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PubMed1992

Effect of testosterone deprivation on expression of the androgen receptor in rat prostate, epididymis and testis.

Adult rats were treated with ethane dimethane sulphonate (EDS) to eliminate the Leydig cells. This treatment resulted in very low levels of testosterone in the blood and in the testis. Furthermore, histological evaluation of spermatogenesis showed no marked differences between control and EDS-treated animals. In the ventral prostate, 5 days after EDS-treatment, a 4.0 +/- 0.3-fold up-regulation of androgen receptor (AR) mRNA was observed, together with a 2.2 +/- 0.2-fold increase in actin mRNA. In the epididymis, a 2.0 +/- 0.5-fold increase in AR mRNA level was observed, without a change in actin mRNA level. In the testes of EDS-treated rats, the AR mRNA level was not changed (1.02 +/- 0.17-fold of controls), and there was also no change in actin mRNA level at 5 days after EDS-treatment. These results indicate that AR mRNA expression in the ventral prostate and epididymis is regulated differentially by testosterone when compared to regulation in the testis. Testicular androgen binding sites were assayed by Scatchard analysis of the binding of 3H-R1881 to a nuclear fraction, that was isolated by a method which involved the use of liquid nitrogen and high sucrose buffer. The number of specific binding sites per testis in EDS-treated rats with testosterone-implants, remained unaltered compared to control rats (9.1 +/- 1.4 pmol/testis). In these rats, 20% of the normal testicular testosterone level was sufficient to maintain the androgen receptor in a tight nuclear binding (transformed) form. In testes from EDS-treated rats without testosterone-implants, the AR did not fractionate into the nuclear fraction; however, the total testicular AR content in these animals was close to control levels, as measured by nuclear 3H-R1881 binding after receptor transformation through injection of a high dose of testosterone (10 mg) 2 h before killing the rats (testosterone pulse). In the different experimental groups, FSH was not required to maintain the total testicular AR content (ligand binding). Immunoprecipitation and Western blotting of the testicular AR using specific monoclonal and polyclonal antibodies indicated that the total testicular amount of immunodetectable AR protein in long-term testosterone deprived rats was very low when compared to that in control rats or rats with testosterone-implants. This is in disagreement with results obtained in the ligand binding assay, and may point to a structural modification of the AR in the testis that possibly occurs in the prolonged absence of androgens.

Animals↗
PubMed1996

Influence of photoinhibition on GABA and glutamic acid levels, and on glutamate decarboxylase activity in the testis and epididymis of the golden hamster.

Gamma-aminobutyric acid (GABA) is found in the gonads and accessory reproductive organs, and a direct effect on steroidogenesis and sperm viability and motility has been described. The golden (Syrian) hamster is a seasonal breeder, and a pattern of regression-recrudescence in their reproductive organs is observed when adult animals are exposed to less than 12.5 h daylight for an extended period of time. The purpose of this study was to investigate: (1) the presence of GABA in the testis and epididymis of golden hamsters undergoing regression and spontaneous recrudescence; (2) glutamic acid levels and glutamate decarboxylase (GAD) activity in both tissues, and (3) testicular and epididymal testosterone, dihydrotestosterone and 5 alpha-androstane-3 alpha, 17 beta-diol concentrations. Adult golden hamsters were exposed to long (LP 14L:10D) or short (SP 6L:18D) photoperiods for 9, 12, 14, 16, 18 or 22 weeks. When animals were exposed to SP for 14-16 weeks, the testis and epididymis reached maximal involution. Testicular and epididymal androgen levels showed a marked decrease (p < 0.05) during the regression period, and after 18-22 weeks, values began to recover. Between 12 and 18 weeks in SP, the testicular and epididymal content of GABA and glutamic acid was reduced significantly. The concentration of GABA in both tissues showed a sharp rise (p < 0.05), while the concentration of glutamic acid diminished during the period of maximal involution (p < 0.05). In the testis, GAD activity was increased (p < 0.001) after 14 weeks in SP, with no change in the epididymis. In conclusion, glutamic acid via GAD activity could be the main source of GABA in the testis, but not in the epididymis. Furthermore, the presence of GABA in testicular cells and its subsequent photoperiodic variations might act as an important autocrine and/or paracrine modulatory signal in gonadal processes.

