Showing papers in "Journal of Andrology in 1986"

Morphometric analysis of spermatozoa in the assessment of human male fertility.

Abstract: A videomicrographic system was developed for measurement of morphometric parameters of human spermatozoa. Contours of the images of spermatozoa on a video monitor are digitized by manually tracing them with the cursor of an electromagnetic digitizer integrated to a microcomputer. The accuracy and precision of the methodology were evaluated. A comparison of human sperm heads in shallow wet preparations and in dried, stained preparations indicated that the latter were smaller in length, width, projected area, and circumference, but that the ratio length/width was not different. An analysis was made of 457 ejaculates from 16 fertile donors. The variation between ejaculates within donors was similar in magnitude to the variation between donors. A study was performed comparing seminal sperm morphometry in single specimens from 30 fertile and 30 infertile men. The sperm head length/width ratio was the parameter that differed the most between these two groups. Moreover, it was the per-ejaculate variability of this parameter, rather than the central tendency, that maximized the difference.

The effect of selective destruction and regeneration of rat Leydig cells on the intratesticular distribution of testosterone and morphology of the seminiferous epithelium.

Abstract: This study was designed to explore the relationship between the intratesticular distribution of testosterone and spermatogenesis by completely destroying the Leydig cells of mature male rats with injection of a single i.p. dose of ethane dimethanesulphonate. After such treatment, testosterone levels in serum, testicular interstitial fluid, seminiferous tubules, and whole testis declined significantly 6 to 24 hours after injection and fell below assay detection limits between 3 and 7 days. At 3 and 7 days, serum LH and FSH levels rose significantly and remained elevated up to 4 and 6 weeks, respectively, in comparison with vehicle-treated controls. Leydig cells disappeared from the interstitium by day 3, but between 2 and 4 weeks postinjection a new generation of fetal-like Leydig cells repopulated the testicular interstitium and, during weeks 6 to 10, were transformed into, or replaced by, Leydig cells with an adult type of morphology. Histologic examination of the seminiferous tubules showed progressive disruption of spermatogenesis between 3 and 14 days post-ethane dimethanesulphonate. The first histologic sign of spermatogenic damage was noted at day 3, with the occurrence of stage-specific degenerating pachytene primary spermatocytes at stages VII to VIII of the spermatogenic cycle. On day 7, these cells and degenerating round, or step 19, spermatids often were observed during stages VII to XI, although qualitatively normal spermatogenesis also was seen in these and all other stages of the cycle. Maximum impairment of spermatogenesis occurred 2 weeks post-ethane dimethane sulphonate, at which time the tubules commonly lacked one or more germ cell generations or, alternatively, showed accumulation of lipid inclusions, extracellular spaces, and variable numbers of degenerating germ cells. Following repopulation of the testis by Leydig cells during weeks 3 and 4, spermatogenesis recovered. By 10 weeks after treatment, qualitatively normal spermatogenesis was seen in the great majority of seminiferous tubules, although a few tubules still remained in which the germ cell complement was severely reduced, and contained only Sertoli cells and spermatogonia.(ABSTRACT TRUNCATED AT 400 WORDS)

Growth and Characterization of Polarized Monolayers of Epididymal Epithelial Cells and Sertoli Cells in Dual Environment Culture Chambers

Abstract: Epididymal epithelial cells isolated from mature rats and Sertoli cells isolated from 10-day-old rats were cultured in serum-free defined media on extracellular matrix impregnated filters maintained in dual environment culture chambers. Epididymal epithelial cells had a polarized appearance only when plated at high density (greater than 1 X 10(6) cells/cm2). Confluent monolayers of these cells formed a permeability barrier to inulin. Sertoli cells were columnar and highly polarized when grown on extracellular matrix-impregnated filters, cuboidal when grown on filters alone, and squamous when grown on plastic. Confluent polarized monolayers of these cells excluded the electron-dense tracer lanthanum nitrate by way of basal-tight junctions. Therefore, polarized monolayers of epididymal epithelial cells and Sertoli cells can be obtained by growing the cells at high density on extracellular matrix-impregnated permeable supports. By maintaining the monolayers in specially constructed culture chambers, the cells can develop a permeability barrier, and are able to achieve the separation of apical from basal compartments so important for their function in vivo.

