Hormonal effects on the activities of glycosidases in the endometrium of rabbit uterus

The activities of several glycosidases in the lysosomal fraction of the uterine endometrium of rabbit were measured using 4-MU-glycosides as substrates. The specific activity of beta-N-acetylglucosaminidase was the highest, which was followed by beta-galactosidase, beta-glucuronidase, and alpha-galactosidase in this order. beta-Glucosidase had the lowest activity among the glycosidases examined. In order to examine the hormonal effects on these glycosidases, the lysosomal fractions were prepared from the uterine endometrium of the control, estrogen-treated, and progesterone-treated rabbits. In all glycosidases examined, except for beta-glucosidase, the specific activity was highest in the lysosome obtained from estrogen-treated rabbit. The specific activity in the lysosome from the progesterone-treated rabbit was between that from the estrogen-treated rabbit and that from control. Hormonal treatments, however, affected neither pH optimum curves nor isozyme patterns of these glycosidases.     

Erectile function and bulbospongiosus EMG activity in estrogen-maintained castrated rats vary with behavioral context.

Electromyographic (EMG) activity in the bulbospongiosus muscles (BS) was recorded to monitor potential castration-induced alterations in muscle activity during copulation and reflexive erections. EMG recordings were made from intact male rats and from castrated rats maintained from 7 to 50 days on estradiol benzoate (300 micrograms/day) or testosterone (200 micrograms/day). Despite a 40-50% postcastration reduction in the weight of the BS and accessory sexual glands in estrogen-treated rats, the pattern of EMG activity during copulation was similar across groups. In estradiol-treated males, the EMG burst frequency during mounts and burst duration during intromissions exceeded the parameters of intact males and of castrated males maintained on testosterone. Between intromissions, and following ejaculatory patterns, estrogen-treated males displayed spontaneous muscle bursts accompanied by visually confirmed erection of the glans penis, but these males quickly lost the capacity for reflexive erections. These data demonstrate that despite castration-induced atrophy of the penile muscles and, presumably, their spinal motor nuclei, the motor output to these muscles is maintained following androgen removal. The capacity for substantial penile erection is retained during copulation long after reflexive erections have diminished.     

Oviduct acid mucus glycoproteins in the estrous rabbit: ultrastructure and histochemistry

Epithelial glycoproteins are likely to be important in many aspects of reproduction. The rabbit oviduct produces mucus glycoproteins. This is indicated both by histochemistry and by gelation of a mucus coat around the rabbit ovum during its tubal transport. We report here that the production of highly acid mucus glycoprotein (apparently of the type that coast the ovum) is confined to the isthmus and, to a lesser extent, the mucosal crypts of the ampullary-isthmic junction; the ampulla is not involved. Using a method of perfusion-fixation that includes the polycation alcian blue in conjunction with glutaraldehyde to precipitate and stabilize glycoproteins, we have demonstrated that this mucus, at least in rabbits in estrus, occupies the isthmic lumen but not the ampullary lumen. Histochemistry shows that it is the electron-lucent secretory granules of the isthmus and ampullary-isthmic junction, but not the denser granules of the ampulla, that exhibit staining characteristics of highly-acid mucus glycoproteins. Important opportunities are likely to exist for interaction of this isthmic mucus with spermatozoa and with fertilized ova during their isthmic transport.     

Effect of ovulation on the ionic and water content of rabbit oviduct

The water content, extracellular space, intracellular water, potassium, sodium and chloride content of oviduct and uterus removed from rabbits in estrus and 24 and 72 h following hCG injection have been determined. Following ovulation, there was an increase in water content of the region of the oviduct corresponding to the ampullary -isthmic junction which, at 24 h, is probably due to increased intracellular water. In the ampullary-isthmic junction, ovulation decreases the potassium content and, at 24 h in the isthmic region of the oviduct, chloride is reduced. The water and ionic content of mucosa and smooth muscle cells has been calculated and mucosa cells have greater intracellular water and chloride and sodium content and less potassium and extracellular space than the smooth muscle cells of the oviduct. The significance of the changes in ionic and water content is discussed.     