Androgens↗
PubMed1977

The amino-acid sequence of trout-testis histone H1.

The complete amino-acid sequence of 194 residues of trout testis histone H1 has been elucidated by automated Edman degradation of large peptides derived from specific cleavages at infrequent amino-acid residues. Trout testis histone H1 has a molecular weight of 19314, is only slighly microheterogeneous, and possesses pentapeptide sequences related to Ala-Ala-Ala-Lys-Lys repeated six times in the complete sequence. Although the N-terminus of histone H1 is known to be blocked, some 10% of intact trout testis H1 contains a free N-terminal alanine residue. Three seryl residues in the C-terminal half (97-194) of the molecule occur in the sequence Lys-Ser-Pro-Lys known to be phosphorylated in trout testis H1. The sequence has been compared to the known partial sequence of rabbit thymus H1. The results are consistent with a role for histone H1 in the maintenance of the higher-order structure of chromatin.

Amino Acid Sequence↗
PubMed1990

Carbonyl reductases from rat testis and vas deferens. Purification, properties and localization.

Three enzyme forms (T1, T2, T3) from rat testis and two from rat vas deferens (V1, V2) of carbonyl reductase have been highly purified to apparent homogeneity. These carbonyl reductases from rat reproductive organs have several similarities in terms of molecular mass (32-33 kDa), isoelectric point (pI 5.9-6.4), immunochemical properties, cofactor requirement (NADPH dependency) and sensitivity to sulfhydryl reagents. The isoenzymes from the vas deferens (V1, V2) have similar catalytic activities, whereas those from the testis (T1, T2, T3) showed different catalytic activities from each other. All enzymes, however, reduced quinones, aromatic aldehydes and ketones, while T3, V1 and V2 were characterized as possessing high affinity towards prostaglandins. An immunoinhibition study using a specific antibody indicated that these enzymes were solely responsible for the overall catalytic activities of 13, 14-dihydro-15-oxo-prostaglandin F2 alpha, 4-benzoylpyridine, and 4-nitroacetophenone reduction and prostaglandin F2 alpha oxidation in both testis and vas deferens cytosol. The immunohistochemical staining revealed a positive immunoreactivity to antibody only in the Leydig cells of the testis, but neither the germ cells nor Sertoli cells in the seminiferous tubule. The staining also showed that the enzymes in the vas deferens were primarily localized in mucosal epithelium cells.

Alcohol Oxidoreductases↗
PubMed1991

[Spatial pattern and microvascularization of the ductuli efferentes testis of bulls (Bos taurus)].

The arrangement, origin, course and opening of the ductuli efferents testis of the bull (Bos taurus) were visualized using scanning electron microscopy. The corresponding capillary structure was also described and documented with the help of corrosive microanatomical casts. The number of the ductuli efferents testis of the bull varies narrowly between 12 to 13. Ductuli removed from the surrounding tissue have a length of up to 78 cm. The duct system is composed of linear as well as tortuous parts, permitting division into three distinct segments. Blind ending ductules are very short. We therefore do not see these structures as being associated with spermiostasis. The capillary density as well as the capillary architecture show regional peculiarities. All segments of the ductuli efferents testis are composed of capillaries of the continuous type. The findings indicate that the increased capillary density in the voluminous initial section of the ductuli efferents testis represents the basis for resorptive as well as secretory activities in the bull.

Animals↗
PubMed2005

Expression and localization of growth factors and their receptors in the mammalian testis. Part I: Fibroblast growth factors and insulin-like growth factors.

It is now well established that normal development and function of testis are mediated by endocrine and paracrine pathways including hormones, growth factors and cytokines as well as by direct cell-to-cell contacts depending on tight, adhering and gap junctions. In the last two decades, several growth factors were identified in the testis of various mammalian species. Growth factors are shown to promote cell proliferation, regulate tissue differentiation, and modulate organogenesis. Interestingly, most of these peptides are expressed not only in the adult mammalian testis during spermatogenesis but also during testicular morphogenesis in prenatal and postnatal life. Our study was launched to provide an overview of the expression, localization, and putative physiological roles of growth factors and their receptors in the mammalian testis. The growth factors considered in this part of our review are fibroblast growth factors and insulin-like growth factors. These factors are found in testicular cells in prenatal, postnatal, and adult animals and are implicated in the regulation of important testicular activities including testicular cord morphogenesis, modulation of testicular hormone secretion and control of spermatogenesis.

