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PubMed2002

Value of testis biopsy in the diagnosis of systemic amyloidosis.

OBJECTIVES: To evaluate testis biopsy as a diagnostic tool in systemic amyloidosis, to assess how amyloidosis affects testicular tissue, and to examine the relationship between testicular amyloidosis and infertility. METHODS: Testicular biopsies from 72 patients with confirmed systemic amyloidosis were examined for amyloid deposition after Congo red and crystal violet staining. A rectal biopsy was also done in each case, and the results were compared with the testicular biopsy findings. RESULTS: Testicular amyloid deposition was detected in 62 (86.1%) of 72 patients. Fifty-one (85%) of 60 patients with secondary amyloidosis, 11 (91.7%) of 12 patients with primary amyloidosis, and 28 (87.5%) of 32 patients with familial Mediterranean fever showed amyloid deposition in the testis. Rectal biopsies were positive in 40 cases (55.6%). Only 4 of the 62 patients with testicular amyloid showed normal spermatogenesis. The remaining 58 exhibited abnormal spermatogenesis, and 77.7% of patients had seconder infertility. Of 62 patients with positive testis biopsies, 30 had serum creatinine levels less than 1.5 mg/dL, and 29 patients with testicular amyloid manifested nephrotic syndrome. CONCLUSIONS: The testis biopsy is a valuable and more sensitive method than rectal biopsy for diagnosing systemic amyloidosis. The results also showed that testicular amyloidosis causes infertility at a higher rate than expected.

Adult↗
PubMed2002

Primary testicular amyloidosis mimicking tumor in a cryptorchid testis.

We present a case of primary testicular amyloidosis mimicking tumor in a cryptorchid testis. Examination revealed the absence of a palpable testis in the left hemiscrotum. The laboratory evaluation showed azoospermia and a low testosterone level. The radiologic investigation demonstrated a heterogeneous left inguinal mass suspicious for tumor in a cryptorchid testis. After radical orchiectomy, the histologic evaluation revealed primary amyloidosis of the testis. The pathogenesis and clinical presentation of testicular amyloidosis are reviewed.

Amyloidosis↗
PubMed1996

Testicular torsion with contralateral vanishing testis.

An infant with a left hydrocele and nonpalpable right testis had an antenatal ultrasonogram showing a left hydrocele and normal right testis. When left inguinal hydrocelectomy was performed, extravaginal torsion of the spermatic cord and a normal testis was discovered. The right spermatic cord ended in a testicular remnant. We propose that this was a case of bilateral antenatal torsion that would have resulted in bilateral vanishing testis syndrome had the left hydrocele not prevented vascular compromise when antenatal testicular torsion occurred. This case gives visible support to the theory that testicular remnants are the result of antenatal torsion.

Humans↗
PubMed1999

Yield and efficacy of biopty gun testis needle biopsy.

OBJECTIVES: To assess whether a preliminary skin incision enhances diagnostic yield of percutaneous testis biopsy and to further evaluate the clinical efficacy of this procedure. METHODS: A total of 45 men (67 testes) underwent testicular biopsy with two passes of a Biopty gun spring-loaded needle. Twenty-seven biopsies were performed without a preliminary skin incision (group 1), and 40 were performed after a small scrotal incision (group 2). In 56 testes, needle biopsy histopathologic diagnosis was compared with that of open biopsy or orchiectomy specimens from the same patient. Needle and surgical specimens were fixed in Bouin's solution and sent separately for independent, blinded, histologic interpretation. RESULTS: Complications of the procedure were negligible. In all 67 needle biopsies, specimen quality was adequate for histopathologic interpretation. The mean number of seminiferous tubules obtained from needle biopsy was 28% higher among patients having a preliminary skin incision (25.9) compared with those without (18.7, P = 0.023). Correlation between needle and open histopathologic diagnosis was excellent (55 of 56, 98%). CONCLUSIONS: A preliminary skin incision made before needle biopsy increases the diagnostic yield of percutaneous testis biopsy. Percutaneous testis biopsy using the Biopty gun needle provides equal diagnostic information when compared with open testis biopsy or orchiectomy specimens. The concomitant reduction in morbidity and cost make this an attractive diagnostic procedure.