The cytosol receptors for progesterone in the different parts of rabbit fallopian tube and uterus during ovum transport

Progesterone receptors were determined in the cytosol from the ampulla, ampullaryisthmic junction and isthmus of rabbit fallopian tube and uterus of estrus and pregnant rabbits. The receptor levels when compared among its various anatomical segments, were the same in ampulla, isthums and uterus but maximum in ampullary-isthmic junction. Significant differences were observed in mated animals at 14, 24, 34, 48, 70 and 144 h after coitus. The receptor concentrations in portions of the fallopian tube showed no significant change between 14 and 24 h after coitus, except for a decrease in ampullary-isthmic junction at 24 h. At 34 h the concentration of receptor further decreased in all parts of the tube. At 48 and 70 h after coitus, receptor concentrations decreased gradually in ampulla and ampullary-isthmic junction, while isthmus showed a gradual increase. At 144 h, the receptor concentration showed no further change in ampulla and ampullary-isthmic junction; however, isthmus showed a decline. The uterine receptor concentration declined steadily from estrus till 70 h after coitus, however, it was increased at 144 h. The dissociation constant (K_d) of cytosol receptor in all the tissues at estrus and during early pregnancy was found similar. The implications of these changes in relation to the normal ovum transport have been correlated in this paper.     

荷包牡丹碱和印防己毒素诱发的脊兔膈神经呼吸样电活动

实验用家兔共71只。乌拉坦浅麻醉,麻痹肌肉,颈_1或颈_2 横断脊髓,记录膈神经电活动。43例中,25例出现紧张性电活动,无一例自行恢复呼吸样节律放电。24例于脊髓蛛网膜下腔(i.s.s.)内给荷包牡丹碱(BCL,20μg/10μl—40μg/20μl i.s.s.),13例给印防已毒素(PIC,20μg/20μl,i.s.s.或3—5mg/kgbt i.v.),能诱发出长串的呼吸样放电(串长在0.25s以上)或短串的抽搐样放电(串长在0.1s 以下)。长串放电,按其募集与去募集特点,可分为三型: Ⅰ型,排放频率平均为23.5±2.3串/min(BCL),出现率为58.5%(BCL)或67.7%,(PIC);Ⅱ型,排放频率平均为33.8±4.7串/min(BCL),出现率为39.3%(BCL)或32.3%(PIC);Ⅱ型,排放频率平均为21.3±2.8串/min(BCL),出现率为2.2%(BCL)或3.3%(PIC)。放电持续时间平均为60.0±18.9min(BCL)或42.0±0.8min(PIG)。Ⅰ型和Ⅱ型放电均具呼吸样放电的特点,Ⅰ型和Ⅱ型的出现率共占长串放电的97.8%。本工作提示:脊髓內源性 GABA 系统对脊髓“呼吸”具有紧张性抑制作用。     

Postnatal development of the oviduct of intact and ovariectomized rabbits

The length of the oviduct, the thickness of its wall, and the height of its mucosal epithelium and cilia were measured in (a) 0-, 2-, 4- and six-month-old rabbits, (b) rabbits ovariectomized at birth and (c) ovariectomized, estrogen-treated rabbits. The length and external diameter of the oviduct increased progressively until four months of age, after which their rates of increase declined. The thickness of the oviductal wall at the uterotubal junction was twice as large as that of the isthmus at two months of age and six times as large at four and six months of age. The height of the mucosal epithelium in the fimbriae was less than that in other oviductal segments at birth, but exceeded that in other segments at six months of age. Ciliated cells and motile cilia were absent 24 hours after birth; they were first observed two months after birth. The cilia of fimbriae were shorter than cilia elsewhere in the oviduct. Neonatal ovariectomy retarded the development of the oviduct and the mesotubarium and caused pyknosis of ciliated and non-ciliated cells of the oviductal mucosa. Cells with scarcely motile cilia were present five and one-half months after neonatal ovariectomy.     

Concentrations of energy substrates in oviductal fluid and blood plasma of pigs during the peri-ovulatory period.

