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Homologous testis transplantation in dogs.

There is growing interest in the possible use of homologous testis transplantation for the treatment of anorchia and male infertility. In order to test the surgical and immunological feasibility of this therapy, three series of experimental studies of homologous testis transplantation were carried out in dogs. In the first pilot study, four beagles from the same litter were transplanted using microsurgical techniques for end-to-end anastomosis of the testicular vessels and the vas deferens. These dogs received cyclosporin A (CyA) for 3 months after transplantation. The longest functional graft survival in this series was 163 days, strongly suggesting that long-term survival of a homologously transplanted testis graft is possible. A second series of operations was performed on ten mongrel dogs. The same surgical technique was employed and the series was divided into three groups. Group 1 received CyA monotherapy, group 2 a combination of CyA and prednisolone, and group 3 received no immunosuppression. The average graft survival time in this series was 28 days, significantly less than the 71 days in the first series. The dogs in group 2, however, had graft survival times that were three times longer than those in the other two groups, suggesting that CyA in combination with prednisolone yields the best graft survival. In the third series, five littermates received a testis graft after castration. Immunosuppression was achieved by administration of CyA and prednisolone for 3 months. In three out of five animals, the graft survived until the immunosuppressive therapy was suspended. Histological biopsies of the graft 3 months after transplantation showed the same maturation of sperm cells as in the control testis of the same dog. The results of the last series suggest that long-term survival of homologously transplanted testis grafts in dogs is, indeed, possible with the aid of CyA and prednisolone.

Animals↗

Prepubertal unilateral spermatic vessel ligation decreases haploid cell population of ipsilateral testis postpubertally in rats.

DNA flow cytometry is a sensitive and rapid technique to evaluate the germ cell maturation of testis. The haploid cell population of testis specifically denotes testicular maturation. High ligation of spermatic vessels (HLSV) and low ligation of spermatic vessels (LLSV) have been described in the surgical treatment of high undescended testis. Estimation of germ cell population of testis after these maneuvers has not been performed so far by DNA flow cytometry; hence this study was designed. Thirty male rats, aged 30 days, were randomized into three groups. Group I underwent sham surgery; group II underwent unilateral HLSV-ligation and division of main spermatic vessels, similar to the Fowler-Stephens technique; and group III underwent unilateral LLSV-ligation and division of main spermatic vessels close to the testis, similar to the Koff and Sethi technique. Thirty days later, the ipsilateral testes were harvested, and haploid, diploid, and tetraploid cells were counted by DNA flow cytometry. The mean (+/-SD) percentage of the haploid cell population was 68.7+/-4.8, 49.3+/-6.5, and 50.8+/-6.4 in groups I, II, and III, respectively. There was a significant (p<0.05) decrease in haploid cell population between groups I and II and between groups I and III. However, there was no significant difference between groups II and III. In conclusion, prepubertal HLSV as well as LLSV decreases the haploid cell population of ipsilateral testis in rats. LLSV has no advantage over HLSV.

Animals↗

The time relationship between ipsilateral testicular ischemia and germ cell apoptosis in the contralateral testis in rat.

Unilateral testicular ischemia-reperfusion (IR) in the rat is followed by histologic damage in the contralateral testis, which has been previously explained on immunologic grounds. There is evidence to suggest that apoptosis in the contralateral testis is involved in germ cell loss following IR injury to the testis. We examined the time-dependent effect of testicular ischemia on germ cell apoptosis in the contralateral testis in a rat. Adult Sprague-Dawley rats weighing 250-280 g, were subjected to testicular ischemia for 1, 2, 3 or 24 h. Twenty-four hours following onset of the ischemic insult, testes were harvested for immunohistochemical studies. Apoptosis was detected using TUNEL immunohistochemical assay. Testicular ischemia in rats led to histological damage, which was related to the duration of the ischemia. In the contralateral testis, the minimal damage included a decrease in number of germ cell layers, mild disorganization, and single cell apoptosis. Apoptosis in the contralateral testes increased significantly after 2, 3, and 24 h of ischemia and showed direct, time-related correlation with the duration of ischemia. We conclude that testicular ischemia causes an increase in germ cell apoptosis in the contralateral testis. The extent of apoptosis increases with the duration of the ischemia.

