The present study was designed to determine whether the treatment with an ethanolic extract of pomegranate (EEP)(Punica granatum) can be useful for the treatment of the deleterious effect of lead acetate (LA) administration on sperm production in rats. The effects of EEP were compared with those of ascorbic acid (AA) that is a strong antioxidant and has been shown to reverse lead-induced damage on the reproductive system. The rats were divided into five different groups: those received distilled water (control group), LA, LA with EEP, LA with AA, and EEP alone, respectively. LA administration inhibited spermatogenesis by reducing the length of the stages related to spermiation (VII and VIII) and onset of mitosis (IX-XI). LA-treated rats also showed a reduction in epididymal sperm number and daily sperm production (DSP). Administration of EEP or AA resulted in longer VIII and IX-XI stages when compared with LA-treated rats. Moreover, EEP and AA administration reduced the deleterious effect of LA on DSP and epididymal sperm number. EEP showed an antioxidant activity similar to that of AA. EEP prevented LA-induced spermatogenic disruption in rats and its antioxidant activity could explain its capacity to reverse the damage produced by LA on spermatogenesis.

Oxidative stress (OS) has been recognized as one of the most important cause of male infertility. Despite the antioxidant activity of seminal plasma, epididymis and spermatozoa, OS damages sperm function and DNA integrity. Since antioxidants suppress the action of reactive oxygen species, these compounds have been used in the medical treatment of male infertility or have been added to the culture medium during sperm separation techniques. Nevertheless, the efficacy of such a treatment has been reported to be very limited. This may relate to:(i) patient selection bias;(ii) late diagnosis of male infertility;(iii) lack of double-blind, placebo-controlled clinical trial; and/or (iv) use of end-points that are not good markers of the presence of OS. This review considers the effects of the main antioxidant compounds used in clinical practice. Overall, the data published suggest that no single antioxidant is able to enhance fertilizing capability in infertile men, whereas a combination of them seems to provide a better approach. Taking into account the pros and the cons of antioxidant treatment of male infertility, the potential advantages that it offers cannot be ignored. Therefore, antioxidant therapy should remain in the forefront of preventive medicine, including human reproductive medicine.

Testicular torsion is a common syndrome that could lead to infertility. We investigated the therapeutic effects of lycopene, an antioxidant caretenoid, on testicular ischemia/reperfusion (IR) injury that resembles testicular torsion. Male Wistar albino rats were divided into three groups: sham (n = 6), IR (n = 18), and ischemia/reperfusion with lycopene (IRL, n = 18). Left testicular artery and vein was occluded for 1 h, followed by reperfusion of 3 h, 24 h or 30 days in IR and IRL animals. Either corn oil (vehicle) or lycopene (4 mg/kg) was administrated once daily by gavage to IR or IRL animals, respectively, 5 min after ischemia. Sham-operated animals were treated with vehicle by gavage 5 min after the operation. IR decreased sperm motility and concentration in both ipsilateral and contralateral testes and increased abnormal sperm rate in ipsilateral testis after 30 days of reperfusion. Treatment with lycopene increased the motility in bilateral testes and decreased the rate of abnormal sperm in ipsilateral testis to the sham level, but did not increase sperm concentration in bilateral testes. IR increased the activities of catalase and glutathione peroxidase and the level of reduced glutathione by 24 h of reperfusion, but malondialdehyde remained unchanged. Lycopene treatment restored the enzyme activities but not the reduced glutathione level. Lycopene treatment also ameliorated the IR-induced tissue damage in bilateral testes. In conclusion, the therapeutic antioxidant effect of lycopene on germ cells could serve as a promising intervention to oxidative stress-associated infertility problems, such as testicular torsion.

OBJECTIVE To determine the effect of melatonin, a pineal secretory product that prevents testicular ischemia/reperfusion (IR) injury through its antioxidative properties, on epididymal sperm quality in a rat testicular IR injury model. DESIGN Experimental study. SETTING University pharmacology laboratory. ANIMAL(S) Fifty-six 8-week-old male Wistar albino rats. INTERVENTION(S) Left testicular artery and vein occluded for 1 hour; before the bilateral orchiectomy, the organ was allowed to reperfuse 30 days. Melatonin (10 mg/kg IP) or vehicle (1% ethanol in saline) was administrated for 10 minutes before reperfusion and for 1 hour after reperfusion. MAIN OUTCOME MEASURE(S) After 24 hours of reperfusion, the rats were decapitated, and the testicular tissue samples were obtained for histologic examination. In addition, after 30 days of reperfusion, the epididymal sperm concentration, motility, and abnormal sperm rates were determined in the sperm collected from the epididymis. RESULT(S) A statistically significant decrease in sperm concentration resulted from IR as well as an increase in sperm abnormalities, but the sperm motility did not change. Melatonin treatment did not prevent the IR-induced reduction in sperm concentration. However, melatonin treatment statistically significantly decreased the sperm abnormalities when compared with the IR injured samples. CONCLUSION(S) Melatonin may improve sperm morphology for a protective effect in IR-induced testicular injury.

