Characterization of plasma progesterone concentrations for two distinct luteal morphologies in mares

Plasma progesterone concentrations in mares were determined in two experiments during the time that the luteal glands were detectable by transrectal ultrasonography. In both experiments, corpora lutea were classified into two tupes of morphologies based on their ultrasonic appearance: centrally nonechogenic luteal glands (fluid-filled) and uniformly echogenic luteal glands (non-fluid-filled). In Experiment 1, daily blood samples were taken from horse mares during August through October and May through July. There were no significant effects of season or luteal morphology on progesterone concentration. There was a significant main effect of day, but no day-by-season or day-by-morphology interactions. Progesterone increased significantly between Days 1 and 3 (mean progesterone concentration, 2.5 vs 5.2 ng/ml, respectively), between Days 3 and 4 (5.2 vs 7.8 ng/ml), and between Days 4 and 5 (7.8 vs 11.0 ng/ml). Progesterone did not decrease significantly until between Days 11 and 15 (11.6 and 6.1 ng/ml). Subsequent decreases occurred between Days 15 and 16 (6.1 vs 3.9 ng/ml), and Days 16 and 17 (3.9 vs 2.5 ng/ml). In Experiment 2, blood samples were obtained from pony mares at 1 2 - h intervals for 3 h before and 2 h after the defined onset of luteal development (end of evacuation of the ovulatory follicle). Additional blood samples were taken at 5, 8 and 12 h after the onset of luteal development, and thereafter at 12-h intervals for 5d. There were no significant differences between centrally nonechogenic luteal glands (n = 7) and uniformly echogenic luteal glands (n = 5) during the first 5 d of luteal development. There was no time-by-morphology interaction, but there was a significant time effect. The first significant increase in progesterone concentration occurred between Hours 12 and 24 (0.5 vs 1.1 ng/ml). Additional increases were detected between Hours 24 and 36 (1.1 vs 2.6 ng/ml), Hours 36 and 48 (2.6 vs 4.3 ng/ml), Hours 48 and 60 (4.3 vs 6.1 ng/ml), Hours 60 and 72 (6.1 vs 9.4 ng/ml), and Hours 72 and 96 (9.4 vs 13.8 ng/ml). The hypothesis was supported that fluid-filled corpora lutea do not differ from non-fluid-filled corpora lutea with regard to progesterone production.     

Ultrasonography for detection of pregnancy and study of embryonic development in heifers

Real-time B-mode ultrasonography was used to evaluate uterine changes associated with the estrous cycle in 22 ovulatory periods in 12 nulliparous heifers. Irregular, nonechogenic (black) areas were seen on the images of uterine horns during the periovulatory period. These nonechogenic areas were presumably due to intraluminal fluids since they coincided with the discharge of clear, viscous mucus preceding ovulation and blood-tinged mucus after ovulation. Eight heifers were bred until five pregnant heifers were obtained for study of the ultrasonic morphology of the conceptus. Ultrasound examinations were done daily to day 50 of pregnancy. Discrete, nonechogenic areas were first visible within the uterus between days 12 and 14, when they were approximately 2 mm in diameter. These discrete nonechogenic structures were identified as the embryonic vesicle, since they were observed only in heifers later confirmed to be pregnant and were always in the uterine horn ipsilateral to the corpus luteum. The presence of an embryo within the embryonic vesicle was confirmed by observing an echogenic (white) area with rhythmic pulsations (heartbeat). The embryonic vesicle gradually increased in length from the day of first observation until day 26 when it extended past the curvature of the horn and began to encroach into the contralateral horn. In all heifers, by day 32 the vesicle extended to the tip of the contralateral horn. The embryo was first visible between days 26 and 29 when the mean length was 10 mm. The embryo increased in length an average of 1.1 mm per day. A heartbeat was detectable in the embryo on the first day observed. In one superovulated heifer, five vesicles were visible in the uterine horns by day 14 and by day 33 seven embryos were observed; two of the seven embryos apparently resorbed by day 43.     

