Epididymal Fat-Derived Sympathoexcitatory Signals Exacerbate Neurogenic Hypertension in Obese Male Mice Exposed to Early Life Stress.

[Figure: see text].

The distribution of nerves supplying the testis, epididymis and accessory sex glands of Suncus murinus.

Chronic testicular pain remains an important challenge for urologists. Investigation of the innervation of male gonads thus becomes essential for deepening our understanding of their regulatory roles in male reproductive physiology and pathophysiology. Studies of testicular innervation are mainly limited to the intratesticular peptidergic nerves of the testis by immunohistochemical and acetylcholinesterase histochemical investigations in some animals. Little is known about the detailed, overall distribution in general experimental animal testis. In this study, the distribution of nerves supplying the testis, epididymis and accessory sex glands of Suncus murinus was investigated by whole mount immunohistochemistry staining using a neurofilament protein antibody. Testicular nerves arose through three routes: nerves deriving from the mesenteric and renal plexuses accompanied the testicular artery, entering into the testicular hilum through the superior ligament of the testis. The nerves originating from the hypogastric plexus then ran along the internal iliac artery, deferential artery, and passed through the mesoductus deferens or mesoepididymis, innervating the cauda and corpus of the epididymis, the vas deferens and the inferior pole of the testis. The third route arose from the pelvic plexus, distributed in the seminal vesicle and the prostate. The density of nerve fibers was higher in the cauda epididymidis than in the testis, and more abundant in the vas deferens. The different origins and distribution densities of testicular nerves in S. murinus may serve different neuronal regulatory functions, and, therefore, S. murinus may be an important model animal for understanding the different characteristics of testicular pain.

Differential sympathetic outflow to adipose depots is required for visceral fat loss in response to calorie restriction.

The sympathetic nervous system (SNS) regulates energy homeostasis in part by governing fatty acid liberation from adipose tissue. We first examined whether SNS activity toward discrete adipose depots changes in response to a weight loss diet in mice. We found that SNS activity toward each adipose depot is unique in timing, pattern of activation, and habituation with the most dramatic contrast between visceral and subcutaneous adipose depots. Sympathetic drive toward visceral epididymal adipose is more than doubled early in weight loss and then suppressed later in the diet when weight loss plateaued. Coincident with the decline in SNS activity toward visceral adipose is an increase in activity toward subcutaneous depots indicating a switch in lipolytic sources. In response to calorie restriction, SNS activity toward retroperitoneal and brown adipose depots is unaffected. Finally, pharmacological blockage of sympathetic activity on adipose tissue using the ß3-adrenergic receptor antagonist, SR59230a, suppressed loss of visceral adipose mass in response to diet. These findings indicate that SNS activity toward discrete adipose depots is dynamic and potentially hierarchical. This pattern of sympathetic activation is required for energy liberation and loss of adipose tissue in response to calorie-restricted diet.

Microsurgical subinguinal varicocelectomy in children, adolescents, and adults: surgical anatomy and anatomically justified technique.

Microsurgical varicocelectomy has become the gold standard in adults because of low recurrence and postoperative hydrocele rates; it is increasingly applied in children and adolescents. This review aims to provide the surgeon with the necessary surgical anatomy of the spermatic cord and with a step-by-step, anatomically justified description of technique, toward clearer comprehension and improved application. The anatomic compartments of the spermatic cord are delineated by the external and internal spermatic fasciae. Venous drainage of testis-epididymis is accomplished by the internal spermatic, deferential, and external spermatic (cremasteric) veins. All 3 anastomose at the caudal pole of testis, and then via gubernacular veins with the posterior scrotal veins. Another anastomosis exists between a cremasteric branch and anterior scrotal veins, which gives the external pudendal vein. Subinguinal approach offers access to varicose spermatic veins and collaterals. Use of surgical microscope offers identification of small veins, preservation of arteries, lymphatics, and nerves, and appreciation of spermatic cord fasciae, which permits the development of two surgical planes. In the surgical plane of internal spermatic vessels, internal spermatic veins are ligated, whereas the testicular artery and innervation, as well as lymphatics, are preserved. In the plane of cremasteric vessels and vas, cremasteric veins are ligated, whereas the cremasteric artery, vas deferens and its vasculature, lymphatics, and the genital branch of genitofemoral nerve are preserved. Delivery of the testis to ligate gubernacular veins is at the discretion of the surgeon. Finally, venous return is effected by deferential and scrotal veins, or, when gubernacular veins are ligated, by deferential veins only.

