Cauda Epididymis Research Articles

Brazilian green propolis doesn't have a beneficial effect on the cryopreservation of domestic cat epididymal spermatozoa

This study evaluated the effect of different concentrations of Brazilian green propolis on the cryopreservation of epididymal sperm from domestic cats. Spermatozoa were collected from cauda epididymis by slicing technique in a TRIS extender, preheated to 37ºC. Then, the sperm were evaluated and divided randomly between the control group (without adding propolis in the extender) and the treatment groups (adding different concentrations of propolis extract in the extender): P1 (0.1 mg/mL), P2 (0.3 mg/mL) e P3 (0.6 mg/mL). Subsequently, sperm were cryopreserved. The evaluated parameters were sperm kinetics by the Computer Assisted Semen Analysis, vigor, viability, membrane functionality, chromatin condensation and morphology. The parameters were measured before and after cryopreservation. Propolis didn’t show toxicity to the sperm in any concentrations, with no changes observed in the motility pattern. There was no significant influence of propolis on motility, vigor, percentage of viable spermatozoa, chromatin quality and other kinetic parameters in the cryopreserved samples. In sperm morphology, it was demonstrated a reduction in the percentage of normal cells in P1 and P2 groups in relation to the fresh sample, control and P3 group. In addition, an interesting effect from the propolis groups was observed on possible growth inhibition of microorganisms in the contaminated samples. Propolis didn’t provide superiority to the sperm parameters evaluated after thawing, except for the values of plasma membrane functionality, which were better. The propolis is an interesting component to be incorporated in the cryoprotectant medium due to its non-toxicity and the potential inhibition of microbial growth.

Deficiency for Lcn8 causes epididymal sperm maturation defects in mice

Lipocalin family members, LCN8 and LCN9, are specifically expressed in the initial segment of mouse caput epididymis. However, the biological functions of the molecules in vivo are yet to be clarified. In this study, CRISPR/Cas9 technology was used to generate Lcn8 and Lcn9 knockout mice, respectively. Lcn8−/− and Lcn9−/− male mice showed normal spermatogenesis and fertility. In the cauda epididymis of Lcn8−/− male mice, morphologically abnormal sperm was increased significantly, the proportion of progressive motility sperm was decreased, the proportion of immobilized sperm was elevated, and the sperm spontaneous acrosome reaction (AR) frequency was increased. Conversely, the knockout of Lcn9 did not have any effect on the ratio of morphologically abnormal sperm, sperm motility, and sperm spontaneous AR frequencies. These results demonstrated the role of LCN8 in maintaining the sperm quality in the epididymis, and suggested that the deficiency of LCN8 leads to epididymal sperm maturation defects.

Protective effects of curcumin on chromatin quality, sperm parameters, and apoptosis following testicular torsion-detorsion in mice.

ObjectiveThe chief outcome of testicular torsion in clinical and experimental contexts is testicular ischemia. Curcumin, a compound with anti-inflammatory and antioxidant properties, has fascinated researchers and clinicians for its promise in the treatment of fertility diseases. MethodsThirty-five fully grown male mice were randomly classified into five groups: control, sham, testicular torsion, treatment group 1 (testicular torsion+short-term curcumin), and treatment group 2 (testicular torsion+long-term curcumin). Thirty-five days later, spermatozoa from the right cauda epididymis were analyzed with regard to count and motility. Toluidine blue (TB), aniline blue (AB), and chromomycin A3 (CMA3) staining assays were used to evaluate the sperm chromatin integrity. In addition, the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) test was used to assess apoptosis. ResultsTreatment group 1 exhibited a remarkably elevated sperm count compared to the testicular torsion group. Additionally, notably lower sperm motility was found in the testicular torsion group compared to the control, treatment 1, and treatment 2 groups. Staining (CMA3, AB, and TB) and the TUNEL test indicated significantly greater testicular torsion in the torsion group compared to the control group (p<0.05). The data also revealed notably lower results of all sperm chromatin assays and lower apoptosis in both treatment groups relative to the testicular torsion group (p<0.05). Significantly elevated (p<0.05) AB and TB results were noted in treatment group 1 compared to treatment group 2. ConclusionCurcumin can compensate for the harmful effects of testicular ischemia and improve sperm chromatin quality in mice.