Animals↗
PubMedDate not supplied

Studies on low molecular weight follicle stimulating hormone receptor binding inhibitor (FSH-RBI) from ovine testis.

Follicle stimulating hormone receptor binding inhibitor (FSH-RBI) has been isolated from the aqueous extracts of ovine testis using Sephadex column chromatography. Sephadex G-75 fraction IV was found to inhibit the binding of (125I) FSH to rat testis receptors. Further purification of Sephadex G-75 fraction on Sephadex G-25 column gave three fractions (I-III). The maximum inhibitory activity to inhibit (125I) FSH binding to rat testis receptor was associated with fraction III only. FSH-RBI exerted a greater inhibitory effect on the formation of the hormone-receptor complex rather than on the dissociation of the preformed complex. FSH-RBI did not inhibit the binding of (125I) LH to rat testis receptors. A significant decrease in the mouse ovarian weight was observed when FSH-RBI was injected to hCG-primed female mice. The molecular weight determination studies show that molecular weight of FSH-RBI is approximately 1400 Daltons.

Animals↗
PubMed1998

A comparative study of mast cells and eosinophil leukocytes in the mammalian testis.

The existence of a physiological integration between the immune and endocrine systems has long been recognized. In spite of the abundant literature data on the presence of cells of the immune system in the testis, mast cells and eosinophil leukocytes have received little attention. We have studied the presence, distribution and numbers of mast cells and eosinophils in the testes of 12 mammalian species. Mast cells were frequently found in equine (stallion, ass and mule) and human testis, whereas eosinophils were nearly absent. On the contrary, eosinophils were abundant in the hare testis, while mast cells were lacking. Both cells types were present in high numbers in swine (wild and domestic boar) testis. Otherwise, mast cells and eosinophils were absent from the testicular parenchyma of several species (rat, dog, cat, bull and deer), although they were present, in most cases, around blood vessels in the tunica albuginea. The presence of high numbers of mast cells and/or eosinophil leukocytes in the testicular parenchyma of some species suggest a role for these cells in local regulatory pathways.

Animals↗
PubMed1978

Age-associated changes in microvasculature of human adult testis.

Age-associated architectural changes of the human testicular microvasculature from 70 autopsy cases were stereoscopically examined with a silicone-rubber injection technique. In the testis of a young subject, the interlobular main arteries run straight. The coiling phenomena of the interlobular centripetal or centrifugal arteries, which are commonly seen in adult testis, have been so far considered as physiological transformation of the vasculature. It was confirmed that the coiling changes in the interlobular main arteries of the human testis appear as an age-dependent alteration of the vasculature closely related to the volume of the gland. The practical importance of the spirallin or coiling of arteries is that it results in a considerable reduction of blood flow. The age-related coiling of the interlobular arteries is virtually accompanied by varying degrees of collapse of the peritubular capillary networks. The reduction of blood supply to the seminiferous tubules plays an active role in promoting aging of the testis. These stereoscopical observations of age-related transfiguration of testicular microvasculature were ascertained also by histometrical examinations.

Adolescent↗
PubMed1996

Transinguinal laparoscopy for nonpalpable testis.

BACKGROUND: About 20% of all undescended testes are nonpalpable. Although surgical exploration was previously the method of choice for management, laparoscopy from the inferior edge of the umbilicus has been established as a useful method of examining a testis that is nonpalpable. However, conventional subumbilical laparoscopy is unnecessary when the testis or its remnant is located below the internal inguinal ring. We evaluated the efficacy of transinguinal laparoscopy after inguinal exploration performed for nonpalpable testis. METHODS: Twelve children with 14 nonpalpable testes (2 children had bilateral nonpalpable testes) were considered for a novel method of laparoscopy. We performed laparoscopy through the internal inguinal ring after opening the inguinal canal. RESULTS: Inguinal exploration revealed the testis or its remnant in 5 patients, and thus prevented unnecessary laparoscopy in these individuals. Transinguinal laparoscopy identified the anatomy in 7 patients. Four of the latter were found to have blind-ending vas deferens and spermatic vessels, while in 3 patients, the testes were located in the peritoneal cavity. CONCLUSION: Transinguinal laparoscopy may become the accepted method for managing nonpalpable testes, as inguinal exploration can eliminate the need for laparoscopy in some cases, and the visualization achieved with transinguinal laparoscopy is just as good as that with conventional laparoscopy.

