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PubMed2004

Polyorchidism presenting with undescended testis.

We report on a case of polyorchidism that presented as undescended testis. Polyorchidism is defined as the presence of more than two testes. As in our case, most cases occur on the left side. The inferior testis is evidently larger than the accessory testis in all of the cases in the literature and, as far as we know, the present case is the first with the small testis located more distally than the larger one. The accessory testis had no reproductive potential due to a lack of attachment to a cord structure; therefore, it was removed because of the high risk of malignancy.

Child, Preschool↗
PubMed2006

Routine contralateral orchiopexy for children with a vanished testis.

AIM: The aim was to determine whether routine contralateral orchiopexy is justified in a child with a vanished testis from intrauterine torsion. METHODS: A retrospective study of thirty-one consecutive boys with a vanished testis who underwent contralateral orchiopexy with the sutureless technique over an eight-year (1995 - 2002 inclusive) period was carried out. Operative findings were analyzed. All patients have been followed to date and interviewed by telephone. RESULTS: Of the thirty-one patients, 22 had testicular abnormalities (71 %). Five patients (16 %) had abnormalities that could predispose them to metachronous torsion. These included bell clapper deformity (3), horizontal lie (1), and ectopic testis (1). Other abnormalities were abnormal epididymal-testicular fusion (2), hydrocele (2), and one testis did not show compensatory hypertrophy. CONCLUSIONS: Sixteen percent of patients had testicular abnormalities that could predispose them to metachronous torsion. There was no morbidity from the operation and no post-orchiopexy torsion on follow-up. Therefore, routine contralateral orchiopexy in a boy with a vanished testis appears to be safe and effective. A large multicenter trial should be done to investigate these preliminary findings. If confirmed, contralateral orchiopexy should be advocated.

Child↗
PubMed1986

The movement of fluids and substances in the testis.

Three aspects of the control of movements of fluids and substances into, out of and inside the testis are discussed: the tubular barrier, the interstitial extracellular fluid and the testicular blood vessels. The functional basis for the tubular barrier is twofold; there are significant differences in the concentration of many substances inside and outside the tubules and marker substances enter or leave the tubular fluid at widely different rates, depending on lipid solubility and the presence of specific carrier systems. The anatomical basis for this barrier appears to be the specialized junctions between adjacent pairs of Sertoli cells. The barrier develops only at puberty, as the first cells undergo meiosis, but the development may not be as sudden as previously believed. The barrier breaks down after efferent duct ligation when spermatogenesis is disrupted. Techniques for measuring the volume, the turnover rate, the composition and fate of the interstitial extracellular fluid are described, and the unsatisfactory features of the presently available techniques for collecting this fluid for analysis are emphasized. There is a relationship between the fluid in the testis and lymph from vessels in the spermatic cord and lymph may be important for the transport of hormones to the general circulation in some circumstances and to other organs close to the testis. The testicular blood vessels display certain unusual features, a very high susceptibility to the toxic effects of cadmium salts, a high level of alkaline phosphatase activity in all endothelial cells but only after puberty and a high level of gamma-glutamyl transpeptidase in the endothelial cells of the arterioles and the testicular artery. These same cells are the site for a specific transport system for leucine and phenylalanine, with kinetic characteristics similar to the system in brain. Flow of blood may limit hormone secretion by the aspermatogenic testis, but diffusion limitation may also be important under some circumstances. A fuller understanding of the ways in which substances move around in the testis, particularly how they cross the endothelial cell layer or penetrate into the tubules, will be important for a better appreciation of testicular function.

Animals↗
PubMed1989

Influence of the immature testis on sexual differentiation in the tammar wallaby, Macropus eugenii (Macropodidae: Marsupialia).