Animals↗

Organotypic culture, a powerful model for studying rat and mouse fetal testis development.

The key role of the fetal testis in the masculinization of genital organs has been known for a long time. More recently, the observed increases in male reproductive disorders has been postulated to be the result of changes in fetal and neonatal testis development in response to increasing environmental pollution. However, few tools are available for studying fetal testis development and the effects of physiological or toxic substances. We have developed an organ culture system in which rat fetal testis is grown on a filter floating on a synthetic medium containing no serum, hormones or biological factors. In this study, we have compared the long-term morpho-functional development of the various testicular cell types in this system with that observed in vivo and have extended this system to the mouse. Rat Leydig, Sertoli and germ cells and macrophages develop normally over a period of 1-2 weeks in this system. Fewer cells are produced than in vivo but the level of differentiated function is similar. Germ cells, which are difficult to culture in vitro, resume mitosis after a quiescent period, at the same time as in vivo. Similar results have been obtained with mouse fetuses, except that Leydig cells dedifferentiate in vitro if the testis is explanted after 13.5 days post conception. Testicular architecture and intercellular communication are sufficiently preserved for the development of the main fetal and neonatal testicular cell types in vitro with no added factors. Our floating-filter organotypic culture system in synthetic medium therefore allows the morpho-functional development of somatic and germ cells in fetal testis explants taken at all developmental stages in rat and at early stages in mouse. This method is potentially useful for studies of the effects of various factors, and of xenobiotics, in particular.

Animals↗

Morphometric studies on the testis of Korean ring-necked pheasant (Phasianus colchicus karpowi) during the breeding and non-breeding seasons.

The purpose of this study was to obtain detailed quantitative information on all cell types in the testis interstitium of Korean ring-necked pheasants and to combine these data with changes in the steroidogenic function of the testis during the breeding and non-breeding seasons. For animals collected during the breeding season, their testis weights, sperm production, serum testosterone levels and leuteinizing hormone (LH)-stimulated testosterone secretion were significantly (p < 0.01) increased compared to the non-breeding season. Testes of the pheasants during the non-breeding season displayed a 98% reduction in testis volume that was associated with a decrease in the absolute volume of seminiferous tubules (98% reduction), tubular lumen (100%), interstitium (90%), blood vessels (84%), lymphatic spaces (97%), Leydig cells (79%), mesenchymal cells (51%) and myoid cells (61%) compared to the breeding season. The numbers of Leydig cells, mesenchymal cells and myoid cells per testis in the breeding season were much higher than in the non-breeding season. Although the mean volume of a Leydig cell was 74% lower in the non-breeding season, the mean volumes of myoid and mesenchymal cells remained unchanged. These results demonstrate that there are striking differences in the testicular structure of the Korean ring-necked pheasant during the breeding and non-breeding seasons. Every structural parameter of the Leydig cell was positively correlated with both testosterone serum levels and LH-stimulated testosterone secretion. The correlation of changes in hormonal status with the morphometric alterations of Leydig cells suggests that the Korean-ring necked pheasant may be used as a model to study structure-function relationships in the avian testis.

Animals↗

Characterization of gonadotropin-releasing hormone (GnRH) binding sites in the pituitary and testis of the frog, Rana esculenta.

