Pubertal development of the boar: age-related changes in testicular morphology and in vitro production of testosterone and estradiol-17 beta

The responsiveness of testicular tissue, in terms of testosterone (T) and estradiol-17 beta (E2) production, to human chorionic gonadotropin (hCG) stimulation in vitro was assessed during pubertal development of the boar. A morphometric investigation was conducted concurrently to quantitate Leydig cell and seminiferous tubule changes in the testes of developing boars. Testicular volume percentage of seminiferous tubules increased from 36% at 40 days of age to a maximum of 72% at 190 days of age. Increases in tubular diameter were from 65 micrometers at 40 days of age to 236 micrometers at 250 days of age. Testicular volume percentage of Leydig cells decreased from 40% at 40 days of age to 10% at 250 days of age. Leydig cell number increased rapidly to 130 days of age, remained constant through 160 days, and then increased steadily to 220 days of age. Volume per Leydig cell changed little from 40 to 130 days of age, increased by 75% at 160 days, and declined thereafter. Total Leydig cell weight increased steadily from 40 to 160 days of age and then declined slightly. The capacity of Leydig cells for T production and testicular tissue for E2 production was greatest (P less than 0.05) after hCG stimulation in boars that were 130 and 160 days of age. In addition, sensitivity, as judged by the regression coefficient of T or E2 production per Leydig cell on log dosage of hCG was greater (p less than 0.05) for T at 130 days of age and for E2 at 160 days of age. The data presented support the hypothesis that one factor in pubertal development of boars is an increased capacity and sensitivity of the testes to gonadotropin stimulation.     

Ultrastructural analysis of a local regulation of Leydig cells in the adult monkey (Macaca fascicularis) and rat

Peritubular Leydig cells located in interstitial areas surrounded by tubules at nearly the same stage of spermatogenesis were analysed. Low-power electron micrographs were used for measurement of cell profile area and higher magnification views provided volume density of SER, Golgi stacks, mitochondria, and lipids. In the adult monkey, no cyclic changes were found in Leydig cells in their size or in the volume density of their organelles. In the adult rat (63 days of age), a comparison limited to stage VII-VIII and stage XI-XII peritubular Leydig cells demonstrated a significantly higher SER content (P less than 0.01) in the former, but no other differences. The study of subadult rats (45 days of age) showed that the full development of spermatogenesis was required to detect significant changes in Leydig cell SER content. The present results provide morphological evidence for an intratesticular control of the Leydig cells of the rat but not for those of the monkey.     

Relationships between Leydig cell morphometry and plasma testosterone during postnatal development of the monkey, Macaca fascicularis

In neonates (0 to 3-4 months), the testis contained a mean number of 4.6 X 10(6) Leydig cells representing 4.2 % of its volume; Leydig cell cytoplasm contained 10.2 % of SER. In infants (up to 45 months), Leydig cells regressed but their number increased; their volume density did not change. Leydig cell cytoplasmic volume (454 microns3 ), which was about 2.5-fold less than in neonates (1 119 microns3 ) or adults (1 170 microns3 ), contained only 8.7% of SER. During meiosis stage (38-52 months). Leydig cell numbers and volume density did not vary but the cells reached a maximal size and an amount of SER comparable with that at birth was measured. When spermatogenesis was complete, the Leydig cells represented no more than 0.8% of testis volume, but their number and SER content were significantly increased. Except for a significant decrease when spermatogenesis was completed, Leydig cell lipid content did not change during development, and the volume density of mitochondria did not vary. The mean level of plasma testosterone was 2 ng/ml in neonates and 0.4 ng/ml in infants; it increased to 3 ng/ml during onset of meiosis and reached 10 ng/ml in adults. The profile of testosterone was positively and significantly correlated with the total volume and total number of Leydig cells (P less than 0.01 and P less than 0.02, respectively) and with changes in their cytoplasmic volume (P less than 0.001). Moreover, plasma testosterone levels were positively and significantly correlated with changes in Leydig cell SER content i.e. SER volume density and mean absolute volume per cell (P less than 0.001), total SER in the whole testis (P less than 0.01).     