Histopathology of Prepubertal Rat Testes Subjected to Various Durations of Spermatic Cord Torsion

Abstract: Sixty prepubertal rats were subjected to unilateral spermatic cord torsion for a duration of 0, 1, 3, 5, 9 or 12 hours. At the end of this period, the ipsilateral testes either were removed for immediate processing or subjected to detorsion and orchiopexy, followed by a six-week recovery period prior to histologic study. Twelve histologic parameters were each scored according to the degree of pathologic findings, thus allowing for a quantitative assessment of testicular damage. The sequence of specific histologic degeneration that occurred with spermatic cord torsion is described. These changes were found to be dependent on the duration of torsion, with the greatest damage occurring after three hours or more. In the animals undergoing detorsion followed by a six-week recovery period, severe degeneration was noted for all durations of torsion studied. The extent of this degeneration was significantly correlated with a reduction in fertility.

Correlation between human sperm swelling in hypoosmotic medium (hypoosmotic swelling test) and in vitro fertilization.

Abstract: Human ejaculates (n = 83) were analyzed for standard sperm parameters (concentration, motility, and morphology), as well as for the ability of the spermatozoa to react (swell) in a hypoosmotic medium (Jeyendran et al, 1984) Subsequently, the fertilizing capacity of the spermatozoa was tested by their ability to fertilize human oocytes in vitro Although the sperm concentration was adjusted for in vitro fertilization, no adjustments were made for sperm motility and morphology Correlation of the in vitro fertilizing capacity of the spermatozoa with the hypoosmotic swelling test (r = 056) was much higher than with standard sperm parameters (r varied from -004 to 025) Complete overlap was noted with standard semen parameters whether the ejaculate did or did not fertilize oocytes and ranged from very low to very high values in both cases By contrast, all the semen samples that fertilized oocytes showed a 60% or higher reaction in the hypoosmotic swelling test, whereas the majority of the "infertile" semen samples showed less than 60% swelling It therefore appears that, under the conditions of our studies, the hypoosmotic swelling test is a more accurate predictor of successful in vitro fertilization outcome than the conventional semen parameters A combination of all parameters, however, is likely to be most useful The hypoosmotic swelling test is simple and economical, and it is recommended that this test be further scrutinized for its value as an additional tool in the assessment of the in vivo fertilizing capacity of ejaculated spermatozoa

Polarized Sertoli cell functions in a new two-compartment culture system.

Abstract: Sertoli cells in vivo are highly polarized and interact with the inner (tubular) and outer (interstitial) fluids. To simulate this condition in vitro we developed a two-compartment culture system in which confluent Sertoli cell monolayers were grown on permeable supports (Millipore filters) separating the inner and outer fluid compartments. Monolayer permeability to (3H)-inulin decreased by 90% after 5 to 7 days of culture, presumably due to formation of tight junctions. This process was influenced by cell plating density. The cells were highly polarized morphologically, resembling their appearance in vivo, and secreted transferrin bidirectionally into both fluid compartments. The amount of transferrin secreted was 166% to 250% of that secreted by the same number of Sertoli cells cultured in plastic dishes. Testosterone (5 X 10(-8) M) doubled and testosterone + FSH (0.1 microgram/ml) increased transferrin secretion 3.6-fold. These results demonstrate that under suitable culture conditions the Sertoli cells remain both morphologically and functionally polarized, reflecting a more physiologic state.

Age-related variation in seminiferous tubules in men. A stereologic evaluation.

Abstract: Tubular boundary tissue and seminiferous epithelia were evaluated stereologically in testes from 28 men aged 20 to 48 years and 28 men aged 50 to 90 years. Testes obtained at autopsy within 15 hours of death were perfused with glutaraldehyde, embedded in Epon (Ladd Research Industries, Inc., Burlington, VT), sectioned at 0.5 micron, and stained with toluidine blue. Volume densities (percentage of the testicular parenchyma) of various parameters determined by point counting and diameter measurements were used to calculate total volumes, length of tubules, and number of cells. Electron microscopy was used to determine the volume density of myoid cells in the boundary tissue. Significant (P less than 0.01) age-related reductions occurred in paired testicular weights, paired parenchymal weights, total volume of seminiferous tubules and of seminiferous epithelium, and length of tubules. The volume density and thickness of boundary tissue increased (P less than 0.01) with age. The volume of boundary tissue per man and the volume density of myoid cells in the boundary tissue did not vary with age. Although the number of myoid cells per man tended to be lower in the older group, the number of myoid cells per cross section of seminiferous tubule was increased (P less than 0.01) in older men. The age-related thickening of the boundary tissue was not due to an increase in boundary tissue but resulted from a reduction in the length of the seminiferous tubules.

Repair of experimental varicoceles in the rat. Long-term effects on testicular blood flow and temperature and cauda epididymidal sperm concentration and motility.