Frog, Rana esculenta, pituitary and testis gonadotropin-releasing hormone (GnRH) receptors were characterized by using 125I-chicken IIGnRH (cIIGnRH) as radiolabeled ligand. At 4 C equilibrium binding of 125I-cIIGnRH to pituitary homogenates was achieved after 90 min of incubation; binding of 125I-cIIGnRH to testis membrane fractions reached its maximum at 60 min of incubation. Binding of the radioligand was a function of tissue concentration, with a positive correlation over the range 0.5-2 tissue equivalents per tube. One pituitary and one testis per tube were used as standard experimental condition. Incubation of the pituitary homogenate with increasing concentrations of 125I-cIIGnRH indicated saturable binding at radioligand concentrations of 1 nM and above while for the testis membrane preparation saturation was achieved using 5 nM 125I-cIIGnRH. The binding of 125I-cIIGnRH was found to be reversible after addition of the cold analog and the displacement curves could be resolved into one linear component for both tissues. Scatchard analysis suggested the presence of one class of binding sites for both pituitary and testis (Pituitary: Kd = 1.25 +/- 0.14 nM and Bmax = 8.55 +/- 2.72 fmol/mg protein; testis: Kd = 2.23 +/- 0.89 nM and Bmax = 26.48 +/- 7.39 fmol/mg protein). Buserelin displaced the labeled 125I-cIIGnRH with a lower IC50 as compared with cIIGnRH cold standard, while Arg-vasopressin (AVP) was completely ineffective, confirming the specificity of binding.

Amino Acid Sequence↗

Epoxide hydrolysis in the cytosol of rat liver, kidney, and testis. Measurement in the presence of glutathione and the effect of dietary clofibrate.

The hydrolysis of trans- and cis-stilbene oxide and benzo[a]pyrene-4,5-oxide was measured in cytosol and microsomes of liver, kidney, and testis of control and clofibrate-fed rats. Significant levels of nonprotein sulfhydryls were detected in cytosol from liver (4.6 mM) and testis (1.5 mM). Glutathione was moderately stable in these fractions and interfered with the partition assays as conjugates were retained in the aqueous phase along with diols. When the products were separated by thin-layer chromatography, significant amounts of glutathione-conjugates were found to have been formed in the cytosol of liver and testis. Overnight dialysis or preincubation of cytosol with 0.5 mM diethylmaleate eliminated conjugate formation without affecting diol production. In dialyzed cytosol from clofibrate-fed rats (0.5%, 14 days), the rates of hydrolysis of trans-stilbene oxide were 506, 171, and 96% of controls for liver, kidney, and testis, respectively, and 126% of controls in liver microsomes. Rates of hydrolysis of cis-stilbene oxide were 149, 172, and 96% of controls in microsomes and 154, 124, and 91% of controls in cytosols from livers, kidneys, and testis of clofibrate-fed rats respectively. Hydrolysis of benzo[a]pyrene-4,5-oxide was similar to that of cis-stilbene oxide. Conjugation of the cis-stilbene oxide with glutathione was detected in cytosols from all three tissues with lesser amounts in the microsomes from liver and kidneys. After clofibrate treatment, the rates of this activity were 200, 173, and 95% of controls in cytosol from liver, kidneys and testis, and 203 and 202% of controls in microsomes from liver and kidneys respectively. These results indicate that epoxide hydrolysis and conjugation in rat liver and kidney are responsive to clofibrate treatment and support other evidence which suggests that hydrolysis of cis- and trans-stilbene oxides in cytosol is catalyzed, in part, by distinct enzymes.

Animals↗

A factor from bursa of Fabricius inhibits in vitro the chorionic gonadotropin response of the chick testis.

The effect of bursa of Fabricius on the endocrine function of the chick testis was studied using an isolated testis cell preparation. Testosterone secretion, both basal and under hCG stimulation, was measured in the incubation medium. The bursal extract inhibited the response of the testis cells to hCG detected as a reduced testosterone secretion. The normal basal secretion of the testis cells was not modified by the bursal extract. To analyze some characteristics of the bursal factor, fractions of approximately known molecular weight were obtained by filtering through Amicon membranes. The active factor was found in the fraction corresponding to 1000-10000 Da. Its activity disappeared after heating of trypsin incubation, suggesting the peptidic nature of the bursal factor. Tissue extracts from gut and spleen did not modify the hCG response of the testis cells. There was inhibition of the hCG response by thymus extracts of the newly hatched chicken. These results add new evidence of a modulatory effect of the immunogenic organs on the endocrine function of the testis in the newly hatched chicken.