Testis morphometry, duration of spermatogenesis, and spermatogenic efficiency in the wild boar (Sus scrofa scrofa)

The wild boar is a natural inhabitant of Europe, Asia, and North Africa and is phylogenetically the ancestor of the domestic pig. Because of its phylogenetic and economic importance, this species is an interesting model for studying testis function in boars. Therefore, the present study was performed to investigate the testis structure, spermatogenic cycle length, and Sertoli cell (SC) and spermatogenic efficiencies in eight adult wild boars. Each spermatogenic cycle lasted 9.05 days, and the total duration of spermatogenesis was estimated as lasting approximately 41 days. The percentages of testis volume occupied by seminiferous tubules and by Leydig cells were 87% and 6%, respectively. The mean number of SCs per gram of testis was 42 million. The SC (round spermatids per SC) and spermatogenic (daily sperm production per gram of testis) efficiencies were 6.6 cells and 28.6 million, respectively. In general, the testis structure, overall germ cell associations at the different stages of the seminiferous epithelium cycle, and duration of spermatogenesis in the wild boar were similar to those in domestic pigs. Probably because of the small size of Leydig cells (400 microm3), their number per gram of testis (157 million) was the highest among investigated mammalian species. Although the SC efficiency in wild boars was low, their spermatogenic efficiency was comparable to that observed in domestic pigs, mainly because of the higher number of SCs per gram of testis in wild boars. These data suggest that SCs became more efficient during evolution, genetic selection, and domestication in pigs.     

Expression of FGF2 and TGFalpha and testis morphology during testicular hypertrophy subsequent to hemicastration in the neonatal boar

The objective was to ascertain fibroblast growth factor-2 (FGF2), epidermal growth factor (EGF), and transforming growth factor-alpha (TGFalpha) mRNA expression and testis morphology during accelerated testicular growth after hemicastration in the neonatal boar. On Day 10 after birth (Day 0), boars were assigned to control (n = 28), no treatment; hemicastrated (n = 28), left testis removed. The right testis in both groups (n = 7) was removed on Days 5, 10, 15, and 20. Expression of mRNA for FGF2, EGF, and TGFalpha was determined by qRT-PCR using TaqMan. Testicular morphology was determined on Day 15. On Day 10, hemicastrated boars had a greater (P = 0.01) testis weight (6.2 +/- 0.8 g; mean +/- SEM) than controls (4.3 +/- 0.4 g) and on Day 15 testis weight in hemicastrated boars (8.8 +/- 0.8 g) was twice (P < 0.01) that of control boars (4.2 +/- 0.3 g). Seminiferous tubule volume was approximately doubled in hemicastrated boars (P < 0.01) and was associated with an increase (P < 0.01) in Sertoli cell number. Interstitial compartment volume was greater (P < 0.01) in hemicastrated boars. Leydig cell numbers were similar (P = 0.14) but volume was greater (P < 0.01) for hemicastrates. There were no differences (P > 0.05) between control and hemicastrated boars in TGFalpha or FGF2 expression on Day 5 or Day 10, and EGF was not detected. It was concluded that upregulation of TGFalpha or FGF2 expression is not a pre-requisite for enhanced testicular growth and increased Sertoli cell proliferation that occurs subsequent to hemicastration in the neonatal boar.     

