Urine Release Research Articles

Behavior of the renal kallikrein in spontaneously hypertensive rats: Influence of sexual hormones and aldosterone-sensitive distal nephron ion channels

The renal kallikrein-kinin system (RKKS) has been related to blood pressure control and sodium and water balance. We have previously shown that female spontaneously hypertensive rats (SHR) have high urinary kallikrein activity (UKa) and lower blood pressure (BP) than males whereas ovariectomy stimulates UKa and diminishes BP. We also showed that high K+ intake and prepuberal gonadectomy (Gx) diminish BP with a concomitant increase in UKa and plasma aldosterone levels. Since kallikrein co-localize in the same distal nephron segments of aldosterone effectors, we explored the effect of pharmacological blockage of aldosterone receptor, epithelial Na+ (ENaC) and the rectifying outer medulla K+ (ROMK) channels in different gonad contexts on the gene expression, renal tissue content and urine release of kallikrein. Klk1 gene expression was determined by real-time PCR and enzymatic activity of kallikrein by the amidolytic method. We found that the inhibition of the aldosterone receptor by spironolactone increases kallikrein renal tissue storage and decreases its urinary activity, especially in Gx rats. Moreover, ENaC blockade by benzamil increases the renal content of kallikrein without affecting synthesis or excretion, especially in females and Gx animals, while the inhibition of ROMK by glibenclamide increases the synthesis and renal content of kallikrein only in intact male animals. We concluded that RKKS regulation showed sexual dimorphism and seemed to be modulated by sex hormones throughout a process involving aldosterone and the aldosterone-sensitive ion channels..

COVID-19 drugs in aquatic systems: a review

The outbreak of the human coronavirus disease 2019 (COVID-19) has induced an unprecedented increase in the use of several old and repurposed therapeutic drugs such as veterinary medicines, e.g. ivermectin, nonsteroidal anti-inflammatory drugs, protein and peptide therapeutics, disease-modifying anti-rheumatic drugs and antimalarial drugs, antiretrovirals, analgesics, and supporting agents, e.g. azithromycin and corticosteroids. Excretion of drugs and their metabolites in stools and urine release these drugs into wastewater, and ultimately into surface waters and groundwater systems. Here, we review the sources, behaviour, environmental fate, risks, and remediation of those drugs. We discuss drug transformation in aquatic environments and in wastewater treatment systems. Degradation mechanisms and metabolite toxicity are poorly known. Potential risks include endocrine disruption, acute and chronic toxicity, disruption of ecosystem functions and trophic interactions in aquatic organisms, and the emergence of antimicrobial resistance.

Social isolation uncovers a circuit underlying context-dependent territory-covering micturition

The release of urine, or micturition, serves a fundamental physiological function and, in many species, is critical for social communication. In mice, the pattern of urine release is modulated by external and internal factors and transmitted to the spinal cord via the pontine micturition center (PMC). Here, we exploited a behavioral paradigm in which mice, depending on strain, social experience, and sensory context, either vigorously cover an arena with small urine spots or deposit urine in a few isolated large spots. We refer to these micturition modes as, respectively, high and low territory-covering micturition (TCM) and find that the presence of a urine stimulus robustly induces high TCM in socially isolated mice. Comparison of the brain networks activated by social isolation and by urine stimuli to those upstream of the PMC identified the lateral hypothalamic area as a potential modulator of micturition modes. Indeed, chemogenetic manipulations of the lateral hypothalamus can switch micturition behavior between high and low TCM, overriding the influence of social experience and sensory context. Our results suggest that both inhibitory and excitatory signals arising from a network upstream of the PMC are integrated to determine context- and social-experience-dependent micturition patterns.

