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PubMed1991

A transferrinlike (hemiferrin) mRNA is expressed in the germ cells of rat testis.

In the testis, germ cells which are separated from the serum by the blood-testis barrier rely primarily on the Sertoli cell to obtain nutrients. For example, transferrin synthesized by the Sertoli cell is important in delivering iron from the serum to the developing germ cells. Because of its role in the testis, Sertoli cell transferrin protein and mRNA have been extensively studied. By using RNA blot analysis of rat testicular tissue, we detected a transcript of 2.6 kb which is attributed to transferrin. In addition, we detected a novel mRNA of 0.9 kb which had sequence similarity to the 3' end of transferrin. This 0.9-kb mRNA was present in germ cells, but not Sertoli cells, liver, or brain. The primary source of this mRNA in the testis was round spermatids. Sequence analysis of a cDNA clone showed that this mRNA encoded a protein with sequence similarity to the carboxy terminus of transferrin. Polysome analysis indicated that this transcript was translated and may therefore have importance in the iron metabolism of germ cells. The evolutionary implications between the transferrinlike mRNA germ cells and the gene duplication event which resulted in the diferric binding of transferrin are discussed.

Amino Acid Sequence↗
PubMed1993

A testis-specific gene encoding a nuclear high-mobility-group box protein located in elongating spermatids.

A cDNA encoding a DNA-binding protein has been isolated by screening a mouse testicular expression cDNA library with a concatemer of a 12-bp putative protein-binding element present in the promoter of the testis-specific gene PGK-2. Sequence analysis of the isolated cDNA indicated the presence of an open reading frame that encodes a protein with two conserved DNA-binding motifs known as the high-mobility-group (HMG) boxes. Northern (RNA) blot analysis demonstrated that expression of the gene is restricted to the postpuberal testis. The DNA-binding activity and sequence specificity of the recombinant HMG protein were confirmed by DNA mobility shift assay using the initial concatemer of the PGK-2 promoter element as a probe as well as the wild-type or mutated versions of the 12-bp element within its natural sequence context. Immunocytochemical staining of adult testis sections with polyclonal antisera recognizing this recombinant HMG protein demonstrated that it is located predominantly in the nuclei of elongated spermatids at steps 9 and 10. These results suggest that this novel HMG box protein gene may be involved in the regulation of gene expression of the haploid male genome. The gene from which the cDNA was derived has been termed testis-specific HMG (tsHMG).

Amino Acid Sequence↗
PubMed1994

The Drosophila micropia retrotransposon encodes a testis-specific antisense RNA complementary to reverse transcriptase.

The micropia transposable element of Drosophila hydei is a long terminal repeat-containing retrotransposon present in both the autosomes and the Y chromosome. micropia expression gives rise to a complex set of sense and antisense RNAs transcribed primarily during spermatogenesis. The most abundant sense RNAs constitute an assortment of heterogeneous high-molecular-weight transcripts expressed as constituents of the Y-chromosomal lampbrush loops of primary spermatocytes. In addition, micropia encodes a full-length RNA that extends between the two long terminal repeats of the element. The major 1.0-kb antisense RNA characterized is complementary to the reverse transcriptase and RNase H coding regions of micropia. It is expressed from a testis-specific promoter during the primary spermatocyte stages and is detectable until spermatid elongation stages. Sequence comparison of this promoter with the 5' region of other testis-specific genes allows the conception of a conserved sequence that is responsible for this pattern of expression. A 284-bp fragment containing this sequence is able to drive testis-specific expression of the Escherichia coli lacZ gene in Drosophila melanogaster. This sequence is conserved in the micropia elements present in other Drosophila species that also encode an antisense RNA. The evolutionary conservation of micropia antisense RNA expression and the sequences responsible for its testis-specific transcription suggests a role for this antisense RNA in the control of germ line expression of the full-length transcript or transposon-encoded proteins.