Effects of T-type calcium channel blockers and thalidomide on contractions of rat vas deferens.

BACKGROUND AND PURPOSE: In rat vas deferens, nerve mediated-contractions to a single electrical stimulus consist of an early purinergic and a later adrenergic component with differing sensitivities to L-type calcium channel blockers. We have investigated the effects of the T-type calcium channel blockers mibefradil and (1S, 2S)-2-[2-[[3-(1H-benzimidazol-2-yl)propyl]methylamino]ethyl]-6-fluoro-1,2,3,4-tetrahydro-1-(1-methylethyl)-2-naphthalenyl cyclopropanecarboxylic dihydrochloride (NNC 55-0396) against contractions in rat vas deferens. In addition, the actions of thalidomide were examined. EXPERIMENTAL APPROACH: Prostatic and epididymal portions of rat vas deferens were stimulated with a single electrical stimulus every 5 min, and mouse whole vas deferens was stimulated with 40 pulses at 10 Hz every 5 min. KEY RESULTS: Both mibefradil and NNC 55-0396 (100 microM) produced inhibition of contractions of epididymal portions (42 +/- 13%, n= 7, and 43 +/- 4%, n= 15, of control respectively). However, both agents produced small inhibitions of responses in prostatic portions, presumably by L-type calcium channel block. Thalidomide (100 microM) inhibited contractions in epididymal (55 +/- 4% of control, n= 17) but not in prostatic portions of rat vas deferens. Thalidomide (10-100 microM) also inhibited contractions in mouse vas deferens. CONCLUSIONS AND IMPLICATIONS: The T-type calcium channel blockers mibefradil and NNC 55-0396 block particularly the adrenoceptor-mediated, nifedipine-resistant response to nerve stimulation in rat vas deferens, and this may suggest that this component involves T-type calcium channels. In addition, thalidomide has actions that resemble those of the T-type calcium channel blockers, in that it blocks nifedipine-resistant contractions in epididymal portions.

Muscarinic acetylcholine receptor subtypes in the male reproductive tract: expression and function in rat efferent ductules and epididymis.

In mammals, at least five different muscarinic acetylcholine receptor subtypes (mAChRs; M(1)-M(5)) are known to be widely expressed and distributed in different tissues from different species. They mediate distinct physiological functions according to their location and receptor subtype. Multiple events are associated with the regulation of intracellular signaling by mAChRs, and a coordinated balance of the molecular mechanisms governing receptor signaling, desensitization, resensitization, and mitogenic signaling is known to occur in various cell types. Most of the actions of acetylcholine (ACh) in the male reproductive tract are induced by its effects on mAChRs, but the role of specific mAChR subtypes on male reproductive function and fertility are still not well understood. The rat efferent ductules and epididymis are androgen-dependent tissues of the male reproductive tract, with important roles in the process to form a viable and fertile sperm. In the present study, aspects of the expression, localization, and potential function of mAChR subtypes in rat efferent ductules and epididymis are reviewed. Furthermore, evidences for the implication of mAChRs in the regulation of protein synthesis and secretion in these tissues are presented. Taken together, the studies contribute to our understanding about physiological aspects of mAChR and mechanisms by which the cholinergic system affects male reproduction.

Regulation of epididymal contractility during semen emission, the first part of the ejaculatory process: a role for estrogen.