Biopsy, Needle↗
PubMed1999

Oxytocin gene expression and action in goat testis.

Gene expression, immunohistochemical localization, binding and effects of oxytocin (OT) on androgen production in the testis of adult goats were studied. Using polymerase chain reaction (PCR) analysis we were able to detect OT gene transcripts in the goat testis. Immunohistochemistry revealed that both OT and neurophysin epitopes were expressed together in the intratubular regions, especially in the Sertoli cells, suggesting the production of OT in these cells. However, enzyme immunoassay found no difference in OT concentration between testicular arterial and venous plasma. Saturable, specific [3H]-OT binding sites were present in membrane fractions of the goat testis. Scatchard analysis indicated an apparent affinity of 42 +/- 7 L/nmol and binding capacity of 24 +/- 4 fmol OT bound/mg DNA. In vitro treatment of goat testes for 6 h with 100 nM OT led to a 3.5-fold increase (P < 0.001) in 5 alpha-dihydrotestosterone (DHT) production in spite of the induced decrease in testosterone, suggesting that OT not only affects testosterone production but also modulates its conversion to DHT. These results indicate that the goat testis produces OT, which may be involved in the local control of androgen biosynthesis.

Animals↗
PubMed1997

Identification and characterization of inositol 1,4,5-trisphosphate receptors in rat testis.

PCR analysis and immunoblotting with isoform specific antibodies was used to identify the presence of type I, II and III inositol 1,4,5-trisphosphate receptors (InsP3Rs) in rat testis. PCR analysis also revealed that rat testis express both forms of the S1 splice variant (S1+ and S1-), but only the S2- from of the S2 splice variant of the type I InsP3 receptor. PCR analysis was also used to identify InsP3R isoform expression at a cellular level using myoid, Sertoli and germ cells derived from the testis of Wistar rats. The extent of [3H]-InsP3 binding was found to be 9 times lower for testicular microsomes than for cerebellar microsomes, with a Bmax of 1.4 pmoles/mg protein compared to 12.5 pmoles/mg protein for cerebellar microsomes. The Kd for InsP3 binding to its receptor in testicular microsomes was 60 +/- 10 nM which was similar to that found for cerebellar microsomes (80 +/- 20 nM). InsP3-induced Ca2+ release (IICR) in testicular microsomes was found to have an EC50 (concentration which causes a half-maximal response) of 0.5 +/- 0.03 microM, also similar to that seen for cerebellar microsomes (0.3 microM). Maximal IICR occurred at about 20 microM InsP3, with up to 4% of total intracellular Ca2+ stores being mobilized as compared to between 10-30% for cerebellar microsomes. Time resolved IICR using stopped-flow spectrofluorimetry, showed the kinetics of IICR for this testis preparation to be monophasic with a maximum rate constant of 0.15 s-1 at 30 microM InsP3. The rate constants are 7 times slower than values for cerebellar microsomes under similar conditions (approximately 1 s-1) and taken together with the binding data support the proposal that the receptor density/Ca2+ store is approximately 8 times lower than seen in cerebellar microsomal vesicles. The pharmacological properties as assessed using heparin and InsP3 analogues also confirmed similar behaviour for testicular InsP3Rs and cerebellar InsP3Rs.

Animals↗
PubMed2000

Structure of the 5' region of the human hexokinase type I (HKI) gene and identification of an additional testis-specific HKI mRNA.

We previously reported the structure of the human hexokinase type I (HKI) gene and provided direct evidence of an alternative red blood cell-specific exon 1 located in the 5' flanking region of the gene. Three unique HKI mRNA species have also been described in human spermatogenic cells. These mRNAs contain a testis-specific sequence not present in somatic cell HKI, but lack the sequence for the porin-binding domain necessary for HKI to bind to porin on the outer mitochondrial membrane. The present study reports a new mRNA isoform, hHKI-td, isolated from human sperm. hHKI-td mRNA contains both a testis-specific sequence at the 5' end common to the three other mRNA isoforms and an additional unique sequence. Screening of a cosmid library and analysis of the cosmids containing the HKI gene revealed that testis-specific sequences are encoded by six different exons. Five of these exons are located upstream from the somatic exon 1 (5.6-30 kb) and one within intron 1. This study shows that a single human HKI gene spanning at least 100 kb encodes multiple transcripts that are generated by alternative splicing of different 5' exons. Testis-specific transcripts are probably produced by a separate promoter that induces the expression of the HKI gene in spermatogenic cells.