Animals↗
PubMed1986

Six mouse alpha-tubulin mRNAs encode five distinct isotypes: testis-specific expression of two sister genes.

Five mouse alpha-tubulin isotypes are described, each distinguished by the presence of unique amino acid substitutions within the coding region. Most, though not all of these isotype-specific amino acids, are clustered at the carboxy terminus. One of the alpha-tubulin isotypes described is expressed exclusively in testis and is encoded by two closely related genes (M alpha 3 and M alpha 7) which have homologous 3' untranslated regions but which differ at multiple third codon positions and in their 5' untranslated regions. We show that a subfamily of alpha-tubulin genes encoding the same testis-specific isotype also exists in humans. Thus, we conclude that the duplication event leading to a pair of genes encoding a testis-specific alpha-tubulin isotype predated the mammalian radiation, and both members of the duplicated sequence have been maintained since species divergence. A second alpha-tubulin gene, M alpha 6, is expressed ubiquitously at a low level, whereas a third gene, M alpha 4, is unique in that it does not encode a carboxy-terminal tyrosine residue. This gene yields two transcripts: a 1.8-kilobase (kb) mRNA that is abundant in muscle and a 2.4-kb mRNA that is abundant in testis. Whereas the 1.8-kb mRNA encodes a distinct alpha-tubulin isotype, the 2.4-kb mRNA is defective in that the methionine residue required for translational initiation is missing. Patterns of developmental expression of the various alpha-tubulin isotypes are presented. Our data support the view that individual tubulin isotypes are capable of conferring functional specificity on different kinds of microtubules.

Amino Acid Sequence↗
PubMed1989

Translational control of germ cell-expressed mRNA imposed by alternative splicing: opioid peptide gene expression in rat testis.

The three genes encoding the opioid peptide precursors (prodynorphin, proenkephalin, and proopiomelanocortin) are expressed in the rat testis. The sizes of the three opioid mRNAs in the testis differ from the sizes of the corresponding mRNAs in other rat tissues in which these genes are expressed. The smaller testicular proopiomelanocortin mRNA has previously been demonstrated to arise from alternative transcriptional initiation. In the present study, we found that the smaller testicular prodynorphin mRNA, expressed in Sertoli cells, results from alternative mRNA processing. Exon 2, which makes up 5' untranslated sequence, is removed from the mature transcript. Polysome analysis of brain and testis RNA indicates that the alteration of the prodynorphin leader sequence in the testis-specific transcript does not affect the efficiency of translation of this mRNA. The larger testicular proenkephalin transcript, expressed in developing germ cells, also results from alternative mRNA processing. Alternative acceptor site usage in the splicing of intron A results in a germ cell-specific proenkephalin transcript with a 491-nucleotide 5' untranslated leader sequence preceding the preproenkephalin-coding sequence. Polysome analysis indicates that this germ cell-specific proenkephalin mRNA is not efficiently translated. Mechanisms by which alternative mRNA splicing may serve to confer translational regulation upon the testicular proenkephalin transcript are discussed.

Animals↗
PubMed1999

A lesson in the management of testicular cancer in a patient with a solitary testis.

Five per cent of patients with germ cell tumours of the testis will develop a further tumour in the contralateral testis. Standard treatment in such cases is a second orchidectomy, resulting in infertility, hormone replacement, and psychological morbidity. In this case report we explore the role of testis conservation in these patients and also show that there is a risk of removing a potentially normal testis if a histological diagnosis is not sought prior to orchidectomy.

Adult↗
PubMed1997

Normal pediatric testis: comparison of power Doppler and color Doppler US in the detection of blood flow.