Large White gilts, 9 to 18 months old, that had exhibited at least two natural oestrous cycles were divided into three groups (phases): unmated pre-ovulatory, unmated post-ovulatory and mated post-ovulatory (n = 16, 20 and 18). Oviductal luminal fluid samples were collected under anaesthesia by micropipette from the ampulla and ampullary-isthmic junction and analysed by an ultramicrofluorometric technique. Glucose concentrations (mmol 1(-1), means combining regions; mean +/- SEM) were significantly higher in blood plasma than in oviductal fluid (4.56 +/- 0.20 versus 0.59 +/- 0.16; P < 0.0001; n = 27), whereas lactate was higher in the oviduct (5.71 +/- 0.53 versus 2.48 +/- 0.24; P < 0.0001; n = 27). No significant differences were found between the ampulla and the ampullary-isthmic junction. However, the concentration of glucose was significantly higher (P < 0.05) in the ampulla of the pre-ovulatory group (0.97 +/- 0.20; n = 13) compared with the mated group (0.25 +/- 0.05; n = 14) and its concentration in the ampullary-isthmic junction in the pre-ovulatory group (1.65 +/- 0.63; n = 13) was significantly greater (P < 0.05) than in the post-ovulatory (0.43 +/- 0.11; n = 11) or mated groups (0.17 +/- 0.02; n = 14). Lactate in the ampulla of mated animals was higher than in the pre-ovulatory group (6.83 +/- 0.70 versus 3.86 +/- 0.38; P < 0.05; n = 15 and 13), but neither was significantly different from the post-ovulatory group. Furthermore, no change was seen at the ampullary-isthmic junction in lactate concentration with phase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Luteal enzymes of the luteinizing hormone and beta-adrenergic signal transduction pathways in hypophysectomized rabbits do not require pituitary hormone support.

Experiments were conducted to determine whether continuous pituitary hormone support is required for expression of the LH and beta-adrenergic cAMP signal transduction pathways in rabbit CL during pseudopregnancy. Parameters of the LH and catecholamine cAMP signal transduction pathways in CL of estrogen-treated hypophysectomized rabbits were compared to those of pituitary-intact rabbits. Results showed that each of the parameters of the LH and beta-adrenergic cAMP signal transduction pathways was retained in CL taken from estrogen-treated pseudopregnant rabbits that had been hypophysectomized for as long as 13 days at levels not significantly different from those of estrogen-treated pituitary-intact rabbits. These included luteal basal, and LH-, epinephrine-, and fluoride-stimulated adenylyl cyclase activities; total luteal cAMP levels; the number and affinity of cAMP-dependent protein kinase regulatory subunit cAMP binding sites; binding activity of the type I and type II regulatory subunits; and the amount of catalytic subunit protein of cAMP-dependent protein kinase. We conclude that expression of the proteins of the cAMP signal pathway for LH and beta-adrenergic hormones in CL of estrogen-treated rabbits does not require pituitary hormone support.     

Spontaneous motility of the oviduct in the anaesthetized pig

Intraluminal pressure microtransducers were placed at the uterotubal junction, the proximal isthmus, the ampullary-isthmic junction and the mid-ampulla. Spontaneous motility occurred throughout the oestrous cycle in all segments. During oestrus there were regular, high amplitude peristaltic waves in all segments, superimposed on basal activity. On Day 1 of the cycle the pattern was mostly antiperistaltic, presumably related to sperm transport. During the periovulatory period the number of peristaltic and antiperistaltic waves became equal, perhaps in relation to the transport of gametes to the fertilization site. During Day 3 there was no peristaltic activity; the motility patterns of the isthmus and ampullary-isthmic junction were similar (regular phasic contractions of high frequency and amplitude) while the ampullary motility was low. On Day 4, when the eggs enter the uterine lumen, the ampullary-isthmic junction and particularly the isthmus showed strong contraction waves (mostly peristaltic) superimposed on the basal phasic activity. This suggests an active role of the smooth muscle of the lower oviducal segments in ovum descent. During the mid- and late-luteal phases, the isthmus remained motile, with an irregular base line, but lost the pattern of basal contractions that dominated the activity during the first 4 days of the cycle. The ampulla showed low levels of spontaneous motility throughout the rest of the cycle.     