Ultrasonic detection of the conceptus and characterization of intrauterine fluid on days 10 to 22 in heifers

Nonbred and pregnant heifers were examined ultrasonically on alternate days from Days 10 to 22 in three experiments. The goal was to define criteria that could be used for identification of the early conceptus. The location and size of circular and elongated nonechongenic structure (apparently free intrauterine luminal fluid) were recorded (Experiment 1). On Day 12, there was a significantly greater number of circular nonechogenic structures in the ovarian half of the uterine horn ipsilateral to the corpus luteum in pregnant heifers than in nonbred heifers (means, 5.1 vs 3.5). Therefore, the embryonic vesicle was apparently detected on Day 12, but was not distinguished from circular nonechogenic structures representing free luminal fluid. The number of circular nonechogenic structures decreased after Day 14, with a corresponding increase in elongate nonechogenic structures associated with luteal regression (nonbred heifers) or elongation of the blastocyst (pregnant heifers). On Day 16, there were more elongated than circular nonechogenic structures in pregnant heifers than in nonbred heifers. In nonbred heifers, there was no evidence of a local utero-ovarian relationship between the amount of intrauterine luminal fluid and the location of either the corpus luteum or the next ovulatory follicle. Uterine luminal fluid in each horn increased several fold between Days 16 and 18 in nonbred heifers; there was significantly more lumina fluid in nonbred than in pregnant heifers on Days 18 and 20. In pregnant heifers, intrauterine luminal fluid area increased first in the ovarian end of the uterine horn ipsilateral to the corpus luteum; this was apparently due to detection of the elongating blastocyst. The length of the longest specular reflector on the endometrial surface of the uterine lumen increased over Days 10 to 22 in nonbred and pregnant heifers, but was not a useful criterion for detection of the conceptus (Experiment 2). The accuracy of a 7.5 MHz transducer for pregnancy diagnosis was not greater than a guess (50%) before Day 16 (Experiment 3). Results indicated that early pregnancy diagnosis in heifers was confounded by the presence of intrauterine fluid.     

Ultrasonic anatomy and pathology of the equine uterus

The morphological and pathological status of the uterus in mares was evaluated using a linear-array ultrasound scanner, and the ultrasonic properties of the uterus were characterized. The uterus was examined each day in 16 mares, beginning at mid-diestrus. The uterus was recorded as having an ultrasonic morphology characteristic of diestrus (endometrial folds not distinguishable), estrus (prominent endometrial folds) or an intermediate stage (folds only moderately distinguishable). The number of mares with an intermediate or estrous image increased gradually between day -7 (2 14 mares; ovulation = day 0) and days -3 (11 16 ) and -2 (10 16 ) and then declined between days -2 and +1 (0 12 ). In another study, a large nonechogenic area (ejaculate) was visible in the uterine lumen immediately after mating in all of six mares. During the course of pregnancy diagnoses and hand-breeding, several pathological conditions of the uterus were first observed by ultrasound examination and confirmed by digital exploration of the uterus or at slaughter. Cysts contained nonechogenic material (fluid) and were usually compartmentalized. The purulent material associated with pyometria was relatively nonechogenic but contained echogenic spots. Mummified fetuses and remnants of fetal bones showed echogenic properties consistent with high tissue density. The results demonstrated that ultrasound technology provides a noninvasive form of visual access to the uterus to evaluate normal, morphological changes and to detect and study certain pathological processes.     

Ultrasonic characterization of ovulatory follicular evacuation and luteal development in heifers.

