ABSTRACT
At present, there is no reliable in vitro assembled prepubertal testis-like biomimetic organ culture system designed to assess the functional effects of human gonadotropins on Sertoli and Leydig cells. Spermatogenesis is regulated by endocrine, paracrine, and juxtacrine factors (testicular cross-talk), mainly orchestrated by gonadotropins such as luteinizing hormone (LH) and follicle-stimulating hormone (FSH) that play a pivotal role by stimulating Leydig and Sertoli cells, respectively. The aim of our study was to set up an in vitro prepubertal porcine bioengineered construct as a new model for experimental studies on reassembled Sertoli and Leydig cells. We have evaluated Sertoli and Leydig cells obtained from 15- to 20-day-old neonatal pig testes in terms of purity and function. Subsequently, purified Sertoli and enriched Leydig cells were subjected to coincubation to obtain an in vitro prepubertal porcine testis-like culture system. We performed enzyme-linked immunosorbent assay (ELISA) for anti-Müllerian hormone (AMH), inhibin B, and testosterone secretion in the medium, and Real-Time PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3β-HSD mRNA expression levels. This in vitro testis-like system was highly responsive to the effects of human gonadotropins and testosterone. AMH mRNA expression and secretion declined, and inhibin-B increased, while FSH-receptor expression was downregulated upon FSH/LH exposure/treatment. Finally, the production of testosterone was increased selectively upon LH treatment. In summary, our proposed model could help to better determine the action of human gonadotropins on Sertoli and Leydig cells. The potential usefulness of the system for shedding light into male infertility-related issues is evident.
Subject(s)
Animals , Male , 3-Hydroxysteroid Dehydrogenases/metabolism , Animals, Newborn , Anti-Mullerian Hormone/metabolism , Aromatase/metabolism , Cell Culture Techniques , Enzyme-Linked Immunosorbent Assay , Follicle Stimulating Hormone/pharmacology , Hormones/pharmacology , In Vitro Techniques , Inhibins/metabolism , Leydig Cells/metabolism , Luteinizing Hormone/pharmacology , Models, Biological , Real-Time Polymerase Chain Reaction , Receptors, FSH/metabolism , Receptors, LH/metabolism , Sertoli Cells/metabolism , Swine , Testis/metabolism , Testosterone/metabolismABSTRACT
The effects of supplementation of selenium at a dose of 10 microg/ kg body weight were investigated on ethanol induced testicular toxicity in rats. In the present study, four groups of male albino rats were maintained for 60 days, as follows: (1) Control group (normal diet) (2) Ethanol group (4g/kg body weight) (3) Selenium (10 microg/kg body weight) (4) Ethanol + Selenium (4g/kg body weight + 10 microg/kg body weight). Results revealed that ethanol intake caused drastic changes in the sperm count, sperm motility and sperm morphology. It also reduced the levels of testosterone and fructose. The activities of 3betaHSD, 17betaHSD in the testis and SDH in the seminal plasma were also reduced. Lipid peroxidation was also enhanced as the lipid peroxidation products were increased and the activities of the scavenging enzymes were reduced. But on coadministration of selenium along with alcohol all the biochemical parameters were altered to near normal levels indicating a protective effect of selenium. These results were reinforced by the histopathological studies.
Subject(s)
17-Hydroxysteroid Dehydrogenases/metabolism , 3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Antioxidants/pharmacology , Central Nervous System Depressants/toxicity , Ethanol/toxicity , Fructose/metabolism , Lipid Peroxidation/drug effects , Male , Rats , Rats, Sprague-Dawley , Selenium/pharmacology , Semen/enzymology , Sperm Motility/drug effects , Spermatozoa/enzymology , Testis/enzymology , Testosterone/metabolismABSTRACT
In order to find out the effect of chronic ethanol administration on testicular antioxidant system and steroidogenic enzyme activity, male rats fed with ethanol 1.6g/kg body weight per day for four weeks were studied. Besides a drastic reduction in body and testis weight, there was decrease in ascorbic acid, reduced glutathione and activities of superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase in the testicular tissue of the treated animals. Simultaneously, there was increase in lipid peroxidation and glutathione S-transferase activity. Activities of 3 beta-hydroxy steroid dehydrogenase and 17 beta-hydroxy steroid dehydrogenase were also found decreased in the treated animals. The results indicate that chronic ethanol administration resulted in increase in oxidative stress and decrease in the activities of steroidogenic enzymes in the rat testes.