Comparison of components of the testis interstitium with testosterone secretion in hamster, rat, and guinea pig testes perfused in vitro

Components of the testis and cytoplasmic organelles in Leydig cells were quantified with morphometric techniques in hamster, rat, and guinea pig. Testosterone secretory capacity per gram of testis and per Leydig cell in response to luteinizing hormone (LH) (100 ng/ml) stimulation was determined in these three species from testes perfused in vitro. Numerous correlations were measured among structures, and between structures and testosterone secretion, to provide structural evidence of intratesticular control of Leydig cell function. Testosterone secretion per gm testis and per Leydig cell was significantly different in the three species: highest in the guinea pig, intermediate in the rat, and lowest in the hamster. The volume of seminiferous tubules per gm testis was negatively correlated, and the volumes of interstitium, Leydig cells, and lymphatic space per gm testis were positively correlated with testosterone secretion. No correlations were observed between volumes of blood vessels, elongated spindleshaped cells, or macrophages per gm testes and testosterone secretion. The average volume of a Leydig cell and the volume and surface area of smooth endoplasmic reticulum (SER) and peroxisomes per Leydig cell were positively correlated, and the volume of lysosomes and surface area of inner mitochondrial membrane per Leydig cell were negatively correlated with testosterone secretion. No correlations were observed between volume and surface area of rough endoplasmic reticulum (RER), Golgi apparatus, and lipid, and volume of ribosomes, cytoplasmic matrix, and the nucleus with testosterone secretion per Leydig cell. These results suggest that Leydig cell size is more important than number of Leydig cells in explaining the difference in testosterone-secreting capacity among the three species, and that this increase in average volume of a Leydig cell is associated specifically with increased volume and surface area of SER and peroxisomes. An important unresolved question is what is the role of peroxisomes in Leydig cell steroidogenesis.     

Morphometry of fetal Leydig cells in the monkey (Macaca fascicularis), correlation with plasma testosterone

The evolution of Leydig cells in Macaca fascicularis fetuses was followed throughout gestation (50-150 d) by morphometric procedures (volume densities of: cells, SER, mitochondria and lipid droplets). Testosterone from umbilical artery plasma was radioimmunoassayed starting on day 57. After predifferentiation and differentiation phases, Leydig cells entered the maturity phase (57-66 d), they occupied 19% of testicular volume, SER and lipid droplets represented 19% and 5% respectively of cytoplasmic volume. Then Leydig cells regressed dramatically (involution phase I: 66-83 d), their volume density decreased to 8%, that of SER to 12% whereas lipids doubled. Leydig cell volume density diminished to 5% during the second half of gestation (involution phase II), but their ultrastructure was not significantly altered. High plasma testosterone level (2.4 ng/ml) was observed during the maturity phase of Leydig cells, decline of testosterone occurred during involution phases I and II (1.13 and 0.58 ng/ml respectively). Its was shown that from day 57 to the end of fetal development the evolution of the plasma testosterone level correlated with the Leydig cell volume density and the SER volume density.     

Morphometric analysis of Leydig cells in the normal rat testis

Leydig cells are thought to be the source of most, if not all, the testosterone produced by the testis. The goal of this study was to obtain quantitative information about rat Leydig cells and their organelles that might be correlated with pertinent physiological and biochemical data available either now or in the future. Morphometric analysis of Leydig cells in mature normal rats was carried out on tissue fixed by perfusion with buffered glutaraldehyde, and embedded in glycol methacrylate for light microscopy and in Epon for electron microscopy. In a whole testis, 82.4% of the volume was occupied by seminiferous tubules, 15.7% by the interstitial tissue, and 1.9% by the capsule. Leydig cells constituted 2.7% of testicular volume. Each cubic centimeter (contained approximatelyy 1 g) of rat testis contained about 22 million Leydig cells. An average Leydig cell had a volume of 1,210 micron3 and its plasma membrane had a surface area of 1,520 micron2. The smooth endoplasmic reticulum (SER), the most prominent organelle in Leydig cells and a major site of steroidogenic enzymes, had a surface area of approximately 10,500 micron2/cell, which is 6.9 times that of the plasma membrane and is 60% of the total membrane area of the cell. The total surface area of Leydig SER per cubic centimeter of testis tissue is approximately 2,300 cm2 or 0.23 m2. There were 3.0 mg of Leydig mitochondria in 1 g of testis tissue. The average Leydig cell contained approximately 622 mitochondria, measuring on the average 0.35 micron in diameter and 2.40 micron in length. The mitochondrial inner membrane (including cristae), another important site of steroidogenic enzymes, had a surface area of 2,920 micron2/cell, which is 1.9 times that of the plasma membrane. There were 644 cm2 of inner mitochondrial membrane/cm3 of testis tissue. These morphometric results can be correlated with published data on the rate of testosterone secretion to show that an average Leydig cell secretes approximately 0.44 pg of testosterone/d or 10,600 molecules of testosterone/s. The rate of testosterone production by each square centimeter of SER is 4.2 ng/d or 101 million molecules/s: the corresponding rate for each square centimeter of mitochondrial inner membrane is 15 ng testosterone/d or 362 million molecules/s.     

