Percentage Of Haploid Cells Research Articles

LOCAL-IGF-1 and GH application IMPROVES germ cell histology, spermatogenesis and fertility after experimental testicular torsion and detorsion

To evaluate the impact of insulin like growth factor-1(IGF-1) and growth hormone (GH) on testis histology, spermatogenesis, and fertility in prepubertal rats exposed to 6h of testicular torsion (TT) and detorsion. Forty-eight male Wistar-albino rats weighing 30-70g and at 3-week age were allocated into six groups involving eight rats in each group as follows: Group 1:Sham, Group 2:Control, Group 3:Gelatin, Group 4:Local-IGF-1, 5: Local-GH, Group 6: Systemic-GH. Right testis was only exposed and sutured in the sham group, and right testes were rotated clockwise, 720°, fixed, and 6h later, detorsion on the testis was done in groups 2-6. Unloaded gelatin, 5μg local-IGF-1 loaded, and 2IU rhGH loaded gelatin were sutured to the right testis after detorsion in groups 3-5. In Group 6, 0.3IU/100gr/d rhGH was given for seven days via subcuticular route after detorsion. Each of the rats cohabited with two female rats five weeks later. Afterward, both right and left testes were removed. Mean diameter of seminiferous tubules (STD), mean biopsy score count of the testis (TBSC), mean percentage of haploid cells (HCP) were assessed, and fertility parameters were evaluated. STD and TBSC of the ipsilateral testes were significantly reduced in control and gelatin groups when compared to sham, local-IGF-1, and local-GH groups. STD and TBSC of the ipsilateral testes of the systemic-GH group were decreased compared to the sham group. HCP of the ipsilateral testes of control, gelatin, and systemic-GH groups were significantly lower than the sham, local-IGF-1, and local-GH groups. STD, TBSC, and HCP of the contralateral testes were significantly reduced in control and gelatin groups when compared separately to sham, local-IGF-1, systemic- GH, and local-GH groups. The difference between groups regarding potency, fertility, fecundity indexes, and mean fetus numbers were not significant. Even though there was significant and permanent histologic germ cell damage and reduced HCP in both ipsilateral and contralateral testes, experimental 6h TT and detorsion in prepubertal rats did not have a negative impact on future fertility. Local-IGF-1and rhGH treatment improved germ cell histology and spermatogenesis in both ipsilateral and contralateral testes of prepubertal rats, subjected to 6h of TT and detorsion.

The Germ Cell-Specific Markers ZPBP2 and PGK2 in Testicular Biopsies Can Predict the Presence as well as the Quality of Sperm in Non-obstructive Azoospermia Patients.

To assess the role of three testis-specific genes including ZPBP2, PGK2, and ACRV1 in the prediction of sperm retrieval result and quality of retrieved sperm by microdissection testicular sperm extraction (micro-TESE) in non-obstructive azoospermia (NOA) patients. This was a case-control study including 57 testicular samples of NOA patients including 32 patients with successful sperm retrieval (NOA+) and 25 patients with failed sperm retrieval (NOA-), and 9 samples of men with normal spermatogenesis in the testes as the positive control (OA). We investigated the expression of candidate genes by RT-qPCR and germ cell population patterns by DNA flow cytometry in testicular biopsy samples. The association between PGK2 expressions with the quality of retrieved spermatozoa was also evaluated. The RT-qPCR data revealed a significantly higher expression of ZPBP2 and PGK2 in the NOA+ in comparison to NOA- group (P = 0.002, and P = 0.002, respectively). Flow cytometry results revealed that the haploid cell percentage was significantly higher in NOA+ vs. NOA- group (P = 0.0001). In samples with a higher percentage of haploid cells, expression levels of ZPBP2 and PGK2 were higher (P = 0.001). The PGK2 expression was significantly associated with retrieved sperm quality (P = 0.01). Our results contribute to the search for the biomarkers for predicting the presence of testicular sperm and would be useful to avoid unnecessary multiple micro-TESE. Overall, the expression pattern of the ZPBP2 and PGK2 may be useful in predicting sperm recovery success and quality of retrieved sperm in NOA patients.

