The pesticide methoxychlor given orally during the perinatal/juvenile period,reduced the spermatogenic potential of males as adults by reducing their Sertoli cell number

Perinatal and juvenile oral treatment of rats with the insecticide, methoxychlor (MXC), reduced testicular size and other reproductive indices including the number of epididymal spermatozoa in those animals as adults 161. The objective was to determine if these males exposed during development had fewer Sertoli cells which might explain these testicular effects. Rat dams were gavaged with MXC at 0, 5, 50, or 150 mg x kg(-1) x day(-1) for the week before and after they gave birth. Resulting male pups (15/group) then were dosed directly from postnatal day 7 to 42. Testes were fixed in Bouin's and in OsO4, embedded in Epon and sectioned at 0.5 microm, stained with toluidine blue, and evaluated stereologically or cut at 20 microm to measure Sertoli cell nuclei with Nomarski optics. Sertoli cell number was calculated as the volume density of the nucleus times the parenchymal weight (90% of testicular weight) divided by the volume of a single Sertoli cell nucleus. Across dose groups, there were no changes in the nuclear volume density, the volume of a single nucleus, or the number of Sertoli cells per g parenchyma. There were highly significant dose-related changes in the volume of Sertoli cell nuclei per testis and the number of Sertoli cells per testis. Reduced testicular weight (r = 0.94) and reduced numbers of epididymal spermatozoa (r = 0.43) were significantly (p < 0.01) correlated to reduced number of Sertoli cells per testis. Hence, perinatal and juvenile oral exposure to MXC can reduce spermatogenic potential of males as adults by reducing their number of Sertoli cells.     

Season but not age affects Sertoli cell number in adult stallions.

To evaluate the effect of age and season on Sertoli cell number per paired testes, ratio of germ cells per Sertoli cell, and daily sperm production, testes were obtained from 184 adult (4-20 yr) stallions at slaughter throughout one year. Numbers of Sertoli cells or germ cells were derived from nuclear volume density, volume of individual nuclei, and parenchymal volume. Germ cell to Sertoli cell ratios were calculated from cell numbers. Regression analysis was used to detect age-related differences in the breeding season (May-Jul) or throughout the year. A two-way analysis of variance was used to evaluate time periods (Nov-Jan, Feb-Apr, May-Jul, and Aug-Oct) and age groups (4-5.5, 6-12.5, or 13-20 yr). Paired parenchymal weight and daily sperm production per horse increased significantly with age. Neither regression nor analysis of variance revealed an effect of age on Sertoli cell number. While season contributed (p less than 0.01) to variation in Sertoli cell number per horse, there was no (p greater than 0.05) age x season interaction or age effect on Sertoli cell number. In testes obtained from adult stallions, age had no effect on the number of Sertoli cells per horse, the ratio of maturation-phase spermatids to Sertoli cells, or the ratio of all stage VIII germ cells to Sertoli cells. Given no age effect within a given season on Sertoli cell number per horse, the number of Sertoli cells in the recrudesced testis of the breeding season probably is not significantly different for a given stallion between 4 and 20 yr of age.     

A study of Sertoli-spermatid tubulobulbar complexes in selected mammals

Tubulobulbar complexes (TBCs) were found in nine mammalian species (opossum, vole, guinea-pig, mouse, hamster, rabbit, dog, monkey and human) primarily originating from the plasma membrane overlying the acrosome of late spermatids. Fewer complexes (4–10) were noted in these species than has been previously reported for the rat (up to 24). TBCs were not seen emanating from round spermatids or those elongated spermatids located within the deep recesses of the Sertoli cell, but they appeared as the spermatids came to reside much closer to the tubular lumen in preparation for release. TBCs developed in areas deficient or lacking in Sertoli filaments and endoplasmic reticulum (ectoplasmic specialization). In general their structural configuration was similar to that shown in the rat, although minor differences were noted. Fine fibrils were observed connecting the distal portion of the spermatid tube with the Sertoli plasma membrane forming a bristle-coated pit. The length of TBCs from most species studied was 1–2 μm, whereas those of the opossum extended 6–8 μm into an apical Sertoli process. TBCs were degraded within the Sertoli cell by its lysosomes prior to sperm release, and for most species there was evidence indicating that formation of more than one generation of TBCs occurred. As sperm release approached, TBCs formed preferentially from the leading edge of spermatids with spatulate heads. The Sertoli cell gradually withdrew from around the spermatid head until only the tip of the head was embedded within the Sertoli cell. This region of contact frequently demonstrated TBCs. The proposed functions of TBCs are reviewed and discussed in light of these findings from other species.     