INTRODUCTION: Ejaculation is an important step of the male sexual response, and consists of three separate phases: emission, ejection, and orgasm. In contrast to the erectile process, whose neurological and vascular mechanisms have been well elucidated, the pathophysiology of the ejaculatory process remains yet to be completely investigated. In humans, the emission and the ejection phases are regulated by an integrated and time-coordinated activity of the parasympathetic and sympathetic systems, which finally leads to sperm propulsion from the urethra. The first step in the ejaculatory process involves the epididymis, where a series of contractile waves begins, allowing sperm progression throughout the duct and toward the vas deferens. Interestingly, along with the complex neurological pathways, some non-neuronal factors (oxytocin [OT] and endothelin-1 [ET-1]) and sex hormones (estrogen) have been demonstrated to take part in the peripheral regulation of epididymal contractility. AIM: This article reviews some of the physiological non-neuronal mechanisms underlying the epididymal contractility, and reports evidences of an estrogenic regulation. METHODS: We reviewed here our and other groups' publications on the role of ET-1/OT and estrogens in modulating the epididymal contractility. MAIN OUTCOME MEASURE: Data were obtained by an extensive examination of the published peer-reviewed literature on this topic. RESULTS: Evidences support that, although the epididymis has a rich innervation, other local, non-neuronal factors participate in the nerve-independent epididymal contractility. ET-1 and OT, along with their cognate receptors, have been demonstrated to act, in an estrogen-dependent autocrine and paracrine loop, to regulate epididymal contractile activity in rabbit, and at least partially, in humans. CONCLUSION: Ejaculation is the result of the complex, and today still not fully elucidated, interplay between neuronal and non-neuronal, sex-steroid-dependent factors.

Effects of heptanol on neurogenic contractions of vas deferens: A comparative study of stimulation frequency in guinea pig and rat.

This study examines the role of gap junctional communication in smooth muscle in relation to the frequency of stimulation and the innervation density of the tissue in the generation of neurogenic contractions. Toward this end the effects of heptanol, a gap junctional blocker, on the neurogenic contractions of guinea pig and rat vas deferens at different frequencies of stimulation (single pulse, 5, 10, 20, 40, 60, and 80 Hz) were studied. In both the prostatic and epididymal halves of these tissues, heptanol abolished the neurogenic contractions at the lower frequencies of stimulation. At higher frequencies, contractions were resistant to heptanol action. The effect of heptanol on the neurogenic contractions was found to decrease with increasing stimulation frequency. The neurogenic contractions of rat vas deferens were more resistant to heptanol than those of guinea pig vas deferens. Our data indicate that gap junctional communication is significant in the generation of neurogenic contractions in both guinea pig and rat vas deferens in a frequency-dependent manner, and we discuss the mechanisms underlying these findings.

Supraspinal connections of the reproductive organs: structural and functional aspects.

Gonadal functions are governed by the hypothalamo-hypophyseal system. Other organs of the reproduction tract are under the regulatory action of gonadal steroids. In the past two decades several data have been accumulated on the involvement of fine-tuning control mechanisms which include autocrine and paracrine effects of biologically active substances produced locally and the regulatory action of nerves innervating the organs of the system. Recent studies using the viral transsynaptic technique have revealed cell groups in the central nervous system that are transneuronally connected with the male and female reproductive organs. This review summarizes neuromorphological data on the supraspinal innervation of reproductive organs and the functional significance of these brain areas in the control of reproduction.

Effect of olfactory stimulation with flavor of grapefruit oil and lemon oil on the activity of sympathetic branch in the white adipose tissue of the epididymis.

It has been mentioned in the field of aromatherapy that the fragrance of grapefruit has a refreshing and exciting effect, which suggests an activation of sympathetic nerve activity. It also can be assumed that the activation of sympathetic nerve innervating the white adipose tissue (WAT) facilitates lipolysis, then results in a suppression of body weight gain. This study addressed the effect of olfactory stimulation with the scent of grapefruit oil and lemon oil on the efferent nerve activity in the sympathetic branch of the WAT of the epididymis in the anesthetized rat. The results of the experiments demonstrated that the flavor of the citron group increased sympathetic nerve activity to WAT in anaesthetized rat, which suggests an increase in lipolysis and a suppression in body weight gain.

Dual effect of agmatine in the bisected rat vas deferens.