Reproduction in the tammar wallaby, Macropus eugenii (Desmarest), is highly seasonal in the females but not the males. This study was designed to determine whether the difference is established during early life as a result of exposure to the developing testes. At day 10 after birth, when the sex can be distinguished externally, testes were removed from males and placed under the flank skin of females, while other groups of males and females were subjected to surgery without interfering with the gonads. The testis grafts remained palpable for 3-6 months. Sex-chromosome constitution was confirmed by karyotyping. At 3 years of age, the body weights and dimensions of the grafted females were not significantly different from those of the sham-operated females, whereas those of the castrated males were significantly larger and were equal to those of the sham-operated males, indicating that there is genetical control of growth independent of the testis in this species of marsupial. During 5 years of observations, none of the grafted females ever produced young, whereas all of the sham-operated females produced young each year from the second year. The grafted females had a mixture of male and female reproductive structures. The pouch and mammary glands developed normally, as did the Mullerian duct derivatives, the vaginal complex, the uteri and the oviducts. The ovaries were either devoid of oocytes and follicles or had reduced numbers, the Wolffian ducts were retained to varying degrees, the urogenital strand had developed into a prostate indistinguishable in size and structure from that of intact males, and the genital tubercle had developed into a normal-sized penis with a crus penis and Cowper's glands. In the castrated males, the scrotum developed normally and contained the gubernaculum and vas deferens. There was no evidence of Mullerian duct derivatives, and the urogenital strand was a simple canal, as in females. There were no Cowper's glands and no penis or erectile tissue. In one hemicastrated male, there was no development of the penis, although the remaining testis occupied the scrotum and showed compensatory hypertrophy. These findings indicate that the testis, at day 10, has a profound influence on the early differentiation of the Wolffian ducts, prostate and penis but cannot influence the differentiation of the Mullerian duct derivatives. The testis does not have any effect on the development of the pouch, mammary glands or scrotum or on somatic growth, all of which are apparently under independent genetical control.

Animals↗
PubMed2003

The endocrine-gland-derived VEGF homologue Bv8 promotes angiogenesis in the testis: Localization of Bv8 receptors to endothelial cells.

We recently identified an angiogenic mitogen, endocrine-gland-derived vascular endothelial growth factor (EG-VEGF), with selective activity for endothelial cells of endocrine tissues. Here we describe the characterization of a highly related molecule, Bv8, also known as prokineticin-2. Human Bv8 shares 60% identity and 75% similarity with EG-VEGF. The human and mouse Bv8 genes share a common structure. Like EG-VEGF, Bv8 is able to induce proliferation, survival and migration of adrenal cortical capillary endothelial cells. Bv8 gene expression is induced by hypoxic stress. Bv8 expression occurs predominantly in the testis and is largely restricted to primary spermatocytes. Adenoviral delivery of Bv8 or EG-VEGF to the mouse testis resulted in a potent angiogenic response. We have localized the expression of the Bv8EG-VEGF receptors within the testis to vascular endothelial cells. The testis exhibits relatively high turnover of endothelial cells. Therefore, Bv8 and EG-VEGF, along with other factors such as VEGF-A, may maintain the integrity and also regulate proliferation of the blood vessels in the testis.

Adenoviridae↗
PubMed1972

Amino-terminal sequences and sites of in vivo acetylation of trout-testis histones 3 and IIb 2 .

The sequences of the first 25 residues of histone III, and the first 22 residues of histone IIb(2), from trout testis have been determined on an automatic protein sequencer. The amino-terminal sequence of trout-testis histone III is identical to the corresponding region of calfthymus histone III, whereas the trout-testis histone IIb(2) sequence differs from that of calf-thymus histone IIb(2) at several positions in the amino-terminal region. Several in vivo sites of acetylation of these trout-testis histones have also been determined, by the same automated procedure. In addition to the two main sites at lysyl residues 14 and 23 acetylated in calf-thymus histone III, a lower degree of acetylation at two other sites, lysyl residues 9 and 18, has been detected. Four sites of acetylation have also been detected in trout-testis histone IIb(2), at lysyl residues 5, 10, 13, and 18. When the amino-acid sequences around the acetylated lysyl residues of different histones are compared, striking similarities are seen. The methods used in these studies should prove useful in elucidation of the locations of chemically stable modifications in other proteins.