The success in the short-distance communication for mating does not depend on chemical signals in the crustacean decapod Procambarus clarkii (Girard, 1852)

Pheromone-driven sex recognition has been widely documented in crayfish and a great deal of evidence supports the involvement of pheromones in their mating behaviour. This study investigates whether sexual interaction and mating success in the red swamp crayfish Procambarus clarkii are dependent on short-distance chemical communication between sexes, mediated by urine-borne pheromones. We compared the mating behaviour of intact animals that could release urine to chemically communicate in a natural way with that of urine-blocked animal pairs, for which chemical communication was precluded. Our results show that urine-borne pheromones are not critical for the reproductive success of P. clarkii, at least over the short-range distance (<1 m) considered in this study, during which the animals were confined in a restricted tank, facing one each other, and thus able to promptly mate. Under these experimental conditions, a lack of urine release neither precluded the occurrence, nor affected the duration of the different phases of mating behaviour. We conclude that short-distance chemical communication in P. clarkii is not a prerequisite for mating, and suggest that it could be affected by alternative sensory modalities, likely vision and/or acoustic signalling.

Profile and quality of life of women in pelvic floor rehabilitation.

Describe the sociodemographic, clinical, and sexual profile, identify profile variables that affect the Health-Related Quality of Life (HRQoL), and evaluate the correlation between two HRQoL questionnaires used in a pelvic floor rehabilitation program. This is an observational, analytical, and cross-sectional study, based on patient records and two questionnaires for HRQoL evaluation. Women presented a mean age of 55.4 years; were married; white; had stress, urge, or mixed urinary incontinence (UI) of moderate to large urine release; and daily or diurnal UI. Only 50.5% had an active sex life and most had sexual complaints. The change in sexual activity and some types of UI affected the HRQoL. The two questionnaires presented a correlation. The profile and correlation between the questionnaires are consistent with the literature. The type of UI and changes in sexual activity affect the HRQoL.

Voluntary urination control by brainstem neurons that relax the urethral sphincter.

Voluntary urination ensures that waste is eliminated when safe and socially appropriate, even without a pressing urge. Uncontrolled urination, or incontinence, is a common problem with few treatment options. Normal urine release requires a small region in the brainstem known as Barrington’s nucleus (Bar), but specific neurons that relax the urethral sphincter and enable urine flow are unknown. Here we identify a small subset of novel Bar neurons that control the urethral sphincter in mice. These excitatory neurons express estrogen receptor 1 (BarESR1), project to sphincter-relaxing interneurons in the spinal cord and are active during natural urination. Optogenetic stimulation of BarESR1 neurons rapidly initiates sphincter bursting and efficient voiding in anesthetized and behaving animals. Conversely, optogenetic and chemogenetic inhibition reveals their necessity in motivated urination behavior. The identification of these cells provides an expanded model for the control of urination and its dysfunction.

Pharmacodynamic evaluation of Lys5, MeLeu9, Nle10-NKA(4-10) prokinetic effects on bladder and colon activity in acute spinal cord transected and spinally intact rats.

The purpose of this study was to determine feasibility of a novel therapeutic approach to drug-induced voiding after spinal cord injury (SCI) using a well-characterized, peptide, neurokinin 2 receptor (NK2 receptor) agonist, Lys5, MeLeu9, Nle10-NKA(4-10) (LMN-NKA). Cystometry and colorectal pressure measurements were performed in urethane-anesthetized, intact, and acutely spinalized female rats. Bladder pressure and voiding were monitored in response to intravenous LMN-NKA given with the bladder filled to 70% capacity. LMN-NKA (0.1-300μg/kg) produced dose-dependent, rapid (<60s), short-duration (<15min) increases in bladder pressure. In intact rats, doses above 0.3-1μg/kg induced urine release (voiding efficiency of ~70% at ≥1μg/kg). In spinalized rats, urine release required higher doses (≥10μg/kg) and was less efficient (30-50%). LMN-NKA (0.1-100μg/kg) also produced dose-dependent increases in colorectal pressure. No tachyphylaxis was observed, and the responses were blocked by an NK2 receptor antagonist (GR159897, 1mg/kg i.v.). No obvious cardiorespiratory effects were noted. These results suggest that rapid-onset, short-duration, drug-induced voiding is possible in acute spinal and intact rats with intravenous administration of an NK2 receptor agonist. Future challenges remain in regard to finding alternative routes of administration that produce clinically significant voiding, multiple times per day, in animal models of chronic SCI.