Effect of seasonal changes in Leydig cell number on the volume of smooth endoplasmic reticulum in Leydig cells and intratesticular testosterone content in stallions

Testes from 47 adult (4-20 years) stallions obtained in November-January (non-breeding season) and 41 adult stallions obtained in May-July (breeding season) were perfused with glutaraldehyde, placed in osmium and embedded in Epon 812. Percentage Leydig cell cytoplasm or nuclei in the testis was determined by point counting of 0.5 micron sections under bright-field microscopy. Testes from 6 randomly selected horses per season were processed for electron microscopy. The volume (ml) of SER/testis was calculated from the % SER in the cytoplasm % Leydig cell cytoplasm, and parenchymal volume. Number of Leydig cells was calculated from the % nuclei, parenchymal volume, histological correction factor, and volume of single nucleus. Intratesticular testosterone content was determined from the contralateral testis by radioimmunoassay. The volume of SER/g and testosterone/g tended to be higher in the breeding than non-breeding season. Leydig cell number/g, volume of SER/testis, testosterone/testis, and Leydig cell number/testis were significantly greater in the breeding than in the non-breeding season. Volume of SER/testis and testosterone/testis were related significantly to the cell number/testis, and SER/testis was related (P less than 0.05) to testosterone/testis. These results emphasize the importance of seasonal changes in the number of Leydig cells on the amount of SER available to produce testosterone and on testosterone content/testis in the stallion.     

Impact of boar exposure on puberty attainment and breeding outcomes in gilts

We examined the most effective method of boar exposure for the attainment of puberty in 89 gilts. At 160 days of age, we allocated gilts to daily direct contact with a vasectomized boar after movement of pen groups of gilts to a detection-mating area (DGB: n = 30); daily direct contact with boars in the gilt home pens (DBG: n = 31); or daily fenceline contact between boars and gilts housed in individual gilt stalls (FBG: n = 28). DGB gilts were younger (P < or = 0.05) than FBG gilts at puberty. Direct boar contact reduced the interval from initial boar contact to puberty in DGB and DBG gilts, compared to fenceline contact in FBG gilts (P < 0.05). There was no difference (P > or = 0.05) between treatment for pubertal weight, backfat, lifetime growth rate, or duration of first pubertal estrus. Backfat depth and leptin concentration at 160 days of age were positively correlated (P < or = 0.05). We detected no relationships between leptin or IGF-1 concentration at 160 days of age and the interval from initial exposure to a vasectomized boar to puberty (P > 0.05). Based on objective criteria, fenceline contact with a boar (BC) during artificial insemination improved the quality of artificial insemination compared to no boar contact (NC) (P < 0.05).     