An ultrastructural and autoradiographic study of the immune response in Hyalophora cecropia pupae

Summary Membrane-bounded spherical vesicles found in rat Sertoli cells have been examined quantitatively during the cycle of the seminiferous epithelium. Most of the vesicles were localized to the basal and columnar portions of the Sertoli cell cytoplasm. The thin lateral projections of the Sertoli cells contained very few vesicles. Morphometric analysis of the basal portion of the Sertoli cell cytoplasm revealed that the volume density (V _v ) of the vesicles changed markedly during the cycle. The V _v was at its minimum (0.036) at stage VII and maximum (0.117) at stages XI-I. The vesicles were also smaller at stage VII compared to the vesicles at stages IX-V. The stage-dependent difference in the size of the vesicles was found both in the basal and the columnar portions of the Sertoli cells. At stage VII some of the vesicles appeared to be elongated much like the tubular elements of the smooth endoplasmic reticulum (SER) from which they are probably derived. The stage-dependent differences in volume density and size of the Sertoli cell vesicles may be related to cyclic biochemical variations in the Sertoli cells, and are further indications of a variation in Sertoli cell function during the cycle of the seminiferous epithelium. Whether or not this is due to an internal cycle of the Sertoli cell or to influences from adjacent germ cells remains to be determined.     

Beyond Testis Size: Links between Spermatogenesis and Sperm Traits in a Seasonal Breeding Mammal

Spermatogenesis is a costly process that is expected to be under selection to maximise sperm quantity and quality. Testis size is often regarded as a proxy measure of sperm investment, implicitly overlooking the quantitative assessment of spermatogenesis. An enhanced understanding of testicular function, beyond testis size, may reveal further sexual traits involved in sperm quantity and quality. Here, we first estimated the inter-male variation in testicular function and sperm traits in red deer across the breeding and non-breeding seasons. Then, we analysed the relationships between the testis mass, eight parameters of spermatogenic function, and seven parameters of sperm quality. Our findings revealed that the Sertoli cell number and function parameters vary greatly between red deer males, and that spermatogenic activity co-varies with testis mass and sperm quality across the breeding and non-breeding seasons. For the first time in a seasonal breeder, we found that not only is the Sertoli cell number important in determining testis mass (r = 0.619, p = 0.007 and r = 0.248, p = 0.047 for the Sertoli cell number assessed by histology and cytology, respectively), but also sperm function (r = 0.703, p = 0.002 and r = 0.328, p = 0.012 for the Sertoli cell number assessed by histology and cytology, respectively). Testicular histology also revealed that a high Sertoli cell number per tubular cross-section is associated with high sperm production (r = 0.600, p = 0.009). Sperm production and function were also positively correlated (r = 0.384, p = 0.004), suggesting that these traits co-vary to maximise sperm fertilisation ability in red deer. In conclusion, our findings contribute to the understanding of the dynamics of spermatogenesis, and reveal new insights into the role of testicular function and the Sertoli cell number on testis size and sperm quality in red deer.     

A coupled morphological and biochemical study on the cellular localisation of the intra-adrenal renin granules in rats.

The effects of prolonged (3-week) sodium restriction on the rat zona glomerulosa (ZG) were investigated by ultrastructural stereological and biochemical techniques. The plasma level of aldosterone was increased, and this was coupled with a notable decrease in the volume density (Vv, microns 3/100 microns 3 of cell) of lipid droplets in ZG cells. Renin-like activity (RLA) underwent a significant rise in ZG, and Vv of dense bodies significantly rose in ZG cells. Since RLA and dense-body Vv displayed a highly significant linear correlation (r = 0.884; n = 22, P less than 0.01), the hypothesis is advanced that part of the dense bodies may be granules of prorenin or renin, and that ZG parenchymal cells are directly involved in the intra-adrenal renin production.     