Central Control Circuit for Context-Dependent Micturition

Urine release (micturition) serves an essential physiological function as well as a critical role in social communication in many animals. Here, we show a combined effect of olfaction and social hierarchy on micturition patterns in adult male mice, confirming the existence of a micturition control center that integrates pro- and anti-micturition cues. Furthermore, we demonstrate that a cluster of neurons expressing corticotropin-releasing hormone (Crh) in the pontine micturition center (PMC) is electrophysiologically distinct from their Crh-negative neighbors and sends glutamatergic projections to the spinal cord. The activity of PMC Crh-expressing neurons correlates with and is sufficient to drive bladder contraction, and when silenced impairs micturition behavior. These neurons receive convergent input from widespread higher brain areas that are capable of carrying diverse pro- and anti-micturition signals, and whose activity modulates hierarchy-dependent micturition. Taken together, our results indicate that PMC Crh-expressing neurons are likely the integration center for context-dependent micturition behavior.

Serotonin and noradrenaline reuptake inhibitors improve micturition control in mice.

Poor micturition control may cause profound distress, because proper voiding is mandatory for an active social life. Micturition results from the subtle interplay of central and peripheral components. It involves the coordination of autonomic and neuromuscular activity at the brainstem level, under the executive control of the prefrontal cortex. We tested the hypothesis that administration of molecules acting as reuptake inhibitors of serotonin, noradrenaline or both may exert a strong effect on the control of urine release, in a mouse model of overactive bladder. Mice were injected with cyclophosphamide (40 mg/kg), to increase micturition acts. Mice were then given one of four molecules: the serotonin reuptake inhibitor imipramine, its metabolite desipramine that acts on noradrenaline reuptake, the serotonin and noradrenaline reuptake inhibitor duloxetine or its active metabolite 4-hydroxy-duloxetine. Cyclophosphamide increased urine release without inducing overt toxicity or inflammation, except for increase in urothelium thickness. All the antidepressants were able to decrease the cyclophosphamide effects, as apparent from longer latency to the first micturition act, decreased number of urine spots and volume of released urine. These results suggest that serotonin and noradrenaline reuptake inhibitors exert a strong and effective modulatory effect on the control of urine release and prompt to additional studies on their central effects on brain areas involved in the social and behavioral control of micturition.

Functional roles of TRPV1 and TRPV4 in control of lower urinary tract activity: dual analysis of behavior and reflex during the micturition cycle

The present study used a dual analysis of voiding behavior and reflex micturition to examine lower urinary tract function in transient receptor potential vanilloid (TRPV)1 knockout (KO) mice and TRPV4 KO mice. In metabolic cage experiments conducted under conscious conditions (i.e., voluntary voiding behavior), TRPV4 KO mice showed a markedly higher voiding frequency (VF; 19.3 ± 1.2 times/day) and a smaller urine volume/voiding (UVV; 114 ± 9 μl) compared with wild-type (WT) littermates (VF: 5.2 ± 0.5 times/day and UVV: 380 ± 34 μl). Meanwhile, TRPV1 KO mice showed a similar VF to WT littermates (6.8 ± 0.5 times/day) with a significantly smaller UVV (276 ± 20 μl). Water intake among these genotypes was the same, but TRPV4 KO mice had a larger urine output than the other two groups. In cystometrogram experiments conducted in decerebrate unanesthetized mice (i.e., reflex micturition response), no differences between the three groups were found in any cystometrogram variables, including voided volume, volume threshold for inducing micturition contraction, maximal voiding pressure, and bladder compliance. However, both TRPV1 KO and TRPV4 KO mice showed a significant number of nonvoiding bladder contractions (NVCs; 3.5 ± 0.9 and 2.8 ± 0.7 contractions, respectively) before each voiding, whereas WT mice showed virtually no NVCs. These results suggest that in the reflex micturition circuit, a lack of either channel is involved in NVCs during bladder filling, whereas in the forebrain, it is involved in the early timing of urine release, possibly in the conscious response to the bladder instability.