Characterization of luteinizing hormone-human chorionic gonadotropin receptor and its relationships to testicular development and steroidogenesis during sexual maturation in boars

Luteinizing hormone-human chorionic gonadotropin (LH-hCG) binding capacity for testes of boars and receptor sites per Leydig cell were estimated during pubertal development from 70 to 250 days of age, and changes in these two traits were correlated with morphological and endocrine parameters. Binding capacity increased linearly from 70 to 160 days of age, remained constant through 250 days of age, and was correlated (P less than 0.05) with paired testes weight, Leydig cell number and weight per paired testes, and serum estradiol-17 beta (E2) concentrations. LH-hCG receptor sites per Leydig cell were constant at all ages except for an increase observed at 160 days of age and were correlated (P less than 0.05) to in vitro maximum testosterone (T) production and sensitivity of E2 production per Leydig cell in response to hCG stimulation. Number of LH-hCG receptor sites was correlated (P less than 0.05) with Leydig cell surface area, and sites per unit surface area increased with age. Equilibrium association constants did not differ with age, and they averaged 8.6 +/- 1.0 X 10(9) M-1. Results from the present study indicate that LH-hCG receptor capacity per paired porcine testes increases throughout pubertal development primarily as Leydig cell numbers increase.     

Ultrastructure and morphometry of testicular Leydig cells and the interstitial components correlated with testosterone in aging rats

The ultrastructure of testicular interstitium in young and aged adult rats was analysed using morphometric methods, and the plasma testosterone concentration was measured. With increasing age there was an augumentation in the volume of collagen fibrils in the intercellular matrix and in blood vessels. During the aging process (approximately two years) the average volume of the Leydig cell decreased from 1364 m³ to 637 m³, but the number of Leydig cells in paired testes increased from 53x10⁶ to 113x10⁶. The absolute volume of smooth surfaced endoplasmic reticulum (SER) per Leydig cell amounted in aged rats to 78% of that in young adult rats. The total amount of SER in paired testes increased by 62% with aging. The present analysis suggests that the ability of SER to maintain peripheral testosterone concentration decreases with age. In young adult rats the absolute volume of peroxisomes per Leydig cell correlated significantly with the concentration of testosterone in blood and also with the absolute volume of SER per Leydig cell. These results combined with ultrastructural observations of close apposition of peroxisomes and SER suggest that peroxisomes have a role in testosterone secretion by Leydig cells.Visiting scientist to Laboratory of Electron Microscopy (Director: Prof. L.J. Pelliniemi)     

Effects of prolonged cyclosporine-A treatment on the Leydig cells of the rat testis

The effects of a prolonged (30-day) treatment with daily therapeutical doses of cyclosporin A (CAS) (20 mg/kg) on testicular Leydig cells were studied in adult rats. CSA administration provoked a significant decrease in both basal and human chorionic gonadotropin (hCG)-stimulated testosterone concentration in the peripheral blood without affecting the volume of the testes or the interstitial space. However, there was conspicuous atrophy of the Leydig cells, due mainly to a decrease in mitochondria and smooth endoplasmic reticulum, the organelles containing the enzymes of testosterone synthesis. Lipid droplets, in which cholesterol is stored, were notably increased. The nuclear volume and the surface area per cell of rough endoplasmic reticulum fell significantly in Leydig cells of CAS-treated animals. In light of these findings, it is concluded that CSA inhibits the growth and steroidogenic capacity of rat Leydig cells, probably by depressing their protein synthesis. Whether the mechanism underlying the action of CSA on Leydig cells is only indirect, by blockade of hypophyseal gonadotropin release, or also direct is unsettled and requires further investigation.     