Increased germ cell degeneration and reduced germ cell:Sertoli cell ratio in stallions with low sperm production

To determine the relationship between germ cell degeneration or germ cell:Sertoli cell ratio and daily sperm production, testes were obtained during the months of May to July (breeding season) and November to January (nonbreeding season) from adult (4 to 20-yr-old) stallions with either high (n = 15) or low (n = 15) sperm production. Serum was assayed for concentrations of LH, FSH and testosterone. Testes were assayed for testosterone content and for the number of elongated spermatids, after which parenchymal samples were prepared for histologic assessment. Using morphometric procedures, the types and numbers of spermatogonia, germ cells and Sertoli cells were determined. High sperm producing stallions had greater serum testosterone concentration, total intratesticular testosterone content, testicular parenchymal weight, seminiferous epithelial height, diameter of seminiferous tubules, numbers of A and B spermatogonia per testis, number of Sertoli cells per testis, and number of B spermatogonia, late primary spermatocytes, round spermatids and elongated spermatids per Sertoli cell than low sperm producing stallions (P < 0.05). The number of germ cells (total number of all spermatocytes and spermatids in Stage VIII tubules) accommodated by Sertoli cells was reduced in low sperm producing stallions (18.6 +/- 1.3 germ cells/Sertoli cell) compared with that of high sperm producing stallions (25.4 +/- 1.3 germ cells/Sertoli cell; P < 0.001). The conversion from (yield between) early to late primary spermatocytes and round to elongated spermatids was less efficient for the low sperm producing stallions (P < 0.05). Increased germ cell degeneration during early meiosis and spermiogenesis and reduced germ cell:Sertoli cell ratio was associated with low daily sperm production. These findings can be explained either by a compromised ability of the Sertoli cells to support germ cell division and/or maturation or the presence of defects in germ cells that predisposed them to degeneration.     

The origin and fate of spermatophores in the viviparous teleost Cymatogaster aggregata (Perciformes: Embiotocidae)

The sperm of the shiner surfperch are packaged into high density aggregations which are introduced into the female genital tract at insemination. Germ cell differentiation occurs within cysts formed by nongerminal Sertoli cells. In late spermiogenesis, spermatozoa within the cysts come to lie parallel to each other and become more densely packed. These sperm packets (spermatophores), containing approximately 600 spermatozoa, then are released into the efferent sperm ducts. The exact nature of the spermatophore binding material is not known, but a major component is proteinaceous and is synthesized in the rough endoplasmic reticulum of the efferent sperm duct epithelial cells. The mechanism by which the spermatophores pass from cysts into ducts is not clear. It appears that whereas many Sertoli cells degenerate causing the cyst wall to break down, many Sertoli cells do not degenerate, but rather assume the configuration of columnar duct cells. The spermatophores remain intact within the testicular ducts, but rapidly dissolve within the female ducts in response to increased pH.     

Quantification of the human Sertoli cell population: its distribution, relation to germ cell numbers, and age-related decline

The human Sertoli cell population was characterized in 14 men by histometric analysis and by direct counts of nuclei in testicular homogenates. Testes obtained at autopsy were perfused with glutaraldehyde and embedded in Epon. Nucleolar and nuclear volumes were determined by the formula of a sphere given the diameter of the nucleoli or average diameter of nuclei measured at the height and width. Nuclear volume was also estimated by adding volumes of nuclear profiles in 0.5-micron serial sections. Sertoli cell number/g was calculated by the product of the percentage nucleoli or nuclei in the parenchyma, parenchymal volume, and histologic correction factor divided by the volume of a single nucleolus or nucleus. Also, Sertoli cell nuclei were counted directly in homogenates of fixed parenchyma. Number of Sertoli cells/g was similar (P greater than 0.05) whether determined by serial sections or in homogenates, but the estimate based on the nucleolar method was higher (P less than 0.01) and the nuclear measurement method was lower (P less than 0.01) than that for serial sections. A group of 37 men aged 20 to 48 yr had significantly (P less than 0.01) more Sertoli cells than did 34 men aged 50 to 85 yr. It is concluded that: 1) the homogenate method is valid for quantification of the Sertoli cell population, 2) Sertoli cells are evenly distributed in different regions of the testis, 3) the average human Sertoli cell supports relatively few germ cells, 4) the human Sertoli cell population declines with age, and 5) there is a significant relationship between sperm production rates and number of Sertoli cells.     