Excitatory effects of neurokinin 2 and bombesin receptor peptide agonists vivo in urinary tract of rats with voiding dysfunctions

Bladder function was examined in anesthetized and awake adult female Sprague Dawley rats. Voiding dysfunction models included STZ‐induced diabetic (streptozotocin; STZ; 65mg/kg i.p., 9–20 weeks post treatment), polyuria (5% sucrose‐fed; &gt;;3 weeks post treatment) and spinal cord injured (SCI; T8‐T9 cut; acute or 3 days – 6 weeks post SCI) rats. Telemetry transmitters were implanted into polyuria group to measure bladder pressure in freely moving rats. [Lys5,MeLeu9,Nle10]‐NKA(4–10) (LMN‐NKA), a neurokinin 2 receptor (NK2R) agonist, gastrin‐releasing peptide (GRP), a bombesin receptor 2 (BBR2) agonist, and neuromedin B (NMB), a BBR1 agonist, were delivered via jugular vein catheters.In urethrane anesthetized STZ and control rats, LMN‐NKA (0.1–100μg/kg), GRP (0.1–300μg/kg) and NMB (0.1–300μg/kg) dose dependently increased bladder contraction (BC, up to 4x) and micturition frequency. In STZ rats, LMN‐NKA reduced residual volume by half. In acute anesthetized SCI rats, LMN‐NKA (&gt;;10μg/kg) induced urine release (voiding efficiency ~40%). In awake polyuria rats, LMN‐NKA and GRP (1–100μg/kg) induced BC that was often associated with voiding. In awake SCI rats, LMN‐NKA (100μg/kg) and GRP (100, 300μg/kg) triggered voiding in 25 – 80% of rats.Activation of NK2R and BBRs enhanced or triggered voiding in naïve rats and in several models of voiding dysfunction including STZ and SCI rats.

Tactical Release of a Sexually-Selected Pheromone in a Swordtail Fish

BackgroundChemical communication plays a critical role in sexual selection and speciation in fishes; however, it is generally assumed that most fish pheromones are passively released since most fishes lack specialized scent glands or scent-marking behavior. Swordtails (genus Xiphophorus) are widely used in studies of female mate choice, and female response to male chemical cues is important to sexual selection, reproductive isolation, and hybridization. However, it is unclear whether females are attending to passively produced cues, or to pheromones produced in the context of communication.Methodology/Principal FindingsWe used fluorescein dye injections to visualize pulsed urine release in male sheepshead swordtails, Xiphophorus birchmanni. Simultaneous-choice assays of mating preference showed that females attend to species- and sex-specific chemical cues emitted in male urine. Males urinated more frequently in the presence and proximity of an audience (conspecific females). In the wild, males preferentially courted upstream of females, facilitating transmission of pheromone cues.Conclusions/SignificanceMales in a teleost fish have evolved sophisticated temporal and spatial control of pheromone release, comparable to that found in terrestrial animals. Pheromones are released specifically in a communicative context, and the timing and positioning of release favors efficient signal transmission.

β-Adrenoceptor-mediated differences in transverse and longitudinal strips from the rat detrusor

Contractions and relaxations of the urinary bladder occur in all directions to facilitate urine release and storage. Transverse and longitudinal contractility of detrusor smooth muscle have been studied before using various pharmacologic stimuli but not β agonists. Given the importance of β-adrenoceptors in mediating bladder relaxation, the effects of isoprenaline (IPNA) in transverse and longitudinal contractility were examined. Pretreatment with a low concentration of IPNA (0.1 or 1μM) suppressed carbachol (CCh)-induced contractions, more in the transverse than longitudinal direction. Increasing the IPNA concentration to 10 or 100μM resulted in greater inhibition of longitudinal contractions. Also in the longitudinal direction, IPNA-induced relaxation was greater than in the transverse direction. When precontracted with a submaximal concentration of CCh (1μM), IPNA increased the phasic activity in the longitudinal direction only. In summary, β-adrenoceptor-mediated differences between transverse and longitudinal contractility were revealed. In testing the relaxant properties of selective β-agonists, the findings here should be considered such that other than the conventional longitudinal contractions, measurements are also made in other directions.