Blockade of the hypothalamic-pituitary-testicular axis with a GnRH antagonist in the neonatal marmoset monkey: changes in Leydig cell ultrastructure

Little is known of the cell biology of Leydig cells during the neonatal activation of the hypothalamic-pituitary-testicular (HPT) axis. The current study examined the effect of blockade of the HPT axis with a GnRH antagonist (antide) on the neonatal population of Leydig cells in the new world primate, the common marmoset. Three sets of twins, age 7 weeks, were studied: in each pair one twin was used as a control, while the other received treatment with GnRH antagonist from the day of birth to suppress pituitary gonadotrophin secretion. Leydig cells of treated animals were dramatically different from those of controls. The cells were atrophic and exhibited very irregular nuclei. The organelles involved in steroid synthesis were reduced to the extent to being barely evident. The smooth endoplasmic reticulum (SER) was greatly diminished in quantity and distribution. The usual form of the SER (anastomosing tubules) was not evident, but, instead, the SER was relatively unbranched. Peroxisomes, organelles involved in transfer of cholesterol to the mitochondria, were greatly reduced in number. Mitochondria were relatively sparse and exhibited a non-typical morphology, as tubular elements of the cristae were rarely evident. Thus, the central apparatus in steroid production, the SER, mitochondria and peroxisomes, was essentially shut down in the GnRH-antagonist-treated animals. Storage of cholesterol, the precursor of steroid biosynthesis, was also not in evidence, as lipid droplets were extremely rare. Two prominent features of control in neonatal marmoset Leydig cells, the membranofibrillar inclusion (MFI) and basal laminae, remain prominent in the Leydig cells of treated animals. Evidence of apoptosis was not observed. These results provide strong support that the gonadotrophic hormones are the primary regulator of neonatal Leydig cell development in primates, and also suggest cell regression, rather than apoptosis, being the mechanism of this inhibition.     

B-mode ultrasonographic evaluation of paired testicular diameter of mature boars in relation to average total sperm numbers

Crossbred, meat-type terminal sire boars (n = 215) were randomly assigned by age group (240-300, 301-360, 361-420, 421-480, 481-540, 541-600, and >721 days). Stud boars were on a once or twice weekly semen collection schedule. Testis diameters, in duplicate, were obtained using B-mode ultrasonography. Summation of average left and right testis diameter within boar gave the paired testicular diameter (PTD). Average ejaculate volume, sperm concentration (sperm/ml), and total sperm numbers for each boar were determined using composite data (average values) obtained from the last four semen collections. There was a <0.5cm difference between left and right testis diameters, with the left testis being the larger of the two testes (P = 0.03). There was no difference in PTD found between age groups in this study. Conversely, a dramatic increase in average total sperm numbers (ATSN) was observed between boars of 240-300 days (57.0+/-27.4 x 10(9) sperm) and up to 420 days (118.6+/-33.6 x 10(9) sperm) of age. The ATSN (127+/-32.5 x 10(9) sperm) remained constant for the 421-480 to >721-day age groups. The correlation between PTD and ATSN was low (r = 0.24) in this study. The results of this study demonstrate that normal boars should exhibit <0.5cm diameter difference between testes. As observed in other studies, the left testis was usually larger than the right testis. Correlation of total sperm numbers in a boar ejaculate using a composite ejaculate score (average values) and PTD measurements obtained using B-mode ultrasonography was poor when used in boars >8 months of age.     

大鼠睾丸间质细胞的自体吞噬活动

本文结合超微结构和细胞化学观察,研究大鼠睾丸间质细胞(Leydig细胞)中溶酶体的结??构与功能。观察结果表明,大鼠睾丸间质细胞中高尔基体非常发达,在高尔基体的成熟面存在着CMP酶阳性反应的GERL系统,说明这种细胞有不断产生溶酶体的能力。细胞化学结果也证实在睾丸间质细胞有较多的初级和次级溶酶体。睾丸间质细胞不仅有较多的溶酶体,而且还有相当数量的自噬小体,存在着活跃的自体吞噬活动。自噬小体的界膜来源于特化的光面内质网或高尔基体膜囊,包围的内容物主要是光面内质网和少量线粒体。当自噬小体与溶酶体融合后即成为自体吞噬泡,由于酶的消化作用,自体吞噬泡内的细胞器有一系列形态变化。根据CMP酶细胞化学反应,可以区分自噬小体和自体吞噬泡,后者是一种次级溶酶体,呈CMP酶阳性反应。睾丸间质细胞是分泌雄性激素的内分泌细胞,其光面内质网和线粒体在类固醇激素分泌中起重要作用,自体吞噬活动的结果是去除部分内质网和线粒体,可能在细胞水平上起着对雄性激素分泌的调节作用。     