Lead increasingly contributes to pollution of the environment and may play a role in the development of adverse effects in the human and animal body. Data concerning its mutagenic, clastogenic, and carcinogenic properties have been conflicting. In this study, we evaluated the frequency of micronuclei in bone marrow erythrocytes of rats treated with lead acetate trihydrate. Outbred Wistar rats were exposed to a daily dose of 100 mg/L drinking water for 125 days. The mean value of the total number of micronuclei observed in polychromatic erythrocytes of female rats was significantly higher than that found in the control group (13.375 +/- 2.722 against 9.625 +/- 3.204 micronuclei/1000 cells; P = 0.024 in ANOVA). In exposed female animals, no significant reduction of the ratio of polychromatic to normochromatic erythrocytes was observed (0.990 +/- 0.228 against 1.208 +/- 0.195; P = 0.060 in ANOVA). The effects of lead acetate trihydrate in male rats are both cytotoxic and genotoxic because of a decrease in ratio of polychromatic to normochromatic erythrocytes (0.715 +/- 0.431 against 1.343 +/- 0.306; P = 0.023, ANOVA followed by Tukey test) and an increase in frequency of micronucleated polychromatic erythrocytes (24.167 +/- 7.859 against 4.0 +/- 4.528 micronuclei/1000 cells; P </= 0.001, ANOVA followed by Tukey test), respectively.

OBJECTIVE: To evaluate whether increasing antioxidant intake in men with high levels of DNA damage or lipid peroxidation improves gestational results in couples with history of recurrent embryo loss. DESIGN: Descriptive study (case series). SETTING: Early recurrent embryo loss program at the University of Antioquia, Medellín, Colombia. PATIENT(S): Seventeen men whose spouses had a history of two or more embryo losses before 12 weeks of gestation. INTERVENTION(S): Male partners with increased DNA fragmentation index (%DFI) or high thiobarbituric acid reactive substances (TBARS) were instructed to consume a diet rich in antioxidants or commercial multivitamins containing beta-carotene, vitamin C, vitamin E, and zinc for at least 3 months. MAIN OUTCOME MEASURE(S): Pregnancy outcome was recorded in the spouses of men with increased %DFI or TBARS who received antioxidant supplementation. RESULTS: Of the 17 men, 9 (53%) presented with an increased %DFI or TBARS. They were started on an antioxidant supplementation regimen. Of these nine men, six of their spouses became pregnant. All couples whose male partners accepted antioxidant supplementation achieved a successful pregnancy. CONCLUSIONS: Our study demonstrates the benefits of an increased intake of antioxidant-rich food or antioxidant supplements by men who show high levels of sperm DNA fragmentation or lipid peroxidation, which could result in an improvement in gestational outcomes in couples with history of recurrent embryo losses.

Lead is highly toxic and persistent in the environment and, thus, a major concern for public health. In this study, the effects of lead chloride (PbCl(2)) on mouse epididymal sperm were evaluated. Male mice were subcutaneously injected with 74 and 100 mg PbCl(2)/kg body weight for four consecutive days. Sperm was collected from the epididymis and several parameters of sperm function, such as sperm density, motility, viability, mitochondrial function, acrosome integrity and morphology, were evaluated. Furthermore, DNA fragmentation was assessed by the terminal deoxylnucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick-end labelling (TUNEL) assay and chromatin integrity was evaluated by sperm chromatin structure assay (SCSA). In order to assess direct effects on existing sperm population, we sacrificed one group for each condition at day 5. The effects of lead upon one entire spermatogenic cycle were evaluated on day 35. Both lead concentrations used in this work affected sperm motility, although no significant differences were observed in sperm viability, mitochondrial function and DNA/chromatin integrity. However, a decrease in the percentage of intact acrosomes was also observed, mirroring a lead-induced premature acrosome reaction. Thus, the results obtained indicate that, together with impaired motility, the effect of lead toxicity on acrosome integrity, leading to premature reaction, may compromise the ability of sperm to fertilize the oocyte.