Child↗
PubMed1981

Immunoreactivity in vitro to human testis in patients with lepromatous leprosy.

Patients with lepromatous leprosy and tuberculoid leprosy, together with normal aged matched controls, were tested for in vitro immunoreactivity against a panel of soluble extracts prepared from normal human tissues. The panel consisted of a soluble homogenate of human testis as well as two partially purified fractions of this extract, and control extracts from other human tissues. Immunoreactivity was assessed by extract induced peripheral blood lymphocyte (PBL) transformation, leucocyte migration inhibition (LMI) and leucocyte adherence inhibition (LAI). Although no difference in PBL transformation, LMI, or LAI was obtained between the groups with the control tissue extracts, significant reactivity was obtained for the patients with lepromatous leprosy in at least one of the in vitro tests, when one of the testis extracts was used (Mann-Whitney test). No such reactivity to the testis extracts was obtained in the patients with tuberculoid leprosy or the normal control subjects. Of the twenty-two patients studied with lepromatous leprosy, thirteen had clinical evidence of testicular disease and nine of these patients had raised PBL transformation and LAI reactivity to the testis extracts. This finding of immunoreactivity against testicular extracts in a significant number of patients with lepromatous leprosy by at least one of the vitro tests used, suggests that the associated in vivo testicular atrophy that occurs in these patients may have an auto-immune basis.

Adolescent↗
PubMed1990

Management of the contralateral testis in patients with testicular germ cell cancer.

Patients with testicular germ cell tumours (TGCT) are at increased risk of developing a tumour in the contralateral testis. Such a tumour may be preceded by carcinoma in situ (CIS), which is more common in patients with infertility, atrophic testis or a history of cryptorchism. Of 1219 patients with TGCT seen at the Royal Marsden Hospital between 1962 and 1984 in whom the contralateral testis was managed by surveillance, 38 (3.1%) developed a second tumour and 8 died of germ cell tumours. Seventeen of 26 assessable patients (65%) exhibited at least one of the known aetiological risk factors for carcinoma in situ. Diagnosis of carcinoma in situ may lead to more appropriate management of the contralateral testis.

Carcinoma in Situ↗
PubMed1999

Molecular cloning and characterization of a novel splicing variant of the Kir3.2 subunit predominantly expressed in mouse testis.

1. One of the features of weaver mutant mice is male infertility, which suggests that Kir3.2, a G-protein-gated inwardly rectifying K+ channel subunit, may be involved in spermatogenesis. Therefore, we have characterized the Kir3.2 isoform in mouse testis using immunological, molecular biological and electrophysiological techniques. 2. Testicular membrane contained a protein that was recognized by the antibody specific to the C-terminus of Kir3.2c (aG2C-3). Its molecular mass was approximately 45 kDa, which was smaller than that of Kir3.2c ( approximately 48 kDa). The immunoprecipitant obtained from testis with aG2C-3 contained a single band of the 45 kDa protein, which could not be detected by the antibody to the N-terminus common to the known Kir3.2 isoforms (aG2N-2). 3. A novel alternative splicing variant of Kir3.2, designated Kir3.2d, was isolated from a mouse testis cDNA library. The cDNA had an open reading frame encoding 407 amino acids, whose molecular mass was calculated to be approximately 45 kDa. Kir3.2d was 18 amino acids shorter than Kir3.2c at its N-terminal end, which was the only difference between the two clones. The 18 amino acid region possesses the epitope for aG2N-2. 4. In heterologous expression systems of both Xenopus oocytes and mammalian cells (HEK 293T), Kir3.2d either alone or with Kir3.1 exhibited G-protein-gated inwardly rectifying K+ channel activity. 5. Prominent Kir3.2d immunoreactivity in the testis was detected exclusively in the acrosomal vesicles of spermatids, while Kir3.1 immunoreactivity was diffuse in the spermatogonia and spermatocytes. These results indicate the possibility that the testicular variant of Kir3.2, Kir3. 2d, may assemble to form a homomultimeric G-protein-gated K+ channel and be involved in the development of the acrosome during spermiogenesis.