Seasonal changes of the Leydig cells of viscacha (Lagostomus maximus maximus). A light and electron microscopy study

The Leydig cells of viscacha (seasonal rodent) show cytoplasmic hypertrophy and regional distribution during the breeding period (summer-autumn). The dominant organelles are smooth endoplasmic reticulum (SER) and mitochondria. A moderately well-developed Golgi, abundant lipid inclusions, dense bodies like lysosomes in different stages, and centrioles are observed. Extensive or focal desmosome and gap-like junctions between neighbouring Leydig cells are present. These cells exhibit an evident hypotrophy and an increase in the number of dense bodies during the gonadal regression in winter (July and August). Cells in different stages of involution are observed in this period. Their nuclei are irregular and heterochromatic. The cytoplasm contains few mitochondria. The vesicular SER is scarse. Irregular and large intercellular spaces with microvilli and amorphous material are present. The junctional complexes are absent. The nuclear and cytoplasmic volume and development of SER and mitochondria increase during the recovery period (spring). The lipid inclusions decrease. Dilatations of the intercellular space with microvilli and limited by focal desmosome-like junctions are observed. In conclusion, the Leydig cells of Lagostomus maximus maximus show deep changes alongside the reproductive cycle. The photoperiod variations, through pineal hypothalamus pituitary axis and the hormone melatonin, are probably responsible for them. Moreover, the fall of serum and tubular testosterone would be one of the factors responsible for gonadal regression.     

Differences in aromatase activity between Leydig cells from the scrotal and abdominal testis in the naturally unilateral-cryptorchid boar

In an earlier study, estrogen production was much lower in Leydig cells from the abdominal than from the scrotal testis in naturally occurring unilateral cryptorchidism in the boar. A more direct assessment of aromatase activity was made in thirty-two mature male pigs to examine this observation further, using nonradioactive androstenedione (delta 4A 1.0 x 10(-6) M - 1.5 x 10(-5) M) and [1 beta, 2 beta-3H] delta 4A as substrates. Purified Leydig cells were prepared from normal boars and from unilaterally and bilaterally cryptorchid animals. Combined estrone sulfate (E1S) and estrone (E1) formation from delta 4A were measured by radioimmunoassay. Little or no estrogen secretion was seen with cells from the abdominal testis in unilaterally cryptorchid boars (n = 7), and E1S formation from delta 4A was 6- to 14-fold higher for scrotal cells (n = 6). Aromatase activity as reflected in percent conversion of substrate to [3H]-labeled water was clearly lower in cells from the abdominal testis (1.10 +/- 0.08 and 11.22 +/- 0.7%, respectively, p less than 0.01, n = 6). No marked reduction was noted for unilaterally cryptorchid boars with an inguinally located testis (10.18 +/- 0.27 and 13.09 +/- 0.58% for inguinal and scrotal testes, respectively, n = 3). Concentrations of E1S in testicular arterial and venous blood (n = 9) gave additional evidence of lower estrogen production by the undescended testis of the cryptorchid boar. It was concluded that lower aromatase activity is present in Leydig cells of the abdominal testis.     