Amino Acid Sequence↗
PubMed2003

Sublethal effects of diazinon on the structure of the testis of bluegill, Lepomis macrochirus: a microscopic analysis.

A histopathological analysis of the testis exposed to diazinon, an organophosphorus pesticide was conducted. The testis from the control fish exhibited a normal histological structure with a round, oval or somewhat rectangular shaped with closely associated seminiferous tubules. The seminiferous tubules were surrounded by a layer of connective tissue with myoid cells, Leydig cells, blood vessels, lymph vessels and nerves. Upon exposure for 24 h, the overall structure of the testis looked disrupted and more often the sperm cells appeared to be somewhat larger, but spermatid were smaller than that of the control group. The 48 h exposure caused more dense and closely associated seminiferous tubules. The 72 h exposure induced dilation of the seminiferous tubules and their lumen. Ninety-six hours exposure induced more loose and disrupted structures. In this group (96 h), there was a significant reduction in both lumen and seminiferous tubule size in comparison with the control, 24, 48, and 72 h groups. After 2 weeks of exposure hardly any lumen was seen, which imparted more dense and compacted structures. The biometric analysis gave a highly significant increase in the diameter of the seminiferous lumen. After 48 h of exposure a significant reduction was seen in diameter in comparison with that of the control group. Seventy-two hour exposure indicated an increase in diameter, whereas 96 h exhibited another decrease. One week caused the diameter to increase whereas 2 weeks of exposure caused them to decrease again. The increase in diameter of the seminiferous tabule may be caused by weakening of the surrounding connective tissue and myoid cells, due to exposure to diazinon. The change in the diameters of the seminiferous tabules was very irregular. No apparent co-relation was found between the size of the fish, body weight and weight of the testis after they have been subjected to different exposure periods. There was significant change in the germ cells' diameters. As the germ cells are the essential first step in the process of spermatogenesis, their size reduction may hinder the production of viable spermatozoa, endangering the population dynamics. In order to find out the pathways of this pesticide effects, we propose to study the histopathology of pituitary gland a vital organ of the reproductive axis.

Animals↗
PubMed2003

Alteration of lactate production and transport in the adult rat testis exposed in utero to flutamide.

Although it is established that in utero exposure to the antiandrogen flutamide induces alteration of spermatogenesis in the adult rat testis offspring, the cellular and molecular mechanisms involved in such an effect remain to be investigated. In the present paper, by using as model adult rats exposed in utero to flutamide (0, 2, 10 mg/kg per day), we have investigated the hypothesis that germ cell alterations could be related to defects of energy metabolism and particularly to defects of the production and transport of lactate. Lactate is a preferential energy substrate produced by Sertoli cells and transported to germ cells by monocarboxylate transporters (MCT). A significant decrease (60%, P<0.001) in lactate production was observed in cultured Sertoli cells from rat testes exposed in utero to flutamide from the dose of 2 mg/kg per day. Such a decrease is concurrent to a decrease in lactate dehydrogenase A (LDHA) mRNA levels (evaluated through semiquantitative RT-PCR) and LDHA4 activity. The decrease in LDHA mRNA levels (to 64 +/- 9% of the control, P<0.05) was observed with the lowest dose (2 mg/kg per day) of flutamide tested. The decrease in LDHA mRNA levels was observed in both the whole testis and in isolated Sertoli cells, suggesting that such a decrease in LDHA expression occurred also in the (Sertoli) cells producing lactate. Lactate is transported from Sertoli cells to germ cells via MCT1 and MCT2. We immunolocalized MCT1 to all the different germ cell types and MCT2 exclusively to elongated spermatids. In the adult testis exposed in utero to flutamide, MCT1 (53 +/- 8%, P<0.02) and MCT2 (52 +/- 9%, P<0.02) mRNA levels were significantly reduced indicating that lactate transport to germ cells could be also altered. Together, these data support (i) the existence of a relationship between the antiandrogen activity and the energy metabolism in the testis and (ii) the concept of an androgen-dependent programming, occurring early in the fetal life in relation to the expression of some of the key genes involved in the production and transport of lactate in the seminiferous tubules.