Acetates↗
PubMed1984

Expression of pro-opiomelanocortin-like gene in the testis and epididymis.

Adrenocorticotropin (ACTH), beta-endorphin, and the melanocyte-stimulating hormones (MSHs), which are products of a common precursor, pro-opiomelanocortin (POMC), are present in a variety of tissues other than pituitary. The recent detection of immunoreactive POMC-derived peptides in the male reproductive tract raised the possibility that these hormones might regulate reproductive function. To determine whether the low concentrations of POMC-derived peptides in the male reproductive tract are synthesized locally and are not contaminants from blood, we have demonstrated POMC-like gene expression in both testis and epididymis. The identification of cells in testis capable of synthesizing POMC mRNA was established by showing the presence of this mRNA in mouse Leydig cell lines (TM3 and I10A). The hybridizing species of POMC-like mRNA in the testis, epididymis, and Leydig cell lines (TM3 and I10A) were approximately 150 bases shorter than those in the pituitary or hypothalamus but were similar in size to that in the amygdaloid nucleus of rat brain. The concentration of POMC-like mRNA in the testis is almost as high as that in the hypothalamus. This finding is quite unexpected because the concentrations of POMC-derived peptides in the testis were 2-3 orders of magnitude lower than those in the hypothalamus. The demonstration of a POMC-like gene expression in male reproductive tissues suggests that POMC-derived peptides are synthesized in Leydig cells and epididymis. These observations are consistent with the postulate that POMC-derived peptides may exert paracrine and/or autocrine effects in these organs.

Animals↗
PubMed1985

Expression of preproenkephalin-like mRNA and its peptide products in mammalian testis and ovary.

The distribution of preproenkephalin mRNA and proenkephalin-derived peptides have been examined in gonadal tissues from rats, hamsters, and cattle. A preproenkephalin mRNA band was detected in the ovaries of all three species and in hamster testis that is identical in size to the 1450-nucleotide mRNA typically found in tissues that express proenkephalin. Rat testis, on the other hand, expresses at least one preproenkephalin-like mRNA that is substantially greater in size (1900 nucleotides). [Met]enkephalin-containing peptides were also detected in each of the gonadal tissues examined. Although the abundance of preproenkephalin-like mRNA in rat testis was comparable to that in rat brain, the testicular content of proenkephalin-derived [Met]enkephalin sequences was less than 4% of the rat brain content. Together these data suggest that preproenkephalin-like mRNA in rat testis is not efficiently translated, proenkephalin-derived peptides undergo rapid turnover in this tissue, or the mRNA in rat testis has a frameshift resulting in an altered coding sequence.

Animals↗
PubMed1989

Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

Angiotensin-converting enzyme (ACE; EC 3.4.15.1) is a zinc-containing dipeptidyl carboxypeptidase widely distributed in mammalian tissues and is thought to play a critical role in blood pressure regulation. Testis contains a unique, androgen-dependent ACE isozyme of unknown function. We have determined the cDNA sequence for human testicular ACE; it encodes a protein that is identical, from residue 37 to its C terminus, to the second half or C-terminal domain of the endothelial ACE sequence [Soubrier, F., Alhenc-Gelas, F., Hubert, C., Allegrini, J., John, M., Tregear, G. & Corvol, P. (1988) Proc. Natl. Acad. Sci. USA 85, 9386-9390]. The full-length human testis ACE cDNA was constructed from a composite of cloned cDNAs, obtained by a combination of (i) immunoscreening and hybridization screening of a human testicular cDNA library in lambda gt11 and (ii) hybridization screening of human testis cDNAs constructed with ACE-specific primers and amplified by the polymerase chain reaction. The protein sequence inferred consists of a 732-residue preprotein including a 31-residue signal peptide. The mature polypeptide has a molecular weight of 80,073. The testis enzyme contains the second of the two putative metal-binding sites (His-Glu-Met-Gly-His) identified in endothelial ACE. This indicates that the functionally active catalytic site is within the C-terminal domain of the endothelial enzyme, accounting for the previous finding that these two structurally dissimilar isozymes are virtually identical catalytically. Of 22 testis ACE cDNAs cloned and sequenced, 3 have unique 5' regions, consisting of inserted, deleted, or substituted sequences up to 328 base pairs long, which have apparently arisen by alternative pre-mRNA splicing.