Cadmium (Cd) is a potential pollutant of the environment. It manifests cyto-toxic effects in different organs in animals. In the present study, intraperitoneal injection of CdCl(2)(1mg/kg body weight) increased lipid peroxidation in Swiss mice testes indicating oxidative stress during 5th to 8th week of post-treatment . The enzymatic activity of superoxide dismutase (SOD), catalase (CT) and peroxidase (PD) were significantly decreased over the post-treatment phase in Cd-treated mice testes compared to vehicle controls. Further, ascorbic acid content also declined significantly in Cd-exposed mice testes. Following Cd treatment, a marked increase in sperm abnormality percentage and significant decrease in sperm count was observed. The purpose of the present study was to evaluate the effect of vitamins C and E supplementation on Cd-treated mice testes. Therefore, Cd-treated mice groups were injected with vitamins C and E, separately, to assess the effect of the vitamins in combating Cd-induced cytotoxicity and other manifestations. Supplementation of vitamin C (10mg/kg body weight) and vitamin E (100mg/kg body weight) to Cd-induced mice groups declined lipid peroxidation, increased sperm count profile, depressed the percentage of sperm abnormality, increased the activity of antioxidant enzymes mentioned above and also increased the concentration of ascorbic acid to a measurable extent. The role of vitamins in reducing oxidative stress-related effects on spermatogenesis in Cd-treated Swiss mice testes have been reported.

Different forms of Aluminium (Al) are environmental xenobiotics that induce free radical-mediated cytotoxicity and reproductive toxicity. Vitamin E (alpha -tocopherol) is an antioxidative agent that has been reported to be important for detoxification pathways. This study was thus aimed at elucidating the protective effects of vitamin E towards aluminium toxicity on the histology of the rat testis. Al (5 mg/kg body weight) was administered intraperitoneally in 2 ml saline, either alone or immediately before vitamin E (500 mg/kg body weight), at a different point of abdomen, and the alterations in the testis tissue were analyzed histologically. Seven treated animals were sacrificed for each group, with the testes removed and examined histologically. In the Al-treated group, the germinal epithelium of the seminiferous tubules was thinner in places and spermatids were almost absent; sperm numbers were low and there were no sperm in the lumen. In the Al plus vitamin E rats, there were large numbers of spermatids and sperm in the seminiferous tubule lumen. In the vitamin E alone group, a normal histology was seen. Electron microscopically, in the Al-treated group there were irregularities in the nuclear membrane, some damaged mitochondria, a decrease in the number of ribosomes, and an increase in the number of lysosomes in the sertoli cell cytoplasm. In the primary spermatocyte cytoplasm, there was an increase in the rough endoplasmic reticulum. In the Al plus vitamin E group, the spermatogeneic cells and the sertoli cell cytoplasm showed an almost normal appearance. The ultrastructure of the testis in the vitamin E alone group showed a normal appearance. In conclusion, vitamin E antagonizes the toxic effects of Al at the histological level, thus potentially contributing to an amelioration of the testis histology in the Al-treated rats. The associated biochemical parameters merit further investigation.

For a long time, superoxide generation by an NADPH oxidase was considered as an oddity only found in professional phagocytes. Over the last years, six homologs of the cytochrome subunit of the phagocyte NADPH oxidase were found: NOX1, NOX3, NOX4, NOX5, DUOX1, and DUOX2. Together with the phagocyte NADPH oxidase itself (NOX2/gp91(phox)), the homologs are now referred to as the NOX family of NADPH oxidases. These enzymes share the capacity to transport electrons across the plasma membrane and to generate superoxide and other downstream reactive oxygen species (ROS). Activation mechanisms and tissue distribution of the different members of the family are markedly different. The physiological functions of NOX family enzymes include host defense, posttranlational processing of proteins, cellular signaling, regulation of gene expression, and cell differentiation. NOX enzymes also contribute to a wide range of pathological processes. NOX deficiency may lead to immunosuppresion, lack of otoconogenesis, or hypothyroidism. Increased NOX activity also contributes to a large number or pathologies, in particular cardiovascular diseases and neurodegeneration. This review summarizes the current state of knowledge of the functions of NOX enzymes in physiology and pathology.

Cadmium (Cd) is a potential pollutant of the environment. It manifests cyto-toxic effects in different organs in animals. In the present study, intraperitoneal injection of CdCl(2)(1mg/kg body weight) increased lipid peroxidation in Swiss mice testes indicating oxidative stress during 5th to 8th week of post-treatment . The enzymatic activity of superoxide dismutase (SOD), catalase (CT) and peroxidase (PD) were significantly decreased over the post-treatment phase in Cd-treated mice testes compared to vehicle controls. Further, ascorbic acid content also declined significantly in Cd-exposed mice testes. Following Cd treatment, a marked increase in sperm abnormality percentage and significant decrease in sperm count was observed. The purpose of the present study was to evaluate the effect of vitamins C and E supplementation on Cd-treated mice testes. Therefore, Cd-treated mice groups were injected with vitamins C and E, separately, to assess the effect of the vitamins in combating Cd-induced cytotoxicity and other manifestations. Supplementation of vitamin C (10mg/kg body weight) and vitamin E (100mg/kg body weight) to Cd-induced mice groups declined lipid peroxidation, increased sperm count profile, depressed the percentage of sperm abnormality, increased the activity of antioxidant enzymes mentioned above and also increased the concentration of ascorbic acid to a measurable extent. The role of vitamins in reducing oxidative stress-related effects on spermatogenesis in Cd-treated Swiss mice testes have been reported.