To signal or not to signal? Chemical communication by urine-borne signals mirrors sexual conflict in crayfish

BackgroundSexual selection theory predicts that females, being the limiting sex, invest less in courtship signals than males. However, when chemical signals are involved it is often the female that initiates mating by producing stimuli that inform about sex and/or receptivity. This apparent contradiction has been discussed in the literature as 'the female pheromone fallacy'. Because the release of chemical stimuli may not have evolved to elicit the male's courtship response, whether these female stimuli represent signals remains an open question. Using techniques to visualise and block release of urine, we studied the role of urine signals during fighting and mating interactions of crayfish (Pacifastacus leniusculus). Test individuals were blindfolded to exclude visual disturbance from dye release and artificial urine introduction.ResultsStaged female-male pairings during the reproductive season often resulted in male mating attempts. Blocking female urine release in such pairings prevented any male courtship behaviour. Artificial introduction of female urine re-established male mating attempts. Urine visualisation showed that female urine release coincides with aggressive behaviours but not with female submissive behaviour in reproductive interactions as well as in intersexual and intrasexual fights. In reproductive interactions, females predominately released urine during precopulatory aggression; males subsequently released significantly less urine during mating than in fights.ConclusionsUrine-blocking experiments demonstrate that female urine contains sex-specific components that elicit male mating behaviour. The coincidence of chemical signalling and aggressive behaviour in both females and males suggests that urine release has evolved as an aggressive signal in both sexes of crayfish. By limiting urine release to aggressive behaviours in reproductive interactions females challenge their potential mating partners at the same time as they trigger a sexual response. These double messages should favour stronger males that are able to overcome the resistance of the female. We conclude that the difference between the sexes in disclosing urine-borne information reflects their conflicting interests in reproduction. Males discontinue aggressive urine signalling in order to increase their chances of mating. Females resume urine signalling in connection with aggressive behaviour, potentially repelling low quality or sexually inactive males while favouring reproduction with high quality males.

Maintenance of dominance is mediated by urinary chemical signals in male European lobsters, Homarus gammarus

We studied the relevance of urine cues in Homarus gammarus dominance maintenance, hypothesising that urinary signals are necessary to mediate recognition of former opponents. Males in size-matched pairs interacted on two consecutive days with or without blocking urine release by adding catheters to both contestants on the second day. European lobsters established dominance in a first fight, and fight duration and aggression levels decreased strongly from first to second day in animals with free urine release, indicating the maintenance of this dominance relationship. If urine was blocked on the second day, fight durations were long in both first and second day interactions. Results demonstrate that urine signals contribute to the maintenance of dominance in H. gammarus males.

Fighting behaviour and the role of urinary signals in dominance assessment of Norway lobsters, Nephrops norvegicus

Norway lobsters, Nephrops norvegicus, live on the bottom of the continental shelf where they construct and defend burrows. Little is known about their agonistic behaviour and potential mechanisms of dominance. This paper investigates fighting behaviour of size-matched male Norway lobsters with a particular emphasis on the role of urinary chemical signals in the assessment of dominance. Norway lobsters were paired in dyadic encounters on two consecutive days. A decrease in fight duration from first to second encounters indicates that N. norvegicus are able to maintain dominance in sequential contests. The main difference between the two encounters is in the behaviour of the loser. Losers strongly reduce their aggression level from first to second encounters. Olfactory sampling behaviour (antennule flicking) of eventual losers is higher than that of the winner indicating that the loser assesses chemical signals of the dominant male. When urine release is blocked for the second encounter, there is no difference in fight duration between first and second encounter. The results suggest that Norway lobsters develop lasting dominance relationships. The study also provides preliminary evidence that urine-borne chemical signals play an important role in mediating dominance. © 2008 Brill.

Social context may affect urinary excretion of 11-ketotestosterone in African cichlids

Summary We previously investigated the androgen responsiveness of males to simulated partner and territory intrusions in five African cichlid species (Neolamprologus pulcher, Lamprologus callipterus, Tropheus moorii, Pseudosimochromis curvifrons, Oreochromis mossambicus; Hirschenhauser et al., 2004). Here we re-analysed data on 11-ketotestosterone (11-KT) levels in holding water to compare the free (presumably from the gills) and conjugated (presumably from urine and faeces) 11-KT fractions. We sampled (i) pre-test baseline control levels from individual males in social isolation and (ii) response levels released after social interactions, either with an ovulating female or a male territory intruder. In four out of five species, conjugated metabolites contributed to the observed total 11-KT responses in water during social context, which was particularly apparent in peak responsive individuals exposed to male intruders. Thus, in water from males sampled in isolation immunoreactive 11-KT seemed to derive both from gills and urine, whereas the urinary 11-KT component apparently increased in the social context, particularly when a male was challenged by a same-sex intruder. These results suggest that (i) the social context may affect urine release patterns of males and (ii) 11KT data acquired by using fish-holding water may not simply reflect the passive transmission of steroid hormones via the gills.