Chronic unpredictable stress disturbs the blood–testis barrier affecting sperm parameters in mice

Research questionDoes chronic stress affect the key proteins and sperm parameters of the blood–testis barrier (BTB)? DesignC57Bl/6 mice were divided into two groups: a non-treated control group and a chronic unpredictable stress (CUS) applied group. The stress status of the animals was confirmed with behavioural tests. Histopathologic evaluation was conducted by haematoxylin and eosin staining and electron microscope. Malondialdehyde, corticosterone and testosterone levels were evaluated in peripheral blood. Expression levels of BTB proteins, namely zonula occludens-1 (ZO-1), claudin-11 (CLDN11) and clathrin in Sertoli cells, were assessed by Western blotting and immunofluorescence techniques. Sperm samples were collected from cauda epididymis, and sperm parameters analysed. ResultsThe stress model was confirmed by behavioural tests. Histopathological evaluation of the testes demonstrated a mild degeneration in seminiferous tubules. Malondialdehyde (P = 0.008) and corticosterone levels increased (P = 0.004) and testosterone levels decreased (P = 0.005) in the CUS group. Electron microscopic evaluation confirmed the damage in BTB integrity in the CUS group. Western blot analysis showed that ZO-1 and CLDN11 levels were significantly decreased, although clathrin levels were unchanged. Although sperm concentration and total motility rate were not significantly different between the groups, progressive motility (P = 0.03), normal sperm morphology (P = 0.04), chromatin integrity (toluidine blue) (P = 0.002) and the acrosomal reaction rate (P = 0.002) were significantly decreased, and acrosomal abnormality rate was dramatically increased (P = 0.04) in the CUS group. ConclusionsIn mice, CUS disrupted BTB integrity and impaired sperm parameters. A decrease in ZO-1 and CLDN11 expression levels may be proposed as the causative factor.

Modulatory Role of Vitamin E on Proton Pump (ATPase) Activity of Cadmium Chloride-Induced Testicular Damage in Wistar Rats.

Proton pumps are membrane-bound enzymes important in generating gradients that help in maintaining cellular ion homeostasis, cell membrane potential, water, and solute transport across the cell surface. This study investigated the modulatory role of vitamin E on proton pump activity and reproductive parameters in cadmium-induced testicular damage. Twenty (20) male Wistar rats weighing between 180 and 200 g were sorted into 4 groups of five rats each. Group I served as the control and was given normal saline orally, Group II rats were treated with a single dose of 2 mg/kg BW cadmium chloride (CdCl2) intraperitoneally, Group III rats were given 100 mg/kg BW of vitamin E orally, and Group IV rats were given 100 mg/kg BW of vitamin E orally for 30 days prior to intraperitoneal administration of single dose of 2 mg/kg BW of cadmium chloride. The rats were anaesthetized with diethyl ether, and blood samples were obtained for sex hormonal analysis; caudal epididymis was dissected for sperm count, motility, and viability, and the testis were homogenized for lipid peroxidation and proton pump (Na+/K+ ATPase, Ca2+ ATPase, and Mg2+ ATPase) activity. Proton pump activity was assayed spectrophotometrically using the Stewart method to determine the inorganic phosphate level. Histopathological changes of the testis were also studied. The group treated with CdCl2 showed a significant (p < 0.05) decrease in proton pump activity, sperm count, and motility and a significant (p < 0.05) increase in malondialdehyde level when compared with the control group. The CdCl2-treated group also showed decrease reproductive organ weights and hormonal levels and cause necrosis of spermatogonia lining the seminiferous tubules. Rats treated with vitamin E orally for 30 days prior to CdCl2 exposure showed improvement in proton pump activity, a significant (p < 0.05) increase in sperm parameters and luteinizing hormonal level, and a decrease in the lipid peroxidation level as compared with the CdCl2 group. This study showed that vitamin E ameliorated the toxic effect of CdCl2 on proton pump activity in the testes, hence improving testicular integrity, structures, and functions.

The current perspective on genetic and epigenetic factors in sperm maturation in the epididymis.