Alternative Splicing↗
PubMed1999

Expression of Fas-Fas ligand in murine testis.

PROBLEM: During spermatogenesis, it has been suggested that the number of germ cells to be matured is regulated and restricted through the apoptotic mechanism. In the present study, we investigated the expression and apoptotic role of Fas and Fas ligand (L) in the murine testis. METHOD OF STUDY: The expression of Fas-FasL in the murine testis was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR)-Southern blot hybridization, in situ hybridization, and Western blot methods. The terminal deoxynucleotide transferase mediated dUTP-nick end label (TUNEL) and DNA fragmentation methods were applied to detect the generation of apoptosis in germ cells. RESULTS: By means of RT-PCR-Southern blot hybridization, we demonstrated the positive expression of Fas in testicular germ cells, and of FasL in testicular cells. supporting the findings with in situ hybridization that Fas was localized in germ cells, whereas FasL was localized in Sertoli cells of murine testis. A specific band at 45 kDa was obtained in the lysates from testis and germ cells with Western blot analysis. Then, the co-incubation of germ cells with Spodoptera frugiperda (Sf9)-FasL cells in vitro resulted in the induction of apoptosis in germ cells detected by the TUNEL method. Furthermore, DNA fragmented ladders were also demonstrated in germ cells co-incubated with Sf9-FasL cells. CONCLUSION: Fas-FasL system seemed to play an apoptotic role in spermatogenesis by the molecular interaction between FasL on Sertoli cells and Fas on germ cells.

Animals↗
PubMed2006

Histological changes of the testis and epididymis in adult rats as a result of Leydig cell destruction after ethane dimethane sulfonate treatment: a morphometric study.

AIM: To quantitatively study the histological changes of the testis and epididymis as a result of a drastic reduction of testosterone secretion. METHODS: Fourteen adult Sprague-Dawley rats were injected intraperitoneally with ethane dimethane sulfonate (EDS, 75 mg/kg) and the same number of animals were injected with normal saline as a control. At days 7 and 12 (after treatment), respectively, half of the animals from each group were killed. The testes and epididymides were removed and tissue blocks embedded in methacrylate resin. The cell number per testis was estimated using the stereological optical disector and some other parameters were obtained using other morphometric methods. RESULTS: The EDS treatment resulted in an almost complete elimination of Leydig cells but had no effect on the numbers of Sertoli cells per testis. At day 7 after EDS treatment, many elongated spermatids were retained in the seminiferous epithelium and many round spermatids could be seen in the epididymal ducts. At day 12, a looser arrangement of spermatids and spermatocytes became evident, with apparent narrow empty spaces being formed between germ cells in an approximately radial direction towards the tubule lumen; the numbers (per testis) of non-type B spermatogonia and spermatocytes were similar to controls, whereas that of type B spermatogonia increased by 59%, and that of early round, elongating and late elongated spermatids decreased by 37%, 72% and 52%, respectively. CONCLUSION: The primary spermatogenic lesions following EDS administration were (i) spermiation failure and (ii) detachment of spermatids and spermatocytes associated with impairment in spermiogenesis and meiosis.

Animals↗
PubMed1984

Penetration of some compounds through blood-brain and blood-testis barriers in chronically hypertensive rats.

