Inequality in function of the right and left ovaries and uterine horns of the mouse

Inequality in function of the left and right ovaries and uterine horns of mice was evaluated in three separate experiments. In Exp. 1, the effect of position in the reproductive tract on various reproductive characteristics was evaluated in 158 pregnant hybrid mice. Ovulation rate, number of fetuses, total fetal weight and total placental weight were higher (P less than 0.05) on the right than the left on Day 18 of pregnancy (vaginal plug = Day 1). In Exp. 2, the effect of previous sham or unilateral ovariectomy (right or left) in mated Swiss-Webster mice was compared with unoperated mated controls (N = 17-24/treatment). In control mice, ovulation rate, total fetal weight and ovarian weight were higher (P less than 0.05) on the right than left side. Surgery (sham or unilateral, ovariectomy) decreased (P less than 0.005) ovulation rates, number of fetuses, ovarian weights, total fetal weight and total placental weight on Day 18 of pregnancy. Unilateral ovariectomy decreased (P less than 0.05) ovulation rates and ovarian weights more than did sham operation. Ovulation rates were higher (P less than 0.01) when the left ovary was manipulated or removed rather than the right ovary. For Exp. 3, pairs of 8 hybrid mouse embryos each (morulae and blastocysts) were surgically transferred to the left and right uterine horns of the same (bilateral, N = 15) or different (unilateral, N = 28) Swiss-Webster recipients. In almost all incidences, embryo survival (to Day 18 of pregnancy) was twice as high (P less than 0.05) in right than left uterine horns. We conclude that the left and right ovaries and uterine horns are not equal in function in Swiss-Webster and a hybrid strain of mice.     

Dissimilarity of corpora lutea within the same ovaries or those from right and left ovaries of pigs during the oestrous cycle

A total of 48 corpora lutea from the right and left ovaries of 2 gilts on Day 9 and 2 gilts on Day 13 of the oestrous cycle were analysed for gonadotrophin binding, progesterone concentration and 3 enzyme activities. The weights of corpora lutea from the right and left ovaries on Days 9 and 13 did not differ, but the values on Day 13 were lower than those on Day 9. The specific binding of 125I-labelled hCG, progesterone concentration, and activities of cytochrome c oxidase (a mitochondrial enzyme), beta-N-acetyl-D-glucosaminidase (a lysosomal enzyme) and glucose 6-phosphate dehydrogenase (a cytosol enzyme) differed, with some exceptions, among the corpora lutea within the same ovaries and those from the right and left ovaries on Days 9 and 13 of the cycle. The gonadotrophin binding differences amongst the corpora lutea appeared to be due to the differences in the total number of available receptors rather than in the receptor affinities. There was no strict correspondence between the magnitude of gonadotrophin binding and luteal progesterone concentration. These data show that porcine corpora lutea within the same ovaries, and those from the right and the left ovary, are quite dissimilar.     

Spermatogenic activities of the left and right testes of hemicastrated and intact cockerels

One hundred cockerels of the Hacco strain were reared in deep litter. At 5 weeks of age the left testes of 30 cockerels and right testes of another 30 cockerels were surgically removed. The remaining cockerels were sham operated. At 10, 15, 20 and 25 weeks of age equal numbers from each group were killed. The rate of testis growth, the spermatid/spermatozoa reserve for each genital tract, and the relative spermatogenic activity (i.e., spermatid/sperm per g of testicular tissue) were determined for the left and right testes at the different ages.At 10 weeks of age only spermatids were found in all the testes, with concomitant very low relative spermatogenic activities. The weights of the testes increased with age and compensatory hypertrophy occurred in the testes of the hemicastrates from 15 weeks of age. The mean of the sperm reserve of the left (7.391 × 10⁹ sperm) and right (8.66 × 10⁹ sperm) genital tracts of hemicastrates contained significantly more (P < 0.05) sperm than the sum of the means of the sperm reserves in the left and right genital tracts (5.19 × 10⁹sperm) of the intact cockerels at 25 weeks of age.The relative spermatogenic activities varied with age but the pattern was similar for all cockerels, with the activities of the testes of the hemicastrates being significantly higher (P < 0.5). In all cases the values for the right testes were higher than those for the corresponding left testes.At 15 weeks of age both spermatids and spermatozoa were found in 71.4% and 100% of the right and left testes of the hemicastrates, respectively, and in 25% of the left testes of the intact cockerels. Only spermatids were found in the right testes of the intact cockerels.Hemicastration caused the right testes of the hemicastrates to produce sperm earlier than those of intact cockerels. The left testes in all cases produced sperm earlier than the corresponding right testes. Hemicastration enhanced sperm numbers per g testes (i.e., relative spermatogenic activity).     