Male infertility affects approximately 30% of infertile couples. As spermatozoa mature in the epididymal lumen, their potential for mobility increases, and their protein, lipid and small RNA (sRNA) content changes, whereas capacitation and fertilisation take place in the female reproductive tract. Both of the latter processes are affected by maturation, because impaired maturation causes premature capacitation and fertilization. The epididymis produces a suitable environment for sperm maturation via ion transport, vesicle secretion and protein matrix formation. The microenvironment for sperm maturation varies in three broad segments: the caput, the corpus and the cauda epididymis. Epididymosomes transfer proteins, lipids and sRNAs from the epididymal epithelium to spermatozoa and genetic alterations of epididymal genes can lead to decreased sperm motility, morphological abnormalities of spermatozoa and subfertility. Genetic factors are involved in all aetiological categories in male infertility. However, studies conducted on the genes involved in epididymal functions are limited. The sRNA content of spermatozoa changes during epididymal migration, and these sRNAs play a role in embryo development and epigenetic inheritance. This review aims to clarify the role of the epididymal epithelium in the maturation of spermatozoa in light of the current molecular genomic knowledge.

Effect of Alpinia officinarum extract on reproductive damages in streptozotocin induced diabetic male rats.

Infertility is one of the systemic problems in diabetic men. The purpose of the present study is investigation of the effects of the Alpinia officinarum (AO) hydro-alcoholic extract on the reproductive system damages in diabetic male rats. Twenty four male rats were randomly assigned into 4 groups (n = 6); i.e., control, diabetic control, and diabetic rats treated orally with AO extract (200 and 500mgkg-1). A single dose (60mgkg-1) of streptozotocin (STZ) was injected intraperitoneally (IP) to induce diabetes. After 8weeks of treatment, blood samples, testis, and cauda epididymis were excised to evaluate specific hormonal changes, sperm parameters, and testis morphology. Diabetic control rats showed remarkably lower body and testicular weights, testicular volumes, and sperm parameters compared with the control group (p <0.05). Diabetic control rats also exhibited significantly decreased serum testosterone and follicle stimulating hormone (FSH). Sperm parameters were considerably enhanced in diabetic animals gavaged with AO extract. Testosterone levels were significantly elevated by administrating 500mgkg-1 AO extract to the diabetic control rats (p <0.05). The morphological assessment of testis of treatment group (500mgkg-1) indicated remarkable differences (p <0.05) by increasing the seminiferous tubules diameter (STD) and thickness of the seminiferous epithelium (TSE) compared with diabetic control rats. As demonstrated by the results, AO extract ameliorated sperm damage and improved sperm morphology besides improving histological damage in the testis in diabetic rats. In addition, the dose of 500mgkg-1 worked more efficiently than 200mgkg-1.

Ameliorative effects of Garcinia hydroxybiflavanonol 1 (GB1) isolated from Garcinia kola seeds on cadmium chloride (CdCl2) induced reproductive toxicity in the testis of the male Wistar rats

A high association between cadmium, a common environmental pollutant, and infertility in males has been established. Studies have established that antioxidants can enhance fertility either directly or indirectly. Garcinia hydroxybiflavanonol (GB1) extracts from bitter kola is known for its natural antioxidant properties. This study is aimed at investigating the ameliorative effects of administration of (GB1) on the reproductive health of cadmium chloride (CdCl2)–intoxicated male Wistar rats. Extraction, fractionation and isolation of Garcinia hydroxybiflavanonol (GB 1) were done from seeds of Garcinia kola. Thirty-six male adult Wistar rats weighing 170–190 g were acclimatized for 2 weeks and randomly divided into three groups. Group A were given distilled water as control, group B 2.5 mg/kg b.w. of CdCl2 and group C 2.5 mg/kg b.w. of CdCl2 and 2 mg/kg b.w. of GB1 dissolved in Tween20. The rats were orally dosed daily for 90 days. Every 30 days, 4 animals from each group were euthanized and blood samples collected for testosterone, GnRH, LH and FSH assay. Sperm qualities were determined using the cauda epididymis. The neutral-buffered formalin-fixed testes were processed for histology. The result showed significantly higher levels of testosterone, GnRH, FSH, LH and sperm quality in the CdCl2 + GB1–treated rats than in the CdCl2-only-treated rats. Histopathology showed progressive distortion, disorganisation, erosion and vacuolation of the seminiferous epithelium in CdCl2-only-treated rats which were not observed in the other groups. These findings indicate a protective effect of the GB1 extract on the testes of Wistar rats.