Abstract: The effects of varicocele and varicocele repair on testicular blood flow, temperature, sperm counts, and sperm motility were assessed in adult male rats. The duration of the experimental varicocele and the varicocele repair were three and two times as long, respectively, as that studied previously. Varicoceles were created by partial ligation of the left renal vein and repairs were accomplished by high ligation of the left spermatic vein. Testicular blood flow was determined by using the radiolabeled microsphere technique. Testicular temperature was taken via needle probe thermometer. Sperm samples were obtained by micropuncture of the cauda epididymidis, and were counted on a hemacytometer and observed for motility under the light microscope. Varicoceles were studied 100 days after their creation. Repairs were performed on varicoceles that had lasted 100 days and the animals were studied 60 days after repair. Mean testicular blood flow (ml/100 g tissue/min) was significantly increased (P less than 0.05) in animals with varicocele (left testis (LT) = 42.2 +/- 1.1, right testis (RT) = 39.1 +/- 1.2) when compared with normal controls (LT = 29.3 +/- 1.6, RT = 29.6 +/- 1.7), animals with varicocele repair (LT = 30.7 +/- 1.3, RT = 30.0 +/- 1.6), or sham-operated animals (LT = 29.7 +/- 1.4, RT = 31.1 +/- 1.4).(ABSTRACT TRUNCATED AT 250 WORDS)

α-Glucosidase as a Specific Epididymal Enzyme Marker Its Validity for the Etiologic Diagnosis of Azoospermia

Abstract: Azoospermic semen was obtained from 39 vasectomized men and 93 patients consulting for infertility. The latter underwent bioclinical investigations including measurement of plasma FSH and testicular biopsies. Carnitine content and alpha-glucosidase activity in semen samples were measured. The activity of alpha-glucosidase was determined systematically by a semiquantitative microtechnique and was verified by a spectrophotometric assay. A positive correlation was observed between carnitine and alpha-glucosidase activity. Both parameters were severely diminished in semen from the vasectomized men and the patients suffering from a complete obstruction of the genital tract. Enzyme activity was the most discriminant parameter in terms of sensitivity and specificity. The measurement of alpha-glucosidase in semen is a simple and sensitive method for determining the origin of azoospermia when used in conjunction with assays for plasma FSH.

Dose and time relationships in the endocrine response of the irradiated adult rat testis.

Abstract: The dose- and time-dependent responses for the interstitial and tubular compartments in irradiated adult rat testes are described. Leydig cell dysfunction, as indicated by increased serum LH (to a maximum of 385% of control after 5 Gy) and decreased serum T (to a minimum of 30% of control after 10 Gy), was observed at 8 weeks postirradiation. Subsequent recovery of Leydig cell function was then observed, so that after 9 months serum T was normal but LH was still marginally elevated. The dysfunction, with a threshold of about 4 to 5 Gy, was associated with a loss of Leydig cells from the testis. Spermatogenic damage was observed; after doses of 3 Gy and above a marked dose-response was recorded as assessed by counts of tubule cross sections exhibiting spermatogenesis. Reduced serum levels of androgen binding protein indicated Sertoli cell dysfunction at 8 weeks after 3 Gy and above, with values of less than one half of those seen in the controls. Serum FSH also was elevated to between 150% and 200% of control, and after 9 months closely reflected androgen binding protein changes. Unlike the Leydig cell, no recovery with time was observed for this aspect of Sertoli cell function.

Transferrin-mediated transcellular transport of 59Fe across confluent epithelial sheets of Sertoli cells grown in bicameral cell culture chambers.

Abstract: The transferrin-mediated transcellular transport of 59Fe across confluent epithelial sheets of Sertoli cells grown on Millipore filters was investigated. These filters had been impregnated with reconstituted basement membrane and suspended in bicameral (two houses) culture chambers. After five days of culture, Sertoli cells from 10-day-old rats formed basally-located tight junctional complexes. Concomitantly, there was an increase in electrical resistance and the epithelial sheet became impermeable to lanthanum nitrate. The rate of passage of [3H]inulin across the epithelial sheet was considerably less than passage across a filter alone, a filter impregnated with reconstituted basement membrane or an epithelial sheet pretreated with 2 mM EGTA. We conclude from these permeability studies that the tight junctional complexes between Sertoli cells formed an effective transepithelial permeability barrier. Following addition of human serum [59Fe]transferrin to media bathing the basal cytoplasm of the cells, rat testicular [59Fe]transferrin was immunoprecipitated from apical media overlying the Sertoli cells. Cross-reactivity of the rabbit anti-rat transferrin antibody with human serum transferrin was less than 0.001%. Substitution of the primary antibody with normal rabbit serum reduced the amount of immunoprecipitable rat testicular [59Fe]transferrin to 20% of normal levels. Prior fixation of the Sertoli cell epithelial sheet in 2.5% glutaraldehyde, addition of a 100-fold excess of holotransferrin to the basal media, and incubation of the Sertoli cell epithelial sheet at 4 C all reduced the immunoprecipitable rat testicular [59Fe]transferrin in apical media to levels below that for the non-specific binding of the primary antibody. From these studies we conclude that 59Fe is shuttled across Sertoli cells by two different forms of transferrin. Serum transferrin delivers the 59Fe to the basal cytoplasm of the Sertoli cells. The 59Fe dissociates from the serum transferrin, is delivered to testicular transferrin, and is subsequently secreted from the apical surface of the epithelial sheet of Sertoli cells as testicular [59Fe]transferrin.