The Drosophila compound eye is a regular structure, in which about 750 units, called ommatidia, are arranged in a highly regular pattern. Eye development proceeds in a stereotypical fashion, where epithelial cells of the eye imaginal discs are specified, recruited, and differentiated in a sequential order that leads to the highly precise structure of an adult eye. Even small perturbations, for example in signaling pathways that control proliferation, cell death, or differentiation, can impair the regular structure of the eye, which can be easily detected and analyzed. In addition, the Drosophila eye has proven to be an ideal model for studying the genetic control of neurodegeneration, since the eye is not essential for viability. Several human neurodegeneration diseases have been modeled in the fly, leading to a better understanding of the function/misfunction of the respective gene. In many cases, the genes involved and their function are conserved between flies and human. More strikingly, when ectopically expressed in the fly eye some human genes without a Drosophila counterpart can induce neurodegeneration, detectable by aberrant phototaxis, impaired electrophysiology, or defects in eye morphology. These defects are often rather subtle alteration in shape, size, or arrangement of the cells, and can be easily scored at the ultrastructural level. This chapter aims to provide an overview regarding the analysis of the retina by various means.

The evolutionary conserved transmembrane protein Crumbs (Crb) regulates morphogenesis of photoreceptor cells in the compound eye of Drosophila and prevents light-dependent retinal degeneration. Here we examine the role of Crb in the ocelli, the simple eyes of Drosophila. We show that Crb is expressed in ocellar photoreceptor cells, where it defines a stalk membrane apical to the adherens junctions, similar as in photoreceptor cells of the compound eyes. Loss of function of crb disrupts polarity of ocellar photoreceptor cells, and results in mislocalisation of adherens junction proteins. This phenotype is more severe than that observed in mutant photoreceptor cells of the compound eye, and resembles more that of embryonic epithelia lacking crb. Similar as in compound eyes, crb protects ocellar photoreceptors from light induced degeneration, a function that depends on the extracellular portion of the Crb protein. Our data demonstrate that the function of crb in photoreceptor development and homeostasis is conserved in compound eyes and ocelli and underscores the evolutionarily relationship between these visual sense organs of Drosophila. The data will be discussed with respect to the difference in apico-basal organisation of these two cell types.

Usher syndrome is the most prevalent cause of hereditary deaf-blindness, characterized by congenital sensorineural hearing impairment and progressive photoreceptor degeneration beginning in childhood or adolescence. Diagnosis and management of this disease are complex, and the molecular changes underlying sensory cell impairment remain poorly understood. Here we characterize two zebrafish models for a severe form of Usher syndrome, Usher syndrome type 1C (USH1C): one model is a mutant with a newly identified ush1c nonsense mutation, and the other is a morpholino knockdown of ush1c. Both have defects in hearing, balance and visual function from the first week of life. Histological analyses reveal specific defects in sensory cell structure that are consistent with these behavioral phenotypes and could implicate Müller glia in the retinal pathology of Usher syndrome. This study shows that visual defects associated with loss of ush1c function in zebrafish can be detected from the onset of vision, and thus could be applicable to early diagnosis for USH1C patients.

The function and integrity of photoreceptor cells are dependent upon the creation and maintenance of specialized apical structures: membrane discs/outer segments in vertebrates and rhabdomeres in insects. We performed a molecular and morphological comparison of Drosophila Pph13 and orthodenticle (otd) mutants to investigate the transcriptional network controlling the late stages of rhabdomeric photoreceptor cell development and function. Although Otd and Pph13 have been implicated in rhabdomere morphogenesis, we demonstrate that it is necessary to remove both factors to completely eliminate rhabdomere formation. Rhabdomere absence is not the result of degeneration or a failure of initiation, but rather the inability of the apical membrane to transform and elaborate into a rhabdomere. Transcriptional profiling revealed that Pph13 plays an integral role in promoting rhabdomeric photoreceptor cell function. Pph13 regulates Rh2 and Rh6, and other phototransduction genes, demonstrating that Pph13 and Otd control a distinct subset of Rhodopsin-encoding genes in adult visual systems. Bioinformatic, DNA binding and transcriptional reporter assays showed that Pph13 can bind and activate transcription via a perfect Pax6 homeodomain palindromic binding site and the Rhodopsin core sequence I (RCSI) found upstream of Drosophila Rhodopsin genes. In vivo studies indicate that Pph13 is necessary and sufficient to mediate the expression of a multimerized RCSI reporter, a marker of photoreceptor cell specificity previously suggested to be regulated by Pax6. Our studies define a key transcriptional regulatory pathway that is necessary for late Drosophila photoreceptor development and will serve as a basis for better understanding rhabdomeric photoreceptor cell development and function.