Apoptosis in epididymis of sand rat Psammomys obesus, Cretzschmar, 1828: Effects of seasonal variations, castration and efferent duct ligation

The aim of this study was to visualize apoptosis throughout the reproductive cycle and after castration, castration then treatment with testosterone, and ligation of efferent ducts. The sand rat, Psammomysobesus, Cretzschmar 1828, is a diurnal rodent belonging to the family Gerbillidae. Its breeding cycle is seasonal with reproduction in autumn, winter and early spring and a short resting period from late spring to early summer. Five groups of males were studied: (1) animals captured during the breeding season; (2) animals captured during the resting season; (3) animals castrated and kept 30 days; (4) animals castrated, kept 30 days, and then treated with testosterone for 15 days; (5) animals subjected to the ligation of efferent ducts and kept 30 days. Epididymis were removed and the presence of apoptotic cells was explored using the "Apostain" immunohistochemical method. Histological results showed cell and tissue remodeling. During the breeding season, a positive apoptotic signal was observed mainly in smooth muscle cells of caput and cauda epididymis. This signal persisted throughout the resting season. The orchiectomy induced apoptosis in almost of epithelial and connective cells. However, this intense cell death was not reversed by treatment with testosterone. In animals that experienced efferent duct ligation, principal cells and smooth muscle cells showed a positive signal for apoptosis. Our results converge to qualify the sand rat epididymis as an excellent model for the study of apoptosis and argue for continued cell death, at least independent of circulating testosterone levels.

Characteristics of reproductive organs and estimates of reproductive potential in Scandinavian male grey wolves (Canis lupus)

Improved knowledge about reproductive patterns and potential in male wolves (i.e., testicular development and size relative to age, pubertal age, and seasonal effects) is needed for evaluation and monitoring of reproductive outcomes in populations. Reproductive organs from 215 male wolves, culled as a result of licensed hunting, protective culling or from carcasses found were examined. The testes and epididymis were weighed and measured. There were biopsy samples collected from the testes and the cauda epididymis for histological determinations if there were spermatozoa in tissues collected. There were reproductive tissue analyses of 197 males while there were separate evaluations of tissues from ten cryptorchid animals. Juvenile wolves (< 1 year, n = 47) had a lesser body mass and mean testes mass than subadult (1–2 years, n = 71) and adult (>2 years, n = 79) males. Season also affected testicular characteristics of structures evaluated with subadult and adult males having a lesser mass during summer months (May–August). Of the 197 males, 70 % had spermatozoa in the seminiferous tubules and the cauda epididymis and were classified as being ‘potentially fertile’ when tissues were collected, while 22 % were classified as being non-fertile (no spermatozoa, including males that were pre-pubertal) and tissues of 8% could not be evaluated. When testes mass was greater, there was a greater likelihood that spermatozoa were present. There were seven of the ten cryptorchid males of the unilateral type. These testicular and epididymal findings will be useful for evaluating the reproductive potential and management of wolves in Scandinavia.

Relation between washed sperm cells and fluid from White Fulani Genitalia in some enzyme activities and lipid contents

Lipid contents and enzyme activities were studied in the fluid and washed sperm (WS) from the genitalia of 26 mature White Fulani (WF) bulls of mean live weight 213.96±4.88kg (range 140 260kg) in a complete randomized design experiment. Tissue homogenization and biochemical assay methods were employed after storing the tissue in deep freeze for 24 hours post slaughter. Total lipid contents did not differ significantly (P&gt;0.05) between WS and fluid of the testis, the caput epididymis and the cauda epididymis but was significantly (P&lt;0.05) higher in WS than the fluid in corpus epididymis and vas deferens. Phospholipid and phospholipid phosphorus contents of WS and fluid did not vary significantly (P&gt;0.05) benveen WS in the testis and epididymis. Alkaline phosphatatse (ALP) activity was higher (P&lt;0.05) in the corpus and cauda edididymal fluid than in the WS. Glutamic Oxalate Transaminase (GOT) activity was significantly (P&lt;0.05) higher in the fluid than the WS in all segments but not in the testis. The generally higher values of lipids in the washed sperm over the fluid the need for lipids as protective cover and as substrates for sperm survival from its formation up to maturation. The generally higher enzyme activity in the fluid is an indicator of damage to sperm acrosome, with corresponding leakage of enzyme into the fluid consequent upon the various stress conditions encountered as sperm traverses the epididymis for the maturation process and finally sojourns in the epididymis.