Female goldfish signal spawning readiness by altering when and where they release a urinary pheromone

Although it is established that many species of fish release hormonally derived sex pheromones in their urine, whether these cues are unspecialized metabolic by-products or specialized communicatory signals is not known. Here, we addressed this question in the goldfish, Carassius auratus (a leading model of hormonal sex pheromone function) by conducting two experiments. We first monitored urine release by sexually receptive females that were placed with a sexually active male, an inactive male, another female, or no other fish. Sexual receptivity was induced using prostaglandin F2α, a hormone also known to evoke release of 15-keto prostaglandin F2α (15K-PGF2α), a potent urinary pheromone. We found that sexually receptive females increased the frequency with which they urinated when placed with males (regardless of male sexual activity) but not females. Furthermore, females showed a strong tendency to urinate when rising into spawning substrate, suggesting they might be using urine as an attractive lure. This hypothesis was confirmed by a second experiment, which found that 15K-PGF2α attracted males to specific locations. We conclude that goldfish, and likely many other fish, control the release of urinary prostaglandin pheromones to advertise their condition and location, perhaps exemplifying an early step in the evolution of a chemical communication system.

Male–Female Communication in the Crayfish Orconectes rusticus: The Use of Urinary Signals in Reproductive and Non‐Reproductive Pairings

AbstractAnimals use sensory communication to locate conspecifics, food, shelter, and avoid predators. Using urine visualization techniques as well as Digital Particle Illumination Velocimetry, we examined the role of urinary signals and current generations during social interactions of male and female crayfish. Both reproductive and non‐reproductive crayfish were paired to gain a better understanding of how reproductive state influences communication. Analyses of agonistic and mating events were paired in time with recorded urine release and current generation, illustrating a correlation of chemical communication with ritualized social behavior. Four treatment groups were run with specific combinations of different reproductive status: (1) both opponents reproductively active, (2) only the male in reproductive, (3) only the female reproductive, or (4) both opponents non‐reproductive. Results showed differences between treatment groups in urine release, current generation, and social behavior. Within reproductive pairings, both the male and female crayfish generated currents and released urine at higher rates than those in other treatment groups. Urine was released most often when opponents were in chelae contact with each other and these releases were often accompanied by anterior current generation. In addition, communication was different in reproductive trials where mating occurred. Overall, the results indicate that the use of hydrodynamic and chemical signals changes as a function of reproductive state and that this change in communication probably indicates readiness to mate.

The impacts of flow on chemical communication strategies and fight dynamics of crayfish

Signal transmission is influenced by the physics of an environment. Consequently, a physical effect on sensory signals can influence how animals send or sample sensory information. Habitat-specific physics may constrain or enhance signal transmission (e.g. sound transmission in a flowing river versus a still pond) and provide a mechanism for the evolution of sensory biases. This study investigated how the transmission of chemically mediated social signals in crayfish is influenced by two different aquatic environments. Agonistic bouts between crayfish were performed under lotic (flowing water) and lentic (nonflowing, still water) conditions. When crayfish (Orconectes rusticus) collected from a lotic system (river) interacted under lotic conditions, we noted that dominant O. rusticus spent more time upstream than subordinate O. rusticus. Orconectes rusticus positioned themselves randomly and spent equal amounts of time with respect to upstream and downstream in the nonflowing environment. We tested another species, Orconectes virilis, collected from a nonflowing environment (lake) and they showed no positional preference when tested in flow. Additionally, both O. rusticus and O. virilis took longer to reach high fight intensities under flow conditions. It was possible to visualize O. rusticus urine release, and they released urine more often when upstream of an opponent in a flow environment during these agonistic bouts. These results suggest that O. rusticus collected from lotic environments release urine to maximize the transmission of chemical cues to a fight opponent. It appears that crayfish may adapt their signalling processes based upon their long-term ambient environments.