We are describing a rhabdom organization of the eye of the chrysanthemum beetle Phytoecia rufiventris that to date has not been described from any other insect. In cerambycid beetles free rhabdomeres, forming a circular, open rhabdom, surround a central rhabdom made up of the rhabdomeres of one or two cells. In Phytoecia rufiventris the central rhabdomeres are missing throughout the eye and the microvilli of the outer 6 rhabdomeres are regularly oriented in three directions. Following the classification of rhabdom types suggested by Wachmann (1979), we suggest to name the rhabdom arrangement seen in the retina of Phytoecia rufiventris "Grundmuster 3". This pattern ought to facilitate polarization sensitivity and movement perception, features that agree with the behavioural repertoire of Phytoecia rufiventris.

Usher syndrome (USH) is a genetically heterogeneous group of autosomal recessive deaf-blinding disorders. Pathophysiology leading to the blinding retinal degeneration in USH is uncertain. There is evidence for involvement of the photoreceptor cilium, photoreceptor synapse, the adjacent retinal pigment epithelium (RPE) cells, and the Crumbs protein complex, the latter implying developmental abnormalities in the retina. Testing hypotheses has been difficult in murine USH models because most do not show a retinal degeneration phenotype. We defined the retinal disease expression in vivo in human USH using optical imaging of the retina and visual function. In MYO7A (USH1B), results from young individuals or those at early stages indicated the photoreceptor was the first detectable site of disease. Later stages showed photoreceptor and RPE cell pathology. Mosaic retinas in Myo7a-deficient shaker1 mice supported the notion that the mutant photoreceptor phenotype was cell autonomous and not secondary to mutant RPE. Humans with PCDH15 (USH1F), USH2A or GPR98 (USH2C) had a similar retinal phenotype to MYO7A (USH1B). There was no evidence of photoreceptor synaptic dysfunction and no dysplastic phenotype as in CRB1 (Crumbs homologue1) retinopathy. The results point to the photoreceptor cell as the therapeutic target for USH treatment trials, such as MYO7A somatic gene replacement therapy.

The structural organization of the eyes belonging to 12 winged male and 12 wingless female Orgyia antiqua moths, exposed for 1 h to UV-radiation (lambda(max)=351 nm) of 1.4 kW/m2, was compared with that of 12 male and 12 female non-irradiated control specimens. Following the UV-exposure, the screening pigments were found in a position indicative of extreme light-adaptation. Extensive formations of vesicles along the perimeter of the cones as well as disintegrating ER in the cone cytoplasm were noticeable, especially in the eye of the female. On the retinal side of the clearzone, the microvilli of the rhabdoms had become affected by the UV in characteristic ways: in the male eye, retinal cell damage in the form of microvillar swellings and disintegrations were largely confined to just two cells per ommatidium, placed opposite to each other. The female eye, once again, exhibited greater vulnerability and more widespread microvillar disruptions that affected all of the ommatidial retinula cells. The greater resistance of the eye of the male to an exposure with UV makes sense, if we consider the consequences of the retinal damage, which would clearly be a more severe handicap for an actively flying individual than for an almost sedentary one like the wingless female.

The pollen-consuming beetle Xanthochroa luteipennis, which belongs to the family Oedemeridae, possesses a nearly spherical eye of approximately 400 microm in diameter. The eye contains 750-800, mostly hexagonal ommatidia, which are of the acone apposition type and have an open rhabdom. A well-developed pupil mechanism controls the light flux to the rhabdom. The pupil is formed with the help of screening pigment translocations, involving primary and secondary (accessory) pigment cells. Cross-sections of rhabdoms reveal that they are developed as ring-like structures, made up of the rhabdomeres of six retinula cells, surrounding a rod-like inner column of two fused rhabdomeres. Rhabdoms of ommatidia in the middle of the eye differ somewhat from those in more peripheral areas. In the former the central rhabdom is circular in cross-section, while in the latter it is spindle-shaped. The rhabdom organization in combination with the distal pupil mechanism is seen as an adaptation to maximize photon capture under a variety of ambient light intensities, for Oedemerid beetles are commonly active during the day as well as the night.

The psocopteran Psyllipsocus ramburi Sélys-Longchamps can render food stuffs unpalatable and may serve as an intermediate host for cestodes. Its two circular compound eyes consist of about 26 facets, capped by strongly convexly curved corneae of 10-18mum in diameter. Corneal nipples or interommatidial hairs are not developed. Beneath each corneal lens a cluster of four cone cells, enveloped by two primary pigment cells, separates an ommatidial group of eight retinula cells from the inner corneal surface. Membrane specializations of the retinula cells, known as the microvilli, measure 60nm in diameter, and collectively make up the rhabdom, which is columnar in shape and has a distal diameter of 4 or 5mum, depending on whether it is day- or night-adapted. Cone cell lengths measure 4.5mum during the day and 8.5mum at night and retinula cell screening pigments closely approach the edge of the rhabdom during the day. A 1-h exposure to UV-A (lambda(max)=351nm) of ca. 1200lx causes an almost total destruction of the photoreceptive membranes of the rhabdom and bleached all retinula cell screening pigments, but not the pigment grains of the primary pigment cells. Calculations, based on the anatomical data, suggest that the eyes are adapted to function under dim light levels, but cannot produce sharp images since their best possible acceptance angles are 22 degrees and 28 degrees in light- and dark-adapted states, respectively. Destruction of vision, likely affecting biorhythm and reproduction, by exposing the insects to UV-A may offer an alternative to the use of chemicals in controlling these insects.

