Luna Samanta

Testosterone‐induced changes in testicular antioxidant system

Summary. In order to investigate the role of testosterone propionate (TP) on the antioxidant system of the rat testis, lipid peroxidation (LPX) and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) of the testis of testosterone‐treated and control rats were compared. The results indicate that TP administration to intact adult rats resulted in a significant decline in protein content of various subcellular fractions. This is accompanied with significant elevation in LPX levels of various sub‐cellular fractions suggesting induction of oxidative stress. Activities of three enzymes related to the metabolism of superoxide radical (SOD) and hydrogen peroxide (CAT and GPx) of testis, were found to be significantly decreased in response to TP treatment. The role of testosterone in regulating testicular spermatogenesis through oxidative stress is discussed.

Comparison of Hexachlorocyclohexane-Induced Oxidative Stress in the Testis of Immature and Adult Rats

1. The acute effect of hexachlorocyclohexane (HCH) administration (i.p.) on testicular antioxidant system and lipid peroxidation (LPX) in immature and mature rats (15- and 90-day-old, respectively) were compared. 2. In both the age groups, the level of LPX in crude homogenate of testis (endogenous, as well as FeSO4, and ascorbic acid-stimulated) was increased after 6 hr of HCH treatment and remained high till 24 hr. However, FeSO4 and ascorbic acid-stimulated LPX was higher in 90-day-old rats in comparison to 15-day-old rats. HCH treatment also resulted in elevation of LPX level in testicular subcellular (nuclear, mitochondrial and microsomal) fractions by 6 hr of treatment. However, the magnitude of increase was greater in case of 90-day-old rats. 3. Activities of testicular cytosolic superoxide dismutases (total and CN(-)-resistant) of rats of 15- and 90-day-old age groups decreased significantly after 6 hr of HCH treatment, and remained decreased till 24 hr of the pesticide treatment. The percentage of decrease was higher in 15-day-old rats than 90-day-old rats. CN(-)-sensitive SOD activity of testis was found to decrease by 12 and 24 hr after the pesticide treatment in 15- and 90-day-old rats, respectively. The activity of catalase decreased 6 hr after the pesticide treatment in both the age groups. However, the magnitude of decrease was similar for both age groups of rats. 4. Testicular glutathione content, as well as levels of glutathione metabolizing enzymes (glutathione peroxidase and glutathione reductase), did not change in response to HCH treatment, whereas ascorbic acid content decreased by 12 and 6 hr after HCH treatment in 15- and 90-day-old rats, respectively. The level of H2O2 was found to be elevated after 6 hr of the pesticide treatment in both age groups. 5. Total epididymal sperm number was comparable in all experimental groups. However, the percentage of dead and damaged spermatozoa was significantly enhanced in HCH treated rats. 6. Acute HCH administration to rats results in induction of oxidative stress in the testis which depends upon the age of the animal.

Changes in rat testicular antioxidant defence profile as a function of age and its impairment by hexachlorocyclohexane during critical stages of maturation

Summary. Age‐related changes in rat testicular oxidative stress parameters were investigated. A biphasic pattern was evident for lipid peroxidation and for the activity ratio of superoxide dismutase to catalase and glutathione peroxidase with increasing age. In the first phase of life (birth‐7 days), a linear fall in lipid peroxidation was accompanied by a gradual increase in the enzyme ratio which was reversed in the second phase (15–600 days). Glutathione and ascorbic acid levels increased from birth to the 45th day and remained unchanged up till 365 days and then reduced at 600 days of age. The maximum level of H2O2 observed at birth gradually decreased till 90 days and remained unchanged up till 365 days of age; thereafter its level was elevated on day 600. The results suggest that an antioxidant defence system plays a crucial role in development and maturation of the rat testis. When the rats were treated with hexachlorocyclohexane during critical stages of testicular development (6th‐30th day) and responses were evaluated on the 46th day of age, elevations in the levels of testicular lipid peroxidation and H2O2 along with reduction in levels of superoxide dismutase, catalase and ascorbic acid were observed. However, no change in glutathione and its metabolizing enzymes was recorded. On the other hand, hexachlorocyclohexane elevated total testicular Ca2+‐Mg2+‐ATPase activity. The results advocate for impairment of testicular functions in adult age as a consequence of some permanent lesions induced by hexachlorocyclohexane during critical stages of sexual maturation.