Amino Acid Sequence↗
PubMed1998

PAGE-1, an X chromosome-linked GAGE-like gene that is expressed in normal and neoplastic prostate, testis, and uterus.

We have used a combination of computerized database mining and experimental expression analyses to identify a gene that is preferentially expressed in normal male and female reproductive tissues, prostate, testis, fallopian tube, uterus, and placenta, as well as in prostate cancer, testicular cancer, and uterine cancer. This gene is located on the human X chromosome, and it is homologous to a family of genes encoding GAGE-like proteins. GAGE proteins are expressed in a variety of tumors and in testis. We designate the novel gene PAGE-1 because the expression pattern in the Cancer Genome Anatomy Project libraries indicates that it is predominantly expressed in normal and neoplastic prostate. Further database analysis indicates the presence of other genes with high homology to PAGE-1, which were found in cDNA libraries derived from testis, pooled libraries (with testis), and in a germ cell tumor library. The expression of PAGE-1 in normal and malignant prostate, testicular, and uterine tissues makes it a possible target for the diagnosis and possibly for the vaccine-based therapy of neoplasms of prostate, testis, and uterus.

Amino Acid Sequence↗
PubMed2000

Spermatogonial stem cell enrichment by multiparameter selection of mouse testis cells.

The spermatogonial stem cell initiates and maintains spermatogenesis in the testis. To perform this role, the stem cell must self replicate as well as produce daughter cells that can expand and differentiate to form spermatozoa. Despite the central importance of the spermatogonial stem cell to male reproduction, little is known about its morphological or biochemical characteristics. This results, in part, from the fact that spermatogonial stem cells are an extremely rare cell population in the testis, and techniques for their enrichment are just beginning to be established. In this investigation, we used a multiparameter selection strategy, combining the in vivo cryptorchid testis model with in vitro fluorescence-activated cell sorting analysis. Cryptorchid testis cells were fractionated by fluorescence-activated cell sorting analysis based on light-scattering properties and expression of the cell surface molecules alpha6-integrin, alphav-integrin, and the c-kit receptor. Two important observations emerged from these analyses. First, spermatogonial stem cells from the adult cryptorchid testis express little or no c-kit. Second, the most effective enrichment strategy, in this study, selected cells with low side scatter light-scattering properties, positive staining for alpha6-integrin, and negative or low alphav-integrin expression, and resulted in a 166-fold enrichment of spermatogonial stem cells. Identification of these characteristics will allow further purification of these valuable cells and facilitate the investigation of molecular mechanisms governing spermatogonial stem cell self renewal and hierarchical differentiation.

Animals↗
PubMed1999

Testis-specific TTF-D binds to single-stranded DNA in the c-mos and Odf1 promoters and activates Odf1.

We recently identified testis-specific nuclear factor binding sites in the testis-specific promoters of the c-mos gene and the Odf1 gene, which are 80% identical. Here we characterize a testis-specific nuclear factor, TTF-D, which is able to complex with both binding sites and stimulates Odf1 promoter activity. TTF-D is detectable in mouse testis as early as day 11 postpartum and contains three peptides of 22, 25, and 35 kDa in size. Surprisingly, TTF-D binds specifically to its cognate double-stranded DNA binding site as well as to its single-stranded DNA binding site. Both double-stranded and single-stranded binding site oligonucleotide DNA can specifically repress Odf1 promoter activity. Our results suggest that TTF-D is involved in positive transcription regulation of a pre-meiotic and a post-meiotic gene in the testis.

Animals↗
PubMed2000

Testis hormone-sensitive lipase expression in spermatids is governed by a short promoter in transgenic mice.