Subject(s)
17-Hydroxysteroid Dehydrogenases/metabolism , 3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Antioxidants/metabolism , Ethanol/administration & dosage , Male , Rats , Rats, Wistar , Reactive Oxygen Species , Testis/drug effectsABSTRACT
Adrenal delta5-3beta-hydroxysteroid dehydrogenase (delta5-3beta-HSD) activity and serum corticosterone level were significantly higher in rats fed with 5% casein or 4% albumin diets after 1 hr of ether anaesthetic stress as compared to the controls, 5% casein and 20% casein (equivalent to 4% albumin) respectively. Ether anaesthesia to 20% casein fed rats caused no change in adrenal delta5-3beta-HSD activity and serum corticosterone level when compared with controls fed 20% casein diet. The results suggest that high milk protein diet may prevent acute stress effects by protecting adrenocortical activity. The present investigation opens up a new area of management of stress.
Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Adrenal Cortex/drug effects , Albumins/pharmacology , Anesthesia , Animals , Caseins/pharmacology , Chelating Agents/pharmacology , Corticosterone/blood , Diet , Ether/chemistry , Male , Rats , Rats, WistarABSTRACT
Feeding 50% ethanolic extract of A. aspera to male rats resulted in reduced sperm counts, weight of epididymis, serum level of testosterone and testicular activity of 3beta-hydroxysteroid dehydrogenase, while motility of the sperm and activity of the HMG CoA reductase were not affected. Cholesterol level in the testis, incorporation of labelled acetate into cholesterol, 17-ketosteroids in urine and hepatic and fecal bile acids were increased. The results suggest that ethanolic extract of A. aspera caused reproductive toxicity in male rats and the action may be by suppressing the synthesis of androgen.
Subject(s)
17-Ketosteroids/metabolism , 3-Hydroxysteroid Dehydrogenases/metabolism , Achyranthes , Animals , Cholesterol/metabolism , Diet , Epididymis/drug effects , Ethanol/pharmacology , Hydroxymethylglutaryl CoA Reductases/metabolism , Infertility, Male , Liver/metabolism , Male , Organ Size/drug effects , Plant Extracts/administration & dosage , Rats , Rats, Sprague-Dawley , Seminal Vesicles/chemistry , Solvents/pharmacology , Sperm Count , Sperm Motility/drug effects , Testis/chemistry , Testosterone/bloodABSTRACT
Adrenal weight, adrenal hydroxysteroid dehydrogenase activity and serum corticosterone level were significantly higher in rats fed with 5% casein diet after 7 days of swimming stress (45 min/day) as compared to their controls. All the parameters were similar to their control levels in rats receiving 20% casein diet and exposed to swimming stress. The results suggest that casein can play an important role in preventing adrenocortical hyperactivity in swimming stressed rats.
Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Adrenal Cortex/drug effects , Animals , Caseins/pharmacology , Corticosterone/blood , Dietary Proteins/pharmacology , Male , Rats , Rats, Wistar , Stress, Physiological/physiopathologyABSTRACT
Young adult male rats, maintained either in an LD 12 : 12 or in continuous illumination (LL) for one week, were given a single injection of 25 microg melatonin/100 g body wt or ethanolic-saline (control) at 17.00 h. Animals from each group were sacrificed at 11.00 h on the following day. The activity of two important steroidogenic enzymes, 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) and delta(5)-3 beta-hydroxysteroid dehydrogenase (delta(5)-3 beta-HSD), and serum concentrations of testosterone, were measured following highly specific and sensitive spectrophotometric techniques and RIA, respectively. A significant decrease in the activity of both the steroidogenic enzymes was noted in the testes of melatonin-treated rats maintained under normal light-dark schedules, but this response was found to be lacking in the LL rats. However, no significant changes in the level of serum testosterone were noted in either group of melatonin-treated rats from the values in respective groups of ethanolic saline-administered LD and LL rats. Exposure of ethanolic saline-injected rats to continuous light also did not cause any change in the steroidogenic activity of the testis from those in LD rats. The study indicates that continuous light as such does not affect the endocrine function of testis but abolishes suppressive effects of melatonin on the steroidogenic activity of the testis in rat.