PURPOSE: To compare power Doppler and conventional color Doppler ultrasound (US) in the detection of blood flow in the normal pediatric testis and to assess the symmetry of blood flow and the spectral Doppler tracing waveforms in the normal pediatric testis. MATERIALS AND METHODS: Testicular blood flow was assessed prospectively in 68 testes in 34 boys (age range, 6 weeks to 13 years; mean age, 4.6 years) with both conventional color and power Doppler US. Intratesticular blood flow was graded as follows: 0, no intratesticular flow; 1, single intratesticular Doppler signal identified; and 2, multiple intratesticular Doppler signals identified. The symmetry of intratesticular flow was assessed both subjectively and objectively by using the same grading system. Spectral Doppler tracings were obtained in 62 testes in 31 patients. RESULTS: Power Doppler US demonstrated intratesticular blood flow in 66 (97%) testes. Color Doppler US demonstrated intratesticular blood flow in 60 (88%) testes. Combined techniques depicted blood flow in all 68 (100%) testes. Testicular blood flow was judged symmetric in all 34 (100%) patients with power Doppler US and in 31 (91%) patients with color Doppler US. Spectral Doppler tracings demonstrated absence of diastolic flow in 20 (32%) of 62 testes. CONCLUSION: In children, power Doppler US is more sensitive than color Doppler US in the detection of intratesticular blood flow. With power Doppler US, testicular blood flow in healthy children is symmetric, underscoring that the asymptomatic testis can be used as a baseline for assessing flow in the symptomatic testis.

Child, Preschool↗
PubMed2002

Coordinated expression of UT-A and UT-B urea transporters in rat testis.

The blood-seminiferous tubule barrier is responsible for maintaining the unique microenvironment conducive to spermatogenesis. A key feature of the blood-testis barrier is selective permeability to solutes and water transport, conferred by the Sertoli cells of the seminiferous tubules (SMTs). Movement of fluid into the lumen of the seminiferous tubule is crucial to spermatogenesis. By Northern analysis, we have shown that 4.0-, 3.3-, 2.8-, and ~1.7-kb UT-A mRNA transcripts and a 3.8-kb UT-B mRNA transcript are detected within rat testis. Western analysis revealed the expression of both characterized and novel UT-A and UT-B proteins within the testis. Immunolocalization studies determined that UT-A and UT-B protein expression are coordinated with the developmental stage of the SMT. UT-A proteins were detected in Sertoli cell nuclei at all stages of tubule development and in residual bodies of stage VIII tubules. UT-B protein was expressed on Sertoli cell membranes of stage II-III tubules. Using in vitro perfusion, we determined that a phloretin-inhibitable urea pathway exists across the SMTs of rat testis and conclude that UT-B is likely to participate in this pathway.

Animals↗
PubMed1999

Fas regulates germ cell apoptosis in the human testis in vitro.

The Fas-Fas ligand (FasL) system has been implicated in maintaining the immune privileged nature of the testis. The present report concerns the role of the Fas-FasL system in regulating germ cell apoptosis, another important function of this system in the human testis. Fas was localized immunohistochemically to the same types of germ cells that were identified as apoptotic, namely spermatocytes and spermatids. Strong expression of Fas was also observed in Western blot analysis of the human testis. Furthermore, an antagonistic antibody to the FasL blocked germ cell apoptosis induced in vitro by incubating segments of seminiferous tubules under serum- and hormone-free conditions (i.e., without survival factors). Thus Fas appears to mediate germ cell apoptosis. A universal caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone, also inhibited germ cell death, suggesting that Fas-associated germ cell apoptosis is mediated via the caspase pathway. The present results suggest an important role for the Fas-FasL system in the regulation of germ cell apoptosis in the human testis.

Aged↗
PubMed1998

Biomechanical properties of the internal spermatic vein in the normal population and patients with left-sided varicocele testis.