Prognosis of subfertility in men with corrected or uncorrected varicocele.

Abstract: To evaluate whether correction of varicocele improves fertility, pregnancy rates were compared in 115 varicocele patients consulting for infertility and having oligozoospermia, asthenospermia, or teratozoospermia, in any combination, and FSH levels within the normal range. Ninety of these patients had corrected, and 25 had uncorrected varicoceles, respectively. The value of clinical and seminal parameters for predicting the eventuality of pregnancy for varicocele patients was also studied. Although both groups were comparable in terms of duration of infertility, mean age, sperm density, motility or fertility index, cumulative pregnancy rates over 12 months were similar, whether or not the varicocele was corrected. The value of clinical or seminal parameters, in any combination, for the prediction of outcome for varicocele patients was poor. The prognosis was poor for men with less than 15% of spermatozoa with normal morphology, FSH levels higher than the mean + 3 SD of those values found in young fathers, and a fertility index below 3. In subjects who achieved pregnancy within one year, pretreatment sperm characteristics were similar in both the corrected and uncorrected groups. Correction of varicocele slightly improved sperm characteristics. It seems likely that in most men with subfertility and varicocele, other factors besides venous reflux are responsible for their infertility.

Factors influencing the success of sperm-cervical mucus interaction in patients exhibiting unexplained infertility.

Abstract: The factors regulating the success of sperm-cervical mucus interaction in 46 patients exhibiting unexplained infertility were analyzed. Within this group of patients, considerable variation in the degree of mucus penetration, which appeared to be related to the properties exhibited by the spermatozoa rather than the quality of the mucus itself, was observed. The ability of the spermatozoa to fuse with zona-free hamster oocytes reflected their capacity for mucus penetration, regardless of whether human or bovine cervical mucus was used as the target. Multiple regression analysis also indicated the importance of the movement characteristics exhibited by the spermatozoa, which alone could account for up to 85% of the variability in mucus penetration. The concentration of motile spermatozoa and their linear velocity of progression influenced the number of spermatozoa penetrating the mucus in unit time, presumably through an effect on the frequency of collisions at the cervical mucus interface. Whether these collisions resulted in mucus penetration depended upon the movement characteristics of the spermatozoa and was positively associated with a "rolling" mode of progression and the amplitude of lateral sperm head displacement.

Laser Light‐scattering Study of the Toxic Effects of Methylmercury on Sperm Motility

Abstract: An in vitro study was designed using the laser light-scattering technique to obtain further information on the dose-effect relationship of methylmercury on sperm motility. The technique provided a quantitative evaluation of sperm swimming speed. Semen samples were collected from normal male Macaca fascicularis monkeys by anal electroejaculation. Methylmercury was added to aliquots of sperm suspensions in BWW medium in doses of 10, 5, 2, and 1 ppm. After 3 hours, the relative speed was 35%, 59%, 69%, and 92% of the corresponding controls at doses of 10, 5, 2, and 1 ppm, respectively. The percentage of motile spermatozoa decreased significantly at 10 ppm. By microscopic observation abnormal motility was detected at 5 and 10 ppm, especially after 20 to 40 minutes. Head movement increased from side to side, and many spermatozoa developed coiled tails. The technique proved useful for defining the dose-effect relationship of methylmercury and sperm swimming speed.

The incidence of spermatic granulomas and their relation to testis weight after vasectomy and vasovasostomy in Lewis rats.