The functional effects of the amine agmatine, the putative endogenous ligand for alpha(2)-adrenoceptors and imidazoline receptors, in rat vas deferens were investigated by using the epididymal and prostatic portions. Tissues were contracted by electrical stimulation or by exogenous drugs. In electrically stimulated portions, agmatine caused a dual effect on contractions. In the epididymal portion an inhibition on twitch contractions was observed, which was partially antagonised by idazoxan and yohimbine, indicating the involvement of at least a presynaptic alpha(2)-adrenoceptor-mediated mechanism, without the interference of imidazoline receptors. In the prostatic portion, agmatine enhanced the amplitude of twitches. In contractions induced by exogenous drugs, agmatine potentiated, only in the prostatic segment, the effects of noradrenaline (norepinephrine) or ATP; it also enhanced the effect of low concentrations of KCl and blocked the maximum effect of the higher concentrations. Effects induced by agmatine on the exogenous ATP in the prostatic portion were blocked by cromakalim, suggesting a blocking action on the postsynaptic K(+) channels, which explains, in part, the potentiation of the twitch amplitude. It was concluded that agmatine interferes with sympathetic neurotransmission, but the physiological relevance of this needs to be better understood because of the high doses employed to induce its effects.

Immunohistochemical investigations of the autonomous innervation of the cervine testis.

The innervation of the cervine testis was studied in 6 roe deers, 7 red deers and 14 fallow deers. The results for the three species are rather similar. With anti-sera to neurofilament (NF) and neuron specific enolase (NSE), all small and large nerve fascicles can be demonstrated, but single fibers are incompletely stained. Immunoreactions against protein gene product-9.5 (PGP-9.5) and GAP-43 (growth-associated protein-43) are better suited to depict the complete innervation pattern. Bundles of the superior spermatic and inferior spermatic nerves reach the testis via three access routes as funicular, mesorchial and caudal nerve contributions. We found no morphological evidence that the nerves in the cervine testis are directly involved in regulating Leydig cell function or seminiferous tubular motility. The majority of the testicular nerves are associated with the testicular arteries, but the musculature in the walls of the venous plexus pampiniformis is also innervated. All vascular nerve fibers represent postjunctional sympathetic axons displaying a strong dopamine-beta-hydroxylase (DBH) activity, mostly co-expressed with neuropeptide Y (NPY). The presence of cholinergic fibers in the testis of the deer is only sporadic and probably of no functional importance. In all three species of deer, a small quantity of myelinated nerve fibers is encountered in spermatic cord and tunica albuginea and regarded as afferent. The viscerosensory quality in the testicular intrinsic innervation is very likely mediated by the CGRP (calcitonin gene-related peptide)-positive fibers that run independently from the testicular vessels and end in the connective tissue of spermatic cord and tunica albuginea. The testis of the red deer contains significantly more VIP (vasoactive intestinal polypeptide)-positive axons than that of roe and fallow deer. The nerve density in the interior of the testicular lobules shows no regional differences, but there are age- and season-related changes that correlate with the developmental and functional state of the seminiferous tubules. Small testes with solid and narrow tubules, as seen in the prepuberal phase and during seasonal reproductive quiescence, are better innervated than large testes with expanded and spermatogenetically active seminiferous tubules.

On the intrinsic innervation of the epididymis of the camel (Camelus dromedarius).

The innervation of the camel epididymis was studied in 26 apparently healthy, sexually mature animals aged between 4 and 12 years. The material was collected during the different seasons of the year. Generally, five samples were taken from each epididymis. To demonstrate the general innervation pattern, immunohistochemical reactions to protein gene product-9.5, neurofilaments and neuron-specific enolase were used, in addition to acetylcholinesterase histochemistry. The nerve supply of the epididymis comes from two sources: (1) The majority of fibers come from the N. spermaticus inferior and accompany the deferent duct. (2) Another contribution stems from the N. spermaticus superior and enters the head region of the epididymis. From the exterior, the nerves penetrate the capsule of the organ to reach the interductular connective tissue. The terminal ramifications are observed directly within the wall of the duct and the wall of the epididymal arteries. The veins of the camel epididymis are not innervated. In the wall of the ductus epididymidis, the nerve fibers form plexuses at the subepithelial level and in the muscular coat. The amount of nerve fibers increases from the head to the tail, paralleling an increase in the intrinsic musculature. The intramural and interductular innervation of epididymal body and tail shows clear seasonal variations: More fibers and stronger reactions are observed during the winter season; the lowest density and the weakest reactions occur during the summer season. All epididymal nerves of the camel are unmyelinated. The majority of the intramural fibers and all in the arterial wall represent postjunctional sympathetic axons, but in the intramural plexuses of the duct a considerable number of cholinergic fibers are also present. Neuropeptide Y is the most frequent peptidergic transmitter and generally co-localized with dopamine-beta-hydroxylase in the sympathetic axons. Vasoactive intestinal polypeptide has a distribution similar to that of the cholinergic fibers. Calcitonin gene-related peptide-positive axons occur in moderate numbers, but never in the arterial innervation. Together with the relatively rare substance P-containing fibers, the calcitonin gene-related peptide-positive axons seem to represent the only sensory nerves in the camel epididymis.