Animals↗

Multiple sites of action of the vitamin D endocrine system: FSH stimulation of testis 1,25-dihydroxyvitamin D3 receptors.

1,25-Dihydroxyvitamin D [1,25(OH)2D] receptors exist in numerous unexpected tissues. These include, for example, rat lung, heart, testis, and uterus, but not prostate and bladder. The issues of 1,25(OH)2D effects on and receptor location in the testis were addressed by (a) physiological and pharmacological manipulations of tubule cell types and (b) histological examination of testes of vitamin D-deficient rats. FSH treatment in hypophysectomized adult rats increased 1,25(OH)2D receptor levels by 135% (P less than 0.01). Busulfan treatment reduced testis receptor levels by 35% (P less than 0.05) after 35 days (maximum effect), and the effect was reversed after recovery (85 d). Cryptorchidism for 5 or 50 days resulted in modest (33%, P less than 0.05) or substantial (79%, P less than 0.001) reductions in receptor levels. Only the FSH treatment and 50 days cryptorchidism reduced receptor levels in the residual tissue. The testes of vit. D-deficient rats showed incomplete spermatogenesis and degenerative changes. Although interpretation is complicated by the intricate communication among testis cell types, these data suggest that the Sertoli cell is a primary site of action of 1,25(OH)2D in the testis. Moreover, these data indicate that 1,25(OH)2D receptor function in the testis relates to germ cell division/maturation, although this may be an indirect effect via the Sertoli cells.

Animals↗

Neurotoxic esterase in rooster testis.

Neurotoxic esterase (NTE) is the putative target protein in the nervous system for the initiation of organophosphorus-induced delayed neuropathy. Here it is reported that NTE activity is present in rooster testis. Complete titration of rooster testis phenyl valerate esterases with paraoxon shows that about 15% of the enzymic activity is resistant to paraoxon. NTE activity after complete mipafox titration accounts for 30% of paraoxon-resistant phenyl valerate esterases and corresponds to 7.93 +/- 0.39 nmol/min/mg of protein (mean +/- SD, n = 7). Testis NTE is inhibited in vitro similarly to brain NTE by several organophosphorus compounds. Subcellular fractionation studies of the testis indicate that most NTE activity is particle bound. Testis NTE is also inhibited in vivo by several organophosphorus esters but to a lesser extent than brain NTE. Birds doses with organophosphorus compounds, causing delayed neuropathy, became grossly ataxic, but no testicular pathology was noted by light microscopy in roosters killed 15 days after administration. Serum testosterone levels also measured 15 days after dosing were not different from those of a control group. Recovery of NTE activity was faster in testis than in brain (4 days vs 6 days to recover to 50% of initial activity) in animals that received a high dose of an organophosphorus ester which cause delayed neuropathy.

Animals↗

Generation of superoxide anion and lipid peroxidation in different cell types and subcellular fractions from rat testis.

Mitochondria and microsomes from whole rat testis, seminiferous tubules and Leydig cells were investigated with respect to their capacity to generate superoxide anion. In addition, lipid peroxidation by whole testis mitochondria and microsomes was measured. In the presence of NADH and various respiratory inhibitors all three mitochondrial preparations catalyzed the formation of superoxide anion at a rate of 0.27-1.67 nmol/min.mg. This formation was concluded to be confined mainly to the NADH dehydrogenase region of the respiratory chain. Addition of NADPH to whole testis or Leydig cell mitochondria, but not tubule mitochondria, caused an additional formation of superoxide anion, which was unrelated to the respiratory chain, accelerated several-fold by menadione, and presumably catalyzed by NADPH-cytochrome c reductase and cytochrome P-450. Microsomes isolated from whole testis, seminiferous tubules, and Leydig cells generated superoxide anion at rates between 0.19 and 0.44 nmol/min.mg. These rates were also strongly stimulated by menadione. It is likely that both NADPH-cytochrome c reductase and cytochrome P-450 were involved in the microsomal generation of superoxide. Free radical scavengers of various types inhibited both the mitochondrial and microsomal formation of superoxide anion. Lipid peroxidation in whole testis essentially paralleled superoxide anion generation. However, the rate of mitochondrial lipid peroxidation was twice that of the microsomal rate. It is concluded that seminiferous tubules and Leydig cells generate superoxide anion at different rates and by different mechanisms. Together with cytochrome P-450-dependent hydroxylases, e.g., BP and DMBA hydroxylases, this superoxide generation may reflect a potential for cell-specific peroxidative damage in the testis.