A Chemical Screen Identifies Compounds Capable of Selecting for Haploidy in Mammalian Cells

SummaryThe recent availability of somatic haploid cell lines has provided a unique tool for genetic studies in mammals. However, the percentage of haploid cells rapidly decreases in these cell lines, which we recently showed is due to their overgrowth by diploid cells present in the cultures. Based on this property, we have now performed a phenotypic chemical screen in human haploid HAP1 cells aiming to identify compounds that facilitate the maintenance of haploid cells. Our top hit was 10-Deacetyl-baccatin-III (DAB), a chemical precursor in the synthesis of Taxol, which selects for haploid cells in HAP1 and mouse haploid embryonic stem cultures. Interestingly, DAB also enriches for diploid cells in mixed cultures of diploid and tetraploid cells, including in the colon cancer cell line DLD-1, revealing a general strategy for selecting cells with lower ploidy in mixed populations of mammalian cells.

Testicular function in cultured postnatal mouse testis fragments is similar to that of animals during the first wave of spermatogenesis.

Previously, we evaluated optimal organ culture conditions to produce elongated spermatids in an in vitro mouse testis culture system. However, differences in testicular function between the cultured testis fragments and animal testis have not been determined. To examine how closely cultured testis fragments in vitro approximates what typically occurs during the first wave of spermatogenesis in vivo, C57BL/6J mouse testis fragments obtained on postnatal day (PND) 5 were cultured in AlbuMAX™ I/ α-Minimal Essential Medium for 15, 23, 30, 35, 42, and 49 days, and compared to mouse testes obtained at PND 5, 14, 20, 24, 28, 30, 35, and 40. At the specified days of culture or PND of mice, the following analyses were conducted: histology, flow cytometry for haploid cell detection, qPCR for spermatid markers, and liquid chromatography/mass spectrometry for testosterone levels. Round spermatids were initially observed at 23 days, and their percentage of the total number of cells continued to increase with culture time, as did gene expression of the spermatid markers and haploid cell percentage in the cultured testis fragments. These results were similar in temporal sequence to those in animals. Testosterone levels in the testis fragments reached a maximum at Day 49. These findings show this in vitro mouse testis organ culture model may be a useful and convenient tool for mechanistic studies. However, because germ cell differentiation in all seminiferous tubules was not observed, improvements in the system/methods are needed to more closely replicate spermatogenesis as observed in animals.

The effects of IGF-1 on mouse spermatogenesis using an organ culture method

Currently available organ culture methods can induce the differentiation of spermatogonial stem cells (SSCs) to spermatids in vitro, but the percentages of haploid cells and elongated spermatids are extremely low. The goal of this study was to test strategies to increase the differentiation rate of SSCs into elongated spermatids in vitro. RNA-seq was performed from forty round spermatids isolated by laser capture microdissection from cultured mouse testicular fragments (MTFs) or 27 days post-partum testes. Gene Ontology (GO) and KEGG analysis of the transcriptome revealed that many cell cycle and apoptosis-associated genes were among the differently expressed genes. Quantitative real-time PCR confirmed that the expression of Ccnd3 decreased and the expression of Trp53, Casp8 and Cyct increased in round spermatids from cultured MTFs. As insulin-like growth factor (IGF-1) can regulate cell cycle and apoptosis of many kinds of cells, the expression of Igf-1 decreased in cultured MTFs and IGF-1 receptor expressed strongly in germ cells, IGF-1 was added to the basal medium. IGF-1 increased the percentages of round and elongated spermatids by decreasing the apoptosis of germ cells and increasing the density of germ cells in cultured MTFs. These results indicate that IGF-1 plays a critical role in spermatogenesis from SSCs.

The Effects of Local and Systemic Growth Hormone Treatment on Germ Cell Population and Fertility in an Experimental Unilateral Testicular Torsion and Orchiectomy Model