Androgen Antagonists↗
PubMed1997

Isolation and comparative studies of mitochondrial F1-ATPase from rat testis and beef heart.

The isolation and properties of F1-mitochondrial ATPase from rat testis are described. The isolation medium involves a chloroform extraction, and it is suitable even with small amounts of starting material that have a relatively low specific activity as in the case of rat testis submitochondrial particles. The isolated enzyme from rat testis had a specific activity of 30-45 mumol Pi/min/mg protein, which could be increased up to 90 mumol Pi/min/mg protein only in the presence of bicarbonate and maleate. The isolated enzyme represented less than 0.6% of the initial membrane proteins. It exhibited a typical five-band pattern in sodium dodecyl sulfate gel electrophoresis. However, it showed a ratio of subunits alpha:beta higher than the heart enzyme; its significance is unknown. The purified enzyme was cold labile and inhibited by natural ATPase inhibitor protein from bovine heart mitochondria and by dicyclohexylcarbodiimide. The results presented suggest that the low ATPase activity of testis submitochondrial particles is due to a reduced content of the F1-ATPase.

Adenosine Triphosphate↗
PubMed2000

Foreign gene expression by in vivo gene electroporation in the quail testis.

To investigate whether or not foreign gene expression is attained in the testis of living Japanese quails, a firefly luciferase reporter gene was transfected by in vivo electroporation (EP), and transcriptional activity of different promoters was compared. In addition, the effect of the Epstein-Barr virus self-replication sequence was also tested. The results showed that luciferase activity in the testis reached almost a plateau value at 50 V. Under this EP condition, no difference was found in transcriptional activity between the simian virus 40 (SV40) and miw promoters. The reporter gene expression in the quail testis was observed over 28 days after in vivo gene EP, although the activity gradually decreased, and the presence of the Epstein-Barr virus self-replication sequence in the SV40 promoter did not significantly prolong the luciferase activity. These results suggest that in vivo gene EP confers strong, though transient, foreign gene expression in the Japanese quail, and it may provide a new powerful approach for studies on transcriptional regulation of genes during proliferation and differentiation of spermatogenic cells in the quail testis.

Animals↗
PubMed2000

Two isoforms of FTZ-F1 messenger RNA: molecular cloning and their expression in the frog testis.

FTZ-F1, a member of the orphan nuclear receptors, is a transcriptional factor regulating the expression of the fushi tarazu gene in Drosophila (Lavorgna et al., 1991. Science 252, 848-851). Previously, we cloned a frog homologue of FTZ-F1 (rrFTZ-F1alpha; GenBank Accession No. AB035498). In this study, we isolated the rrFTZ-F1beta cDNA encoding a protein of 469 amino acids. Then, expressions of two types (alpha and beta) of rrFTZ-F1 mRNAs were examined during development of embryos and gonads in the frog Rana rugosa. They were expressed in the embryo at stage 12. Expressions of both the alpha and beta mRNAs became stronger in the testis of frogs at stage XXV and were most prominent in that of frogs 2months after metamorphosis. In the former testis, spermatogonia were the only germ cells in the seminiferous tubules, whilst postmeiotic cells were observed in the latter testis. Expression of the typealpha mRNA was more prominent. In addition, we cloned the regions with either exon I or II of the rrFTZ-F1 gene. Genomic structure analysis revealed that rrFTZ-F1beta is a partial exon I-truncated variant of rrFTZ-F1alpha. The results suggest that rrFTZ-F1alpha and -beta are expressed from the same gene by alternative splicing and that they may play an important role(s) in differentiation of premeiotic germ cells in the testis of the frog R. rugosa.

Alternative Splicing↗
PubMed2000

Molecular cloning and characterization of a protein tyrosine phosphatase enriched in testis, a putative murine homologue of human PTPMEG.