The characteristics of follicle evacuation during ovulation and the development of the corpus luteum until day 5 (day 0 = ovulation) were studied in seven nulliparous Holstein heifers using real-time ultrasonography. Ovulation was induced and synchronized with a single injection of PGF2 alpha followed in 36 h by GnRH. Continuous scanning and videotaping was performed from apparent stigma formation until antral fluid was no longer detected. The beginning of follicular evacuation (second 0) was defined, retrospectively, after the antral area decreased 10% or more in 1 s. The completion of evacuation was defined as the inability to detect the antrum (the beginning of luteal development, 0 h). Corpora lutea development was monitored at 0, 4 and 20 h, and every 24 h thereafter until day 5. Changes in the maximal cross-sectional area of the antrum, luteal tissue, and central luteal cavities and in the pixel intensity of luteal tissue were determined using a computerized image program. The initial antral fluid evacuation occurred in two patterns that could be readily separated: (1) rapid, means of 58 and 89% evacuation in 1 and 4 s, respectively (four heifers); and (2) slow, means of 17 and 35% in 1 and 4 s, respectively (three heifers). The initial loss that distinguished the two patterns involved about 4 and 20 s for rapid and slow evacuation, respectively. Thereafter, the loss patterns were similar for the two types. The time from the beginning to the completion of evacuation ranged from 6 s to 14.5 min. Mean luteal tissue area increased (P < 0.05) between completion of evacuation (91.2 +/- 6.5 mm2) and day 3 (164.4 +/- 13.7 mm2) and between day 3 and day 4 (263.4 +/- 26.6 mm2). The growth rate of the luteal tissue area between day 3 and day 4 (103.2 +/- 16.0 mm2 day-1) was greater (P < 0.05) than that between day 2 and day 3 (41.9 +/- 12.4 mm2 day-1) and between day 4 and day 5 (49.7 +/- 22.0 mm2 day-1). In contrast to increasing luteal tissue area, mean pixel intensity decreased (P < 0.05) progressively between the completion of evacuation (78.4 +/- 6.3) and day 2 (60.4 +/- 2.5) and did not change significantly thereafter. In conclusion, initial follicular fluid loss during ovulation occurred in two patterns, involving about 4 and 20 s, respectively. The most intensive luteal tissue growth occurred between day 3 and day 4, and the echogenicity of the luteal tissue decreased between day 0 and day 2.     

Laparoscopic evaluation of the reproductive organs and abdominal cavity content of the lowland gorilla

Laparoscopy was used to examine the abdominal cavity and evaluate reproductive competence in six adult female gorillas which had not demonstrated copulatory activity within 12 months of examination. Blood samples were obtained on the day of laparoscopy and analyzed for serum estradiol-17β and progesterone. The uterus and oviducts of each gorilla appeared morphologically normal and free of lesions. The ovaries of three animals contained anatomical evidence of activity as indicated by the presence of luteal scars and a prominent corpus hemorrhagicum, corpus luteum, or developing follicles. No visible luteal or follicular tissue was observed on the ovaries of three gorillas. In the eldest female (age, 22 years) the ovaries were flaccid in texture and demonstrated irregularities in ovarian surface integrity, similar to that observed in women during reproductive senescence. Serum concentrations of estradiol-17β and progesterone on the day of laparoscopy corresponded with observed ovarian activity. All six females had extensive adhesions in the abdominal cavity exclusive of the reproductive organs. Numerous gut, omentum, and liver to peritoneum adhesions were observed. The results indicated that laparoscopy was a safe and effective method for evaluating anatomical competence in the gorilla. Uterine and ovarian integrity appeared morphologically normal, but only three of the six gorillas showed evidence of ongoing ovarian activity.     

Ultrasonic morphology of corpora lutea and central luteal cavities during the estrous cycle and early pregnancy in heifers