We evaluated the effects of local and systemic growth hormone on the germ cell population of the contralateral testes of pubertal rats subjected to unilateral testicular torsion and orchiectomy 24 hours later. A total of 40 male Wistar-Albino rats at age 3 weeks were divided into 5 groups. In the sham operated group the right testis was sutured and orchiectomy was performed 24 hours later. In groups 2 to 5 orchiectomy was performed 24 hours later following testicular torsion. In groups 3 and 4 unloaded and growth hormone loaded gelatin films, respectively, were sutured on the contralateral testes. In group 5 systemic growth hormone was administered for 7 days. Five weeks later each rat was cohabited with 2 female rats and the left testes were removed for evaluation. Mean seminiferous tubular diameter, mean testicular biopsy score and the mean haploid cell percentage were calculated. Mating studies were performed and fertility parameters were assayed. Mean seminiferous tubular diameter, mean testicular biopsy score and the mean haploid cell percentage of the contralateral testes were significantly decreased in the control and gelatin groups compared with the other groups. There was no difference between the local and systemic growth hormone groups regarding the haploid cell percentage. There were no differences between the groups in mean fetus numbers, mating or fertility and fecundity indexes except in the gelatin group, in which the mean fetus number was significantly lower. Fertility is not affected in rats after 24 hours of testicular torsion and orchiectomy, although there is germ cell injury and a decrease in the percent of haploid cells. Growth hormone administration resulted in the restoration of germ cell histology and an increase in the haploid cell percentage of the contralateral testes. Growth hormone may improve fertility after unilateral testicular torsion and orchiectomy.

Development of retroviral vectors for insertional mutagenesis in medaka haploid cells

Insertional mutagenesis (IM) by retrovirus (RV) is a high-throughput approach for interrogating gene functions in model species. Haploid cell provides a unique system for genetic screening by IM and prosperous progress has been achieved in mammal cells. However, little was known in lower vertebrate cells. Here, we report development of retroviral vectors (rvSAchCVgfp, rvSAchCVpf and rvSAchSTpf) and establishment of IM library in medaka haploid cells. Each vector contains a modified gene trapping (GT) cassette, which could extend the mutated cell population including GT insertions not in-frame or in weakly expressed genes. Virus titration determined by flow cytometry showed that rvSAchSTpf possessed the highest supernatant virus titer (1.5×105TU/ml) in medaka haploid cell, while rvSAchCVpf produced the lowest titer (2.8×104TU/ml). However, quantification of proviral DNAs in transduced cells by droplet digital PCR (ddPCR) demonstrated that the “real titer” may be similar among the three vectors. Furthermore, an IM library was established by FACS of haploid cells transduced with rvSAchCVgfp at a MOI of 0.1. A single copy RV integration in the majority of cells was confirmed by ddPCR in the library. Notably, there was a significant decrease of haploid cell percentage after FACS, suggesting potential trapping for survival/growth essential genes. Our results demonstrated successful development of retroviral vectors for IM in medaka haploid cells, serving for haploid genetic screening of host factors for virus infection and genes underlying certain cellular processes in fish model.

Proanthocyanidin as a cytogenetic protective agent against adverse effects of plant growth regulators supplementation in rats.

The aim of the present study was to investigate the protective role of grape seed extract (containing proanthocyandin) against the adverse effects of plant growth regulators (GA3 (gibberellic acid) and IAA (indoleacetic acid)). The present data showed that the administration of either GA3 and IAA caused undesirable changes in both hepatic and testicular structure. This was evidenced by a disturbed hepatic strands, pyknotic nuclei, central vein with collapsed endothelium, dilatation in bile sinusoids, congested blood vessel, binucleatd hepatocytes, lymphocytic infiltration, vacuolation, giant hepatic cells, increased Kupffer cells and karyoryxis. Additionally, it was shown that degenerative changes in the testis, spermatogenic arrest, moderate tubular necrosis, Leydig cell degeneration and reduction in the number and size of the seminiferous tubules with some spermatogonia detached from the basement membrane. Concerning flow cytometric study of the liver a significant decrease in G0/1% and a significant increase in S phase %, G2/M %, P(53)% and apoptosis % (sub G1) were detected. However, in testis the data recorded a significant decrease in the percentage of mature sperm (percentage of haploid cells) and a significant increase in the percentage of spermatide, diploid cells, P(53)and of apoptotic cells. On the other hand, a distinct recovery of the mentioned hepatic and testicular histopathological and cytogenetic disorders was observed when proanthocyanidin was supplemented to rats administered either of the plant growth hormones (GA3 and IAA).