Detection of steroidogenic acute regulatory protein in equine ovaries

A steroidogenic acute regulatory (StAR) protein has been identified in several species as a probable important rate-limiting step in steroidogenesis. This protein is believed to be responsible for transporting cholesterol from the outer to the inner mitochondrial membrane. It is known that equine chorionic gonadotrophin (eCG) stimulates steroidogenesis in the corpora lutea of early pregnant mares and that eCG also upregulates StAR mRNA in bovine ovaries. In the present study, ovarian tissue from cyclic and early pregnant mares was immunostained to detect the distribution of the StAR protein. Western blot analysis was performed, followed by phosphor imaging to establish whether the onset of eCG secretion in pregnancy was associated with increased expression of the StAR protein. Immunostaining for StAR was confined to the theca interna of growing and preovulatory follicles, but 24 h after treatment with hCG, some granulosa cells were positively stained. Positive staining was confined to the large luteal cells of the equine corpus luteum. There was no difference in the distribution of immunostaining before or after onset of eCG secretion in pregnant mares, but increased amounts of StAR were detected in corpora lutea from mares at day 40 or day 41 of pregnancy compared with non-pregnant mares and mares at days 20-30 of pregnancy.     

Intraovarianism and differences in ovulation frequency between left and right ovaries in Bos taurus heifers

Sides of ovulation at beginning and end of an interovulatory interval (IOI) were studied for 10 successive ovulations in each of 100 heifers (1000 ovulations, 900 IOI). The frequency of side for 1000 ovulations was less (P < 0.0002) for the left ovary (LO or L; 44.4 %) than for the right ovary (RO or R; 55.6 %). Number of observed ratios of L to R was determined within each of 100 sets of 10 ovulations per set (eg, L2 to R8, L6 to R4). Expected ratios were calculated by combinatorics using the criterion that side of ovulation was an independent event. Differences in ratios between observed and expected were significant supporting the hypothesis that side of ovulation is dependent on the side of the previous ovulation. Number of pairs of ovulations for each end of the 900 IOI was significantly less for the LL pair (144, 16.0 %) than for the pairs of RR (255, 28.3 %), LR (247, 27.5 %), and RL (254, 28.2 %). The hypothesis was supported that the frequency of the pairs of ovulations at the beginning and end of an IOI differed among LL, RR, LR, and RL pairs. Novel observations were that frequency was lowest for the LL pair and similar among the the RR, LR, and RL pairs. These observations indicated that the greater frequency of RO ovulation was mathematically and functionally related to lower frequency of LO ovulation. The interpretation was that intraovarianism in the LL pair accounted for the lower frequency of LO than RO ovulation.     

Interactive roles of progesterone, prostaglandins, and collagenase in the ovulatory mechanism of the ewe

Interrelationships between production of progesterone (P4), prostaglandin (PG) E2 and PGF2 alpha, and collagenase by periovulatory ovine follicles and their possible involvements in the ovulatory process were investigated. Follicles were isolated from ovaries at intervals (0 to 24 h) after the initiation of the preovulatory surge of luteinizing hormone (LH). Progesterone and PGs within follicles were determined by radioimmunoassay. Digestion of radioactive collagen during coincubation with tissue homogenates was used to assess the production of a bioactive follicular collagenase(s). Follicular accumulation of PGs and P4 increased at 12 and 16 h, respectively, after the onset of the surge of LH; PGE2 then decreased at 20 h. Collagenolytic activity of follicular tissue increased at 20 h and was maximal at 24 h (during the time of follicular rupture). An inhibitor of synthesis of P4 (isoxazol) or PGs (indomethacin) was injected into the follicular antrum at 8 h. Isoxazol did not prevent the initial rise in PGs, but inhibited synthesis of PGF2 alpha at 16 h and therafter. Isoxazol negated the decline in PGE2 and increase in collagenolysis. Indomethacin did not influence synthesis of P4; however, it suppressed collagenolytic activity of follicular tissue. Ovaries with treated follicles were left in situ and observed for an ovulation point at 30 h. Isoxazol or indomethacin was a potent inhibitor of ovulation. The blockade of ovulation by isoxazol was reversed by systemic administration of P4 or PGF2 alpha, but not by PGE2. Reversal of the blockade by indomethacin was accomplished with PGE2 or PGF2 alpha. Collagenolytic activity of follicular tissue was likewise restored by such treatments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Correlation between steroids and plasmin in rat ovaries during ovulatory process.