Distribution patterns of rabbit embryos during preimplantation stage

The pattern of transport and distribution of rabbit embryos in the oviduct and uterus was studied 15 to 168 hours post coitum (p. c.). The reproductive tract was frozen in liquid nitrogen, thawed, and cleared in benzyl-benzoate solution using Orsini's technique. The location of the eggs and the ampullary-isthmic junction were identified using transmitted light from a dissecting microscope. Accumulation of the eggs in the oviduct occured in two phases. In the first phase the eggs were retained above the ampullaryisthmic junction, 3–12 hours after ovulation. In the second phase, the eggs were retained 36–60 hours after ovulation, above the uterotubal junction (at a distance approximately 12 % of the oviductal length). The rate of transport of individual eggs in the oviduct, and the time of the entry of eggs into the uterus were variable. Au 78 hours p. c. most blastocysts occupied the proximal half of the uterine horn, although some appeared very close to the internal os of the cervix. Spacing of blastocysts in the uterus, 114 to 120 hours p. c., involved movement of blastocysts away from the cervix. Unfertilized eggs remained in the uterus, along with developing blastocysts 168 hours p. c. Few eggs were retained in the oviduct at 108 and 115 hours p. c.     

Evidence of the origin of the gelatinous masses in the oviducts of mares

The gelatinous masses known to occur in the mare oviduct were examined by light and scanning electron microscopy. Many (44%) of the masses which occurred in 16 of the 24 oviducts studied were connected with the mucosa of the ampulla near the ampullary-isthmic junction. The masses consisted of lobules of fine fibres probably derived from the fibrous connective tissue of the lamina propria in the oviductal mucosa.     

Estrogen modulates the expression of myosin heavy chain in detrusor smooth muscle

The effect of low serum estrogen levels on urinary bladder function remains poorly understood. Using a rabbit model, we analyzed the effects of estrogen on the expression of the isoforms of myosin, the molecular motor for muscle contraction, in detrusor smooth muscle. Expression of myosin heavy chain (MHC) isoforms, which differ in the COOH-terminal (SM1 and SM2) and the NH(2)-terminal (SM-A and SM-B) regions as a result of alternative splicing of the mRNA at either the 3'- or 5'-ends, was analyzed in age-matched female rabbits that were sham operated, ovariectomized (Ovx), and given estrogen after ovariectomy (4 rabbits/group). Ovx rabbits showed a significant decrease in the overall MHC content per gram of wet detrusor smooth muscle compared with controls (P < 0.04), which was reversed by estrogen replacement (P < 0.02). MHC content, as a proportion of total milligram of protein in the bladder tissue extracted, was also increased in estrogen-treated Ovx rabbits. Quantitative competitive RT-PCR revealed 1.72-, 2.63-, and 5.82 x 10(6) copies of MHC mRNA/100 ng total mRNA in Ovx, control, and estrogen-treated rabbits, respectively (P < 0.01). RT-PCR analysis using oligonucleotides specific for the region containing the SM1/SM2 MHC alternative splice sites indicated a lower SM2-to-SM1 ratio in estrogen-treated compared with control and Ovx rabbits (P < 0.05). Similarly, SDS-PAGE analysis of extracted myosin from estrogen-treated rabbits revealed a significantly lower SM2-to-SM1 isoform ratio compared with control and Ovx rabbits (P < 0.05). Expression of the SM-A and SM-B isoforms was not affected. These results indicate that myosin content is increased upon estrogen replacement in Ovx rabbits and that the abundance of SM1 relative to SM2 is greater in estrogen-treated rabbits compared with normal and Ovx rabbits. These data suggest that estrogen affects alternative splicing at the 3'-end of the MHC pre-mRNA to increase the proportion of SM1 vs. SM2.     

Oxytocin receptors in rat oviduct.