Cadmium is a toxic heavy metal that is widely distributed in the environment. As a critical process, oxidative toxicity mediates the morphological and functional damages in germ cells after cadmium exposure. In this study, the protective effect of quercetin on cadmium-induced oxidative toxicity was investigated in mouse testicular germ cells. After oral administration of cadmium chloride at 4 mg/kg body weight for 2 weeks, damages in spermatozoa occurred in the early stage of spermatogenesis. Cadmium treatment significantly decreased the testicular antioxidant system, including decreases in the glutathione (GSH) level, superoxide dismutase (SOD), and GSH peroxidase (GSH-Px) activities. Moreover, exposure to cadmium resulted in an increase of hydrogen peroxide production and lipid peroxidation in testes. In addition, cadmium provoked germ cell apoptosis by upregulating expression of the proapoptotic proteins Bax and caspase-3 and downregulating expression of the antiapoptotic protein Bcl-XL. However, combined administration of a common flavonoid quercetin at 75 mg/kg body weight significantly attenuated cadmium-induced germ cell apoptosis by suppressing the hydrogen peroxide production and lipid peroxidation in testicular tissue. Simultaneous supplementation of quercetin markedly restored the decrease in GSH level and SOD and GSH-Px activities elicited by cadmium treatment. Additionally, quercetin protected germ cells from cadmium-induced apoptosis by downregulating the expression of Bax and caspase-3 and upregulating Bcl-XL expression. These results indicate that quercetin, due to its antioxidative and antiapoptotic characters, may manifest effective protective action against cadmium-induced oxidative toxicity in mouse testicular germ cells.

This study was aimed to evaluate the effect on spermatogenesis of a 62 kDa protein (Rp) isolated from 50% ethanolic extract of the root of Ricinus communis in mice. A dose response study in mice revealed that 25mg/kg body weight/day was the most effective dose. Swiss strain mature male mice of 30 days old were divided into two group namely control and Rp treated (25mg/kg body weight/day). The study showed that sperm motility and count were decreased significantly in the treated group as compared to the control. The fertility index of the treated groups was reduced by 100%. The activity of HMG Co A reductase and cholesterol were increased significantly in the treated group. The testicular activities of 3βHSD, 17βHSD, glucose 6-phosphate dehydrogenase and malic enzyme and the level of serum testosterone were decreased significantly in the treated group. The expression of 3βHSD and 17βHSD were decreased and the expression of StAR increased significantly in the treated group as compared to the control. Proteolytic digestion of the native protein with trypsin and chymotrypsin showed that the proteolytic cleavage did not affect the spermicidal action of Rp. Hence this study can be concluded that Rp impaired spermatogenesis in vivo by suppressing the production of testosterone.

A number of animal studies as well as human epidemiological studies have demonstrated that exposure of males to various agents could result in abnormal reproductive toxicity. Acyclovir (ACV) is known to be toxic to gonads, but fails to provide the in-depth analyses of timing of damage, the types of germ cells affected, dose and the duration of damage and timing of reversal of fertility. Hence this study on sperm morphology, sperm count and sperm motility. Doses of 4 mg, 16 mg, 32 mg and 48 mg/kg body weight of ACV were administered to 9-12 weeks old male swiss albino mice by intraperitoneal route for 15 days continuously. One hundred and eighty animals were segregated into 30 groups (N=6). Twenty four groups were injected with acyclovir (4 mg, 8 mg, 16 mg and 32 mg/kg body weight) and the rest six groups served as control. After the last treatment, the animals were sacrificed on 7, 14, 21, 28, 35 and 70 days sample times and the sperm parameters were estimated. ACV causes increased incidence of abnormal sperms on most of the dose ranges from day 21 to 35 indicating the effect on spermatocytes and spermatogonial cells. ACV is cytotoxic to the testis. It causes oligospermia from day 7 to day 35 after the last exposure. It also decreased the sperm motility on same time points. All these effects were reversible by day 70. ACV exerts reversible genotoxic and cytotoxic effect on germ cells. ACV does not affect stem cell lines of spermatogenesis since all sperm parameters return to control level on day 70.

