Enhancement of aphrodisiac activity in male rats by ethanol extract of Kaempferia parviflora and exercise training.

This study aimed to investigate the effects of Kaempferia parviflora extract (KD) and exercise training on reproductive function in male rats. Sexually mature males were assigned to four groups: control, KD70 (received 70 mg kg(-1) day(-1) for 4 weeks), Ex (exercise training for 4 weeks), Ex + KD70 (exercise training with KD 70 mg kg(-1) day(-1)). At the end of treatment regimes, sexual behaviours including mount latency (ML), mount frequency (MF), ejaculation latency (EL), post-ejaculation latency (PEL), number of mount within 30 min (MF(30)) and number of ejaculation (NEL) were assessed by a video camera, and fertility was tested by natural mating. Results showed that KD had no effect on the weights of reproductive organs, liver, kidneys and levator ani muscle. On the other hand, the weights of epididymis, seminal vesicles, prostate gland and levator ani muscle were significantly increased in the Ex and Ex+KD70 groups. ML and EL were shortened in all treatment groups, but PEL was decreased only in KP70 group. Only Ex and Ex + KD70 groups exhibited lower MF and higher NEL whilst MF(30) were not changed in all groups. None of the treatments altered male fertility. It is concluded that KD enhanced sexual motivation whereas exercise training promoted both sexual motivation and performance.

Effects of Kaempferia parviflora extracts on reproductive parameters and spermatic blood flow in male rats.

Krachaidum (KD, Kaempferia parviflora Wall. Ex. Baker), a native plant of Southeast Asia, is traditionally used to enhance male sexual function. However, only few scientific data in support of this anecdote have been reported. The present study investigated the effects of feeding three different extracts of KD (alcohol, hexane, and water extracts) for 3-5 weeks on the reproductive organs, the aphrodisiac activity, fertility, sperm motility, and blood flow to the testis of male rats. Sexual performances (mount latency, mount frequency, ejaculatory latency, post-ejaculatory latency) and sperm motility were assessed by a video camera and computer-assisted sperm analysis respectively, while blood flow to the testis was measured by a directional pulsed Doppler flowmeter. The results showed that all extracts of KD had virtually no effect on the reproductive organ weights even after 5 weeks. However, administration of the alcohol extract at a dose of 70 mg/kg body weight (BW)/day for 4 weeks significantly decreased mount and ejaculatory latencies when compared with the control. By contrast, hexane and water extracts had no influence on any sexual behavior parameters. All types of extracts of KD had no effect on fertility or sperm motility. On the other hand, alcohol extract produced a significant increase in blood flow to the testis without affecting the heart rate and mean arterial blood pressure. In a separate study, an acute effect of alcohol extract of KD on blood flow to the testis was investigated. Intravenous injection of KD at doses of 10, 20, and 40 mg/kg BW caused dose-dependent increases in blood flow to the testis. The results indicate that alcohol extract of KD had an aphrodisiac activity probably via a marked increase in blood flow to the testis.

Mediation of contraction in rat cauda epididymidis by alpha-adrenoceptors.

Subtypes of alpha-adrenoceptors responsible for contractions of the rat cauda epididymidis were studied in vivo by micropuncture using a servo-nulling pressure transducer system. Administration of both non-selective and selective alpha-adrenoceptor agonists in doses of 1-40 microg noradrenaline kg(-1) body weight (BW), 1-100 microg clonidine kg(-1) BW, or 100-800 mg methoxamine kg(-1) BW enhanced contractions of the proximal cauda epididymidis in a dose-response manner. The potency of the agonists were noradrenaline > or = clonidine>methoxamine. Pre-treatments with selective alpha(1)-adrenoceptor antagonist (prazosin) and alpha(2)-adrenoceptor antagonist (yohimbine) at the doses of 400 and 800 microg kg(-1) BW, respectively, had very little effect on spontaneous contractions, but effectively blocked the responses to the maximal doses of methoxamine and clonidine. The responses could not be explained by the systemic effects of agonists and antagonists. The results suggest that contraction of the proximal cauda epididymidis of rats is mediated by both alpha(1)- and alpha(2)-adrenoceptors. The latter appears to be more abundant.