Testicular testosterone: Estradiol ratio in domestic cats and its relationship to spermatogenesis and epididymal sperm morphology

The phenomenon of teratozoospermia in felids is not fully understood. In this study, we investigated the testicular androgen:estrogen balance in domestic cats and correlated these data with epididymal sperm morphology and the degree of spermatogenic activity. During spring and summer, testes and blood samples were obtained from 37 mixed-breed domestic cats (12 to 48 mo). The epididymal sperm were harvested and evaluated for sperm counts, motility, and morphology. Distal cytoplasmic droplets were not considered a defect, and samples were considered normozoospermic if they contained more than 60% normal sperm (N = 25) or teratozoospermic if they contained less than 45% normal sperm (N = 12). The testicular and serum concentrations of testosterone (T) and 17β-estradiol (E2) were determined with an enzyme immunoassay. The gonadosomatic index and epididymal sperm numbers and motility did not differ between groups. The percentage of normal sperm was higher in normozoospermic (74.3 ± 2.0, mean ± SEM) than in teratozoospermic samples (43.1 ± 1.4). The most prevalent sperm defects in the teratozoospermic group were abnormal acrosomes (9.7 ± 2.0) and bent midpieces (12.2 ± 2.0) or tails (24.0 ± 2.7) with cytoplasmic droplets. Histomorphometric data were similar between groups, although there was a lower Leydig cell nuclear volume in teratozoospermic samples. Normozoospermic samples contained a higher percentage of haploid cells and had a higher index of total spermatogenic transformation than teratozoospermic samples. Serum concentrations of T (0.5 ± 0.1 vs. 0.8 ± 0.4 ng/mL) and E2 (9.5 ± 1.2 vs. 11.4 ± 2.3 pg/mL) and testicular T concentrations (471.6 ± 65.3 vs. 313.4 ± 57.6 ng/g) were similar between groups. However, compared with normozoospermic samples, teratozoospermic samples had higher testicular E2 concentrations (8.5 ± 3.6 vs. 5.4 ± 0.5 ng/g) and a lower T:E2 ratio (31.8 ± 4.1 vs. 87.2 ± 11.6). There were significant correlations between testicular E2 values and percentages of normal sperm (r = −0.55) as well as those with primary sperm defects (r = 0.58) or abnormal acrosomes (r = 0.64). The T:E2 ratio was also correlated with meiotic index (r = 0.45) and percentage of normal sperm (r = 0.58). In conclusion, a high testicular E2 concentration and a reduced T:E2 ratio were significantly associated with higher ratios of abnormal sperm types, suggesting that the balance between androgens and estrogens is an important endocrine component in the genesis of teratozoospermia in felids.

Effects of Local and Sustained Release of FGF, IGF, and GH on Germ Cells in Unilateral Undescended Testis in Rats

To evaluate the effects of the local release of fibroblast growth factor (FGF), insulin-like growth factor (IGF), and growth hormone (GH) on a germ cell population of ipsilateral undescended and contralateral descended testes of rats with a surgically created unilateral abdominal testis for 12 weeks and after the application of growth factors after orchidopexy. Forty 4-week old male Wistar albino rats were divided into 5 groups as follows: group 1, sham; group 2, gelatin; group 3, FGF; group 4, IGF; and group 5, GH. In the sham group, the right testis was exposed and sutured with 5-0 silk sutures. In groups 2-5, a right intra-abdominal testis was surgically created. After 12 weeks, orchidopexy was performed, and 1 cm(2) gelatin films were sutured to the right testes, either unloaded (group 2), or containing 2.5 microg FGF (group 3), 5 microg IGF (group 4), or 5 microg GH (group 5). After 30 days, both testes were removed for histopathologic investigation and DNA flow cytometry. The mean seminiferous tubular diameters (MSTDs), mean testicular biopsy scores (MTBSs), and percentages of haploid (1n) cells were calculated. Ipsilateral MSTD and MTBS significantly decreased in the gelatin and FGF groups compared with the sham, IGF, and GH groups. Contralateral MSTDs and MTBSs did not differ among groups. The haploid cell percentage significantly decreased in the ipsilateral and contralateral testes of the gelatin group compared with the sham, FGF, IGF, and GH groups. Local release of IGF and GH resulted in the preservation of germ cell histology in the ipsilateral testes of rats with a surgically created unilateral undescended testis for 12 weeks and after orchidopexy. The administration of IGF, GH, and FGF increased the haploid germ cell population in both ipsilateral undescended and contralateral descended testes.

Flow cytometric DNA analysis demonstrates contralateral testicular deterioration in experimental unilateral testicular torsion of prepubertal rats.