Administration of cypermethrin (CYP), orally by gavage (3 doses: 1.38, 2.76, and 5.52 mg/kg body weight) to mice either for 6 (D1) or 12 (D2) weeks caused a significant reduction in epididymal spermatozoa count and an increase in abnormal spermatozoa count when compared to controls. These counts returned to normal levels 6 weeks after cessation of 1.38 or 2.76 mg/kg body weight (BW) treatment either after D1 or D2. In 5.52 mg/kg BW treated mice the counts returned to normal levels following D1 but not after D2. Mice in all the treatment groups showed normal fertility. Weight of the litter born to mice mated with CYP treated (all three doses) males either in D1 or D2 was significantly lower than controls whereas gestation period and litter size did not significantly vary from controls. This is the first report revealing that CYP as low as 1.38 mg/kg BW adversely affects spermatogenesis and that the effect is reversible up to 2.76 mg/kg BW/kg BW exposure for 3 months. The results further reveal that despite reduction in sperm count and increase in proportion of abnormal spermatozoa, normal fertility is possible. Hence, in reproductive toxicity evaluation of pesticides, fertility test alone is misleading.

Testicular torsion is associated with damage to the testicular tissue as a result of ischemia-reperfusion injury (IRI) and induction of apoptosis leading to progressive damage to spermatogenesis. Survivin is suggested to be an important regulator in the control of the mitochondrial apoptotic pathway, although its role in torsion-induced IRI is unknown. Therefore, we sought to evaluate testicular survivin expression after long term IRI induced by testicular torsion. Survivin expression was measured by real-time PCR in 6-12 month old New Zealand white rabbits divided into three groups (4 animals/group): group (A) sham control, group (B) ischemia alone for 60 min and group (C) ischemia for 60 min followed by reperfusion for 6 months. Germ cell apoptosis was evaluated by TUNEL assay, Bax/Bcl-2 ratio and DNA fragmentation. The Johnsen score was used to assess testicular morphological damage, while lipid peroxidation was used as an indicator for oxidative stress. Survivin expression was detected in all testicular tissue samples. The rate of survivin expression after IRI was significantly higher (p<0.05) compared with ischemic only and sham control testes. Its expression in IRI samples was inversely correlated with the significant increase (p<0.05) in apoptosis, oxidative levels and spermatogenic damage. In conclusion, down-regulation of testicular survivin expression after long term IRI to the testis and its association with apoptosis induction suggests its involvement in the regulation of this apoptotic pathway. These findings also identify survivin as a potential new target for the prevention of germ cell death during testicular torsion.

p-tert-Octylphenol (OP) is a degradation product of alkylphenol ethoxylates. OP is an endocrine disruptor known to bind to the estrogen receptor; however, effects on males are controversial. The objective of this study was to evaluate the effects of chronic exposure to OP on male reproduction. Adult Sprague-Dawley rats were administered OP for 60 d, representing 1.5 cycles of spermatogenesis. Experimental groups included a vehicle control, and three doses of OP (25, 50, or 125 mg/kg body weight [bw]) administered daily by gavage. There was a significant decrease in body weight in the 125-mg/kg group after 60 d of treatment. Both testicular and epididymal weights and histology were not altered by treatment with OP at any of the doses administered. There were no marked differences in cauda epididymal sperm counts at any doses; however, total percent sperm motility was significantly lower in rats exposed to the intermediate dose (50 mg/kg bw). There was an increase in percent static sperm cells in all OP-treated groups, with the intermediate dose (50 mg/kg) displaying a significantly higher proportion of static cells relative to untreated controls. Caput epididymal sperm motility was unaltered by OP treatment. Gene expression profiles of testes from control and high-dose-exposed rats indicate that 14 genes were modulated by at least twofold, although these changes were not statistically significant. Taken together, results from this study indicate that OP treatment of adult rats does not appear to exert major effects on male reproductive endpoints at relevant environmental exposure doses.