A testicular form of hormone-sensitive lipase (HSL(tes)), a triacylglycerol lipase, and cholesterol esterase, is expressed in male germ cells. Northern blot analysis showed HSL(tes) mRNA expression in early spermatids. Immunolocalization of the protein in human and rodent seminiferous tubules indicated that the highest level of expression occurred in elongated spermatids. We have previously shown that 0.5 kilobase pairs of the human HSL(tes) promoter directs testis-specific expression of a chloramphenicol acetyltransferase reporter gene in transgenic mice and determined regions binding nuclear proteins expressed in testis but not in liver (Blaise, R., Grober, J., Rouet, P., Tavernier, G., Daegelen, D., and Langin, D. (1999) J. Biol. Chem. 274, 9327-9334). Mutation of a SRY/Sox-binding site in one of the regions did not impair in vivo testis-specific expression of the reporter gene. Further transgenic analyses established that 95 base pairs upstream of the transcription start site were sufficient for correct testis expression. In gel retardation assays using early spermatid nuclear extracts, a germ cell-specific DNA-protein interaction was mapped between -46 and -29 base pairs. The DNA binding nuclear protein showed properties of zinc finger transcription factors. Mutation of the region abolished reporter gene activity in transgenic mice, showing that it is necessary for testis expression of HSL(tes).

Animals↗
PubMedDate not supplied

Human sperm membrane protein (hSMP-1): a developmental testis-specific component during germ cell differentiation.

Serum was obtained from an infertile woman having antibodies with sperm agglutinating activity. The antibodies interacted with a human sperm membrane protein (hSMP-1) with an estimated Mr of 55 kD. The gene (HSD-1) coding hSMP-1 was isolated from a human testis cDNA expression library and assigned the accession number U12978. The cDNA was conjugated to a prokaryotic expression vector to construct the recombinant vector, pRSET-HSD-I, which was expressed in Escherichia coli. The recombinant hSMP-1 was isolated and used to immunize rabbits to raise polyclonal antibodies. Usingan immunocytochemical technique, hSMP-1 protein was immunolocalized in germ cells of human testis at all stages of spermatogenesis. mRNAs were prepared from 16 different human tissues and analyzed by Northern blot using HSD-1 as probe. A positive reaction was elicited only with testis mRNA. The present findings suggest that the expression of hSMP-1 gene is testis-specific and occurs during the early stages of germ cell differentiation. In a comparative study, the location of the hSMP-I protein in sperm and in germ cells of the seminiferous tubules of rats was determined. The target antigen was immunolocated on the head and tail of rat sperm and in late spermatids and spermatozoa of rat testis. These results suggest that, in the rat, the HSD-1 gene is expressed during spermiogenesis.

Animals↗
PubMedDate not supplied

Effects of testicular fixation on oxidative stress-related parameters in prepubertal rat testis.

The goal of orchiopexy is to provide adequate scrotal fixation to prevent recurrent torsion of the testis and the spermatic cord or ascent of the testis, to be achieved with minimal trauma to the testis. Forty-two prepubertal male Sprague-Dawley rats were equally distributed into 6 groups (1 SHAM + 5 Operation). In operational groups, two lateral sutures were placed near the upper and lower pole of the testis passing through the tunica albuginea with 4-zero polyglactin suture in the transverse axis of the testis. At the 1st-3rd-7th-15th and 30th postoperative days, bilateral orchiectomy was performed, then malondialdehyde (MDA), nitrite, superoxide dismutase (SOD) and glutathion peroxidase (GPx) values were measured in testicular tissue homogenates. MDA and nitrite levels increased, while GPx and SOD levels diminished, statistically significant in both ipsilateral and contralateral testicular tissue homogenates. Prepubertal testicular fixation causes overproduction of the lipid peroxidation end product and free radicals while heavy consuming anti-oxidant reserve systems in both ipsilateral and contralateral testicular tissue. These results clearly reveal the extent of the testicular deterioration due to suture fixation technique.