Particles from rat oviduct homogenates sedimenting between 1,000 × g for 10 min and 48,000 × g for 30 min bound [³H]oxytocin $\text{in vitro}$. The apparent K_d for oxytocin binding to high affinity sites in particles prepared from estrogen-treated rats was 1.8 × 10^?9 M. About 215 fmoles of oxytocin were bound per mg of particulate protein. Oviducal preparations from untreated rats had about 25% the affinity for oxytocin of preparations from estrogen-treated rats. Oxytocin analogues were bound to oviducal particles in the same rank order as their uterotonic potencies: (desamino)oxytocin > (4-threonine)oxytocin > oxytocin > (8-lysine)vasopressin ? desaminotocinol. No oxytocin binding could be shown with the particulate fractions from rat ovary. The binding of oxytocin to the oviduct and uterus are similar in affinity, number of binding sites, ligand specificity, and the increase in response to estrogen treatment.     

The effects of progesterone on follicular growth in the rabbit ovary

Studies were undertaken to measure the growth of follicles in the rabbit ovary during periods of elevated blood levels of progesterone. The progestin was increased in the blood by pregnancy or by implantation of progesterone pellets, which raised blood progesterone to near the levels measured during pregnancy. After 1, 2, 3, or 4 weeks of pregnancy or progesterone-pellet treatment, follicles of 1.0 mm external diameter or greater were dissected out of the ovaries and their external diameters were measured; then, each follicle was extracted for measurement of estradiol content. Blood levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured in these animals as well. Follicles up to 2.5 mm in diameter were found in the ovaries of nonpregnant and untreated animals while 1.8 mm was the maximal size found during pregnancy or progesterone-pellet treatment. Furthermore, both in pregnant and in progesterone-treated rabbits, the follicular estradiol content and concentration were significantly suppressed compared to follicles from untreated rabbits. The progesterone pellets had no major effect on the levels of LH and FSH in the blood; the concentration of these gonadotropins in the progesterone-treated rabbits was virtually identical to levels previously measured in the blood of pregnant animals. The results of these studies indicate that progesterone exerts an inhibitory action on follicular development and steroidogenic function in the rabbit ovary. The progesterone action appears to be exerted directly on the ovary and is not indirect, by way of an inhibition of gonadotropin secretion.     

Distribution of progestin-binding cells in estrogen-treated and untreated neonatal mouse uterus and oviduct: autoradiographic study with [125I]progestin

Summary Progestin-binding sites in uteri and oviducts of estrogen-treated and untreated 8-day-old mice were studied by thaw-mount autoradiography with [¹²⁵I]progestin. In the untreated uteri, nuclear concentration of radiolabelled progestin was observed in all tissues of the uterus, with strongest nuclear labelling in luminal and glandular epithelia and in stroma. In the estrogen-treated uteri, the degree of labelling was markedly augmented in stroma and muscle, but much reduced in the luminal and glandular epithelia, compared to untreated uteri. In untreated oviducts, nuclear labelling was observed in stroma and muscle in all regions and in epithelium in the isthmic and uterine regions. The epithelium in infundibular and ampullar regions was only scarcely labelled. The estrogen-treatment augmented the labelling in stroma and muscle of the oviduct as in the uterus, but the labelling in epithelium was not affected. These results indicate that estrogen-treatment induces progesteron receptor differentially among tissue compartments both in the uterus and oviduct.     

Distribution of progestin-binding cells in estrogen-treated and untreated neonatal mouse uterus and oviduct: autoradiographic study with [125I]progestin

Progestin-binding sites in uteri and oviducts of estrogen-treated and untreated 8-day-old mice were studied by thaw-mount autoradiography with [125I]progestin. In the untreated uteri, nuclear concentration of radiolabelled progestin was observed in all tissues of the uterus, with strongest nuclear labelling in luminal and glandular epithelia and in stroma. In the estrogen-treated uteri, the degree of labelling was markedly augmented in stroma and muscle, but much reduced in the luminal and glandular epithelia, compared to untreated uteri. In untreated oviducts, nuclear labelling was observed in stroma and muscle in all regions and in epithelium in the isthmic and uterine regions. The epithelium in infundibular and ampullar regions was only scarcely labelled. The estrogen-treatment augmented the labelling in stroma and muscle of the oviduct as in the uterus, but the labelling in epithelium was not affected. These results indicate that estrogen-treatment induces progesterone receptor differentially among tissue compartments both in the uterus and oviduct.     