9,10-Dimethyl-1,2-benzanthracene↗

Testis temperature, infertility, and the varicocele paradox.

The author presents arguments which favor elevated temperature of the testis and adnexae as an important factor in patients with poor semen seen in varicocele and "idiopathic" infertility. This appears to be primarily the result of disturbed thermoregulation due to internal spermatic vein retrograde blood flow. Elevated testis temperature without varicocele may be present in as high as 54% of all patients with "idiopathic" infertility. The prevailing theory in the United States is that the varicocele's bilateral effect is due to a substance noxious to spermatogeneis which is carried to both testes by retrograde internal spermatic vein blood flow and via crossover to the contralateral testis. A review of the published evidence for the crossover does not confirm its presence in the deep circulation of the contralateral testis, but rather its restriction to the superficial contralateral scrotal venous drainage. A simpler explanation for the bilateral effect of varicocele on spermatogenesis and sperm maturation by the epididymis is a gradient to the contralateral testis from the affected testis which results in demonstrable elevated temperature in both.

Body Temperature↗

Identification, localization and developmental studies of rat prepro thyrotropin-releasing hormone mRNA in the testis.

Thyrotropin-releasing hormone (TRH) plays the central regulatory role in the hypothalamic-pituitary-thyroid axis, but is also present in many extra-hypothalamic loci. The adult rat testis has been identified previously as a source of hypothalamic neuropeptides including TRH. To investigate whether the TRH gene is transcribed in testis, the identification and localization of prepro(pp) TRH mRNA and TRH were studied. Northern blot analyses of ppTRH mRNA in the adult rat testis showed a 2.0 kb band, hybridized with a ppTRH cRNA probe. This band was 0.4 kb greater than the 1.6 kb hypothalamic band. The concentration of ppTRH mRNA in the adult testis was approximately 13% of that found in the hypothalamus. Developmental studies of testicular ppTRH mRNA revealed that no ppTRH mRNA could be detected at the earliest stage (day 8). However, hybridization signals were detected on day 20 and increased progressively on days 35, 45 and 70 by 5.8, 6.4, and 9.8-fold, respectively. In addition, ppTRH mRNA was determined in Leydig cells by Northern analyses of elutriated testicular cell fractions. TRH was also measured in the rat testes at different developmental stages by RIA. TRH concentrations paralleled ppTRH mRNA during development. TRH was localized to Leydig cells by immunohistochemistry. These results indicate that ppTRH mRNA and TRH are present in the rat testis, especially in the Leydig cells. The changes of ppTRH gene expression and the concentration of TRH in the rat testis are developmentally dependent. TRH may function as a new paracrine or autocrine regulator of testicular function.

Animals↗

Poly(ADP-ribosylation) of nuclear proteins in rat testis correlates with active spermatogenesis.