Animals↗
PubMed1975

Electrophoretic analysis of liver and testis histones of the frog Rana pipiens.

Histones were extracted from frog livers and testes and analyzed by electrophoresis on long polyacrylamide gels and on sodium dodecyl sulfate (SDS)-containing polyacrylamide gels. Frog histones were found to be similar to those of calf thymus except that frog histone fraction F2A2 showed a marked dependence on the temperature at which the long gels were run, and frog histone fraction F3 could be separated from frog F2B on SDS-containing gels. Comparisons between frog liver and frog testis histones indicated that the testis contains as its major F1 component a fast migrating species not found in liver. Testis histones also showed less microheterogeneity of fractions F3 and F2A1 than liver histones. These were the only differences observed between liver and testis histones, even when testis histones were prepared from sperm suspensions that were rich in cells in the late stages of spermiogenesis. Thus it seems that, in Rana, the electrophoretic properties of the basic proteins of sperm differ from those of somatic cells only in the nature of histone F1 and in the degree of microheterogeneity of fractions F2A1 and F3.

Animals↗
PubMed1987

Isolation of rat testis cDNAs encoding an insulin-like growth factor I precursor.

We have characterized rat testis cDNAs encoding insulin-like growth factor I (IGF-I) precursor to facilitate studies of IGF-I expression in the male reproductive system. Two clones, P2 and P3, with inserts of 786 and 1200 bp, respectively, were isolated from a lambda gt11 library of rat testis cDNAs. The longest open reading frame of cDNA P2 predicts a 153-amino-acid residue IGF-I precursor that has only 11 amino acid substitutions compared with a human IGF-IA precursor encoded by a human liver mRNA. Three substitutions are within the predicted rat IGF-I sequence: a Pro for Asp in the B domain, an Ile for Ser in the C domain, and Thr for Ala in the D domain. Only two substitutions distinguish the predicted rat sequence from a mouse liver IGF-IA precursor: Thr for Ala in the signal peptide and Ala for Ser in the D domain. P2 hybridizes with poly(A)+ mRNAs of 7.5, 4.7, 1.7, and 1.2-0.9 kb in rat liver and testis. The other testis cDNA, P3, appears to represent a partially processed rat IGF-I mRNA precursor. By comparing the sequence of cDNA P2 with that of cDNA P3 and a 2.3-kb rat IGF-I genomic fragment, we predict exon splice sites within the codon for residue 26 and between residues 86-87 of the rat IGF-I precursor. Both of the predicted splice sites align with exon-intron junctions in the human IGF-I gene. We conclude, therefore, that IGF-I is synthesized as a precursor in the rat testis and that the structure of IGF-I genes, mRNAs, and precursors are highly conserved across species.

Animals↗
PubMed2006

Organizational and expressional uniqueness of a testis-specific mRNA transcript of protooncogene c-kit receptor in water buffalo Bubalus bubalis.

Protooncogene c-kit receptor is implicated with spermatogenesis, melanogenesis, and hematopoeisis, and undergoes tissue/stage specific alternate splicing. We have isolated 2973-bp full-length cDNA sequence (CDS) of this gene from testis and other tissues of water buffalo Bubalus bubalis. Upon comparison, the c-kit sequences showed tissue specific nucleotide changes resulting in novel truncated peptides. These peptides lacked intracellular and/or transmembrane domains in all the tissues except testis. Other alternately spliced tissue-specific transcripts were also detected, which are the integral parts of the open reading frame and have been reported in other mammals. Phylogenetic analysis of the sequences revealed unique tyrosine kinase domain in buffalo. Copy number calculation and expressional analysis of c-kit using real-time PCR established its single copy status and highest expression (137-177 folds) in testis compared to that (least) in liver. c-kit expression was detected in semen samples although 10 times lesser compared to that in testis. The highest expression of c-kit in testis and the presence of mRNA transcript in sperms substantiate its predominant role in spermatogenesis. This study establishes unequivocal involvement of an autosomal gene c-kit receptor in testicular function.

Amino Acid Sequence↗