Progesterone (0.5 mg/rat) and estradiol-17 beta (10 micrograms/rat) injected(im) to adult female albino rats on the morning of proesterous significantly enhanced the ovarian plasmin activity as measured by the fibrin plate method at the time of ovulation which was confirmed to be at 2.30 a.m. by estimation of ovarian plasmin activity at definite intervals before and after ovulation. The ovarian plasmin activity showed a gradual increase towards ovulation and reached a maximum level at 2.30 a.m. and again decreased after ovulation. However, the nonsteroidal estrogen antagonist, tamoxifen induced inhibitory effects on the ovarian plasmin activity as compared to control and estradiol-17 beta treatment. Thus these studies reveal a positive relationship between steroids and the ovarian plasmin activity during ovulation.     

Uniformity in dynamic baroreflex regulation of left and right cardiac sympathetic nerve activities

Functional laterality of cardiac sympathetic nerve stimulation in chronotropic and inotropic effects is well known. Whether left (LSNA) and right (RSNA) cardiac sympathetic nerve activities show laterality during dynamic baroreflex activation remains to be determined. In nine anesthetized, vagotomized, and aortic-denervated rabbits, we randomly perturbed intracarotid sinus pressure (CSP) in both carotid sinus regions while simultaneously recording LSNA and RSNA. The baroreflex neural arc transfer function from CSP to LSNA and from CSP to RSNA revealed derivative characteristics, i.e., the magnitude of LSNA and RSNA responses became greater as the input frequency of CSP perturbation increased. The average slope of increasing gain in the frequencies between 0.03 and 0.3 Hz showed no difference between LSNA and RSNA responses (9.7 +/- 2.9 vs. 9.7 +/- 3.1 dB/decade, means +/- SD). The amplitude ratio and phase difference between LSNA and RSNA approximated unity and zero radians, respectively, in the frequencies from 0.01 to 1 Hz. In addition, the LSNA-RSNA relationship during stepwise CSP perturbation from 40 to 160 mmHg showed a straight line (r(2) ranged from 0.969 to 0.999). These findings indicate no laterality in the dynamic as well as static baroreflex regulation of LSNA and RSNA as far as grouped axonal activity is concerned.     

Cytochrome P-450 and steroidogenic activities of the human placental mitochondria

The relationship between cytochrome P-450 concentration, cholesterol side-chain cleavage and 3 beta-hydroxysteroid dehydrogenase/isomerase activity, in different density mitochondrial preparations from human term placenta has been studied. The heavy mitochondrial fraction shows a higher cytochrome P-450 concentration and cholesterol side-chain cleavage activity as compared to the light mitochondrial fraction, it has however lower cytochrome P-450AROM level and 3 beta-hydroxysteroid dehydrogenase/isomerase activity.     

Ovarian steroidogenic enzyme activities during the rat estrous cycle

We have correlated the concentrations of serum LH, estradiol and progesterone with the activities of 2 ovarian steroid biosynthetic enzymes during the rat estrous cycle. Ovarian 3 β-hydroxysteroid dehydrogenase isomerase (3-βHSD) activity decreased from 29 ± 6 nmol/mg protein/ min (mean ± SEM) in diestrus, to 7 ± 0.4 nmol/mg protein/min in late proestrus (p < 0.005), and subsequently increased to 36 ± 9 nmol/mg protein/min in metestrus (p < 0.01). Ovarian 17-hydroxylase (17-OH) activity decreased from early to late proestrus (3.3 ± 0.2 vs 2.2 ± 0.2 nmol/mg protein/min, p <0.0025), and subsequently increased to 3.9 ± 0.2 in metestrus (p<0.001). Serum LH, estradiol and progesterone peaked during proestrus, and reached a nadir during estrus. We conclude that the activities of 3-βHSD and 17-OH in the rat ovary vary markedly during the estrous cycle. These changes may underlie the pattern of steroid secretion characteristic of this process.     