OBJECTIVE: The aim of the study was to investigate a possible decreased strength of the wall of the left internal spermatic vein between patients with varicocele testis and controls. MATERIALS: From 14 patients with varicocele testis, 2 cm of the vein was obtained during operation per varicocele and compared to samples of the left internal spermatic vein taken from the same anatomical localization of 19 patients with no varicocele testis or other conditions with relation to the scrotum; additionally, samples from the right internal spermatic vein similar to those described above were taken from 12 patients. The biomechanical properties of ring-shaped venous specimens were investigated by loading the specimens at a constant deformation rate until rupture. MAIN RESULTS: The regression coefficient, standard error and p value revealed no significant differences in ID (diameter), UC (unit collagen/ID), E-max (ultimate extensibility), Max (maximum strength), Tan-a (maximum stiffness), E-fail (relative failure energy). The trend to significance was shown in ID between the varicocele and left-sided veins (p = 0.05) and between left- and right-sided veins (p = 0.05). CONCLUSIONS: These biomechanical tests of the spermatic veins from healthy subjects and patients operated for varicocele showed that biomechanical differences were associated with age and could play a part in the development of varicocele testis in an age-matched group.

Adult↗
PubMed2000

Monoclonality of asynchronous bilateral lymphoma of the testis.

OBJECTIVES: Lymphoma is the most frequent testicular malignancy in men over 60 years of age. Even though patients present initially with localized disease, the high incidence of bilateral involvement, synchronous or not, and early systemic dissemination are characteristic of these neoplasms. Sometimes the interval between tumor involvement of both testes is long. The question is raised whether either the patient has a predisposition to present new clones of transformed lymphocytes, or the same disease using the same pathway from a systemic reservoir infiltrates the contralateral testis. METHOD: Polymerase chain reaction and DNA sequencing were used to detect immunoglobulin heavy chain (IgH) rearrangement in paraffin-embedded specimens from asynchronous tumors affecting the right and left testis of a 85-year-old man with an interval period of 13 months. RESULTS: Both tumors showed the same IgH rearrangement. CONCLUSIONS: The lymphoma affecting the left and right testis derived from the same clone. It makes a strong case that lymphoma of the testis is the first manifestation of a systemic disease and should be treated aggressively early at the beginning of the disease.

Aged↗
PubMed2005

In vivo study to evaluate the protective effects of amifostine on radiation-induced damage of testis tissue.

OBJECTIVE: To investigate the early protective effects of amifostine against radiation-induced damage on rat testis tissue. METHODS: Eighty adult male Wistar rats were randomized to 4 groups: Saline solution was given to group A for control, 200 mg/kg amifostine (WR-2721) to group B, a single fraction of 6 Gy local irradiation to testes in group C and 200 mg/kg amifostine 15-30 min before 6 Gy testicular irradiation to group D. Animals were sacrificed 3 weeks after treatment and their testes were removed for macroscopic, microscopic and ultrastructural histopathological examination. RESULTS: The weights, widths and lengths of testes in the last 3 groups had decreased significantly when compared with the control group, but the decrease in widths after irradiation was found to be significantly less only in the amifostine plus radiation group. There was a significant reduction of testis weights in relation to the individual body weights in the irradiated testes compared with the other groups (p < 0.005), while there was no significant change of testis weight/total body weight ratio in amifostine plus irradiation group. Spermatogonium A and primary spermatocyte counts were also less in the treatment groups, and primary spermatocyte numbers were significantly higher in amifostine plus radiation group when compared with radiation alone group (p < 0.005). Pretreatment with amifostine reduced the decrease of primary spermatocyte counts by a factor of 1.28. Electron microscopic analysis did not show any cytotoxic effect of amifostine alone, and furthermore, ultrastructural findings were normal with the addition of amifostine prior to irradiation, though there was damage in the radiation exposure group. CONCLUSION: Amifostine when given alone by itself appears to cause adverse alterations in testis tissue; however, it has a radioprotective effect on spermiogenetic cells when used prior to radiation.

Amifostine↗
PubMed1985

Descent of the testis in the fetal calf. A summary of the anatomy and process.