Sperm production rate, gonadal and extragonadal sperm reserves in slaughtered White Fulani (Bunaji) bulls in a lowland tropical environment

Reproductive tract morphometry, sperm production rates, gonadal and extragonadal sperm reserves in Bunaji bulls were evaluated using samples from a total of 13 mature animals. The morphometric characteristics of the reproductive organs had generally low values but showed normal development. The daily sperm production (DSP) averaged 1.23 ± 0.16 x 109, while the daily sperm production (DSG/g) was 0.62 ± 0.11 x 107. Gonadal sperm reserve was 2.17 ± 0.73 x 109 while extragonadal sperm reserve was 3.82 x 109 for caput, 2.68 x 109 for corpus, 4.36 x 10 (for cauda epididymis and 0.62 x 109 for the ductus deferens. The relative contributions of the epididymal segments and the ductus deferns to the total extragonadal sperm reserves were 33.28%, 23.34%, 37.98% and 5.40% respectively. Testicular morphomeric characteristics correlated highly with themselves and with sperm production parameters as well as extragonadal sperm reserve. While these results provided information that would be useful in the determination if male/female ratio during mating and artificial insemination programmmes, the study also shows that good sires could be obtained from these animals for breeding and improvement programmes in this breed.

MORPHOLOGICAL CHANGES IN EPIDIDYMAL SPERMATOZOA OF RED SOKOTO (MARADI) BUCKS

Morphological changes in the caput, corpus and cauda epididymides were studied in this experiment. The objective was to study morphological changes in goat spermatozoa using the Red Sokoto goat. Progressive motile spermatozoa were first observed in the caput epididymis (5-7%) and was highest at the cauda epididymis (60%). Progressive motility for the corpus epididymis ranged between 20-30%. Cytoplasmic droplets were lost as the spermatozoa moved up the epididymis. Proximal cytoplasmic droplet was significantly (P&lt;0.05) lowest at the cauda epididymis. Other morphological changes did not follow a particular trend.

Male Reproductive Tract Origin Sperm-quiescent Proteins: Prospects in Semen Cryopreservation of Livestock Species

Cauda epididymis is known to preserve mature sperm in a quiescent state for several weeks without causing potential decline to fertilizing capacity of the sperm. This unique property of cauda epididymis has been attributed to the unique biochemical composition of cauda epididymal plasma and the presence of certain proteins having decapacitating and motility–inhibitory activities. Cryo-capacitation and low progressive motility are two common changes in cryopreserved semen across species and have been considered the underlying causes for low fertility rate of cryopreserved semen. Considering their potential roles in reversible inhibition of sperm capacitation and motility these naturally-occurring proteins may be suitable tools for minimizing cryo-capacitation and improving post-thaw progressive motility of cryopreserved semen by sparing available energy in sperm during cryopreservation. The present review attempts to bring out the status of purification, functional and mechanistic characterization of these sperm-quiescent proteins across species. In addition the status of recombinant production of these proteins by using different host systems and the scope for application of these proteins in semen cryopreservation of livestock species has also been described.

Melatonin-Caffeine Combination Modulates Gamma Radiation-induced Sperm Malformations in C57BL/6 Male Mice at Sublethal Dose of Gamma Radiation.

Aims:The aim of this study was to assess the protective effect of the melatonin–caffeine combination against γ radiation-induced alterations in the morphological characteristics of sperms.Settings and Design:C57BL/6 male mice (n = 30) were randomly divided into five groups: control, radiation (2 Gy), melatonin (100 mg/kg body wt.) + radiation (2 Gy), caffeine (30 mg/kg body wt.) + radiation (2 Gy), melatonin–caffeine (100–30 mg/kg body wt.) + radiation (2 Gy).Materials and Methods:All the mice were sacrificed 24 h postirradiation, and cauda epididymis was collected. In this study, sperm concentration along with any abnormality in their morphology (amorphous heads, pinheads, hookless, coiled tails, midpiece defect, and tail-less) was observed in the control and treatment group of animals.Results:Radiation exposure (2 Gy) considerably decreases the sperm count when compared with the control group. However, pretreatment with melatonin and melatonin–caffeine combination before gamma irradiation increased the sperm count (P < 0.05), but with caffeine alone could not produce a significant difference. The higher rate of abnormal sperms was observed in the γ-irradiated mice when compared with the control group (P < 0.05). Besides, the numbers of sperm that are hookless and coiled tails were significantly increased after irradiation. Melatonin significantly reduced the number of sperm with amorphous heads and coiled tails. Melatonin–caffeine combination further reduced sperm malformations when compared with the melatonin + 2 Gy radiation and caffeine + 2 Gy radiation group.Conclusions:This study suggests that caffeine exerts a protective effect when given in combination with melatonin against gamma irradiation in sperms of C57BL/6 mice and could be a potent combination for the development of radioprotector.