It has been postulated that unilateral testicular torsion causes damage to the contralateral testis and reduces fertility. However, in animal studies such an effect has not been fully proven by histopathologic examination or other conventional assays of spermatogenesis. We investigated the effect of unilateral testicular torsion on contralateral spermatogenesis in prepubertal rats using quantitative flow cytometric DNA analysis. Male rats were divided into three groups which underwent sham-operation, simple hemiorchiectomy or unilateral testicular torsion. Five weeks after these operations, fertility and spermatogenesis by flow cytometry were evaluated. No significant differences were observed in body weight, contralateral testicular weight or serum testosterone concentration among the three experimental groups. In the torsion group, mean seminiferous tubular diameter, number of foetuses, fertility rate and percentage of haploid cells were all significantly decreased compared to the other two groups. These results suggest that unilateral testicular torsion causes damage to the contralateral testis and consequently can reduce the future fertility of prepubertal rats.

Pathogenesis of Varicocele: Experimental Study Using Flow Cytometric DNA Analysis: Zur Pathogenese der Varikocele: Experimentelle Studie unter Verwendung der Flow-Cytometrie-DNA-Analyse

Despite extensive investigation, a cause-and-effect relationship between varicocele and male infertility has not been fully proven. We investigated the effect of a unilateral varicocele on both testes. Spermatogenesis in the rats with a surgically induced left varicocele was evaluated 5 weeks postoperatively by measurement of testicular weight, flow cytometric DNA analysis and evaluation of the mean seminiferous tubular diameter (MSTD). In rats with varicoceles, the testicular weight, the percentage of haploid cells and the MSTD were decreased in both testes in contrast to sham-operated rats. There was a greater decrease on the left side. These results suggest that unilateral varicocele may impair spermatogenesis in both testes, with the impairment being greater on the ipsilateral side.

Apoptosis is not the cause of seasonal testicular involution in roe deer

Apoptosis is involved in the regulation of spermatogenesis. The involution of testes in seasonal breeders might be expected to involve enhanced apoptotic cell elimination. We have compared seasonally changing testicular apoptosis in roe deer with that in non-seasonally breeding cattle. Apoptotic cells were detected as TUNEL-positive cells by both flow-cytometric analysis and in situ localisation of fragmented DNA in tissue sections. Apoptosis-induced DNA fragments were also assessed by enzyme-linked immunosorbent assay (ELISA) in homogenised testicular parenchyma. As expected, the testis mass and the percentage of haploid cells in roe deer showed a seasonal pattern with a significant maximum during the rut (August), whereas no annual variation of these parameters was found in bulls. All three methods for determining apoptosis showed similar findings. Roe deer exhibited significant seasonal fluctuation of total apoptotic activity (ELISA, apoptotic cells per tubule cross section) with a maximum during the breeding season. However, the seasonal differences in the number of apoptotic cells corresponded to the variable total numbers of spermatogonia and spermatocytes per tubule cross section. Thus, the percentages of TUNEL-positive cells related to the combined number of both germ cell types showed no seasonal variance, as confirmed by percentages of apoptotic cells analysed flow-cytometrically. The maximum level of apoptosis during the rut in roe deer was similar to the values obtained during the invariably high spermatogenic activity in cattle. These results suggest that, in roe deer, apoptosis is not the cause of the seasonal involution of testes.

Long-Term Influence of Prepubertal Testicular Torsion on Spermatogenesis

Purpose: To study the influence of prepubertal unilateral testicular torsion on spermatogenesis postpubertally. Methods: Sixty prepubertal SD male rats were divided into 6 groups. In each group, animal suffered different courses of unilateral testicular torsion including sham operation, 2- and 6-hour-long torsion, and permanent torsion. Salvia miltiorrhiza was injected as a remedy to release the I/R injury in the 2- and 6-hour-long torted groups. Postpubertally, the percentage of DNA content of haploid cells in the testes was determined individually. Results: The percentage of haploid cells in the sham operation group was 76.5 ± 1.9%. The number decreased in every other group (p < 0.01). Comparing with the same course of torsion, Salvia miltiorrhiza injection could improve the percentage of haploid cells (p < 0.01). In the permanent unilateral testicular torted group, the percentage was nearly zero (0.4 ± 0.2%). Conclusions: Prepubertal unilateral testicular torsion induces decreased spermatogenesis postpubertally. The result takes place in the bilateral testes. Salvia miltiorrhiza, as an antioxidant remedy, could relieve the injury which manifests improved spermatogenesis.