Effects of sulphapyridine on sperm transport through the rat epididymis and contractility of the epididymal duct.

This study was undertaken to investigate the effects of sulphapyridine on the transport of spermatozoa through different regions of the epididymis and on the contractility of the epididymal duct in the rat. Sperm transport was investigated by labelling testicular spermatozoa with [3H]thymidine and measuring intraluminal pressures of the epididymis by micropuncture, using a servo-nulling pressure transducer system. In control rats, the transit times of epididymal spermatozoa from the initial segment to the caput, from the caput to the proximal cauda, and from the proximal cauda to the distal cauda were 2, 6 and 3 days, respectively, giving a total transit time of 11 days. The total transit time was shortened to 8 days after treatment with sulphapyridine at a dosage of 450 mg kg-1 for 38-52 days. The rate of sperm transport was most affected in the caput epididymidis. Measurements of intraluminal pressures showed that sulphapyridine had no effect on spontaneous contractions in any regions of the epididymis. However, the frequency of contraction of the corpus and cauda epididymides in response to administration of 10 micrograms noradrenaline kg-1 in the sulphapyridine-treated rats was significantly higher (P < 0.05) than it was in the controls. Methacholine, at a dose of 20 micrograms kg-1, produced a smaller increase in basal pressure in the caput epididymidis of sulphapyridine-treated rats (P < 0.05) compared with controls. The results led to the conclusion that sulphapyridine increases the rate of sperm transport from the caput through the cauda epididymidis, in part, by changes in the responsiveness of the epididymis to the autonomic nervous system.

H+ transporters in the main excretory duct of the mouse mandibular salivary gland.

1. We used microspectrofluorimetry with the pH-sensitive fluoroprobe 2',7'-bis(2-carboxyethyl)-5(and-6)-carboxyfluorescein (BCECF) to study the regulation of cytosolic pH (pHi) in the isolated, perfused main excretory duct of the mouse mandibular gland. 2. In nominally HCO3(-)-free solutions, removal of Na+ from the lumen alone caused pHi to decline whereas removing it from the bath alone did not. 3. Readmission of Na+ to the lumen of ducts studied under zero-Na+ conditions caused pHi to recover fully. This recovery was blocked by 5-(N-ethyl-N-isopropyl)-amiloride (EIPA) with a half-maximum concentration of 0.5 mumol l-1, indicating the presence of an apical Na(+)-H+ exchanger. 4. Readmission of Na+ to the bath of ducts studied under zero-Na+ conditions also caused pHi to recover. This recovery was blocked by 100 mumol l-1 EIPA, indicating the presence of a basolateral Na(+)-H+ exchanger. 5. Measurements of H+ fluxes indicated that the apical Na(+)-H+ exchanger was approximately four times more active than the basolateral Na(+)-H+ exchanger. 6. In three sets of experiments (in the absence of Na+, in the presence of Na+, and in the presence of Na+ plus 100 mumol l-1 EIPA), the effects of changing luminal K+ concentration on pHi were examined. We found no evidence for the presence of K(+)-H+ exchange or Na(+)-coupled K(+)-H+ exchange in the apical membranes of duct cells. 7. pHi recovery under nominally HCO3(-)-free conditions following acidification with an NH4Cl pulse was abolished by removal of Na+ from the bath and luminal solutions, indicating that no Na(+)-independent systems such as H(+)-ATPases were present. 8. A repeat of the above experiments in the presence of 25 mmol l-1 HCO3- plus 5% CO2 did not reveal any additional H+ transport systems. The removal of luminal Cl-, however, caused a small rise in pHi. This latter effect was blocked by 500 mumol l-1 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulphonic acid (H2-DIDS), suggesting that a Cl(-)-HCO3- exchanger in the apical membrane might contribute in a minor way to pHi regulation. 9. We conclude that the predominant H+ transport systems in the mouse mandibular main excretory duct are Na(+)-H+ exchangers in the apical and the basolateral membranes. The model we postulate to account for electrolyte transport across the main duct in the mouse mandibular gland is quite different from that previously developed for the rat duct but is similar to that developed for the rabbit duct. The difference is in concordance with the known ability of the mandibular gland of the rat, but not the rabbit or the mouse, to secrete a HCO3(-)-rich final saliva.