Poly(ADP-ribosylation) of nuclear proteins has been investigated in rat testis under different experimental conditions to determine whether it is associated with somatic or germinal cells. Isolated, intact nuclei were incubated with [14C]NAD and extracted sequentially with 5% HClO4 and 0.25 M HCl, and labelled soluble proteins were analysed by reverse-phase high-performance liquid chromatography and acetic acid-urea polyacrylamide gel electrophoresis (pH 2.9). Results show that in normal adult testis a major acceptor protein for poly(ADP-ribose) in HClO4 extracts is the tissue-specific histone, H1t. Core histones and three proteins (alpha, beta and gamma) with low mobility on acetic acid-urea gels were the major acceptors identified in HCl extracts. Poly(ADP-ribosylation) of all the aforementioned proteins is very low in isolated intact nuclei of testis from 8-day-old animals (only spermatogonia present in seminiferous tubules), increases significantly by 16-day (pachytene spermatocytes appear) and reaches adult proportions by 32 days (condensing spermatids present). In the nuclei from cryptorchid testes, poly(ADP-ribosylation) of nuclear proteins resembles 8-day-old testis. It is concluded that (a) poly(ADP-ribosylation) of nuclear proteins in rat testis is closely correlated with spermatogenesis and can be inferred that is particularly active in the early stages of meiosis; (b) testis-specific proteins (histone H1t and low mobility proteins, alpha, beta and gamma) are poly(ADP-ribosylated) to higher specific radioactivity than somatic histones.

Age Factors↗

Ontogeny of the inhibitory guanine nucleotide-binding regulatory protein in the rat testis: mRNA expression and modulation of LH and FSH action.

The ontogeny of function and mRNA expression of the inhibitory guanine nucleotide-binding regulatory protein (Gi) was studied in the rat testis. Dispersed testis cells of animals aged 8, 15, 20 and 30 days were cultured with or without 100 micrograms/l pertussis toxin (PT) for 24 h. The cells were then cultured for another 24 h with medium only, cholera toxin (CT), PT, or their combination, and the amount of testosterone and cAMP production was measured. PT preincubation increased CT-stimulated cAMP production at all ages, thus indicating the presence of a functional Gi-protein in the postnatal testis. However, when testosterone production was measured, the enhancing effect of PT was absent at the age of 8 days only, indicating that Leydig cells at this age did not have functional Gi-protein. We then cultured 2-day-old and 8-11-day-old testis cells, after 24 h pretreatment with PT, in the presence of ovine follicle-stimulating hormone (FSH) (1 mg/l). The FSH-stimulated cAMP production was enhanced at both ages, thus indicating the presence of a functional Gi-protein in neonatal Sertoli cells. In Northern blot analyses, fetal and postnatal testis tissue had very similar levels of G alpha i2 and G alpha i3 mRNAs; the mRNA level of Gi1 in Northern blots remained low compared to those of Gi alpha 2 and Gi alpha 3. In conclusion, the Gi protein appears in the developing rat testis in utero but the activity first seems to be confined to non-Leydig cells including the Sertoli cell. In Leydig cells, the functional Gi-protein appears between days 8-15 post partum. This finding may be related to the fact that the fetal-neonatal population of Leydig cells possesses a high steroidogenic capacity and an apparent lack of the ability to respond to high gonadotropic stimulation with LH receptor down-regulation and steroidogenic enzyme desensitization.

Aging↗

Follitropin receptors in rat testis. Characterization with enzymatically 125I-labeled human follitropin.

The interaction between enzymatically radioiodinated human follitropin and the follitropin receptors in testis homogenate was investigated in immature and adult rats. The 125I-labeled human follitropin exhibited high binding activity with specific binding of up to 17% in the presence of an excess of testis homogenate. Approx. 50% of the bound hormone could be eluted at pH 5, and the receptor purified tracer exhibited a 3.6-fold increase in binding activity when compared with the original tracer preparation. Quantitative analysis of equilibrium binding data was performed with corrections for the measured specific activity and maximum binding activity of the tracer hormone. The equilibrium association constants (Ka) determined 24 degrees C were not significantly different in immature and adult rat testis, and the mean value for Ka was 3.9 . 10(9) M-1. At 37 degrees C, the Ka value obtained using immature rat testis was 1.3 . 10(10) M-1. The association of 125I-labeled human follitropin with immature rat testis homogenate was time and temperature dependent. In the presence of an excess of unlabeled hormone, 30--60% of the preformed hormone . receptor complex was dissociated after 24 h incubation. A specific and sensitive radioligand-receptor assay for follitropin was developed using immature rat testis homogenate. The minimum detectable dose of purified human follitropin was 0.6 ng, and human urinary and pituitary follitropin, ovine follitropin and pregnant mare serum gonadotropin reacted in the assay with equivalent slopes. The potencies of highly purified pregnent mare serum gonadotropin and highly purified human follitropin were similar in the radioligand-receptor assay, consistent with the follitropin bioactivity of the equine gonadotropin.