Oxidative Stress and Heart Failure in Altered Thyroid States

Increased or reduced action of thyroid hormone on certain molecular pathways in the heart and vasculature causes relevant cardiovascular derangements. It is well established that hyperthyroidism induces a hyperdynamic cardiovascular state, which is associated with a faster heart rate, enhanced left ventricular systolic and diastolic function whereas hypothyroidism is characterized by the opposite changes. Hyperthyroidism and hypothyroidism represent opposite clinical conditions, albeit not mirror images. Recent experimental and clinical studies have suggested the involvement of ROS tissue damage under altered thyroid status. Altered-thyroid state-linked changes in heart modify their susceptibility to oxidants and the extent of the oxidative damage they suffer following oxidative challenge. Chronic increase in the cellular levels of ROS can lead to a catastrophic cycle of DNA damage, mitochondrial dysfunction, further ROS generation and cellular injury. Thus, these cellular events might play an important role in the development and progression of myocardial remodeling and heart failure in altered thyroid states (hypo- and hyper-thyroidism). The present review aims at elucidating the various signaling pathways mediated via ROS and their modulation under altered thyroid state and the possibility of antioxidant therapy.

Age-related changes in rat testicular oxidative stress parameters by hexachlorocyclohexane

Abstract Effect of repeated oral administration of hexachlorocyclohexane (HCH; 10 and 20 mg/kg body weight per day for 7, 15 and 30 days) on antioxidant defence system and lipid peroxidation (LPX) in the testis was compared between immature (15-day-old) and mature (90-day-old) rats. In both age-groups of rats, the pesticide elicited a significant decrease in the activities of cytosolic superoxide dismutase (SOD; total and CN−-resistant) and catalase, and ascorbic acid content together with an increase in the levels of LPX (both in crude homogenate and subcellular fractions) and H2O2. Testicular glutathione peroxidase (GPx; total and non-selenium-dependent) activity was enhanced in both the age-groups of rats while the testicular glutathione content as well as glutathione reductase activity remained unaltered. HCH treatment resulted in a decrease of total epididymal sperm number with a higher incidence of dead and damaged spermatozoa, and sperms having anomalous head. Statistical analyses suggest that the alterations in the testicular antioxidant defence profile in the rat are not only dependent on the duration of pesticide treatment, but also influenced by age.

Hexachlorocyclohexane-induced behavioural and neurochemical changes in rat

Effect of chronic oral exposure (10 and 20 mg kg−1 body wt. for 7, 15 and 30 days) to hexachlorocyclohexane (HCH) on open‐field behaviour and activities of cerebral Na+,K+‐ATPase, Mg2+‐ATPase and acetylcholinesterase (AChE) of rat was evaluated. Motor and grooming activities were altered, whereas vertical exploratory activity was unaffected by HCH. Activities of Na+,K+‐ATPase, Mg2+‐ATPase and AChE were inhibited significantly by the pesticide. The results suggest that HCH induces impairment of the enzymes involved in synaptic activity, resulting in behavioural alterations. Copyright

Thermosensitive ion channel TRPV1 is endogenously expressed in the sperm of a fresh water teleost fish (Labeo rohita) and regulates sperm motility

Sperm cells exhibit extremely high sensitivity in response to slight changes in temperature, osmotic pressure and/or presence of various chemical stimuli. In most cases throughout the evolution, these physico-chemical stimuli trigger Ca2+-signaling and subsequently alter structure, cellular function, motility and survival of the sperm cells. Few reports have recently demonstrated the presence of Transient Receptor Potential (TRP) channels in the sperm cells from higher eukaryotes, mainly from higher mammals. In this work, we have explored if the sperm cells from lower vertebrates can also have thermo-sensitive TRP channels. In this paper, we demonstrate the endogenous presence of one specific thermo-sensitive ion channel, namely Transient Receptor Potential Vanilloid family member sub type 1 (TRPV1) in the sperm cells collected from fresh water teleost fish, Labeo rohita. By using western blot analysis, fluorescence assisted cell sorting (FACS) and confocal microscopy; we confirm the presence of this non-selective cation channel. Activation of TRPV1 by an endogenous activator NADA significantly increases the quality as well as the duration of fish sperm movement. The sperm cell specific expression of TRPV1 matches well with our in silico sequence analysis. The results demonstrate that TRPV1 gene is conserved in various fishes, ranging from 1–3 in copy number, and it originated by fish-specific duplication events within the last 320 million years (MY). To the best of our knowledge, this is the first report demonstrating the presence of any thermo-sensitive TRP channels in the sperm cells of early vertebrates as well as of aquatic animals, which undergo external fertilization in fresh water. This observation may have implications in the aquaculture, breeding of several fresh water and marine fish species and cryopreservation of fish sperms.