Role of androgens in survival of spermatozoa in epididymis of tammar wallaby (Macropus eugenii).

Studies of undiluted micropuncture samples of luminal fluid from the cauda epididymidis of the tammar indicated that spermatozoa are immotile in situ and spontaneously activate during collection or subsequent incubation in vitro. The suppression of sperm motility was related to the androgen status of the tammars and when this was increased by the use of Silastic implants of testosterone propionate, the spontaneous activation of samples was delayed for up to 2 h during incubation in vitro. Spermatozoa survived for up to 9 weeks when isolated in the cauda epididymidis between ligatures around the ductus. However, even after isolation for 3 weeks their viability was reduced compared with samples from the contralateral, unligated duct. Isolation of a length of ductus between ligatures also reduced the concentration of spermatozoa in the lumen of the duct and reduced the concentration of some proteins in the epididymal plasma. However, it did not affect the electrophoretic pattern of detergent extracts of spermatozoa. A study of the effects of orchidectomy and testosterone therapy indicated that sperm survival in the epididymis is androgen dependent. Orchidectomy reduced the concentration of spermatozoa in the luminal fluid and the volume of luminal fluid, and resulted in an increase in the concentration and a change in the electrophoretic pattern of protein in the fluid. The effects of orchidectomy were reduced or prevented by testosterone therapy. It is concluded that the cauda epididymidis of the tammar is at least as well adapted for sperm storage as it is in the eutherian mammals that have been studied.

Protein synthesis and secretion by the epididymis of the tammar wallaby, Macropus eugenii (Macropodidae: Marsupialia).

The objectives were to assess the following in a marsupial: which proteins are synthesized by the different regions of the epididymis and secreted into the lumen of the ductus; the effect of the experimental method on the detection of protein secretion; the role of the testis in regulating the protein synthesis and secretion; and whether any of the secreted proteins may associate with spermatozoa. Samples from untreated animals were collected for examination by perfusing Krebs-bicarbonate through the ductus epididymidis in vivo (microperfusion), and after incorporation of [35S]methionine during incubation of minced duct in vitro. Electrophoresis of the samples showed that the caput and corpus epididymidis (initial segments) secreted most of the proteins that were synthesized and secreted by the epididymal mucosa, and that the cauda epididymidis secreted mainly blood proteins. Also, many more proteins were secreted in vitro than into the microperfusates in vivo, or were found by Jones (1987) in micropuncture samples of epididymal plasma. The synthesis and secretion of five proteins was androgen dependent (M(r) 75,700, 30,000, 18,700, 17,400 and 12,800). Also, the luminal fluids from the testis stimulated the secretion of two proteins (M(r) 46,300 and 36,100) and inhibited the secretion of three proteins (M(r) 43,000, 32,300 and 21,400). Examination of detergent extracts of spermatozoa indicated that they lose three proteins (M(r) 28,000, 30,000 and 47,000) and gain one (M(r) 30,400) during passage through the epididymis. The method of determining protein secretion affected the findings. Protein secretion, its control and its association with spermatozoa are broadly similar in the tammar wallaby to the processes described in eutherian mammals.

Morphometry of the epididymis of the tammar wallaby, Macropus eugenii, and estimation of some physiological parameters.