Effects of level of food intake on ovarian follicle number,size and steroidogenic capacity in the ewe

The effect of high (H) or low (L) levels of food intake, during the preceding 4 weeks, on ovarian follicle numbers and steroidogenic capacity were investigated in groups of 12 adult Scottish Blackface ewes. Ewes of the two treatments had similar levels of body condition at the time of study but there was a twofold difference in levels of food intake. Ovaries were surgically removed on day 11 or 12 of the oestrous cycle (luteal phase; n=6 per nutritional treatment) or at 30 h after injection (i.m.) of prostaglandin F_2α analogue on day 11 or 12 of the cycle (follicular phase; n=6 per nutritional treatment). Ovarian follicles >1 mm diameter were dissected out and incubated individually for 2 h at 37°C, in 1 ml of medium 199 which was then assayed to determine concentrations of oestradiol and testosterone. There were significantly more small follicles (1–2.5 mm diameter) in (H) than (L) ewes (P<0.05) but no treatment difference in the numbers of large follicles (>2.5 mm diameter) during either phase of the cycle and no difference in the mean diameters (mm) of the two largest follicles in each animal. However, although there were higher rates of synthesis of both oestrogen (P<0.05) and testosterone (P<0.01) in the large follicles of (L) ewes as compared with (H) ewes, there was a lower oestrogen/testosterone ratio in (L) than (H) follicles which may indicate a lower level of aromatase activity in (L) follicles. It is concluded that the effects of level of food intake on ovulation rate are expressed through differences in late stages of follicle development, probably through effects on the intrafollicular steroid milieux.     

Complex interrelationships among CL,preovulatory follicle,number of follicular waves,and right or left ovaries in heifers

The diameter of the dominant follicle (DF) of wave 1 was studied on Days 9 to 17 (Day 0 = ovulation) in a survey of the ipsilateral and contralateral relationships between the location of the DF and CL, and number of follicular waves per interovulatory interval (IOI). For contralateral relationships, regardless of number of waves the diameter of the DF of wave 1 decreased (P < 0.03) between Days 11 and 13 when referenced to the follicle–CL relationship of wave 1 and decreased (P < 0.008) between Days 9 and 11 when referenced to the preovulatory follicle (PF)–CL relationship. For wave 2 in two-wave IOIs, the CL ovary of ipsilateral relationships had more (P < 0.05) follicles that reached at least 6 mm than the non-CL ovary. In three-wave IOIs, frequency of IOIs with the DF in the CL ovary was greater (P < 0.02) for wave 2 than for wave 3. In wave 3, the preovulatory and the largest subordinate follicles were located more frequently (P < 0.005) in the contralateral ovary. Ovulation in two-wave IOIs occurred more frequently (P < 0.0009) from the right ovary. In three-wave IOIs with a contralateral relationship ovulation occurred more frequently (P < 0.003) from the left ovary; a negative intraovarian effect of the CL on location of the PF may account for more ovulations from the left ovary and a reported greater frequency of the contralateral relationship. The hypothesis was supported that the ipsilateral versus contralateral relationship between the PF and CL is affected by the DF–CL relationship during the previous follicular waves and by the number and identity of waves per IOI.     