The effect of modified eros centre, outdoor raising or conventional group housing on breeding gilts and its effects on reproductive performance over four parities

The present study was conducted in a large Croatian "built up unit". The objective of the study was to determine if an indoor modified eros centre (MEC) compared to indoor or outdoor group housing of gilts, influenced the onset of puberty of gilts and the reproductive performance of the evaluated females (n = 783) over four parities. The gilts were from the same nucleus herd. Gilts of same age (140-150 days of age), body condition (body condition score of 3-4) and similar genetics (four-way cross females), during the same season (January to April 1999), were randomly divided at arrival into three groups and treated as follows:MEC gilts (n = 279): These were placed into indoor MEC pens in groups of 8-10. The gilts had continuous fenceline contact to boars (one boar to two groups of gilts, boars were changed daily) and to shortly weaned oestrous sows. Gilts were regrouped and dislocated at 10-day intervals. Outdoor gilts (n = 263): These were kept in groups of 8-10 on a large pasture (80-100 m2 per group). The animals had fenceline contact to mature boar for 5-10 min daily. Control indoor gilts (n = 241): These were housed indoors in large pens in groups of 8-10. The animals had fenceline contact to mature boars for 5-10 min daily. Each outdoor group had an insulated hut with straw bedding. All gilts were fed ad libitum with the same commercial diet. Housing gilts in MEC resulted in earlier (P < 0.001) onset of estrus (MEC: 174.8 +/- 2.4 days, indoor group housing: 207.6 +/- 4.1 days, outdoor group housing: 187.4 +/- 2.1 days) and lower (P < 0.001) farrowing rate to first service (MEC: 70.97%, indoor group housing: 89.73%, outdoor group housing: 89.62%). Farrowing rate of regularly returning MEC gilts to second service was 95.00%. First total-born litter size, first liveborn litter size, first wean-to-estrus interval (WEI), percent of sows bred after first weaning, second total-born litter size, second liveborn litter size, average third and fourth total-born and liveborn litter size, number of sows having four litters, number of litters per sow, total number of pigs per sow, total number of liveborn pigs per sow showed no significant differences between the groups. More (P < 0.05) sows were culled in outdoor group. Compared to MEC and outdoor housing, indoor housed sows suffered higher (P < 0.05) percentage of anoestrus.     

Peroxisomes and intracellular cholesterol trafficking in adult rat Leydig cells following Luteinizing hormone stimulation

The present study was designed to explore the intracellular cholesterol trafficking in Leydig cells of adult rats following Luteinizing hormone (LH) injection. Histochemical techniques were used to demonstrate distribution of free cholesterol in Leydig cells of control and LH-injected rats. Two groups of sexually mature male Sprague Dawley rats (n=4/group) were used. Fifteen min following an injection of 200 microl of either saline (control) or luteinizing hormone (LH, 500 microg in saline) testes of rats were fixed by whole body perfusion using 0.5% glutaraldehyde and 4% paraformaldehyde in 0.1 M cacodylate buffer for 20 min. Fixed testes were cut into 3 mm3 and kept immersed in the fixative for further 15 min. Tissue cubes were then incubated at 37 degrees C in a medium containing cholesterol oxidase, 3,3'-diaminobenzidine tetrahydrochloride, horseradish peroxidase and dimethyl sulfoxide to histochemically localize free cholesterol in Leydig cells and processed for electron microscopy. Thin sections of these tissues were stained with aqueous uranyl acetate and lead citrate and examined with a Philips 201C electron microscope. In Leydig cells of control rats, free cholesterol was detected primarily in lipid droplets and plasma membrane. In the majority of Leydig cells, peroxisomes were unstained for free cholesterol, but occasionally few stained ones were present. Staining was not detected in mitochondria and smooth endoplasmic reticulum (SER) in Leydig cells of control rats. In LH-injected rats, lipid droplets, many peroxisomes, inner and outer mitochondrial membranes and some cisternae of SER in Leydig cells showed staining for free cholesterol. Fusion of Leydig cell peroxisomes with lipid droplets and mitochondria was also observed in the LH treated rats. These findings suggested that peroxisomes in adult rat Leydig cells participate in the intracellular cholesterol trafficking and delivery into mitochondria during LH stimulated steroidogenesis. Lipid droplets are used as one source for cholesterol for this process.