The effects of orchidopexy and orchidectomy on testes of rats subjected to ipsilateral abdominal testis with vas deferens obstruction

An experimental study was planned to evaluate and compare the effects of orchidopexy and orchidectomy on the testes of rats subjected to unilateral abdominal testis with vas deferens obstruction. Four groups were established. Rats in the control group underwent a sham procedure. While the testis was maintained in the abdomen with the vas deferens ligated for 8 weeks in group 2, rats in groups 3 and 4 underwent orchidopexy or orchidectomy after 4 weeks. Remaining testes were harvested at the end of the 8-week period. Testis and body weights were obtained during harvest. Samples were evaluated through DNA flow cytometry, and percentages of haploid cells were determined. Groups were compared through unpaired t-test, and p-values less than 0.05 were considered significant. All three treatments had decreased testis weight over body weight values of ipsilateral testes. Ipsilateral orchidectomy increased the value among contralateral testes. However, none of the groups had a contralateral testicular value less than the sham-operated group. All three treatments decreased the percentage of haploid cells among ipsilateral testes, but only an abdominal testis was associated with a decrease in the percentage of haploid cells among contralateral testes. Maintaining a testis with an obstructed vas deferens in the abdomen for 8 weeks damages both ipsilateral and contralateral testes. Orchidopexy, while showing amelioration of the ipsilateral testis, spares the contralateral testis as well as orchidectomy. Orchidopexy for an undescended testis with vas deferens obstruction is a rational approach.

Evaluation of developing rat testis by phosporus 31P magnetic resonance spectroscopy and DNA flowcytometry.

For the assessment of germ-cell maturation of the seminiferous tubules, DNA flowcytometry is a rapid and sensitive method. Phosporus 31P magnetic resonance spectroscopy (MRS) is a non-invasive alternative that demonstrates the metabolic status of the testis, reflecting the type and relative proportion of germ cells in the testis. This study was designed to evaluate the utility of 31P MRS in reflecting the haploid-cell population of the testis as measured by DNA flowcytometry. A single testicle was evaluated in male Wistar pre-pubertal rats at 30, 40, 50, and 60 days of age. In order to minimize the contamination of signals from the contralateral testis, scrotum, and tail, the technique was modified and the testis was evaluated ex-vivo with an intact blood supply. At 30 days of age the percentage of haploid cells was 43.6 +/- 1.8, and this increased to 72.7 +/- 1.4 at 60 days of age. During this period, the testicular phosphomonoester/adenosine triphosphate (PM/ATP) ratio changed from 1.70 +/- 0.21 to 0.32 +/- 0.08. There was a significant (P < 0.001) linear correlation between the proportion of haploid cells evaluated by DNA flowcytometry and the PM/ATP ratio evaluated by 31P MRS. 31P MRS is thus a reliable, noninvasive technique for accurately assessing the status of the seminiferous epithelium.

The impact of ofloxacin on rat testicular DNA: application of image analysis

Ofloxacin induces its antibacterial action mainly by inhibition of DNA gyrase which is equivalent to topoisomerase II in mammalian cells. The present study was focused on the impact of ofloxacin on rat testicular DNA ploidy in a dose–response relationship using an image analysis technique on testicular sections following Fuelgen DNA staining. Sperm count and motility as well as sperm head abnormality tests were also investigated. Ofloxacin was given p.o. in doses of 36, 72 and 360 mgkg−1day−1for 15 consecutive days. The animals were then left to live safely to day 60 from starting the experiment to give them a chance to complete the cycle of spermatogenesis. Results revealed that ofloxacin significantly decreased the percentage of haploid cells in a dose-dependent manner with concomitant increase in the percentage of diploid cells. Sperm count and motility was also markedly decreased in a dose-dependent fashion. Sperm head abnormality showed no marked change following ofloxacin treatment. In conclusion, ofloxacin induced a marked disturbance in rat testicular DNA ploidy, which may be explained on the basis of cross-reactivity to topoisomerase II.