The associations among semen quality,oxidative DNA damage in human spermatozoa and concentrations of cadmium,lead and selenium in seminal plasma

To explore the associations among semen quality, oxidative DNA damage in human spermatozoa and concentrations of cadmium, lead and selenium in seminal plasma, 56 non-smoking subjects were asked to collect semen by masturbation into a sterile wide-mouth metal-free plastic container after 3 days of abstinence. The conventional semen parameters were analysed. The concentrations of Cd, Pb and Se in seminal plasma were detected using atomic absorption spectrophotometer. 8-OHdG levels in sperm DNA were measured using HPLC-EC. The results showed that the geometric mean concentrations of Cd, Pb and Se were 0.78, 7.8 and 51.4 microg/l, respectively. The geometric mean of 8-OHdG/10(6) dG was 51.4 (95% CI: 21.5-123.0). A significant inverse correlation exists between Cd and sperm density (r=-0.28, P<0.05), and between Cd and sperm number per ejaculum (r=-0.27, P<0.05). In contrast, there was a significantly positive correlation between Se and sperm density (r=0.50, P<0.01), between Se and sperm number (r=0.49, P<0.01), between Se and sperm motility (r=0.40, P<0.01), and between Se and sperm viability (r=0.38, P<0.01). No statistically significant correlation was observed between Pb and semen quality. A significant inverse correlation was observed between 8-OHdG and sperm density (r=-0.34, P<0.01), between 8-OHdG and sperm number per ejaculum (r=-0.30, P<0.01), and 8-OHdG and sperm viability (r=-0.24, P<0.05). 8-OHdG was significantly correlated with Cd in seminal plasma (r=0.55, P<0.01). A significant but weak positive correlation was found between 8-OHdG and Pb concentration in seminal plasma (r=0.28, P<0.05). In contract, a significant inverse correlation was observed between 8-OHdG and Se concentration in seminal plasma (r=-0.40, P<0.01). The results indicate that Cd in seminal plasma could affect semen quality and oxidative DNA damage in human spermatozoa. Se could protect against oxidative DNA damage in human sperm cells. Pb did not appear to have any association with the semen quality when concentration of Pb in seminal plasma was below 10 microg/l.     

The role of sertoli cells in spermatid maturation in the testis of the crayfish, Procambarus paeninsulanus

With the onset of spermiogenesis, many changes become apparent in the crayfish spermatid during its transition to mature sperm. The nucleus passes through a series of stages, excess cytoplasm is removed, the acrosome develops, and nuclear arms form and become wrapped around the sperm prior to its enclosure in a capsule. Changes are also apparent in the Sertoli cells surrounding the germ cells in the crayfish testis. The amount of cytoplasm of individual Sertoli cells appears to increase in quantity and changes in the intracellular organelles become apparent. As spermiogenesis commences, the cytoplasm along one side of Sertoli cells adjacent to the spermatids is devoid of obvious organelles. Numerous finger/like projections of Sertoli cytoplasm penetrate into the spermatid and appear to isolate portions of the sperm cytoplasm. During later stages of spermiogenesis, several vesicles in the Sertoli cells which appear to contain droplets of this isolated sperm cytoplasm. appear to undergo lytic changes, As the amount of cytoplasm of the spermatid is reduced, contact is maintained between the spermatid and Sertoli cell in the area of the acrosome. The nuclear arms of the sperm extend into the Sertoli cell during their formation and later become wrapped around the acrosomal area of the sperm. At this time, very little space exists between the Sertoli cell and its many sperm. Large vesicles of electron dense material appear to be released by the Sertoli cells into the space between the sperm and Sertoli cell. This material completely surrounds the sperm and forms the sperm capsule. Spermiation involves the gradual dissolution of the points of contact between the sperm capsule and the Sertoli cell.     

Structural characteristics of immature rat Sertoli cells in vivo and in vitro.