T3 fails to restore mitochondrial thiol redox status altered by experimental hypothyroidism in rat testis

Oxidative stress impaired sperm function might lead to infertility. The objective of this study was to evaluate the effects of altered thyroid hormone levels on regulation of mitochondrial glutathione redox status and its dependent antioxidant defense system in adult rat testis and their correlation with testicular function. Adult male Wistar rats were rendered hypothyroid by administration of 6-n-propyl-2-thiouracil in drinking water for six weeks. At the end of the treatment period, a subset of the hypothyroid rats was treated with T(3) (20 μg/100g body weight/day for 3 days). Mitochondria were isolated from euthyroid, hypothyroid and hypothyroid+T(3)-treated rat testes, and sub-fractionated into sub-mitochondrial particles and matrix fractions. Mitochondrial respiration, oxidative stress indices and antioxidant defenses were assayed. The results were correlated with daily testicular sperm production and epididymal sperm viability. Increased pro-oxidant level and reduced antioxidant capacity rendered the hypothyroid mitochondria susceptible to oxidative injury. The extent of damage was more evident in the membrane fraction. This was reflected in higher degree of oxidative damages inflicted upon membrane lipids and proteins. While membrane proteins were more susceptible to carbonylation, thiol residue damage was evident in matrix fraction. Reduced levels of glutathione and ascorbate further weakened the antioxidant defenses and impaired testicular function. Hypothyroid condition disturbed intra-mitochondrial thiol redox status leading to testicular dysfunction. Hypothyroidism-induced oxidative stress condition could not be reversed with T(3) treatment.

Multivariate analysis of potential biomarkers of oxidative stress in Notopterus notopterus tissues from Mahanadi River as a function of concentration of heavy metals.

In this study, investigation were done on the Mahanadi River water and health of dwelling Indian Knife fish Notopterus notopterus from three sites along the course of the river in an around Cuttack city (Odisha). Oxidative stress biomarker assays such as thiobarbituric acid reactive substances, protein carbonyls, protein and non-protein thionyls, reduced glutathione, metallothionein, superoxide dismutase, catalase, glutathione peroxidase/reductase couple, glutathione-S-transferase, and tissue metal (Fe, Cu, Ni, Cd, Pb and Zn) levels along with water quality assessments were assayed to measure the impacts on fish health. Results indicate that except Fe all other metals studied were within approved limits for fish liver and gill as approved by FAO/WHO. However, the muscle tissue do not have any metal beyond the permissible limit. A site and tissue specific response of the above mentioned oxidative biomarkers as well as metal accumulation in the fish tissues were noticed. Lipid peroxidation and protein carbonylation were increased gradually in the fish tissues collected from experimental sites along the course of the River in comparison to upstream reference site. Glutathione-S-transferase, glutathione peroxidase/reductase couple, reduced glutathione and non-protein thiol content were significantly decreased in fish tissues from experimental sites. An increase in metallothionein content was observed while superoxide dismutase and catalase showed tissue specific responses. Multivariate (Discriminant Function) analysis revealed that lipid peroxidation, protein carbonylation and superoxide dismutase have highest association as predictors of impact in the muscle and liver while that for gill is protein carbonylation, superoxide dismutase and glutathione reductase.

Overlapping dose responses of spermatogenic and extragonadal testosterone actions jeopardize the principle of hormonal male contraception

Testosterone (T), alone or in combination with progestin, provides a promising approach to hormonal male contraception. Its principle relies on enhanced negative feedback of exogenous T to suppress gonadotropins, thereby blocking the testicular T production needed for spermatogenesis, while simultaneously maintaining the extragonadal androgen actions, such as potency and libido, to avoid hypogonadism. A serious drawback of the treatment is that a significant proportion of men do not reach azoospermia or severe oligozoospermia, commensurate with contraceptive efficacy. We tested here, using hypogonadal luteinizing hormone/choriongonadotropin receptor (LHCGR) knockout (LHR–/–) mice, the basic principle of the T‐based male contraceptive method, that a specific T dose could maintain extragonadal androgen actions without simultaneously activating spermatogenesis. LHR–/– mice were treated with increasing T doses, and the responses of their spermatogenesis and extragonadal androgen actions (including gonadotropin suppression and sexual behavior) were assessed. Conspicuously, all dose responses to T were practically superimposable, and no dose of T could be defined that would maintain sexual function and suppress gonadotropins without simultaneously activating spermatogenesis. This finding, never addressed in clinical contraceptive trials, is not unexpected in light of the same androgen receptor mediating androgen actions in all organs. When extrapolated to humans, our findings may jeopardize the current approach to hormonal male contraception and call for more effective means of inhibiting intratesticular T production or action, to achieve consistent spermatogenic suppression.—Oduwole, O. O., Vydra, N., Wood, N. E. M., Samanta, L., Owen, L., Keevil, B., Donaldson, M., Naresh, K., Huhtaniemi, I. T. Overlapping dose responses of spermatogenic and extragonadal testosterone actions jeopardize the principle of hormonal male contraception. FASEB J. 28, 2566–2576 (2014). www.fasebj.org