Higher testicular activity in laboratory gerbils compared to wild Mongolian gerbils (Meriones unguiculatus)

The Mongolian gerbil has been used as laboratory animal since 1935. Breeding gerbils as an isolated laboratory population for decades may have led to a domestication process whose effects include changes in brain size. Quantitative changes in testicular activity could be assumed. Comparative intraspecific measurements were performed in 34 adult males of the laboratory strain (LAB) and in males raised as offspring of wild Mongolian gerbils (WILD) caught in central Mongolia (F1, n= 16; F2, n= 17). LAB and WILD were examined in January. Testicular spermatozoa were counted, proportions of different cell types were analysed using DNA flow cytometry, and mitotic and meiotic activity was calculated from DNA histograms. Intratesticular testosterone concentrations were measured with an enzyme immunoassay. In the WILD, testicular activity was lower and varied more. The overall weight, the efficiency of spermatogenesis (sperm/g testis) and resulting total sperm per testis were significantly less in offspring of wild gerbils. This corresponded with lower levels of haploid cells, total germ cell transformation of diploid cells to spermatids and meiotic transformation of spermatocytes to spermatids. The most profound difference was found in testicular testosterone concentration: the mean level was 405.7 ± 41.2 ng/g testis in LAB vs 6.4 ± 2.0 ng/g in WILD F1. All parameters changed in WILD F2 generation compared with F1 and diminished the differences with LAB. Differences between F1 and F2 were significant for testis mass, testis/body weight ratio, percentages of haploid cells and cells in G2/M phase, both germ cell transformations and testosterone concentration. The results suggest rapid, adaptive changes of male reproductive physiology in the early offspring generations from wild populations under laboratory breeding conditions. The breeding of Mongolian gerbils in the laboratory has influenced the testicular function resulting in increased spermatogenic activity and highly stimulated testosterone production.

Higher testicular activity in laboratory gerbils compared to wild Mongolian gerbils (Meriones unguiculatus)

AbstractThe Mongolian gerbil has been used as laboratory animal since 1935. Breeding gerbils as an isolated laboratory population for decades may have led to a domestication process whose effects include changes in brain size. Quantitative changes in testicular activity could be assumed. Comparative intraspecific measurements were performed in 34 adult males of the laboratory strain (LAB) and in males raised as offspring of wild Mongolian gerbils (WILD) caught in central Mongolia (F1, n= 16; F2, n= 17). LAB and WILD were examined in January. Testicular spermatozoa were counted, proportions of different cell types were analysed using DNA flow cytometry, and mitotic and meiotic activity was calculated from DNA histograms. Intratesticular testosterone concentrations were measured with an enzyme immunoassay. In the WILD, testicular activity was lower and varied more. The overall weight, the efficiency of spermatogenesis (sperm/g testis) and resulting total sperm per testis were significantly less in offspring of wild gerbils. This corresponded with lower levels of haploid cells, total germ cell transformation of diploid cells to spermatids and meiotic transformation of spermatocytes to spermatids. The most profound difference was found in testicular testosterone concentration: the mean level was 405.7 ± 41.2 ng/g testis in LAB vs 6.4 ± 2.0 ng/g in WILD F1. All parameters changed in WILD F2 generation compared with F1 and diminished the differences with LAB. Differences between F1 and F2 were significant for testis mass, testis/body weight ratio, percentages of haploid cells and cells in G2/M phase, both germ cell transformations and testosterone concentration. The results suggest rapid, adaptive changes of male reproductive physiology in the early offspring generations from wild populations under laboratory breeding conditions. The breeding of Mongolian gerbils in the laboratory has influenced the testicular function resulting in increased spermatogenic activity and highly stimulated testosterone production.

Assessment of testicular function in experimental varicocele rats by phosphorus-31 magnetic resonance spectroscopy.

To assess testicular function in experimental varicocele rats, we used 31P magnetic resonance (MR) spectroscopy and compared MR spectroscopic parameters with flow cytometric DNA analysis. In vivo 31P MR spectroscopy and flow cytometric DNA analysis of testes were performed in 10 sham-operated and 9 induced varicocele rats. Although the testicular phosphomonoester (PM)/ATP ratio did not differ between sham-operated and induced varicocele rats, the phosphodiester (PD)/ATP ratio was significantly reduced and the inorganic phosphate (Pi)/ATP ratio was significantly increased in induced varicocele rats. There was no difference between the right and left sides. DNA flow cytometry showed a decrease in the percentage of haploid cells in induced varicocele rats, regardless of the side. This study indicates that in vivo 31P MR spectroscopy provides valuable information for assessment of testicular function.