Abstract: The occurrence of spermatic granulomas of the vas deferens was studied in Lewis rats at intervals up to 7 months after vasectomy or vasectomy followed 3 months later by vasovasostomy. The incidence of granuloma progressed with time to involve one or both tracts in 100% of vasectomized rats. In addition, the majority of animals developed new granulomas after vasovasostomy, even though fluid flow through the reconnected vas deferens was demonstrated in vitro. When individual tracts were analyzed, the weight of the testis was related to ipsilateral spermatic granuloma formation in both vasectomy and vasovasostomy groups at 3 and 4 months after initial operation. Testes were small in the absence of a granuloma but similar to those of sham-operated rats if a granuloma was present. The possible protective effect of spermatic granuloma formation on the testis is discussed.

Intratesticular factors and testosterone secretion. Effect of treatments that alter the level of testosterone within the testis.

Abstract: The authors recently have reported the presence of a nongonadotropic polypeptide factor in rat testicular interstitial fluid that can exert marked stimulatory effects on Leydig cell testosterone production. To assess the potential physiologic significance of this factor, its effective levels in rat interstitial fluid have been investigated in response to treatments that either markedly reduce interstitial fluid testosterone concentrations (anti-LH treatment; transient or chronic experimental cryptorchidism; destruction of Leydig cells with ethane dimethanesulphonate) or that significantly elevate testosterone levels in interstitial fluid by injection of hCG. The possible relationship between this factor and changes in testicular weight, serum LH and FSH, and interstitial fluid volume also were monitored. When testosterone levels in interstitial fluid were decreased by 75 to 99% either acutely (5-72 hours) or chronically (20-75 days), there was an accompanying increase (P less than 0.001) in the levels of the interstitial fluid factor(s), as determined by the ability of charcoal-stripped interstitial fluid from individual rats to enhance hCG-stimulated testosterone production by Percoll-purified Leydig cells in vitro. Anti-LH treatment increased the levels of the interstitial fluid factor(s) over the ensuing 5 to 48 hours. In abdominal testes from rats made unilaterally or bilaterally cryptorchid for 20 or 55 days, a decrease in interstitial fluid testosterone levels was associated with increased levels of the interstitial fluid factor(s). The same inverse relationship was found 72 hours after treatment with ethane dimethanesulphonate in which Leydig cells had disappeared from the testis.(ABSTRACT TRUNCATED AT 250 WORDS)

Markedly Delayed Puberty or Kallmann's Syndrome Variant

Abstract: A diagnosis of Kallmann's syndrome was made in a 25-year-old man After 21 months of treatment with parenteral T, spontaneous puberty occurred at the age of 27

Impaired estrogen production by Leydig cells of the naturally retained testis in unilaterally cryptorchid boars and stallions.

Abstract: Estrogen production in vitro was compared for Leydig cells from cryptorchid and scrotal testes in boars and stallions. Animals with natural and experimental cryptorchidism were used. Purified Leydig cells were prepared from testes of mature animals by collagenase treatment and Percoll density gradients. After incubation for 3 hours (1 X 10(6) cells), estrone sulfate and estrone in the media were measured by direct radioimmunoassay. Androstenedione and testosterone in media extracts also were determined. Cells from the abdominal testis of unilateral cryptorchid boars and stallions showed impaired estrogen production compared with that of the contralateral scrotal testis. Surgical translocation of the scrotal testis to the abdominal cavity in four unilaterally cryptorchid, prepubertal boars did not result in a reduced capacity for estrogen secretion by Leydig cells examined after puberty. Cells from the naturally retained testis in each of these four animals produced practically no estrogen. In a naturally bilateral cryptorchid stallion, there was a high rate of estrogen secretion by both testes. It was concluded that the scrotal testis of a unilaterally cryptorchid animal exerts a suppressive influence on estrogen formation by the abdominal testis.

Accessory sex gland function in normal young (20-25 years) and middle-aged (50-55 years) men.

Abstract: A comparison of various parameters of prostatic and vesicular secretory function was made between the seminal plasma of young (20-25 years; n = 23) and middle-aged (50-55 years; n = 19) male volunteers These parameters included prostatic acid phosphatase, zinc, citric acid, spermine, spermidine, putrescine (prostatic origin), fructose, and prostaglandin E (vesicular origin), in addition to protein and testosterone Spermatozoa were counted and monitored for abnormalities The concentration in the ejaculate of the majority of the parameters investigated did not change with age, although the total contribution to the ejaculate from the prostate and seminal vesicles was reduced significantly in the older men The concentration of three constituents was significantly altered in the older age group: putrescine (P less than 0001) and prostaglandin E (P less than 001) were reduced, while zinc levels were elevated (P less than 005) These changes are discussed in relation to possible disturbances of prostate function and pathology in the middle-aged man

Spectrum of the pulsatile characteristics of LH release in normal men.