The descent of the testis in the fetal calf is reviewed, and the role in that process of the swelling reaction of the gubernaculum testis is discussed. The testes of 30 Dutch Friesian fetuses were examined by dissection and light microscopy of sections prepared from chemically and frozen-fixed specimens. The gubernaculum remains unattached to the scrotal fasciae until descent is completed. Shortening of the intra-abdominal gubernaculum and displacement of the testis begins at fetal week 11; the swelling reaction of the gubernaculum occurs between weeks 14 and 15. The testis is at the deep inguinal ring by week 15, and by week 20 it is in the scrotal position and the gubernaculum has regressed. It is proposed that the swelling of the gubernaculum dilates the vaginal ring and enlarges the inguinal canal. The clinical importance of these anatomical relationships and changes is discussed.

Animals↗
PubMed1995

Distribution pattern of F-actin, vimentin and alpha-tubulin in the bovine testis during postnatal development.

The distribution of F-actin, vimentin and alpha-tubulin was studied immunohistochemically in bovine seminiferous and straight testicular tubules, rete testis and intertubular tissue during postnatal development. Sites of antigenicity were detected by ABC immunoperoxidase technique and visualized by metal-enhanced deposition of diaminobenzidine. Within the seminiferous epithelium, F-actin appears at 20 weeks and is found in adult Sertoli cells as part of specialized cell contacts. In peritubular cells, F-actin increases gradually from 4 to 30 weeks when the adult concentration is achieved. After 20 weeks, subepithelial fibroblasts of the mediastinum testis start to express F-actin and at 52 weeks, a thick layer of positive myofibroblasts is seen beneath the epithelia of rete testis and straight testicular tubules. Testicular macrophages and light intercalated cells (LIC) are also characteristically decorated following F-actin immunoreaction. Vimentin is localized in perinuclear position in pre-Sertoli cells of 4-20 weeks and in adult Sertoli cells. During the period of transformation from pre-Sertoli to Sertoli cells, the perinuclear vimentin coat is absent. The epithelia of rete testis and straight tubules exhibit a strong vimentin immunoreaction in their basal parts. This specific pattern does not change from 4 weeks to adulthood. Alpha -tubulin is absent in 4-week-old seminiferous tubules. At 8 weeks, the perinuclear area of pre-Sertoli cells reacts positive. The alpha-tubulin content increases in these cells continuously, and from 30 weeks on nearly the entire supranuclear cytoplasm of Sertoli cells is heavily decorated. The epithelial of rete and straight tubules display a growing number of alpha-tubulin-positive cells from 4 to 40 weeks. From then on, nearly all epithelial cells contain alpha-tubulin, particularly in a narrow zone beneath their lateral cell borders.

Actins↗
PubMed1989

Electrophoretic heterogeneity of alkaline phosphatase isozymes in seminoma and normal testis.

Electrophoretic patterns of seminoma- and normal-testis-derived alkaline phosphatase isozymes, the placental alkaline phosphatase (PLAP)-like enzyme and the tissue-nonspecific (liver) alkaline phosphatase (LAP), were studied on starch gel and isoelectric focusing (IEF). Different migration patterns of the PLAP-like enzyme were observed with respect to both seminomas and normal testes on starch gel electrophoresis. On IEF, seminomas showed different staining patterns among different tumors; however, a common main activity was focused at pIs of 4.3-4.6, corresponding to pIs of PLAP. Normal testes showed two enzyme-staining regions, at pIs of 4.1 and 5.0-5.2, which were discriminated from pIs of PLAP and the PLAP-like enzyme in seminoma. The PLAP-like enzyme in seminoma was differentiated from PLAP by digestion with neuraminidase. Neuraminidase treatment simplified the distribution patterns of the PLAP-like enzyme in normal testis, but did not alter the pattern of microheterogeneity in seminoma. Two factors other than sialylation, namely structural modification of the carbohydrate moiety and variation of hydrophobicity, were shown to contribute to the microheterogeneity of the PLAP-like enzyme in seminoma. LAP in seminoma and in normal testis also showed marked electrophoretic heterogeneity and differences in pI distributions from LAP of liver. However, the migration patterns after desialylation were very similar to each other. The findings imply that electrophoretic heterogeneity demonstrated in LAP in seminoma and in normal testis is caused by a difference in sialic acid content in the molecule, and the heterogeneity of the PLAP-like enzyme in seminoma is considerable.