Subject(s)
17-Hydroxysteroid Dehydrogenases/metabolism , 3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Circadian Rhythm/physiology , Drug Administration Schedule , Housing, Animal , Injections, Subcutaneous , Light , Lighting , Male , Melatonin/administration & dosage , Organ Size/drug effects , Rats , Testis/anatomy & histology , Testosterone/biosynthesisABSTRACT
La adrenarca se caracteriza por el aumento de secreción de andrógenos adrenales. Ocurre entre los 6-8 años de edad en humanos. Con el objetivo de evaluar el posible rol regulatorio de la 3beta HSD en la síntesis de andrógenos en la adrenal humana, se analizó la abundancia del ARNm de 3beta HSD (Dot blot y RT-PCR semicuantitativa(S)) en 11 tejidos adrenales humanos normales en dos grupos (Gr) de edades, Gr1: <8 años (a) (n= 6, r: 0.1-2.5) y Gr2: si 8a (n = 5, r: 8.0-13.0); en tejido de un tumor adrenal con Síndrome de Cushing (TSC) y de 2 T virilizante (TV) y en células adrenales del TSC y 1 TV al 3er día de cultivo (basal y con ACTH e IGF-1). El ARNm de 3beta HSD fue más alto en el Gr1 que en el Gr2 (Dot blot: 4.65 + 2.70 y 0.28 + 0.27 UA, p = 0.006; RT-PCRS: 21.50 + 12.50 y 6.77 + 3.78 UA, p = 0.039 resp). Por otra parte, en el tejido TSC (8.74 + 1.74) fue más alto que en los TV (0.47 + 0.02, 0.87 + 0.08) p = 0.001. En cultivo del TSC el ARNm basal (0.82 + 0.10) disminuyó con ACTH (0.55 + 0.06), p = 0.005 y se incrementó con IGF-1 (2.36 + 0.07) p = 0.006. En el TV no hubo cambios. Al 3er día de cultivo la DHEAS y androstenediona basales fueron en TV 1170.0 + 210.0 y 335.0 + 29.0, TSC 17.1 + 3.5 y 73.7 + 11.7 pmol/10(6) células en 24 hs respectivamente y se incrementaron bajo ACTH en TV (2006.0 + 360.0 y 525.0 + 76.0) y en TSC 29.8 + 5.4 y 366.8 + 129) p<0.05 y disminuyeron con IGF-1 sólo en TSC (7.9 + 2.4 y 43.7 + 6.1) p<0.05. Estos datos sugieren que la adrenarca humana podría ser secundaria a una disminución de la abundancia del ARNm de 3beta HSD. El hecho de que bajo ACTH aumenta la secreción de andrógenos y disminuye el ARNm de 3beta HSD, mientras que el IGF-1 ejerce un efecto inverso, aporta nuevas evidencias del rol regulador ejercido por la 3beta en la síntesis de andrógenos adrenales.
Subject(s)
Child , Humans , Male , Adolescent , Child, Preschool , Infant , 3-Hydroxysteroid Dehydrogenases/metabolism , Adrenal Gland Neoplasms/enzymology , Adrenal Glands/enzymology , Androgens/metabolism , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger/analysis , RNA, Messenger/metabolismABSTRACT
Administration (i.m.) of synthetic mammalian LHRH (40 ng/frog, alternate day for 30 days) to adult male intact frogs R. cyanophlyctis resulted in activation of the interrenal cells as shown by increases in nuclear diameters of the interrenal cells, activity (histochemical demonstration) of delta 5-3 beta-hydroxysteroid dehydrogenase and glucose-6-phosphate dehydrogenase and concomitant decrease in sudanophilic lipid droplets in the interrenal gland when compared to those of controls. These changes were not observed in the interrenal gland of LHRH treated hypophysectomized frogs. The results indicate that LHRH stimulates secretory activity of the interrenal cells and action may not be directly on the adrenal gland.
Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Glucosephosphate Dehydrogenase/metabolism , Gonadotropin-Releasing Hormone/pharmacology , Hypophysectomy , Interrenal Gland/cytology , Male , Pituitary Gland/physiology , RanidaeABSTRACT
Role of dietary energy and local ovarian hormones in conceptus development in the rat was evaluated by giving ip injections of estrogen and progesterone under diet restricted and/or dextrose supplemented conditions, using the experimental model of ovarian compensatory hypertrophy. Replacement of estrogen on day 2 and progesterone on day 3 was necessary for conceptus development in the ipsilateral uterine horn after unilateral ovariectomy on day 2. Continuation of progesterone injection up to day 9 improved the percentage of conceptus development. Conceptus development was normal when unilateral ovariectomy was performed after day 2 of pregnancy. Compensatory increases in corpus luteum (CL) volume and the activity of delta 5 3 beta-hydroxy steroid dehydrogenase (3 beta-HSD) in contralateral ovary were observed after unilateral ovariectomy on day 2-4 of pregnancy. However, 70% diet restriction reducing the energy intake (from 90 to 27 kcal/rat/day) abolished the single dose hormone therapy effects in ipsilateral horn conceptus development and also reduced the effects of continued progesterone, percentage of contralateral horn conceptus development, compensatory changes in the CL volume and delta 5 3 beta-HSD activity of the contralateral ovary. However, dietary dextrose supplementation (increasing calorie intake from 27 to 56 kcal/rat/day) significantly improved/restored hormone response for conceptus development, compensatory changes of the CL volume and enzyme activity in contralateral ovary. The results suggest that day 2-3 of pregnancy is the critical time when local ovarian supply of estrogen and progesterone are obligatory for establishment and conceptus development in the rat. During this period adequate mobilizable source of energy is necessary for execution of hormonal message.
Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Corpus Luteum/drug effects , Embryo Implantation/drug effects , Energy Metabolism , Estradiol/pharmacology , Female , Gonadal Steroid Hormones/physiology , Ovary/physiology , Progesterone/pharmacology , RatsABSTRACT
Changes in the adrenal weight, adrenal 5-ene-3 beta-hydroxysteroid dehydrogenase (5-ene-3 beta-HSD) activity and serum levels of corticosterone were observed in male wistar rats after the treatment of lithium chloride in the doses of 100, 200 and 400 micrograms/100 g of body weight/day for 7, 14 and 21 days. The experiments indicate that 200 and 400 micrograms/100 g.b.w. administered for 14 and 21 days caused a significant stimulation in the activities of adrenal 5-ene-3 beta-HSD along with elevation of adrenal weights and serum levels of corticosterone. 100 micrograms of lithium chloride was not able to modulate the adrenal activity. Moreover, plasma levels of lithium remain in therapeutic range in this experiment at the doses of 200 and 400 micrograms/100 g body weight. Therefore, our data suggest that lithium can alter the adrenal activity within its therapeutic range according to the duration of treatment.
Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Adrenal Cortex/drug effects , Animals , Chlorides/pharmacology , Corticosterone , Dose-Response Relationship, Drug , Lithium/blood , Lithium Chloride , Male , Organ Size/drug effects , Rats , Rats, Inbred Strains , Time FactorsABSTRACT
Activities of delta 5-3 beta- and 17 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-HSD and 17 beta-HSD), Leydig cell nuclear area (LCNA) and spermatogenesis in the testis were observed after injection of lithium chloride in the 'antiserum to luteinizing hormone (LH)' treated toad. A significant decrease in the activities of steroidogenic enzymes, LCNA and spermatogenesis were noticed after the injections of 'antiserum to LH' to toads. Further decrease in the activities of the above parameters was observed in the lithium chloride--'antiserum to LH' treated toad. It is suggested that lithium chloride may inhibits testicular function without modulating the pituitary activity.
Subject(s)
17-Hydroxysteroid Dehydrogenases/metabolism , 3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Bufonidae , Chlorides/pharmacology , Immune Sera , Lithium/pharmacology , Lithium Chloride , Luteinizing Hormone/immunology , Male , Spermatogenesis/drug effects , Testis/drug effectsABSTRACT
An LH-RH analog (des-Gly10, [D-Trp6]-LH-RH A) was administered to adult male cats for 67 days (20 microng/Kg,sc) in order to study its inhibitory effects on the structure of Leydig cells, as deter,mined by histological and histochemical-morphometric techniques. Histological examination showed that LH-RH A promotes a decrease in the volume of the interstitial tissue. In addition, Leydig cell nuclei exhibited marked structural alterations. Morphometric analyses utilizing histochemistry of the enzyme 3-ß-hydroxysteroid dehydrogenase (3-ß-HOST-D) as a marker of Leydig cells also demostrated a signifcant decrease of the relative volume occupied by the Leydig cells isn the testis
Subject(s)
Cats , Animals , Male , 3-Hydroxysteroid Dehydrogenases/metabolism , Leydig Cells/ultrastructure , Gonadotropin-Releasing Hormone/analogs & derivatives , Testis/drug effectsABSTRACT
The microsomal membranes isolated by sucrose density gradient centrifugation from developing toad ovary have been found to differ significantly in lipid composition and various enzyme activities in different seasons. All the enzymes studied, viz. Na+, K(+)-ATPase, delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta HSD) and prostaglandin synthetase, exhibited maximum activity during the breeding season (July-September) at all stages of development (a,b,c & d). The activities of Na+, K(+)-ATPase and delta 5-3 beta HSD increased with development while that of prostaglandin synthetase followed the reverse order. The total phospholipid, cholesterol and fatty acid contents also varied with season and development. The increase in Na+, K(+)-ATPase and delta 5-3 beta HSD activities in the microsomal membranes of toad ovary at breeding season is accompanied with concomitant increase in phospholipid and unsaturated fatty acid contents at different stages in this season, thereby suggesting some correlation between them.
Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Bufonidae/metabolism , Female , Membrane Lipids/metabolism , Microsomes/metabolism , Ovary/growth & development , Prostaglandin-Endoperoxide Synthases/metabolism , Seasons , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
This is the first report on the histochemical assessment of delta 5-3 beta hydroxysteroid dehydrogenase activity in all the preimplantation embryonic stages in the rhesus monkey (Macaca mulatta). An apparent stage dependent increase in enzyme activity was obtained, however, distinctively a high degree of non-specificity in enzyme reaction was noted primarily in morulae and blastocysts. Such marked non-specificity in the histochemical enzyme reaction for delta 5-3 beta hydroxysteroid dehydrogenase activity was not found in mouse blastocysts. High amounts of endogenous steroids present within rhesus embryos, or the participation of non-specific dehydrogenases could account for the observed non-specificity. Furthermore, the present report documents the pattern and degree of association (r = 0.9; P less than 0.01) between developmental stage and gestational age of preimplantation rhesus embryos, and thus provides a normal in situ cell cleavage rate of preimplantation embryo in the rhesus monkey.
Subject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Blastocyst/enzymology , Histocytochemistry , Macaca mulattaABSTRACT
Endosulfan was studied for its effect on rat testicular toxicity in relation to the enzymes of androgen biosynthesis, viz. 3 beta-hydroxysteroid dehydrogenase (EC 1.1.1.145, 3 beta-HSD) and 17 beta-hydroxysteroid dehydrogenase (EC 1.1.1.64, 17 beta-HSD); cytosolic conjugation enzyme, glutathione-S-transferase (EC 2.5.1.18); and testicular as well as serum testosterone levels at the dose levels of 2.5, 5.0, 7.5 and 10 mg/kg body weight fed orally for 7 and 15 days. Organ and body weights of the treated animals did not change significantly, however, the testicular protein contents were found to be increased appreciably after 7 days treatments. The activity profile of cytosolic conjugation enzyme showed much remained low during 7 days treatment, however, the two steroidogenic enzymes showed much individual variations in response to endosulfan treatments. An overall varied response with respect to testosterone biosynthesis and its secretion to serum was observed suggesting nevertheless, a profound hormonal imbalance caused by this insecticide to male gonads on short term chronic exposures.
Subject(s)
17-Hydroxysteroid Dehydrogenases/metabolism , 3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Body Weight/drug effects , Endosulfan/toxicity , Glutathione Transferase/metabolism , Male , Organ Size/drug effects , Rats , Rats, Inbred Strains , Testis/analysis , Testosterone/analysisSubject(s)
3-Hydroxysteroid Dehydrogenases/metabolism , Animals , Antispermatogenic Agents/pharmacology , Dinoprost , Estradiol/pharmacology , Glucose/analogs & derivatives , Gossypol/pharmacology , Hydrogen-Ion Concentration , Injections, Subcutaneous , Kinetics , Male , Prostaglandins F/pharmacology , Rats , Rats, Inbred Strains , Testis/drug effectsABSTRACT
Fez-se cuidadosa revisäo bibliográfica, procurando mostrar a importância do estudo histoquímico da placenta humana nas diversas idades gestacionais. Vislumbram-se novos passos que poderäo ser dados quando do funcionamento do Laboratório de Histoenzimologia, a qual será instalado em futuro próximo no Departamento de Patologia da Faculdade de Medicina da Universidade Federal do Ceará
Subject(s)
Pregnancy , Humans , Female , Placenta/metabolism , Umbilical Cord/metabolism , Glycogen/metabolism , 3-Hydroxysteroid Dehydrogenases/metabolism , L-Lactate Dehydrogenase/metabolismABSTRACT
The effect of different grades of hypobaric hypoxia for 48 hours was studied on spermatogenesis, Leydig cells and delta 5-3 beta-hydroxysteroid dehydrogenase activity in toad (Bufo melanostictus). Maximum inhibition of testicular activity was noted in 7,315 m exposed animals. The impairment of testicular function at high altitude is possibly due to inhibition of gonadotrophin secretion.