Androgen regulation of the motor copulatory pattern in the male New Zealand white rabbit

The effect of castration and testosterone propionate (TP) treatment on the motor copulatory pattern was studied by an accelerometric technique in five sexually experienced New Zealand white male rabbits. This technique permits the oscillographic recording of thrusting frequency, rhythmicity, and amplitude of the pelvic movements occurring during copulation. Castration resulted in a marked decrease in sexual activity in all rabbits. Mounting, including occasional intromissions, was retained by four of the rabbits for periods ranging from 2 to 15 weeks. Castration did not affect mount duration, but decreased strength and frequency of pelvic thrusting. Diminution in the frequency of pelvic thrusting was mainly due to intercalation of pauses within the mounting trains. The change in the rhythm of pelvic thrusting was related to the failure of most mounts performed by castrated rabbits to stimulate lordosis in the female. Testosterone propionate (TP, 10 mg daily for 15 days) restored mounting activity and increased strength and frequency of pelvic thrusting in all rabbits. It is concluded that TP, besides stimulating sexual motivation, regulates the vigor and rhythm of pelvic movements during copulation in the rabbit.     

Maturation of spontaneous fetal diaphragmatic activity and fetal response to hypercapnia and hypoxemia

The electromyogram (EMG) of the diaphragm, lateral rectus, and nuchal and hindlimb muscles were studied during spontaneous activity and during hypercapnia or hypoxemia in eight fetal sheep from 0.5 to 0.8 gestation (73-128 days). At the earliest gestational age, diaphragmatic EMG activity was mainly tonic and associated with tonic activity of somatic muscles. The stimulus for the diaphragmatic activity originated centrally. Brief periods of a rapid-eye-movement (REM) state characterized by phasic lateral rectus and diaphragmatic activity and absence of nuchal activity were recognized. Furthermore, from 0.5 to 0.7 gestation onward, activity of all muscles increased. Thereafter increased specificity of activity in relation to the apparent REM and non-rapid-eye-movement (NREM) state occurred. With maturation, phasic diaphragmatic activity increased at the expense of tonic activity. The most striking effect of maturation on apnea was a greater proportion of apnea lasting greater than 1 min, but the total duration of apnea as a percent of a total recording remained unchanged. The quantitative response to hypercapnia during maturation was independent of the pattern of spontaneous diaphragmatic activity. Hypercapnia at 0.5 gestation changed the pattern of diaphragmatic EMG activity from mainly tonic to phasic. Thus the central chemoreceptors and appropriate neuronal pathways are present and functional as early as 0.5 gestation. Hypercapnia at 0.5 gestation caused a shift in diaphragmatic EMG power to lower frequencies similar to that found during control conditions in the older fetus. This might suggest that during maturation there is increased recruitment of phrenic motoneurons. Hypoxemia abolished tonic somatic activity at 0.5 gestation and decreased phasic diaphragmatic activity at more advanced gestational ages. Therefore the central inhibitory mechanisms of hypoxemia are developed by 0.5 gestation.     

The action of prostacyclin (PGI2) on the contractility of the isolated circular and longitudinal muscle layers of the human oviduct

Small strips from the circular and longitudinal muscle layers of the ampullary-isthmic portion of the human oviduct were mounted in organ chambers for recording of their spontaneous contractility. Concentrations in the order of 1–300 ng/ml of PGI₂ were tested and compared with similar concentrations of PGE₂ and PGF_2α. It was found that PGI₂ contracted the longitudinal muscle layer in the same manner as did PGE₂. The spontaneous activity of the circular layer was markedly suppressed by PGE₂ but only moderately inhibited by PGI₂ even at high concentrations.