The present study was undertaken to search out the effect of sodium fluoride, a water pollutant noted throughout the world, including India, on oxidative stress induction in reproductive tissues, sperm pellet, and metabolic tissues like the liver and kidney. The protective effects of testosterone or vitamin-E coadministration were also observed on oxidative stress in the above mentioned samples. A significant diminution was noted in the activities of catalase and peroxidase, important antioxidant enzymes in testicular tissue, sperm pellet, prostate, and epididymis in sodium fluoride-treated rats at the dose of 20 mg/kg body weight/day (the level noted in drinking water in fluoride intoxicated areas) for 30 days by oral gavage. Coadministration of testosterone by intraperitoneal injection at the dose of 40 mug/100 g body weight/alternate day, 3 hours after fluoride treatment, resulted in a significant protection in the above mentioned parameters of all these samples. Moreover, fluoride treatment also resulted a significant elevation in the level of malondialdehyde and conjugated dienes, indicators of oxidative stress, in all the above mentioned samples, which were resettled toward the control level after testosterone coadministration. Testiculo-somatic, prostato-somatic, and epididymo-somatic indices were decreased significantly in the fluoride-treated group when compared to the vehicle-treated control group. Testosterone coadministration resulted in significant restoration of these indices to the control level. We also measured the above parameters for the evaluation of oxidative stress in the liver and kidney, important metabolic organs, and noted that there was also a significant elevation in malondialdehyde and conjugated dienes along with diminution in catalase and peroxidase activities in the fluoride treated-group, with respect to the vehicle treated control group. Testosterone coadministration resulted a significant protection in these parameters toward the vehicle-treated control level. There was no significant change in hepato-somatic and reno-somatic indices among fluoride-treated, testosterone coadministered, and vehicle-treated rats. Body weight of the animals among these three groups were not changed significantly. To find out the antioxidative property of testosterone compared to vitamin E, one group of fluoride-treated animals were subjected to coadministration of vitamin E at the dose of 20 mg/100 g body weight. It was noted that in reproductive organs and in metabolic organs, oxidative stress parameters were recovered toward the control level. The results of our experiment suggests that fluoride at the dose noted in drinking water in contaminated areas may induce oxidative stress in reproductive and metabolic organs that can be ameliorated significantly by testosterone or vitamin E coadministration. Moreover, as there was no significant variation in body weights among these groups, it may be predicted that this effect of fluoride on reproductive and metabolic organs is specific and is not due to general effect of fluoride.

PURPOSE To investigate protective effects of vitamins E and C against 1.5 Tesla static magnetic fields in magnetic resonance imaging (MRI) on spermatogenesis parameters was the main goal of the present study. MATERIALS AND METHODS Ninety-two mature male rats were exposed to 1.5 T MRI static magnetic fields for 30 min with or without vitamins C and E alone or in combination. Animals were sacrificed and the testicular tissues were anatomically sectioned, stained, and the number of germ cells and the diameters of sperm ducts were measured and compared with sham and controls. RESULTS Results showed that compared to sham, static magnetic fields may reduce the germ cell count (P = 0.000) and sperm ducts diameters (P = 0.020), and vitamins C and E could modify the reduction in germ cell count (P = 0.019) but they did not show any protective effect on sperm duct diameter reduction (0.647). CONCLUSION The protective effects of vitamins C and E are different, and depend on the type of effects. It seems that the modifying effects of vitamins are to be additive, but vitamin E plays a more important role than vitamin C against the static magnetic field on spermatogenesis parameters in clinical MRI.

BACKGROUND Apoptosis involves in testicular germ cell loss in animals and humans, and plays an important role in male fertility. Previous studies have reported neuroprotective and antiapoptotic effects of minocycline. AIM In this study the protective effect of Minocycline on testicular germ cell apoptosis arising from dexamethasone (Dex) has been evaluated. Dex is a widely used as a glucocorticoid (GC) agent that its apoptotic effect has been shown. MATERIALS AND METHODS Experimental groups of 8 male mice received one of the following treatments daily for 7 days: 100 mg/kg Minocycline, 7 mg/kg Dex and 7 mg/kg Dex +100 mg/kg Minocycline. Control group was treated with 0.5 ml saline given orally for a week. Then the mice were sacrificed, and their testes processed for assessment of germ cell apoptosis (TUNEL method), the quality of spermatogenesis (Johnsen score system) and testicular sperm counts. RESULTS Germ cell apoptosis were significantly increased in Dex treated mice compared with control (P < 0.01). Spermatogenesis and the number of sperms head were significantly reduced in Dex treated mice compared with those of the control group (P < 0.01). Treatment with Dex and Minocycline resulted in an inhibition effect on germ cell apoptosis and a significant increase in the Johnsen score and the number of head sperm compared with Dex treated mice (P < 0.05). CONCLUSION The application of Minocycline may serve as a beneficial medication to protect germ cells against apoptosis.

Effect of arsenic was studied on the testicular tissue of Swiss albino mice. Sodium-meta-arsenite (NaAsO2) was administered to adult mice (25 +/- 30 g) at a dose level of 30 mg/L and 40 mg/L through drinking water for 30, 45 and 60 days. After the treatment, the testicular organ was removed, weighed and processed for histopathological observation. No change in the body weight was recorded in treated groups after arsenic exposure but significant decrease in the relative testicular weight was observed in comparison with the control. The result showed that arsenic-treated mice exhibited dose dependent gradual reductions in seminiferous tubular diameter and various gametogenic cell population i.e. resting spermatocyte, pachytene spermatocyte and step-7-spermatid except spermatogonia. Leydig cell atrophy was significantly increased in dose dependent manner indicating a definite effect of arsenic on the spermatogenesis in mice. These observations were supported by gradual reduction in Leydig cell population in the above treated groups. In conclusion, the above results confirm the toxic effect of arsenic in testis of mice.