Identification and localization of G protein subunits in human spermatozoa.

Antibodies to alpha and beta subunits of guanine nucleotide regulatory proteins (G proteins) were used to identify which G proteins are present in mature human spermatozoa and to determine their subcellular localization. Immunoblots of membranes from spermatozoa demonstrate the presence of Galphai2, Galphai3, Galphaq/11 and Gbeta35 and the absence of Galphai1, Galpha0, Galphas, Galpha12, Galpha13, Galpha16, Galpha and Gbeta36. Indirect immunofluorescence demonstrates the presence of Galphaq/11 in the acrosome, with the highest proportion in the equatorial segment. Galphai2 is present in the acrosome, midpiece and tailpiece and Galphai3 in the postnuclear cap, midpiece and tailpiece. The Gbeta35 subunit is found mostly in the midpiece, with marginal labelling of the head, tailpiece and the equatorial segment of the acrosome. The distinct pattern of distribution of G proteins suggests that they may couple to receptors or effectors which also have discrete regions of localization in spermatozoa. These highly localized signal transduction pathways may regulate discrete functions, such as activation of the acrosome reaction, fusion with the oocyte and motility.

[Mixed secretory and obstructive azoospermia. Transseptal vasovasostomy by vas deferens transposition: a case report and review of the literature]. / Azoospermie mixte sécrétoire et obstructive. Vasovasostomie trans-septale par transposition des déférents: à propos d'un cas et revue de la littérature.

The authors report a case of complex azoospermia in a 35-year-old man, due to a combination of right testicular atrophy and an iatrogenic lesion of the contralateral vas deferens. Vasovasostomy with transseptal transposition of the vasa deferens restored fertile ejaculation.

[Prognostic sperm factors in intra-uterine insemination with partner's sperm]. / Facteurs pronostiques spermatiques des inséminations intra-utérines avec sperme du conjoint.

We studied the prognostic value of sperm characteristics for the outcome of intra-uterine insemination with partner sperm (IUIPS). A total of 712 cycles of IUIPS following induction of ovulation with gonadotrophin (hMG/hCG) for 277 sterile couples attending the assisted reproductive technology centre of Poissy Hospital (78300-France) between January 1991 and December 1994 was studied retrospectively. Ninety-two clinical pregnancies were obtained giving an overall rate of 12.9% per cycle. None of the characteristics of the sperm as assessed initially correlated with outcome. In contrast, the number of motile spermatozoa given (n) affected outcome: for n < 1 x 10(6) the pregnancy rate was 2%; for n = 5 to 8 x 10(6) the rate was 19%. However, for +/- 8 x 10(6) the proportion of biochemical pregnancies and miscarriages was 40% which was significantly higher than for smaller concentration. The resort of IVF following 4 IUIPS failures leads to a pregnancy rate per cycle of only 6.7%.

[Retrospective evaluation of an intra-uterine insemination program with donor]. / Evaluation rétrospective d'un programme d'insémination intra-utérine avec donneur.

The results of 109 intra-uterine inseminations with cryopreserved prepared donor semen (IUId) in stimulated cycles, performed among 42 couples from November 1987 to June 1991, were analysed. The single IUId was performed 39 +/- 3 hours after the injection of human chorionic gonadotrophin. For each IUID 1,25 +/- 1.4 x 10(6) motile spermatozoa (mean +/- standard deviation) were inseminated. 27 pregnancies were obtained and the pregnancy rate per cycle was 24.8%. The number of motile spermatozoa inseminated was significantly higher in the group of pregnant women than in the population without pregnancy (1.64 x 10(6) x 10(6)/IUI versus 1.13 x 10(6)/IUI). The sperm count and the number of motile spermatozoa inseminated were significantly higher when the semen was prepared on discontinuous Percoll gradients than after swim-up in Menezo B2 medium (8.2 x 10(6)/ml versus 4.3 x 10(6)/ml and 1.7 x 10(6) versus 0.77 x 10(6)). Among the 27 clinical pregnancies, 3 were twins and 2 were triplets. The ovarian hyperstimulation associated with the use of a prepared semen of donor, generate a high risk of multiple pregnancies. To improve the results without increasing the risk of multiple pregnancies, we propose to inseminate about 1.5 x 10(6) motile spermatozoa with a maximum plasma estradiol level of 1 100 pg/ml, on the day of hCG injection, associated with 2 preovulatory follicles. This technique may augment the rate of pregnancy and represents an appreciable time gain. These results must be confirmed by randomized prospective studies.