Relationships of changes in ultrasonographic image attributes to ovulatory and steroidogenic capacity of large antral follicles in sheep

Ovarian steroidogenesis and antral follicular development in ewes, following the treatment with medroxyprogesterone acetate (MAP) and equine chorionic gonadotrophin (eCG), are affected by the reproductive season. The objective of this study was to compare the ultrasonographic attributes of large antral follicles between cyclic (December) and seasonally anovular (June–July) ewes, after a 12-day treatment with MAP-soaked intravaginal sponges, with or without the administration of 500 IU of eCG at sponge removal, and to determine whether there is a correlation between the ultrasonographic attributes of the follicular wall and serum concentrations of oestradiol. Digital images of ovulatory follicles from cyclic ewes and eCG-treated anoestrous ewes (n = 34 follicles), and of anovulatory follicles attaining ≥5 mm in control anoestrous ewes (n = 8 follicles), were analysed using the spot and line techniques designed to determine the echotextural characteristics of the follicular antrum (central and peripheral), follicular wall and perifollicular ovarian stroma. The mean diameter of ovulatory follicles was greater (P < 0.001) in cyclic than anoestrous ewes, with or without the eCG treatment. The mean pixel heterogeneity (SD of numerical pixel values) of the follicular antrum (P < 0.05), as well as mean pixel intensity and heterogeneity of the peripheral antrum, follicular wall proper and perifollicular ovarian stroma (P < 0.05), were consistently greater in anoestrous than cyclic ewes at the time of sponge removal and 24 h after the treatment with MAP sponges or MAP/eCG. Mean oestradiol concentrations were greater (P < 0.05) in cyclic compared to anoestrous ewes in both MAP- and MAP/eCG-treated animals, from 1 to 2 days after sponge withdrawal. There was a moderate negative correlation (r² = 0.12, P < 0.05; Pearson's Product Moment and r² = 0.23, P < 0.05; ANCOVA) between mean pixel heterogeneity (standard deviation of mean pixel values) of the follicular wall proper (all follicles ≥5 mm in diameter) and serum concentrations of oestradiol after sponge withdrawal. Our results indicate that large antral follicles from cyclic and seasonally anovular ewes exhibit distinctive ultrasonographic characteristics. The differences in follicular echotexture appear to be related mainly to seasonal variations in ovarian follicular morphology and oestradiol production.     

Effects of ketoconazole on rat testicular steroidogenic enzymatic activities

Ketoconazole (K) is an antifungal imidazole derivative which has been shown to be a potent inhibitor of testosterone (T) biosynthesis in rodents and humans. To study the effect of K on rat testicular steroidogenesis we measured the activities of five testicular microsomal steroidogenic enzymes in K-treated rats and controls. Thirty male adult rats were given either 2 mg K or water every 12 hours by mouth during 5 days. Mean testicular weight was similar in both groups of animals. The K-treated group had a T serum concentration of 83 +/- 14 ng/dL whereas it was 94 +/- 16 ng/dL in the control group (NS). The K-treated animals had decreased activities of the 3 beta-hydroxysteroid dehydrogenase (830 +/- 48 vs 2,245 +/- 109 pmol/mg protein/min, P less than 0.001), 17-hydroxylase (243 +/- 5 vs 676 +/- 17 pmol/mg protein/min, P less than 0.001), 17-ketosteroid reductase (31 +/- 2 vs 169 +/- 7 pmol/mg protein/min, P less than 0.001), and aromatase enzymes (92 +/- 6 vs 123 +/- 7 pmol/mg protein/min, P less than 0.01). The 17,20-desmolase activity was similar in both groups of animals (210 +/- 4 vs 171 +/- 18 pmol/mg protein/min). We conclude that K given orally to rats inhibits the activity of several testicular steroidogenic enzymes.     

Comparisons of Peptide hydrolase activities in cereals

Carboxypeptidase activity (hydrolysis of N-carbobenzoxy-l-phenylalanyl-l-alanine) is high in a number of temperate zone cereals, originating in Asia Minor (wheat, barley, oats, wild oats, rye, triticale) compared to other cereals originating in central America or Asia (maize, sorghum, rice). However, endopeptidase activity (hydrolysis of azocasein or hemoglobin) is relatively much higher in the latter group. Comparison of trichloroacetic acid (TCA)-soluble products derived from the hydrolysis of hemoglobin showed that carboxyterminal amino acids (histidine, arginine, and tyrosine), are released when extracts from wheat and barley endosperms are used. With extracts from corn endosperms, much more TCA-soluble ultraviolet- absorbing material is released, but very little is released as free amino acids within the first 2 hours and the expected C-terminal amino acids of hemoglobin are not detected in significant amounts. These results suggest that the method of hydrolysis of the storage proteins may be significantly different in these two classes of cereals.