Segment-specific decrease of both catecholamine concentration and acetylcholinesterase activity are accompanied by nerve refinement in the rat cauda epididymis during sexual maturation.

In the present work, histochemical and biochemical studies were conducted to analyze changes in the pattern of autonomic innervation during sexual maturation, using the rat epididymis as a model. Glyoxylic acid histochemistry and immunohistochemical studies against dopamine beta-hydroxylase (DbetaH) and acetylcholinesterase (AChE) indicated a reduction in the amount of catecholaminergic and AChE-positive neurons, fibers, and puncta detected in the cauda epididymis of adult rats (120 days old), when compared to immature (40 days) and young adult (60 days) animals. No obvious age-related variations were detected in the few catecholaminergic and AChE-positive fibers and puncta present in the caput region. AChE-positive fibers were found sorting out among epithelial cells and ending free upon the epithelial surface or into the tubular lumen of the cauda region of adult rats. Furthermore, a positive staining for AChE in epithelial cells was also detected in the caput and cauda epididymis in all ages studied. Biochemical analysis confirmed a significant decrease in noradrenaline concentration as well as AChE activity in the cauda epididymis with sexual maturation. Immunohistochemical studies against microtubule-associated protein 1B (MAP 1B), a neuronal cytoskeletal marker, further substantiated the quantitative changes observed in catecholaminergic and AChE-positive neuronal elements in the cauda epididymis. Thus, our results documented segment-specific variations in noradrenaline concentration and AChE activity during epididymal sexual maturation and suggest that such variations result, at least in part, from the refinement of the autonomic innervation pattern with age.

Identification of CNS neurons involved in the innervation of the epididymis: a viral transneuronal tracing study.

Cell groups of the spinal cord and the brain transsynaptically connected with the epididymis (caput, cauda) were identified by means of the viral transneuronal tracing technique. Pseudorabies virus was injected into the caput or the cauda epididymidis, and after survival times 4 and 5 days, the spinal cord and brain were processed immunocytochemically. Virus-labeled neurons could be detected in the preganglionic sympathetic neurons (lower thoracic and upper lumbar segments) and following virus injection into the cauda epididymidis, also in the sacral parasympathetic nucleus (L6-S1). Virus-infected perikarya were present in several brain stem nuclei (lateral reticular nucleus, gigantocellular and paragigantocellular nucleus, A5 noradrenergic cell group, caudal raphe nuclei, locus coeruleus, Barrington's nucleus, nucleus of the solitary tract, periaqueductal gray) and in the diencephalon (hypothalamic paraventricular nucleus, lateral hypothalamus). At the longer survival time, some telencephalic structures also exhibited virus-labeled neurons. The distribution of infected neurons in the brain was similar after virus injection into the caput or cauda epididymidis; however, earlier onset of infection was observed after inoculation into the cauda. The present findings provide the first morphological data on a multisynaptic circuit of neurons innervating the epididymis and presumably involved in the control of epididymal functions. reserved.

Smooth muscle does not have a common P2x receptor phenotype: expression, ontogeny and function of P2x1 receptors in mouse ileum, bladder and reproductive systems.