Protein tyrosine phosphorylation is regulated by protein tyrosine kinase and protein tyrosine phosphatase activities. These two counteracting proteins are implicated in cell growth and transformation. Using polymerase chain reaction with degenerate primers, we have identified a novel mouse protein tyrosine phosphatase (PTP). This cDNA contains a single open reading frame of the predicted 926 amino acids. Those predicted amino acids showed significant identity with human megakaryocyte protein-tyrosine phosphatase by 91% in nucleotide sequences and 94% in amino acid sequences. We have identified that expression of this PTP is highly enriched in the testis in mouse and human and has been termed here as a 'testis-enriched phosphatase' (TEP). Northern analysis detected two mRNA species of 3.7 and 3.2kb for this PTP in mouse testis and the expression of TEP is regulated during development. The recombinant phosphatase domain possesses protein tyrosine phosphatase activity when expressed in Escherichia coli. Immunohistochemical analysis of the cellular localization of TEP on mouse testis sections showed that this PTP is specifically expressed in spermatocytes and spermatids within seminiferous tubules, suggesting an important role in spermatogenesis.

Amino Acid Sequence↗
PubMed1996

Ubiquitous expression and testis-specific alternative polyadenylation of mRNA for the human Ran GTPase activator RanGAP1.

RanGAP1 is the activator of the Ras-related nuclear GTPase Ran, which is involved in the nucleo-cytoplasmic transport of both, proteins and mRNAs, and also in cell cycle regulation. Here, we report a 2970-bp cDNA clone of RanGAP1 isolated from a HeLa lambda gt11 cDNA library. It contains a 215-bp 5' untranslated region (UTR) with a G + C-content of 68%, followed by a 1764-bp open reading frame and a 989-bp 3' UTR preceding a 77-bp poly(A)+ tail. RanGAP1 shows differential patterns of expression in human tissues. In addition to the 3.5-kb transcript present in all tissues and highly expressed in brain, thymus and testis, we found a second transcript of 2.8 kb in testis. In order to analyze this shorter transcript, we screened a human testis lambda gt10 cDNA library and cloned an alternatively polyadenylated RanGAP1 transcript. Taking the 3' UTR of RanGAP1, which lies downstream of the first polyadenylation signal, as a probe in Northern blot analysis, we confirmed that this second transcript found in testis results from a distinct 3' UTR.

Amino Acid Sequence↗
PubMed2001

Hormones and testis development and the possible adverse effects of environmental chemicals.

Development of a fetus into a phenotypic male depends, first, on testis formation and second, on hormone production by the fetal testis. Disorders of testicular hormone production or action can lead in severe cases to phenotypic abnormalities or can predispose towards impaired reproductive health. Evidence for deteriorating human male reproductive health, especially an increase in testicular cancer, points to disturbed (hormonal) development of the fetal testis. By comparison of testicular dysgenesis in humans and exposure to certain phthalates in fetal rats, the similarities in outcomes and testicular cell-cell disruption are highlighted as are the pathways via which oestrogenic and (especially) anti-androgenic environmental chemicals might act to induce such changes. The susceptibility of sperm production in adulthood to 'hormonal' disruption in fetal and neonatal life is also discussed. Though it is concluded that no direct evidence links human exposure to environmental chemicals and male reproductive disorders that stem from disturbed testis development, this is based mainly on lack of information. Using the example of phthalates, for which new data have emerged, it is argued that until the appropriate in vivo studies are undertaken, the safety of hormonally active environmental chemicals, especially in mixtures, will continue to give cause for concern as far as testicular development is concerned.

Animals↗
PubMed2002

Defining the cellular and molecular mechanisms of toxicant action in the testis.