The structural properties of pelleted prepubertal Sertoli cells (pre-culture pelleted cells) from 19-day-old rats and of similar cells cultured for 7 days were compared with Sertoli cells from the intact animal (testis tissue from 19- and 26-day-old rats, the in vivo groups). Sertoli cells from freshly isolated pellets and those cultured for 7 days were similar in cell and nuclear volumes to their in vivo counterparts. Cell volumes, organelle volumes, and organelle volume densities of newly isolated Sertoli cells were similar to those of sectioned cells taken from the 19-day-old in vivo group, indicating that the procedure for isolation does not grossly alter Sertoli cells. Mean height of cells cultured for 7 days was significantly lower than that of cells from intact animals at 19 and 26 days of age. In vivo, Sertoli cells of 26-day-old animals displayed increased organelle volumes and organelle surface areas compared with those from 19-day-old animals; volume densities and surface densities remained relatively constant, indicating that in vivo, organelle growth is in proportion to growth of the cell. Most organelle volume and surface densities were not significantly different when 19-day-old in vivo cells and pre-culture pelleted cells were compared. Many organelle volume and surface density values were significantly less in cells grown in culture for 7 days as compared to freshly isolated pelleted cells. After 7 days of culture, most Sertoli cell organelles were significantly less in both volume density and surface density, as compared to the in vivo cell groups (19 or 26 day). This indicates that in vitro the organelles do not develop in proportion to the growth of the cell. After 7 days in culture, the absolute volumes and surface areas of the organelles remained generally unchanged as compared to cells from 19-day-old animals. The data show that Sertoli cells grow in volume in vitro like their in vivo counterparts; however, their subcellular features, although well maintained, do not develop in proportion to the cell. This suggests that short-term cultures are a more ideal system in which to study biochemical responses. Also, cultured prepubertal Sertoli cells are most appropriately used to study prepubertal Sertoli cell function. This is the first study to quantify developmental changes in Sertoli cell structure in vivo as well as to compare them with cellular changes occurring in vitro.     

Bull sperm surface "craters" and other aspects of semen quality

Semen from 200 Holstein bulls in an artificial insemination center was examined for the frequency of craters on the surface of sperm heads, as visualized with the aid of differential interference contrast microscopy. Semen from 100 of these bulls was examined in more detail in 2 experiments by staining with eosin-aniline blue to determine the relationship of unstained spermatozoa, and spermatozoa with normal acrosomes with apical ridges to the incidence of craters and fertility. Only 3 of 100 bulls had a substantial incidence of craters (15 to 23%), whereas the average of the other 97 bulls in 2 experiments was 1 to 3%. The percentage of sperm cells with craters was correlated (P < 0.05) with the percentage of unstained spermatozoa (r = -0.29 and sperm cells with normal acrosomes (r = -0.52) but was not significantly correlated (r = -0.24) with the nonreturn rate. One bull with many sperm cells with craters was slaughtered, and the epididymal spermatozoa were examined. The high incidence of sperm cells with craters was limited to one side, with the testis on that side having 2 Sertoli cell tumors. The remaining 2 bulls as well as one other that produced 16% of sperm cells with craters did so only temporarily. Within a few months crater sperm production had decreased and semen quality increased. The condition usually appears to be transitory, presumably due to temporary stress.     

Annual cycle of the Sertoli cell population in adult stallions

Stereological methods were employed in two experiments with adult stallions: to confirm seasonal variation in number of Sertoli cells and to characterize the annual cycle of the Sertoli cell population. In the first experiment, testes from 28 adult (4-20 years old) horses obtained in the non-breeding season (December-January) were compared to testes from 28 adult horses in the breeding season (June-July). Sertoli cell numbers were calculated from the nuclear volume density, parenchymal volume, and volume of an individual Sertoli cell nucleus determined by reconstruction of serial sections or from average height and width measurements. The number of Sertoli cells per testis was significantly greater in the breeding season. In a second experiment involving 43-48 adult horses in each 3-month period, the Sertoli cell population was higher (P less than 0.05) in May-July than other periods and higher (P less than 0.01) than in November-January. These combined studies confirm seasonal differences in the Sertoli cell numbers per testis and define the annual cycle of the Sertoli cell population in adult stallions.     

Quantity rather than quality in teratospermic males: a histomorphometric and flow cytometric evaluation of spermatogenesis in the domestic cat (Felis catus)