The distribution, ontogeny and role of P2x1 receptors were examined in the smooth muscle of the mouse intestine, bladder, and male and female reproductive tracts using P2x1 receptor subtype selective antibodies and contraction studies. P2x1 receptor immunoreactivity showed a heterogeneous distribution in smooth muscle with high levels expressed in adult vas deferens, bladder, arteries and male reproductive organs. In contrast, P2x1, receptors were below the level of detection in the smooth muscle of the ileum and female reproductive tract. P2x1 receptor immunoreactivity was detected at adult levels from birth in the bladder. However, in the vas deferens, immunoreactivity was only detected from 10 days after birth and reached adult levels by approximately 1 month old. A similar pattern of expression was seen in the vesicular seminalis, epididymis, gland of the vas deferens and coagulating gland. Sensitivity to the P2x1 receptor agonist alpha,beta-methylene ATP (alpha,beta-meATP) and P2x1 receptor-deficient mice were used in functional studies to determine the role of P2x1 receptors in the control of smooth muscle. alpha,beta-meATP (100 microM) failed to evoke contractions of the epididymis, or seminal vesicle and P2x1 receptors did not contribute to the control of uterine smooth muscle. In the ileum, alpha,beta-meATP (100 microM) evoked a transient relaxation followed by a contraction. These responses were abolished by the P2 receptor antagonist iso-pyridoxalphosphate-6-azophenyl-2'-5'-disulphonate (iso-PPADS) (30 microM). Relaxant responses were abolished by the adenosine A1 receptor antagonist 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) (1 microM). Contractile responses were reduced by > 80% in the ileum from P2x1 receptor-deficient mice. alpha,beta-meATP-evoked contractions were reduced by approximately 35% by TTX (1 microM) and were unaffected by atropine (10 microM). These studies indicate that P2x1 receptors are not expressed throughout all smooth muscles and that their expression is developmentally regulated. In addition, they provide evidence to suggest that P2x1 receptors are present on pre-synaptic nerve terminals in the enteric nervous system.

Spontaneous reconstruction of the canine hypogastric nerve over a long period after removing half of its length.

Spontaneous reconstruction of the sympathetic pathway projecting to the seminal tract after serious injury has not been well understood. Multiple cross-innervation mechanisms from the spinal cord via the hypogastric nerve to the seminal tract have been demonstrated currently. This study was undertaken to explore long-term spontaneous reconstruction of the canine hypogastric nerve (HGN), which controls ejaculation, after removing half of its length. Further, the study tried to identify the crossed-pathway(s) reconstructed. Responses of the vas deferens/epididymis and bladder neck to electrical stimulation of the lumbar splanchnic nerve (LSN) or the HGN were examined. In six dogs whose hypogastric nerve was injured bilaterally as described above 4 years before, corresponding to more than 20 years in human, nine (43%) and 13 (57%) of the 21 LSNs stimulated elicited elevation of vasal and bladder neck pressure, respectively. By combining re-transection of a HGN, the following pathways to the vas deferens/epididymis were identified to have been reconstructed: (1) to the ipsilateral was deferens/epididymis without crossing to the other side; (2) to the contralateral vas deferens/epididymis by crossing to the other side at the caudal mesenteric plexus (CMP); (3) to the contralateral vas deferens/epididymis by crossing to the other side from the ipsilateral HGN at the commissural branches between the right and left pelvic plexuses (CBPP); and (4) to the ipsilateral vas deferens/epididymis by crossing twice at the CMP to the other side and at the CBPP again from the contralateral HGN to the ipsilateral side. The similar patterns of reconstruction were also observed in the bladder neck. The above results indicate that the sympathetic pathways via the HGN to the canine seminal tract can be reconstructed spontaneously in a high rate over a long period after serious injuries and that their cross-innervation system can be repaired.

On the innervation of the donkey testis.