Abstract: To assess the spectrum of LH pulse characteristics in normal men, blood samples from 36 individuals were drawn at 20-minute intervals for 8 hours. The subsequent immunoactive LH concentrations were analyzed by computer algorithms to delineate the frequency and amplitude characteristics of pulsatile LH secretion. The absolute range for LH pulse frequency estimated by a modified threshold method was 1–6 pulses/8 hr, with a mean (± SEM) of 3.36 ± 0.17 (median − 3) pulses/8 hr. The distribution differed significantly from a Gaussian pattern. The mean LH pulse amplitude expressed as a percent increase above nadir was 92.1 ± 6.1 (median-91.5%). When LH pulse amplitude was defined as an increment (mIU/ml) above nadir, the mean value was 5.13 ± 0.4 (median − 4.8) mIU/ml. These two expressions of amplitude were positively correlated (P < 0.01), while the incremental (mIU/ml) pulse amplitude correlated inversely with pulse frequency (P < 0.01). To examine the influence of more intensified rates of venous sampling on the spectrum of LH pulse properties, blood was sampled at 4-minute intervals for 8 hours in a subgroup of 13 men. Under these conditions, estimated LH pulse frequency was significantly higher, with a mean of 10.31 ± 1.87 (median − 9) pulses/8 hr compared with 20-minute sampling in the same individuals (P < 0.001). Although the estimates of LH pulse frequency at 4-minute and 20-minute sampling intervals were significantly correlated (P < 0.01), the dispersion of the LH pulse frequency estimates was considerably larger at more rapid rates of sampling. There was an absolute range of 2–20 pulses/8 hr for the 4-minute sampling, and 1–6 pulses/8 hr for the 20-minute sampling in the same individuals. This increase in LH pulse frequency and the broader dispersion of the range of frequencies estimated at 4-minute compared with 20-minute sampling intervals were confirmed using either another pulse detection algorithm, or separate criteria designed to adjust false-positive error rates in relation to sampling intensity. It was concluded that eugonadal men exhibit a broad spectrum of pulsatile LH characteristics, and the range of LH pulse attributes is even greater at more intensive rates of venous sampling. The results of this study in normal men demonstrate that a wider dispersion of physiologic LH pulse characteristics must be recognized in man. Such information is particularly important whenever possible departures from normal patterns of episodic gonadotropin secretion are sought in various clinical disorders. Moreover, these observations indicate that a wide range of LH pulse signal characteristics will maintain testosterone production effectively in the normal male.

Effects of neonatal estrogen administration on rat testis development with particular reference to Sertoli cells.

Abstract: An ultrastructural and morphometric study of the testes in 15-, 22-, 45-, and 90-day-old neonatally estrogenized rats was performed. At 45 days of age, the Sertoli cells appeared immature in estrogenized rats, whereas they were fully mature in the controls. This finding might be related to a deficiency in gonadotropins and androgens during the postnatal period. In 90-day-old estrogenized rats, however, Sertoli cell maturation had occurred, which might be attributed to a recovery of hormone levels. Cytoplasmic alterations, however, such as vacuolation, were present at this age. The morphometric study revealed decreased testicular and tubular volumes as well as decreased mean tubular diameters in the estrogenized animals. In contrast, the absolute tubular length increased more in these animals than in the controls during the period from 15 to 90 days of age. This lengthening process might be related to the large number of hypercurved tubules in the estrogenized rats.

Sertoli Cell Junctional Complexes in Gossypol‐treated Neonatal and Adult Guinea Pigs

Abstract: The effect of gossypol, an experimental male contraceptive agent, on the development and maintenance of the blood-testis barrier was determined by feeding gossypol daily to prepubertal and adult guinea pigs, and then examining their testes by electron microscopy of thin sections and freeze-fracture replicas. In guinea pigs of 10 to 30 and 10 to 40 days of age that were fed gossypol, impermeable continuous junctional zones did not develop between adjacent Sertoli cells. Compartmentalization of germ cells in the seminiferous epithelium, therefore, was nonexistent. These findings were obtained by use of the sterol-binding polyene, filipin, used as a low molecular weight tracer in combination with freeze-fracture. In general, the seminiferous tubules lacked lumina and spermatogenesis did not progress beyond the pachytene spermatocyte stage. In adult guinea pigs fed gossypol daily for five weeks, continuous zonules at the base of the seminiferous epithelium appeared intact and were impermeable to filipin. Discontinuous zonules found higher in the epithelium showed distensions between interrupted junctional strands and were permeated by filipin. In addition, vacuolated spaces between Sertoli cells and clumps of heterochromatin were conspicuous in some of the Sertoli cell nuclei. Spermatogenesis was disturbed in about 10% of the seminiferous tubules examined. These perturbations included exfoliation of round and elongated spermatids with concomitant formation of multi-nucleated giant cells. Spermatozoa from these adult male guinea pigs were immotile. These findings suggest that, in neonatal animals, gossypol appears to prevent the maturation of Sertoli cells and this effect is expressed as the failure of focal Sertoli cell tight junctional strands to assemble into continuous zonules. In adult animals, gossypol appears to have no effect on the maintenance of the blood-testis barrier.

