explain the developmental steps involved in these anomalies, as well as provide theories for the variations that may be encountered. Vas deferens development is reviewed, and two cases of vas ectopia are presented to illustrate and lend support to current theories of normal and abnormal vasal embryology.

Child, Preschool↗

Morphologic variations along the length of the monkey vas deferens.

Gross and microscopic morphology of the vas deferens of sexually mature macaque monkeys were studied with special reference to the changes and modifications occurring throughout its entire length. The vas deferens was subdivided into four regional segments designated proximal, middle, distal, and ampulla. Each segment showed morphologic variations and it seemed to correspond roughly to the region located in a different part of the body which it traverses. The longitudinal folds of the epithelium are simple in the proximal region but gradually become more complex towards the distal region. In the ampulla, they often branch and form crypts. Stereocilia of epithelial cells are taller and more uniform in the proximal portion in comparison with the short, irregular stereocilia found in the more distal segment of the vas deferens. The luminal epithelial cells of the proximal part apparently have no granules or cytoplasmic blebbings, while the principal cells in the distal portion contain large numbers of cytoplasmic granules and apical blebs. Cells in the middle segments possess cytological features intermediate to the proximal and distal regions. These structural variations along the length of the vas deferens sug gest that this segment of the excurrent duct of the testis performs functions other than just as a passageway for spermatozoa.

Animals↗

Further evidence for adrenergic transmission in the human vas deferens.

1. Isolated portions of human vas deferens responded to field stimulation of the intramural nerve fibres or to exogenously applied noradrenaline with rhythmical contractions of both longitudinal and circular muscle layers. 2. In guinea-pig and rabbit vas deferens field stimulation produced an initial rapid 'twitch' response which was not found with human vasa. 3. Responses of the human vas to field stimulation were depressed by the alpha-adrenoceptor blocking agents phentolamine and yohimbine. 4. It is concluded that the motor innervation of the human vas deferens is adrenergic and the relevance of this to the physiological operation of the tissue is discussed.

Adrenergic Fibers↗

Functional characterisation of alpha(1)-adrenoceptors in denervated rat vas deferens.

We investigated whether or not surgical denervation of the rat vas deferens changes the alpha(1)-adrenoceptor subtypes involved in the contractions to noradrenaline. Denervated vas deferens was approximately 22 times more sensitive to noradrenaline (pD(2)=7.35+/-0.04) than control vas (pD(2)=6.01+/-0.03). This difference in noradrenaline potency was eliminated when cocaine (6 microM) was added to control vas (pD(2)=7.22+/-0.04). The noradrenaline-induced contractions of control and denervated vas deferens were insensitive to the alpha(1B)/alpha(1D)-adrenoceptor alkylating agent chloroethylclonidine (100 microM, 45 min). The concentration-response curves to noradrenaline in control and denervated vas deferens were competitively antagonised by prazosin (pA(2) approximately equal to 9.6), WB-4101 (pA(2) approximately equal to 9.5), 5-methyl urapidil (pA(2) approximately equal to 8.4), phentolamine (pA(2) approximately equal to 8.7), yohimbine (pA(2) approximately equal to 6.9), BMY 7378 (pA(2) approximately equal to 6.9) and indoramin (pA(2) approximately equal to 8.7). After the treatment of control and denervated vas deferens with phenoxybenzamine, the partial agonist oxymetazoline antagonised competitively the concentration-response curves to noradrenaline showing pA(2) values approximately 7.4 in both groups. We conclude that noradrenaline-induced contractions in control and denervated rat vas deferens are mediated by alpha(1A)-adrenoceptors and that surgical denervation of the rat vas deferens is not able to change the alpha(1)-adrenoceptor subtypes involved in the contractions to noradrenaline.

Adrenergic alpha-Agonists↗

Scanning electron microscopy of the human vas deferens epithelium following oestrogenic treatment.

Human vas deferens from 4 subjects submitted to a prolonged oestrogenic therapy was observed with SEM. The microvilli, characterizing the cell apices in the normal cases, are considerably reduced in number and length. Some cells show thin laminar folds (microplicae) arranged in a complicated labyrinth in the central zone of the apocal surface. The globular apical extrusions characterizing the normal vas deferens secretory cells are no longer present. These observations indicate the decrease of the absorbent capacity and the fall of the secretory function in the vas deferens epithelium following oestrogenic treatment.

Epithelium↗

The inactivation of [Met5]-enkephalin by bestatin-sensitive aminopeptidase, captopril-sensitive peptidyl dipeptidase A and thiorphan-sensitive endopeptidase-24.11 in mouse vas deferens.

The enkephalin-hydrolyzing peptidases in mouse vas deferens were studied and compared with those in guinea pig ileum which had been characterized in the previous study. The present results showed that three distinct peptidases, bestatin-sensitive aminopeptidase, captopril-sensitive peptidyl dipeptidase A, and thiorphan-sensitive endopeptidase-24.11, played a critical role in the inactivation of enkephalin in mouse vas deferens, being consistent with the previous results obtained with guinea pig ileum. However, the data in both previous and present studies showed that the activity of the bestatin-sensitive aminopeptidase relative to that of either the captopril-sensitive peptidyl dipeptidase A or the thiorphan-sensitive endopeptidase-24.11 in guinea pig ileum was higher than that in mouse vas deferens, while the activity of either peptidyl dipeptidase A or endopeptidase-24.11 relative to that of aminopeptidase in mouse vas deferens was higher than that in guinea-pig ileum. In contrast to these three enzymes, both L-tyrosyl-L-tyrosine-sensitive dipeptidyl aminopeptidase and D-phenylalanine-sensitive carboxypeptidase were suggested not to be involved significantly in the inactivation of enkephalin in mouse vas deferens as well as guinea pig ileum.

Amino Acids, Sulfur↗