The innervation pattern of the adult donkey testis was investigated by immunohistochemistry and acetylcholinesterase histochemistry. Autonomous nerves reach the testis by three access-routes as funicular, mesorchial and caudal contributions. From these, the funicular contribution accompanying the testicular artery and pampiniform plexus is the strongest and most important one. Testicular innervation in the donkey is not uniform. The spermatic cord as well as the epididymal region, cranial and caudal poles (tunica albuginea and adjacent parenchyma and stroma) are well innervated, mostly by vascular nerves. Towards the free border of the testis, the nerve density in the tunica albuginea decreases continuously. In the interior of the gonad, approximately one third of the testis, situated between the free border and the central mediastinum, is practically devoid of any innervation. The great majority of the testicular nerves demonstrated by the present techniques are non-myelinated vascular nerves which react positive for dopamine-beta-hydroxylase and tyrosine hydroxylase, thus representing postjunctional sympathetic fibers. Many of these also contain neuropeptide Y. The testicular innervation of the donkey testis is free of cholinergic fibers. Calcitonin gene-related peptide-containing nerves are found as solitary varicose axons in the wall of blood vessels, but also in stromal connective tissue of the spermatic cord, tunica albuginea and septula testis.

Ultrastructural study of the luminal surface of the ducts of the epididymis of gallinaceous birds.

The various ducts of the epididymides of four gallinaceous birds, the turkey (Meleagris gallopavo), domestic fowl (Gallus gallus), guinea-fowl (Numida meleagris) and Japanese quail (Coturnix coturnix japonica) were studied at the scanning and transmission electron microscopy levels. The tissues were fixed either by immersion or vascular perfusion, for comparative purposes. Each duct system, save for a few details, presented similar morphological features in all species. The epithelial surface of the rete testis was regular and each cell bore a single cilium, as well as numerous, or in some parts, very few, short, regular microvilli. Each of the Types I and II non-ciliated cells of the proximal and efferent ducts displayed abundant, moderately long and regular microvilli, and a solitary cilium. The ciliated cells exhibited tufts of cilia. The Type III non-ciliated cell of the connecting and epididymal ducts exhibited a solitary cilium, and numerous microvilli which were intermediate in length between those of the rete testis and those of the efferent ducts. Vascular perfusion of the avian epididymal tissue was the superior method of fixation because it minimised the developments of fixation artefacts. Apocrine secretion did not appear to occur in the epididymis of these birds as the apical blebs of Types I, II and III cells, which have previously been reported, only manifest in this study in inadequately fixed tissues, and were therefore viewed as being artefacts. The present findings suggest that the current terminology, as applied to the avian epididymis, be retained.

Investigation of the subtypes of alpha1-adrenoceptor mediating contractions of rat vas deferens.

1 The subtypes of alpha1-adrenoceptor mediating contractions of rat vas deferens to endogenous and exogenous noradrenaline and to the exogenous agonists methoxamine, phenylephrine and A61603 have been examined. 2 The effects of antagonists on the shape of concentration-response curves, both tonic and phasic, to the four agonists were analysed. Prazosin produced parallel shifts in all cases. Particularly for RS 17053 against noradrenaline, there was some evidence for a resistant component of the agonist response. High concentrations of RS 17053 (1-10 microM) virtually abolished tonic contractions but phasic contractions were resistant. 3 A series of nine antagonists (the above and WB4101, benoxathian, phentolamine, BMY 7378, HV 723, spiperone) were investigated against contractions to noradrenaline. The correlation with the potency of the series of alpha1-adrenoceptor antagonists against contractions to noradrenaline was significant only for the alpha1A-adrenoceptor ligand binding site (r=0.88, n=9, P<0.01). 4 In epididymal portions (nifedipine 10 microM), the isometric contraction to a single electrical pulse is alpha1-adrenoceptor mediated. The correlation with ligand binding sites for 11 antagonists (the above plus ARC 239 and (+)-niguldipine) was significant only for the alpha1D-adrenoceptor subtype (r=0.65, n=11, P<0.05). 5 In conclusion, tonic contractions of rat vas deferens produced by exogenous agonists are mediated predominantly by alpha1A-adrenoceptors, although a second subtype of receptor may additionally be involved in phasic contractions. Nerve-stimulation evoked alpha1-adrenoceptor mediated contractions seem to predominantly involve non-alpha1A-adrenoceptors, and the receptor involved resembles the alpha1D-receptor.