A symposium was held at the 41st annual meeting of the Society of Toxicology with presentations that emphasized novel molecular and cellular pathways that modulate the response to testicular toxicants. The first two presentations described cellular alterations after exposure to the Sertoli cell toxicant, mono-(2-ethylhexyl) phthalate (MEHP). The expression of flamingo1, a G protei coupled receptor family member that may couple cell-cell adhesion to G protein-dependent signaling in Sertoli cells, was rapidly altered after MEHP exposure. Sertoli cell associated flamingo1 immunostaining was redistributed early (within 2 h) after MEHP exposure and disappeared by 12 h, suggesting that flamingo1 is a proximal phthalate target. MEHP was also described to alter the expression and activity of the newly identified death receptors DR4, 5 and 6 in the testis. The differential cellular changes in the levels of DR4, 5 and 6 after phthalate exposure suggested that they may act as surrogates or in concert with the widely described Fas-signaling pathway in the initiation of germ cell apoptosis after MEHP exposure. The next two presentations focused on revealing mechanisms that may explain the persistent post-exposure testicular atrophy that is observed in rodents after a variety of chemical or physical insults (radiation, chemotherapeutics, toxicants) and possible strategies to re-initiate spermatogenesis in the atrophic testis. Hormonal manipulations that lower testosterone and serum FSH levels allow for re-initiation of spermatogonial development. Recent investigation of additional models of persistent atrophy such as mutant mice, the aged Brown Norway rat, EDS-induced Leydig cell deficient rat, and primates, have broadened insight into the mechanisms responsible for persistent atrophy. The last presentation described the use of cDNA arrays in the investigation of cellular elements and mechanisms responsible for disruption of spermatogenesis by the drinking water disinfectant bromochloroacetic acid (BCA). A custom mouse testis cDNA array interrogating 950 genes was used for analysis of testis mRNA. BCA altered the expression of 53 of the 950 genes, including two encoding sperm proteins known to be significant for male fertility, and other genes involved in spermatogenesis, stress response, and cell communication/adhesion. These observations strengthen the hypothesis that BCA disrupts spermatogenesis by altering the process of spermiogenesis.

Animals↗
PubMed2000

Testis developments and puberty in the male Tokara (Japanese native) goat.

In male Tokara (Japanese native) goats, testis development and the onset of puberty were studied histologically and observing sexual behaviour. Testes weight increased from 36+/-5.4 (S.E.) g (n=5) at 3 months of age to 126+/-6.3 g (n=6) at 12 months. The degree of the testis development differed among kids at 3 months of age and only one of four had produced spermatozoa in the testis. Large number of spermatozoa were always present in seminiferous tubules and epididymal ducts from 4 months of age. The diameter of seminiferous tubules increased from 133+/-9.9 microm (n=4) at 3 months to 198+/-1.0 microm (n=3) at 6 months with little increase thereafter. Mounting and pelvic thrusts onto a teaser doe started at from 9 to 14 weeks of age. Ejaculated semen contained sperm with good motility for the first time from 17 weeks. The male Tokara goat reaches puberty at around 4 months of age but testis development continues to 12 months.

Age Factors↗
PubMedDate not supplied

The effect of zinc supplementation on the effects of lead on the rat testis.

With increasing concerns about environmental pollution, the interaction of micronutrients with toxic metals is of great interest. The present study was designed to investigate testicular effects of lead following concomitant administration of zinc. Lead was administered orally as lead acetate (50 mg/kg b.w.) daily for 3 months to male Portan rats with or without zinc (1 mg/kg b.w. as zinc sulphate). The control group was given the same volume of distilled water. Endpoints included lead concentration in various body organs as well as in the reproductive system, including testicular subfractions; the testicular enzymes superoxide dismutase (SOD) and catalase; the marker enzyme delta-aminolevulinic acid dehydratase (delta-ALAD); and testicular histoarchitecture. The concentrations of lead in bone, kidney, prostate, testis, liver, epididymis, spleen, seminal vesicles, and blood were significantly higher in lead-treated rats. Lead deposition was reduced in animals that received supplemental zinc. There was a 30% reduction in lead deposition in the testis when zinc was coadministered. At the subcellular level, there was differential accumulation of lead; the nucleus preferentially took up the metal after lead treatment alone, while zinc coadministration shifted lead accumulation to the mitochondria. A significant decrease in delta-ALAD and in SOD activity was seen in the testis with lead treatment. Coadministration of zinc prevented these decreases, at least partially. Zinc coadministration did not prevent the inhibition of catalase observed with lead treatment. Histologically, the alterations in the testis with lead treatment alone were more pronounced compared to animals in which zinc was supplemented. Improvement in the inhibition of delta-ALAD and in the ubiquitous cellular enzyme SOD suggests less testicular tissue damage due to detoxification of free radicals. In conclusion, zinc supplementation ameliorates lead-induced testicular damage both at the cellular and subcellular level. The protective effect may be due to differential distribution of lead, either because of competition between lead and zinc or displacement of lead by zinc.

Animals↗