Heterogeneity of sperm density profiles following 16-week therapy with continuous infusion of high-dose LHRH analog plus testosterone.

Abstract: LHRH agonist analogs have been investigated as potential male contraceptives. It has been shown that the LHRH agonistic analog [D-Trp6,Pro9-NEt] LHRH (LHRHA) given to men in single doses up to 500 micrograms daily for up to 20 weeks with the coadministration of testosterone enanthate produces reversible oligozoospermia. Individual responses to the treatment, however, were variable. In this study, we gave the same analog to eight normal male volunteers as a continuous infusion of 500 micrograms daily for 16 weeks. Testosterone enanthate, 100 mg, was given by injection every second week. Six of the subjects became oligozoospermic but the other two retained sperm counts that were greater than 20 million/ml, although their treatment continued for 20 weeks. The reasons for this variability of response are not clear. Serum immunoreactive LH values increased during the infusion period whereas testosterone declined. FSH values fell during treatment in all subjects except the two non-responders. The acute pituitary response to LHRHA during the treatment or shortly thereafter (48 h) was completely abolished, and bioactive LH values were suppressed totally. FSH, LH, testosterone and sperm counts returned to normal in all subjects following discontinuation of LHRHA infusion. Continuous infusion of 500 micrograms of LHRHA daily for 16 weeks with 100 mg of testosterone enanthate every 2 weeks induced desensitization of the pituitary, loss of LH bioactivity, and decreases of FSH and testosterone. This mode of administration, however, did not improve sperm density results obtained earlier by single daily injections of the analog. Heterogeneity of sperm density profiles still persists for reasons that are not yet clear.

The Presence of a Motility Inhibitor Within Spermatozoa May Explain the Poor Sperm Motility of Some Infertile Men

Abstract: The presence of motility inhibitors in seminal plasma and within spermatozoa from control and infertile men with poor sperm motility was investigated using demembranated reactivated human spermatozoa No difference was found in the inhibitory capacities in seminal plasma of patients with poor sperm motility (less than 50%) when compared with that of fertile controls with motility above 50% No correlation was observed between inhibitory capacity and sperm motility However, when extracts of spermatozoa from these patients were tested for the presence of inhibitor, it was observed that three of nine patients had an inhibitor in their sperm extract By contrast, all sperm extracts from fertile control subjects were devoid of inhibitor It was concluded that the presence of a motility inhibitor in seminal plasma does not explain the poor sperm motility observed in patients The presence of a motility inhibitor within spermatozoa, however, may represent an important factor in the etiology of the poor sperm motility observed in some patients

The heterogeneity of rat androgen binding protein (rABP) in the vascular compartment differs from that in the testicular tubular lumen. Further evidence for bidirectional secretion of rABP.

Abstract: Fractionation of testicular extracts and serum on a Concanavalin A-Sepharose column resolved two peaks of immunoreactive rat androgen binding protein. The rat androgen binding protein in the first peak, designated Form I, was present in the void volume; the other, designated Form II rat androgen binding protein, was bound by the column and specifically eluted by alpha-methylmannoside. In the course of studying the heterogeneity of rat androgen binding protein on Concanavalin A-Sepharose, it was observed that the distribution of the two forms of this protein was similar in the fluid obtained by micropuncture from the seminiferous tubule and the rete testis, that is, the ratios of Form I to Form II were 1:1 and 1:1.8, respectively. By contrast, Form I rat androgen binding protein in blood, interstitial fluid, and thoracic duct lymph of adult rats was reduced relative to Form II; the ratios of Form I:Form II in these fluids were 1:4.4, 1:3.1, and 1:4.6, respectively. since previous studies indicated that the reduced amount of Form I relative to Form II observed in the blood of adult rats was not the result of more rapid clearance of Form I, these results suggest that Form I rat androgen binding protein is preferentially secreted into the lumen of the seminiferous tubule rather than into the interstitial fluid and blood. We conclude that Sertoli cells in adult rats may partition rat androgen binding protein between the interstitial and luminal compartments of the testis based on the carbohydrate composition of this protein.