Teratozoospermia (ejaculation of <40% morphologically normal sperm) commonly occurs within the Felidae, including certain domestic cats, but the cellular and molecular mechanisms that give rise to this phenomenon remain unknown. This study quantified spermatogenesis to identify differential dysfunctions in teratospermic versus normospermic (>60% normal sperm/ejaculate) domestic cats. Sperm used were from electroejaculates and cauda epididymides. Testes from 10 normo- and 10 teratospermic males were obtained by castration and then evaluated by histomorphometry, flow cytometry, and testicular testosterone enzyme immunoassay. Some morphometric traits (tubular diameter, epithelium height, interstitial area, number of Leydig cells, and blood vessels per cross-section) as well as testicular testosterone concentrations were similar between groups, but testicular volume was greater in teratospermic males. Stage frequencies differed also between both cat populations, suggesting possible dysfunctions in spermiation. Quantification of cell populations in most frequent stages revealed more spermatogenic cells and fewer Sertoli cells per tubule cross-section as well as per tissue unit in teratospermic donors. Hence, the ratio of spermatogenic cells per Sertoli cell was elevated in the teratospermic cat. DNA flow cytometry confirmed higher total spermatogenic and meiotic transformations in teratospermic males. In summary, compared with normospermic counterparts, teratospermic cats have a higher sperm output achieved by more sperm-producing tissue, more germ cells per Sertoli cell, and reduced germ cell loss during spermatogenesis. Gains in sperm quantity are produced at the expense of sperm quality.     

Testis morphometry,seminiferous epithelium cycle length,and daily sperm production in domestic cats (Felis catus)

There is very little information regarding the testis structure and function in domestic cats, mainly data related to the cycle of seminiferous epithelium and sperm production. The testis weight in cats investigated in the present study was 1.2 g. Compared with most mammalian species investigated, the value of 0.08% found for testes mass related to the body mass (gonadosomatic index) in cats is very low. The tunica albuginea volume density (%) in these animals was relatively high and comprised about 19% of the testis. Seminiferous tubule and Leydig cell volume density (%) in cats were approximately 90% and 6%, respectively. The mean tubular diameter was 220 microm, and 23 m of seminiferous tubule were found per testis and per gram of testis. The frequencies of the eight stages of the cycle, characterized according to the tubular morphology system, were as follows: stage 1, 24.9%; stage 2, 12.9%; stage 3, 7.7%; stage 4, 17.6%; stage 5, 7.2%; stage 6, 11.9%; stage 7, 6.8%; and stage 8, 11 %. The premeiotic and postmeiotic stage frequency was 46% and 37%, respectively. The duration of each cycle of seminiferous epithelium was 10.4 days and the total duration of spermatogenesis based on 4.5 cycles was 46.8 days. The number of round spermatids for each pachytene primary spermatocytes (meiotic index) was 2.8, meaning that significant cell loss (30%) occurred during the two meiotic divisions. The total number of germ cells and the number of round spermatids per each Sertoli cell nucleolus at stage 1 of the cycle were 9.8 and 5.1, respectively. The Leydig cell volume was approximately 2000 microm3 and the nucleus volume 260 microm3. Both Leydig and Sertoli cell numbers per gram of testis in cats were approximately 30 million. The daily sperm production per gram of testis in cats (efficiency of spermatogenesis) was approximately 16 million. To our knowledge, this is the first investigation to perform a more detailed and comprehensive study of the testis structure and function in domestic cats. Also, this is the first report in the literature showing Sertoli and Leydig cell number per gram of testis and the daily sperm production in any kind of feline species. In this regard, besides providing a background for comparative studies with other fields, the data obtained in the present work might be useful in future studies in which the domestic cat could be utilized as an appropriate receptor model for preservation of genetic stock from rare or endangered wild felines using the germ cell transplantation technique.     

妊娠期和哺乳期大鼠胰岛生长抑素免疫反应阳性细胞的变化

本文用免疫细胞化学和体视学方法,观察妊娠和哺乳期间大鼠胰岛生长抑素（ＳＳ）免疫反应阳性细胞（细胞）的变化。结果显示妊娠期和哺乳期胰岛Ｄ细胞面数密度（ＮＡ）、数密度（Ｎｖ）和体密度（Ｖｖ）值升高,且染色深度加强,提示妊娠期和哺乳期胰岛Ｄ细胞合成ＳＳ增加。关键词     

Spermatozoon head size – the main differentiating feature between spermatozoa of blue and white Arctic fox (Vulpes lagopus)