[A hormone assay as a therapeutic choice in male sterility]. / Apport de l'enquête hormonale au choix thérapeutique d'une stérilité masculine.

In order to study the function of the hypothalamic-pituitary-testicular axis in men referring for severe oligospermia, the hormonal pattern of 57 oligospermic men was compared to those of 19 healthy volunteers. Fourteen patients had plasma gonadotrophin levels in the normal range contrasting with low plasma testosterone (T) levels. An hyperprolactinemia was found in 2 of these men who were treated with bromocriptine. A dramatic increase in sperm count was obtained on month 9 to 12 of the therapy and 5 pregnancies were obtained. Two men with hypogonadotrophic hypogonadism and azoospermia were treated with gonadotrophins. Such a treatment induced a desquamation of immature germinal cells in the sperm on month 6 and the maturation et spermatozoa on month 18. By contrast to the latter patients, 8 men had a decrease in plasma T levels without clinical signs of hypoandrogenism. The spermocytogram showed numerous immature germinal cells. On month 7 of a treatment using gonadotrophins, the sperm count rose and 4 pregnancies were obtained after 3 to 12 months of therapy. In 2 patients an isolated FSH deficiency was suspected on the basis of undetectable FSH levels unresponsive to the infusion of GnRH. These patients were treated with hMG. This treatment induced a sharp increase in sperm count on month 6. Forty-three patients had an increase in either LH and/or FSH: 24 men had plasma testosterone and LH levels in the normal ranges, contrasting with an increase in plasma FSH level. In such men, the mean of testosterone level was significantly (p less than 0.001) lower than in controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemical exploration of azoospermia: interest of the seminal transferrin assay.

Seminal transferrin is considered a good index of Sertoli cell function. In this study, including 19 control subjects, four vasectomized subjects and 65 non-vasectomized subjects with azoospermia, transferrin was measured concomitantly with other classical biochemical seminal markers such as L-carnitine, fructose and zinc. This parameter should provide additional information for differentiation between obstructive and non-obstructive azoospermia. A threshold value of 85 micrograms/ejaculate for transferrin was defined in the control and the vasectomized groups. The 65 subjects with azoospermia were divided into five groups according to L-carnitine and transferrin threshold values; the nature of azoospermia (obstructive or non-obstructive) was considered in each group. In the biological investigation of sterility, the measurement of transferrin provided information as to the aetiology of azoospermia. However, the determination of L-carnitine and fructose remains important in the localization of any obstruction.

[Labeling of testicular biopsies with lectins. Value of Evonymus europaeus agglutinin]. / Marquage des biopsies testiculaires par les lectines. Intérêt d'Evonymus europaeus agglutinin.

EEA was used to label testicular biopsies with 11 other lectins and three monoclonal antibodies anti blood group A, B, H substances. Although EEA's receptor chemical structure is still unknown and an electron microscopic control necessary, it is possible to use this lectin to label proacrosomic vesicles of spermatids in man and animal, allowing useful comparisons between species. The intensity of the reaction makes it interesting in the standard study, even automatized, of every testicular biopsy.

[Morphologic parameters of sperm. 1) Syndromes]. / Paramètres morphologiques du sperme. 1) Syndromes.