Mouse CD52 is Predominantly Expressed in the Cauda Epididymis, Regulated by Androgen and Lumicrine Factors.

Background:Sperm maturation takes place through contact between sperm and proteins produced in the epididymal lumen. CD52 had been characterised in the sperm; however, the expression and its regulation in the epididymis are mostly unknown.Aim:This study aimed to analyse the expression and regulation of CD52 in the mouse epididymis.Setting and Design:Experimental design was used in this study.Materials and Methods:Epididymis tissues from mice strain Deutch Democratic Yokohama were used as sources of total RNA. Bioinformatic tool was used to predict signal peptides. Quantitative real-time reverse transcription–polymerase chain reaction was used to analyse tissue distribution, androgen, testicular factors dependency and postnatal development.Statistical Analysis:One-way analysis of variance was used to analyse differences between treatment and control untreated group. P < 0.05 was determined as a significant difference.Results:CD52 amino acid sequence contains a signal peptide, indicating it is a secretory protein. CD52 exhibited region-specific expression in the epididymis, with the highest level being in the cauda. CD52 expression was regulated by androgen indicated by a significant downregulation at day 1 and day 3 following a castration (P < 0.05). Dependency on androgen was confirmed by injection of exogenous testosterone which prevented downregulation by 50%. Moreover, lumicrine factors also influenced CD52 expression indicated by ligation of efferent duct which also reduced expression at day 1 to day 5 following the ligation (P < 0.05). CD52 expression was developmentally regulated. This was shown by increase in the level of expression starting at day 15 postnatally.Conclusion:CD52 shows characteristics of genes involved in sperm maturation in the epididymis.

Localization and expression of ADAM2 in the dromedary camel testis, epididymis and sperm during rutting season.

ADAM2 (fertilin β) is a sperm surface protein reported in several mammalian species. However, the presence of ADAM2 in the male reproductive system and sperm of the camel is not well known. The present study was to clarify the localization and expression of ADAM2 in the dromedary camel testis, epididymis and spermatozoa during rutting season using immunohistochemistry (IHC) and the quantitative real-time polymerase chain reaction (qPCR). Tissue samples were obtained from the testis (proximal and distal) and epididymis (caput, corpus, and cauda) from eight mature male camels. Epididymal and ejaculated sperms were collected from four other fertile camels. IHC analysis clearly showed the localization of ADAM2 protein in the spermatocytes and the round and elongated spermatids of the testis, in the epithelial cells along the epididymis tract, on the posterior head of the sperm within the cauda epididymis, and on the acrosomal cap of both the epididymal and ejaculated sperm. The expression of camel ADAM2 mRNA was significantly higher (P < 0.05) in the testis when compared with the epididymis. These findings may suggest an important role of ADAM2 in the fertility of male dromedary camels.

Testicular oxidative stress and apoptosis status in streptozotocin-induced diabetic rats after treatment with rooibos (Aspalathus linearis), honeybush (Cyclopia intermedia), and sutherlandia (Lessertia frutescens) infusions