Possible significance of transferrin levels in seminal plasma of fertile and infertile men.

Abstract: Human seminal plasma contains large amounts of transferrin, which is a protein secreted mostly by Sertoli cells. It has been suggested that the concentration of transferrin may serve as a possible clinical marker of Sertoli cell function. Therefore the concentration of this protein in human seminal plasma from fertile and infertile men has been evaluated in order to find a relationship between transferrin concentrations and human semen parameters and plasma FSH levels. Findings show that seminal transferrin in subjects with oligozoospermia or azoospermia is significantly lower than in controls, and that it is strongly related to sperm count. Results also indicate that transferrin secretion can be impaired when plasma FSH levels are still normal, suggesting that seminal transferrin is an early and specific marker of Sertoli cell function. These results, however, do not clarify whether impairment of transferrin secretion by Sertoli cells is due to an organic dysfunction or to an organic secretory alteration.

Germ cell degeneration in the contralateral testis of the guinea pig with unilateral torsion of the spermatic cord. Quantitative and ultrastructural studies.

Abstract: This study evaluated the long term effects of unilateral torsion of the spermatic cord on the contralateral testis of guinea pigs, employing both fine structural and quantitative studies. Young, adult Hartley strain guinea pigs were divided into six experimental groups (12 animals per group). The first three groups consisted of 36 animals in which unilateral torsion was surgically induced. In group I (torsion maintained), unilateral torsion of the spermatic cord was maintained until the day of sacrifice; in group II (torsion and untwist), torsion of the spermatic cord was maintained for 8 to 12 hours, then the spermatic cord was untwisted and the testis was retained until the day of sacrifice. In group III (torsion and orchiectomy), testes were removed after 8 to 12 hours of spermatic cord torsion. The second three groups consisted of 36 animals: group IV (unilateral orchiectomy), group V (unilateral sham operation), and group VI (pentobarbital injection alone), which served as controls. One half of the animals from each group were killed after 4 months and the other half were killed after 8 months. The most frequently observed histologic changes in the contralateral testes of the experimental animals were focal disorganization and exfoliation of immature germ cells into the lumen. Severe damage, with almost complete absence of germ cells, was noted only in an occasional tubule. Quantitative evaluation of the germ cells of the contralateral testis revealed significant loss of germ cells in groups I, II, and III after 4 months, and in groups I and II after 8 months. Germ cell degeneration was progressive in groups I and II, as demonstrated by the lower germ cell count in the testes of animals of the 8-month group in comparison with the 4-month group. However, in group III animals, a higher germ cell count was recorded at 8 months, which was similar to those of control values. The present study confirms our earlier findings that unilateral torsion of the spermatic cord has adverse effects on the contralateral testis. However, since these effects are subtle and inconsistent, a systematic germ cell quantitation is needed for critical assessment of the deleterious effect of unilateral spermatic cord torsion on the contralateral testis.

Sex steroid-induced changes in collagen of the prostate and seminal vesicle of rats.

Abstract: The effects of androgen alone or in combination with estrogen or prolactin on the collagen of the prostate and seminal vesicles were studied in prepubertal and adult rats. Castration decreased the collagen content of the male accessory sex organs of both prepubertal and adult rats. Androgens showed stimulatory effects in castrated rats irrespective of the age. However, in intact animals, the stimulatory effects of androgens were evident only before puberty. Only the seminal vesicle of prepubertal rats responded to the stimulatory effect of estrogen given along with androgens. Prolactin did not elicit any appreciable effect either in the prostate or the seminal vesicles when administered along with androgens.

Radioautographic Localization of β‐Adrenergic Receptors in the Rat Ventral Prostate

Abstract: Previous studies performed with crude homogenates have demonstrated the presence of beta-adrenergic receptors in the rat ventral prostate. The precise localization of these receptors in prostatic tissue, however, has not been determined. The present study describes the localization of beta-adrenergic receptors using in vitro radioautography. beta-adrenergic receptors are present exclusively in the epithelial cells, while no receptors could be detected in the stromal cells. The silver grains mostly are associated with the apical pole of the glandular cells and are much less concentrated in the basal nuclear region of the epithelium. Very low concentrations of grains are found in the lumen of the acini. Castration caused a dramatic fall in the receptor concentration, while treatment with dihydrotestosterone reversed these effects. The specific presence of beta-adrenergic receptors in prostatic epithelial cells and their control by androgens suggest that they could play a physiologic role in androgen action in prostatic tissue. Moreover, the level and activity of beta-adrenergic receptors could be used as parameters of prostatic activity.