In this study we have been able to pull out 23 case histories of husbands of sterile wives out of a total of 839 patients who consulted us for diminished fertility. These 23 have the characteristics of "normal" male male population and serve as a reference group to estimate normal values of different parameters. The steps taken in this way give figures that are very little different from those obtained by other methods but once a still larger group have been analysed it would be possible by this method to work out the incidence of the different pathological states that can be found in the general population. An analysis of the differences found in our groups shows that the most significant parameter is the percentage of spermatozoa that are moving in straight lines. This means that future efforts should be directed to this parameter when automation or standardization is considered. There are normally random fluctuations in the parameters of healthy subjects but pathological pressures in subjects with lowered fertility mean that some of them vary in parallel coordinated directions. Because of this, seven different profiles can be worked out for different syndromes: Where the head alone is affected: macrocephalic spermatozoa, broken spermatozoa. Where the intermediate portion alone is damaged: a very thin intermediate portion. Where the tail alone is affected: spermatozoa with a short or absent tail. Where several parts of the sperm are affected: hook-shaped spermatozoa, double-headed spermatozoa, and rolled-up spermatozoa.(ABSTRACT TRUNCATED AT 250 WORDS)

Male sterility associated with familial translocation heterozygosity: t(8;15) (q22;p11).

During the investigation of the family of a subject consulting for primary sterility, the same oligoteratospermia was found in two of his brothers. The three karyotypes of these subjects exhibited an equilibrated reciprocal autosomal translocation t(8;15) (q22;p11), which was also detected in their mother. The karyotypes of the remaining siblings, one brother and one sister, were normal. The semen analysis of the sterile subjects suggests that the block of gamete production occurs at the beginning of spermiogenesis. The chromosomal anomaly, which has no effect on the reproduction of the mother, leads to sterility of the male offspring bearing it.

[Total asthenospermia with an ultrastructural anomaly of the flagellum in two sterile brothers (author's transl)]. / Asthenospermie totale avec anomalie ultrastructurale fu flagelle chez deux frères stériles.

The two sterile brothers were found to have the same sperm abnormalities. The flagella were short or absent. The constituent elements of the flagellum were disorganized, and the axone structure was disturbed. These abnormalities give rise to a problem of explaining the genetic origin of certain sperm defects similar to those found particularly in mice.

[Ultrastructural study of anomalies of the acrosome in spermatozoa with irregular heads]. / Etude ultrastructurale des anomalies de l'acrosome dans les spermatozoïdes à tête irrégulière

The study of the ultrastructure of spermatozoa having an irregular head, encountered in large number in semen of men supposed to be sterile, shows that the acrosome is often modified in its shape or its texture. In several samples the spermatozoa have either no acrosome or a small one, at a certain distance from the nucleus. In other cases the ratio between the segments of the acrosome is modified and the parallelism between the edges is not conserved. These acrosomes are often covered with a cytoplasmic velum the post acrosomal cape is missing. Finally the acrosome can be deformed by local expansions of the subacrosomal space, or by large gaps. Other anomalies which are less common have been observed. The elimination of possible artefacts and the comparison between these human anomalies and similar acrosomal anomalies of mammalian semen, are discussed in this paper.

[Inhibition of fertilization in the rabbit using an antisperm serum]. / Inhibition de la fécondation par un sérum antisperme chez le lapin.

PIP: The action of antisperm immune serum on fertilization was tested in rabbits in 2 ways: by incubating sperm with immune serum and complement and inseminating females with the mixture, and by introducing immune serum and complement into the vagina before mating. Immune serum was produced by injecting mixtures of homologous sperm and complete Freund's adjuvant. The first 50 million sperm were incubated with .3 ml antiserum and 3 drops complement for 20 minutes at 37 degrees C, and inseminated in females given 50 U. human chorionic gonadotropin. 28-32 hours later 5 control rabbits (normal serum) had 51 rupture points on the ovaries, and 46 of 50 eggs flushed from the tubes were fertilized, but 5 experimental rabbits had 50 repture points and 50 unfertilized eggs. In the second trial, 1, 2 or 2.4 ml antiserum with an appropriate amount of complement placed intravaginally before mating produced 44 rupture points, 40 eggs, collected and 30 fertile eggs in 6 controls, but 50 rupture points, 42 eggs all unfertilized in 6 test rabbits. This antisperm cytotoxicity in vivo, shown by absence of sperm in the oviducts, opens new possiblities for contraceptive research.^ieng