Objective: To investigate the testicular oxidative stress and apoptosis status, as well as the sperm functional parameters in streptozotocin (STZ) induced diabetic rats following treatment with rooibos (Aspalathus linearis), honeybush (Cyclopia intermedia) and sutherlandia (Lessertia frutescens) infusions. Methods: Diabetes was induced by injecting fourteen-week-old adult male Wistar rats (250-300 g) with a single intraperitoneal injection of STZ (45 mg/kg body weight). Fifty rats were randomly divided into five groups: the vehicle group received 0.1 M citrate buffer, the diabetic control group received 45 mg/kg STZ, the diabetic+rooibos group received 45 mg/kg STZ + 2.0% rooibos, the diabetic+honeybush group received 45 mg/kg STZ + 4.0% honeybush, and the diabetic+sutherlandia group received 45 mg/kg STZ + 0.2% sutherlandia. Rats were sacrificed 7 weeks after induction of diabetes mellitus. The testes and epididymides were harvested and weighed after induction. Spermatozoa were retrieved from the cauda epididymis for motility, concentration, and morphology analysis, and the testis was used for all biochemical assays. Oxidative stress was determined by measuring malondialdehyde levels, catalase, and superoxide dismutase activities, while apoptotic biomarkers were evaluated by Western blotting assays. Results: After induction of diabetes, rats in the diabetic control group, diabetic+rooibos group, diabetic+honeybush group, and diabetic+sutherlandia group presented with significantly elevated blood glucose levels as compared with the vehicle group (P&lt;0.001). Rats in the diabetic control group had a reduction in sperm progressive motility, while rats in the diabetic+rooibos group and the diabetic+sutherlandia group displayed an increase in progressive motility as compared with the diabetic control group. The diabetic control animals showed a 40.0% decrease in sperm concentration when compared to the vehicle group, and there were no significant differences in sperm kinematic and speed parameters between the groups. In addition, the percentage of morphologically normal spermatozoa was increased by 13.0%, 16.0%, and 15.0% after treatment with rooibos, honeybush, and sutherlandia, respectively and the rats in the diabetic+infusion groups also displayed an increase in superoxide dismutase activity when compared to the diabetic control group. Conclusions: Rooibos, honeybush and sutherlandia infusions may partly alleviate diabetes-induced sperm function impairment by reducing oxidative stress.

Qualitative and quantitative analysis of protein expression in the caput and cauda regions of the rat epididymis

Background: The aim of this study was to identify the differentially expressed proteins in the sperm isolated from the caput and the cauda region of the rat epididymis. This is the first study on the quantitative nongel-based proteomics to have identified differentially expressed proteins in the cauda epididymal sperm. Material and Methods: This was achieved by isolation of sperm from the caput and the cauda of the rat epididymis followed by the tryptic digestion of the proteins and the resulted peptides were subjected to isobaric tags for relative and absolute quantitation-label and mass spectrometry (MS)/MS analysis. With the help of quantitative proteomics, we have been able to elucidate some of the major proteins involved in the process of sperm maturation. Results: In total, 999 proteins from the spermatozoa of caput and cauda region of the epididymis were identified. We have reported about 10 downregulated proteins and 15 upregulated proteins that have been in the sperm from the cauda region of the epididymis. HongrES1, the membrane of the SERPIN family specifically expressed in the principal cells of the cauda epididymis, have been found to be expressed three-fold higher. Conclusion: This study contributes to the understanding of the importance of different proteins at the different stages of the maturation during the transit of the spermatozoa. The higher and lower expression of different proteins in the epididymal region depicts their roles in priming the spermatozoa for normal fertilizing ability. Thus, the target proteins can be further studied for the possible development of male contraception.

In Vitro Contraction of Isolated Cauda Epididymal Duct Smooth Muscle as a Complimentary Approach to Physiological, Pathological, Toxicological, and Pharmacological Studies on Epididymal Function.

Contraction of cauda epididymal duct (CE) smooth muscle is one of the very first events of the seminal emission phase of ejaculation. The contraction of CE smooth muscle is governed by a complex interaction of hormones, autacoids, and by the neurotransmitters released from the epididymal intramural nerve endings, and any impairment in the CE smooth muscle contraction has the potential to impair male fertility. Apart the obvious pathophysiological and toxicological importance of CE smooth muscle contraction, modulation of CE contraction has pharmaceutical interest offering a druggable target to development of drugs to improve/impair male fertility. The in vitro contraction experiments constitute a valuable approach to an in-depth evaluation of functional and molecular changes resulting from pathologies or drug exposure. Therefore, this chapter consists in a description of in vitro pharmacological reactivity contractility of the epididymal duct in a controlled medium, maintained at 30°C of temperature and continuously bubbled with 95% O2 and 5% CO2 to obtain cumulative concentration-response curves that has been fundamental to some of our investigations on epididymal physiology, toxicology, and pharmacology.