About 14 ductuli efferentes (mean length 48 cm) leave the testis of the tammar. The caput, corpus and cauda epididymidis constitute 37%, 42% and 21% respectively of the total length of the ductus epididymidis (estimated to be 34.9 m long). The initial segments of the ductus epididymidis are longer, relative to body or testis mass, in the tammar than in eutherian mammals such as the rat. The main morphometric features of the male excurrent duct system of the tammar are a high ratio of surface area of luminal border:luminal volume of the ductuli efferentes (which reabsorb most of the fluid leaving the testis), a high ratio of epithelial volume:luminal volume in the caput and corpus epididymidis (which are involved in sperm maturation) and a low ratio of epithelial volume:luminal volume in the cauda epididymidis (which is involved in sperm storage). Estimates of fluid reabsorption by the ductuli efferentes and protein secretion by the caput epididymidis were respectively 8.9 microL cm-2h-1 and 2.8 micrograms cm-2h-1. Other estimates for the ductuli efferentes, caput, corpus and cauda epididymidis respectively were: sperm velocity (4.5, 4.8, 2.2, and 0.9 mm min-1), duration of sperm transit (107 min, 1.9 days, 4.7 days, and 6.3 days), total number of spermatozoa (4950 x 10(6)) and distribution of extragonadal spermatozoa (0.6, 14, 36 and 49% of the total). The values are within the ranges estimated for eutherian mammals.

Stimulation of protein secretion in the initial segment of the rat epididymis by fluid from the ram rete testis.

Zone 1A of the ductus epididymidis was perfused with ovine rete testis fluid (nRTF) and modifications of it, and a synthetic medium (sRTF) based on the inorganic composition of nRTF. There was little fluid transport by the duct mucosa and nRTF stimulated protein secretion. The secretagogue activity was not extracted by charcoal, was sensitive to protease digestion and was present in a portion of nRTF with a molecular weight of greater than 10,000. The addition of bovine serum albumin to the sRTF stimulated protein secretion, but not to the same extent as equal amounts of protein in nRTF. Polyacrylamide gel electrophoresis of the perfusates showed that proteins with molecular weights of 19,000 (all rats studied), and 22,000, 30,000 and 60,000 (at least half the rats studied) were secreted into the perfusion fluids as well as some blood proteins, but the pattern of secretion was not affected by the composition of the perfusion fluid.

Evidence for blood myo-inositol as a source of the epididymal secretion in the perfused cauda epididymidis of the rat.

The movement of radioactive inositol and glucose across the epididymal epithelium have been studied by perfusion of fluid, which has the major electrolyte compositions resemble those in the native epididymal fluid, through the lumen of a sperm free tubule of the distal cauda epididymidis of intact and nephrectomized rats. During intravenous infusion of (3H)-myo-inositol into the intact rats, labelled myo-inositol and its metabolites entered the lumen. However, only labelled inositol was found in the luminal perfusate when radioactive inositol was infused into the nephrectomized rat. On the other hand, after infusion of (14C)-glucose no trace of labelled inositol appeared in the lumen. Instead, the luminal radioactivity could be accounted for by labelled glucose. The results indicate that the myo-inositol present in the luminal fluid of the rat cauda epididymidis originates, in part, from blood inositol and, in part, from blood glucose.

Effect of sodium and potassium concentrations and pH on the maintenance of motility of rabbit and rat epididymal spermatozoa.

Spermatozoa were collected from the caput and cauda epididymidis of rabbits and rats and diluted in Hank's solution containing BSA, with various concentrations of Na+ and K+. Ionic strength and osmolarity were kept constant. Motility was assessed at various intervals during incubation at 25 degrees C. In the pH range 7.05--7.20, the motility of rabbit spermatozoa was not affected by changes in the ratio of K+ to Na+. Similarly, the motility of rat cauda spermatozoa was not altered, but that of caput spermatozoa was slightly depressed by a high K+/Na+ ratio. In the pH range 5.45--5.85, rabbit cauda and caput spermatozoa had much greater motility in media with a high K+/Na+ ratio. The reverse result was obtained for the rat. These findings indicate that the motility of epididymal spermatozoa is influenced by external Na+ and K+ concentrations and that this phenomenon is pH-dependent.