Acute hypertension impairs the function of biological barriers, e.g. that of blood-brain barrier. We tested the hypothesis that chronic hypertension might influence the penetration of compounds through blood-brain and blood-testis barriers. With this in mind, the penetration of relatively small radioactive compounds (14C-2-methyl-4-chlorophenoxyacetic acid = 14C-MPA, 14C-sucrose and 14C-antipyrine, 5 microCi/kg intravenously) into the brain, liquor space and testis was compared in male normotensive (Wistar and chronically hypertensive SH rats (11-22 months old). In chronic hypertension after 14C-MCPA and 14C-sucrose administration the penetration of radioactivity into the brain, cerebrospinal fluid and testis had significantly decreased or showed evidence of decrease, while after 14C-antipyrine administration the penetration remained unchanged. Penetration of Evans blue-albumin complex into the brain was studied by giving intravenous injection of Evans blue (5% solution) which is firmly bound to serum albumin. Extravasation of dye-protein complex into the brain showed no difference between Wistar and SH rats. The results suggest that chronic hypertension does not increase the penetration of compounds into the brain and testis but rather, reduces it.

2-Methyl-4-chlorophenoxyacetic Acid↗
PubMed1994

Micropuncture and cannulation studies of fluid composition and transport in the ductuli efferentes testis of the rat: comparisons with the homologous metanephric proximal tubule.

Luminal fluids were collected in vivo by micropuncture and cannulation from the rete testis, efferent ducts and ductus epididymidis of the rat to determine the composition of efferent duct fluids and the rates of reabsorption of water and solutes by the efferent ducts. The concentration of spermatozoa increased by a factor of about 25 from 2.42 x 10(4) microliters-1 in the fluid from the rete testis to 6.00 x 10(5) microliters-1 in fluid at the end of the efferent ducts, indicating that 96.2% of the fluid leaving the testis is reabsorbed from the lumen of the efferent ducts. Most of this reabsorption (70.9% or 33.4 microliters h-1) occurs in the region between the rete testis and the middle of the coni vasculosi, with only 25.1% (11.8 microliters h-1) occurring between the coni and the beginning of the ductus epididymidis. However, reabsorption across the epithelium occurs at about the same rate in both regions, with the proximal region reabsorbing 17.2 microliters cm-2 h-1 (70.9% of fluid entering the region) and the distal region reabsorbing 12.2 microliters cm-2 h-1 (86.1% of fluid entering the region). Consequently, the fluid reabsorption rate for the whole efferent duct system (15.6 microliters cm-2 h-1) is similar to the values for individual regions. The principal solutes in luminal fluids from the efferent ducts are Na+ (137-144 mM) and Cl- (113-130 mM). The estimated sum contribution of Na+, Cl- and K+ to the osmotic pressure of luminal fluids was approximately 80% at each site sampled in the efferent ducts. The osmotic pressure of luminal fluid samples (301-307 mosmol kg-1) did not vary significantly along the ducts or differ significantly from that of blood plasma. The results demonstrate that there is a net reabsorption in the efferent ducts of nearly all the testicular output of water and inorganic electrolytes, and most of the protein, and that, in comparison, the ductus epididymidis is a negligible site of net fluid reabsorption. The results indicate that the ductus epididymidis, rather than the efferent ducts, is the site of accumulation of high concentrations of specific organic compounds like inositol. The efferent ducts are similar to the homologous proximal tubules of the metanephric kidney in that the luminal electrolyte composition (principal solutes Na+ and Cl-) and osmotic pressure remain relatively stable and that fluid reabsorption is close to isotonic and occurs at the same rate as the reabsorption of Na+.

Absorption↗
PubMed1990

A murine fer testis-specific transcript (ferT) encodes a truncated Fer protein.

A cDNA for a potential tyrosine kinase-encoding mRNA was isolated from a mouse testis cDNA library. In a survey of eight mouse tissues, a transcript of 2.4 kilobases restricted to testis tissue was found. The mRNA encodes a 453-amino-acid protein of 51,383 daltons, the smallest tyrosine kinase protein ever described. RNA synthesized from the cDNA template directs the synthesis of a 51,000-Mr protein in a cell-free translation system. The carboxy-terminal 409 amino acids are 98 and 90% identical to the carboxy halves of the rat and human Fer proteins, respectively. This suggests that the cDNA represents an alternatively spliced testis-specific fer mRNA and is therefore termed by us ferT. On the basis of the appearance time of the fer mRNA in the testis of maturing neonatal mice, we speculate on the role played by this protein in the development of this organ.

Amino Acid Sequence↗