Morphology and sperm morphometry, this is an important determinant of male reproductive capacity. Morphometric data may provide relevant information in studies focused on evolutionary biology, sperm quality assessment, including prediction of the potential fertility, semen cryopreservation, or the effect of reprotoxicants. The paper presents the morphometric analysis of spermatozoa from two colour morphs of Arctic fox (Vulpes lagopus), and attempts to determine the relationship between selected quality indicators and dimensions and shape of spermatozoa. The research material consisted of ejaculates collected once by manual stimulation from 20 one-year-old Arctic foxes (10 individuals of the blue morph and 10 of the white morph). Ejaculates were analysed for standard parameters (volume, sperm concentration, total number of spermatozoa in the ejaculate) and used for the preparation of microscopic specimens. It was found that, the dimensions of spermatozoa from Arctic foxes depend on the male colour morphs. Spermatozoa from white Arctic foxes were significantly longer (by 1.82 µm) and had larger heads (0.32 µm longer and 0.15 µm wider) compared to spermatozoa from blue Arctic foxes (P<0.05). The interactions between particular sperm dimensions indicated the occurrence of gametes differing in shape. The all correlation coefficients between the morphometric traits of spermatozoa were statistically significant. Our research proved that in the blue Arctic foxes, sperm dimensions (tail length and total sperm length) can be related to the percentage of spermatozoa with primary changes (respectively: r = -0.68 and r = -0.75; at P <0.05). However, in the case of white Arctic foxes, these characteristics depend on the ejaculate volume (respectively: r = 0.65 and r = 0.68; at P <0.05).     

Fine needle aspiration biopsy of the breast. Value of nuclear morphometry after different sampling methods

OBJECTIVE: To study the potential of nuclear morphometry in supporting the interpretation of fine needle aspiration biopsy (FNAB) samples of the breast fixed in 50% ethanol and centrifuged on slides. STUDY DESIGN: Computerized morphometry was used to outline the nuclei of breast epithelial cells in breast cancer, fibroadenoma and fibrocystic disease. The diagnoses were histologically confirmed. We applied 2 different sampling methods (measurements done on cell groups and on free cells). RESULTS: The mean nuclear area of cell groups of malignant samples (23) varied from 42 to 125 microns 2, in fibroadenomas from 30 to 50 microns 2 and in fibrocystic disease from 26 to 57 microns 2. The mean nuclear area of free cells varied as follows: cancer, 66-181 microns 2; fibroadenoma, 33-70 microns 2; fibrocystic disease, 35-60 microns 2. Apocrine metaplasia was excluded from comparison on a morphologic basis. CONCLUSION: The study suggests that if the mean nuclear area of cell groups is < 42 microns 2, the lesion is probably benign; if > 57 microns 2, and apocrine metaplasia is excluded, malignancy should be considered. The differential diagnosis between carcinoma and fibroadenoma could be based on free cells: mean area of free cell nuclei < or = 65 microns 2 suggested a benign lesion, and of > or = 71 microns 2 suggested a malignant lesion. Morphometric nuclear size features (exemplified by nuclear area) appeared efficient in distinguishing between malignant and benign lesions when measured from free cells and cell groups.     

Seasonal differences in equine spermatocytogenesis

Spermatocytogenesis plays a pivotal role in regulation of spermatogenesis; however, its details remain relatively obscure in nonrodent species. The equine testis contains approximately 100% more spermatogonia in summer than in winter and appears to be a good model to identify the flexible components of spermatocytogenesis that cause seasonal changes in daily sperm production. Testes were taken from horses in the winter (n = 47) and in summer (n = 43). Tissues were fixed by glutaraldehyde-perfusion and submission in osmium, embedded in Epon or methacrylate, sectioned at 0.5 micron or 5 microns, stained with toluidine blue, and observed using bright-field microscopy. The combined total number of A1, A2, A3, and B1 (A plus B1) spermatogonia/testis and the numbers of B2 spermatogonia or early primary spermatocytes were determined by stereology of Epon sections involving testicular volume density and volume of spermatogonial nuclei. In a subset of horses, different spermatogonial subtypes (A1, A2, A3, and B1) were counted per 100 Sertoli cells in each of the 8 spermatogenic stages and expressed as percentage of all A plus B1 spermatogonia. The number of each spermatogonial subtype/testis for the large series of horses was calculated by multiplying the number of A plus B1 spermatogonia/testis (determined for each horse) by the percentage of that given spermatogonial subtype. Season did not significantly affect the number of any given subtype per 100 Sertoli cells in any stage or percentages of different subtypes of spermatogonia. Numbers of A1 (p less than 0.05), A2, A3, B1, or B2 spermatogonia (p less than 0.01) were greater in the breeding season.(ABSTRACT TRUNCATED AT 250 WORDS)