Animals↗

Spermatogenesis in testis xenografts grafted from pre-pubertal Holstein bulls is re-established by stem cell or early spermatogonia.

Xenografting of testis explants into recipient mice has resulted in successful restoration of spermatogenesis in several species. Most studies have utilized neonatal donor tissue, although a few have used prepubertal testes. In Holstein bulls, prepubertal development of the testis occurs between 16 and 32 weeks of age. The purpose of the present study was to determine the optimal age during prepubertal development of Holstein bulls for testis grafting. Explants of testis tissue from Holstein bulls between 12 and 32 weeks of age (2 bulls/age; 6 ages) were subcutaneously grafted into castrated or intact immunocompromised mice (n=8/age), then recovered after 75 and 173 days (n=4 mice/grafting period) and evaluated histologically for spermatogenic progression. Seminiferous tubules were assigned a score based on the most advanced type of germ cell present within the tubule and the average for all tubules scored (n=25) within an explant was calculated. Scores for all explants per mouse (n=6) were averaged to give a single spermatogenic progression score per mouse. No difference in spermatogenic progression of grafts between intact and castrated recipients was observed. Spermatocytes were observed in testis grafts from bulls of all ages 75 days post-grafting. At 173 days, the spermatogenic progression score for explants derived from 20 weeks bulls was greater than all ages except 12 weeks donors (p<0.05), with 8% of tubules containing spermatids. Donor material from bulls older than 20 weeks had lesser spermatogenic progression scores largely attributed to the greater number of atrophic tubules in grafts from older donors. Grafts from 28 and 32 weeks donors showed signs of degeneration by 75 days post-grafting, with 30 and 55% atrophic tubules, respectively, and lesser spermatogenic efficiency scores. By 173 days post-grafting, 72% of tubules in explants from 32 weeks donors were atrophic. The results of the present study suggest that the early stages of prepubertal development are optimal for testis grafting while advanced spermatogenesis in the donor tissue prior to grafting had a negative effect on graft development. Spermatogenesis within the grafts apparently needs to be re-established by spermatogonial stem cells or early spermatogonia.

Age Factors↗

Cloning of chicken lanosterol 14alpha-demethylase (CYP51) cDNA: discovery of a testis-specific CYP51 transcript.

A lanosterol 14alpha-demethylase (CYP51) cDNA, which consisted of a 1419 bp open reading frame encoding 472 amino acids and a 918 bp 3'-untranslated region, was isolated from the chicken testis. The sequence corresponding to exon 1 of this cDNA was completely different from those of CYP51 cDNAs in other tissues, including the liver. The expression level of the CYP51 gene with the testis-specific exon 1 was much higher in mature (2-year-old) male chickens than in immature (5-week-old) chickens. In addition, a CYP51 transcript common to several tissues, including the liver, adrenal gland, cerebellum, cerebrum, lobus opticus, kidney, lung, heart, muscle, spleen, small intestine and ovary, was also detected in the testis by RT-PCR. Furthermore, testis-specific shorter transcripts, which have been observed in mammals such as humans, rats and pigs, were not detected in the chicken testis. The results of this study demonstrate for the first time the presence of a chicken testis-specific CYP51 transcript and its sexual maturation-related expression, although its biological significance remains unclear.

Amino Acid Sequence↗