Alkaline Phosphatase↗
PubMed1996

Pathological changes of traumatic dislocated testis.

We present a 17-year-old patient with a right testicular dislocation. In this case, dislocated testis had been considered as hematoma and therefore remained untreated for 4 months. The patient underwent right orchiopexy 4 months after the injury. Intraoperative testicular biopsy revealed impaired spermatogenesis and existence of many alternative Sertoli cells, but no atrophy of seminiferous tubules. Six months after the operation, semen analysis of the patient revealed the deformity of most sperms. Eight months after the operation, rebiopsy of the right testis disclosed slight improvement in the spermatogenesis of the testis. Damage of the testis appears to be not severe in dislocation and may be recovered by repositioning.

Adolescent↗
PubMed1997

The vanishing testis: anatomical and histological findings.

OBJECTIVES: To review anatomical and histological findings in 105 vanishing testes. METHODS: Records of 2,509 boys with 3,064 cryptorchid testes treated at our hospital between 1969 and 1995 were reviewed. RESULTS: 691 (23%) testes were clinically impalpable. Exploration in 691 impalpable testes revealed absent testis in 144 (21%). In 39 (27%) of the 144 absent testes, there was complete agenesis of testis along with the epididymis and vas deferens whereas 105 (73%) were associated with blind-ending cord structures-the vanishing testis. The site of blind-ending cord structures in 105 vanishing testes was intra-abdominal in 22 (21%), inguinal canal in 62 (59%), superficial inguinal ring in 19 (18%) and scrotum in 2 (2%). Histological information was available in 47 vanishing testes and revealed vas, epididymis, or both in 32 (68%), fibrous/vascular tissue in 11 (23%) and testicular cords in 4 (9%). Dystrophic calcification and/or haemosiderin were present in 7 (15%). CONCLUSIONS: Our data show that the incidence of vanishing testis in boys with non-palpable testes is over twice the incidence of testicular agenesis. The most common site of blind-ending cord structures is distal to the internal inguinal ring. The finding of viable testicular tissue at the end of the attenuated cord structures in 4 of our patients, and also reported in other series, suggests that inguinal exploration should be carried out in all patients who on laparoscopy are found to have cord structures entering the internal ring.

Child, Preschool↗
PubMed1995

Is ipsilateral testis mandatory for the occurrence of contralateral intratesticular biochemical changes indicative of hypoxia after unilateral spermatic cord torsion?

Experimental unilateral spermatic cord torsion has been shown to induce both ipsilateral and contralateral intratesticular biochemical changes indicative of hypoxia. An experimental study was conducted to see whether the presence of the ipsilateral testicular tissue is necessary for the occurrence of the biochemical changes in the contralateral testis. Male, adult, albino rats were divided into four groups each containing 10 rats. One group served to determine basal values of biochemical parameters indicative of tissue hypoxia, another group was subjected to unilateral spermatic cord torsion in the presence of ipsilateral testis; a further group was subjected to unilateral spermatic cord torsion after ipsilateral subepididymal orchiectomy, and the last group underwent unilateral subepididymal orchiectomy alone as control. Lactic acid, hypoxanthine and lipid peroxidation product levels which are biochemical indicators of tissue hypoxia were determined in testicular tissues and kidneys. All three parameters increased significantly in contralateral testes but not in kidneys after unilateral spermatic cord torsion both when the ipsilateral testis was present and absent (p < 0.05). It is concluded that the ipsilateral testis does not play a role in transmitting these contralateral changes after unilateral spermatic cord torsion. Impulse-triggering contralateral changes may arise from spermatic vessels or nerves.

Animals↗