Ultrasonography was used once daily to quantify corpora lutea, central luteal cavities, and luteinized tissue during interovulatory intervals (n = 66) and during Days 0 to 60 of pregnancy (n = 14) in nulliparous Holstein heifers (ovulation = Day 0). The corpus luteum of the estrous cycle was detectable by ultrasonography in most heifers from the day of ovulation (mean, Day 0.5) and extending into the regressive phase beyond the next ovulation (mean, Day 1.4 +/-0.2 after the next ovulation). During pregnancy, the corpus luteum was detected until Day 60 (end of study). Maximal central luteal cavity area detected on Days 0 to 20 was used retrospectively to group luteal glands into four cavity categories: no, small, medium, and large. These categories corresponded to approximate cavity diameters of <2 mm, 2 to 5 mm, 6 to 10 mm, and >10 mm, respectively. The incidence of each cavity category was similar between interovulatory intervals and pregnancies (combined incidence, 17 80 , 8 80 , 33 80 , and 22 80 for no, small, medium, and large cavities, respectively; total with cavities, 63 80 , 79%). Mean day of first detection of a central cavity was earliest for large cavities during interovulatory intervals (means, Days 4.7, 4.4, and 3.0 for small, medium, and large cavities, respectively; P<0.04) and during pregnancies (means, Days 5.5, 4.2, and 3.3, respectively; NS). However, the day that the cavities reached maximum size (range of means, Days 5.5 to 7.0) did not differ among categories. Mean day of last detection of the central cavity was significantly different among cavity categories during interovulatory intervals (means, Days 9.3, 11.1, and 17.4 for small, medium, and large cavities, respectively) and pregnancies (means, Days 7.0, 8.8, and 20.2, respectively). Time of loss of central cavities was similar between nonbred and pregnant heifers, and there was no significant difference among cavity categories in the length of the interovulatory interval (mean, 20.1 d). Luteal tissue area was not significantly different among cavity categories during interovulatory intervals. There were no indications that cavities were functionally important. Luteal tissue area increased linearly in pregnant heifers on Days 21 to 60 (mean slope, 2.6 mm(2)/day).     

Relationship between blood flow and steroidogenesis in the rabbit corpus luteum

Blood flow in the corpus luteum of the pseudopregnant rabbit was measured with tracer-labelled microspheres before and at 1 and 3 h after saline treatment (N = 8) or after inhibition of progesterone synthesis with aminoglutethimide (N = 10). Before treatment luteal blood flow (29.5 +/- 3.9 ml/min.g-1 (mean +/- s.e.m.] was much higher than blood flow to other tissues (ovarian stroma = 2.9 +/- 0.6; uterus = 0.5 +/- 0.1; adrenal gland = 2.6 +/- 0.2 ml/min.g-1). Aminoglutethimide reduced serum progesterone by 60% within 1 h but luteal blood flow was unchanged (26.2 +/- 3.5 ml/min.g-1). At 3 h after aminoglutethimide, serum progesterone remained low and luteal blood flow was slightly reduced to 22.5 +/- 3.4 ml/min.g-1. This reduction was associated with a significant decline in mean arterial blood pressure which resulted in luteal vascular resistance being unaltered by aminoglutethimide treatment. Further analysis of these data indicated that serum progesterone concentration was not significantly correlated with blood flow to the corpora lutea or with blood flow to other tissues. In contrast, mean arterial blood pressure was highly correlated with blood flow to the corpus luteum (r = 0.80; P less than 0.001) but not to the ovarian stroma (r = 0.04), or adrenal gland (r = 0.06). These results indicate that luteal blood flow is not acutely responsive to changes in luteal progesterone production and suggest that luteal blood flow changes passively with changes in arterial blood pressure.     

Mammogenesis and changing prolactin receptor concentrations in the mammary glands of the tammar wallaby (Macropus eugenii)

All 4 mammary glands of the tammar wallaby showed a steady increase in weight and prolactin receptor concentration during the luteal phase of the oestrous cycle to reach a peak at oestrus. Removal of the corpus luteum abolished this mammogenesis , while pregnancy, which in this species is a day or so shorter than the oestrous cycle, had no effect. This provides an explanation for the previous finding that pregnancy is not a necessary pre-requisite for lactation in marsupials and that nonpregnant animals will lactate very successfully, provided the suckling stimulus is applied at the correct stage of the oestrous cycle. During lactation, only the gland supplying the teat to which the pouch young was attached developed and showed any further increase in prolactin receptors; the other 3 glands remained small and inactive. These results indicate the importance of the suckling stimulus and milk withdrawal on the initiation and maintenance of lactation.     

Progesterone and LH variations after LH-RH administration in the luteal phase of the menstrual cycle

We have investigated the pituitary and luteal responses to LH-RH and their related changes. 11 normal women were studied during the luteal phase (day +4/+11). Blood samples were collected every 15 min for a basal period of 180 and 120 min after the intravenous administration of 25 micrograms of LH-RH. Progesterone (P) and LH were assayed by radioimmunoassay. Data were analyzed as maximum peak and its percent increase (delta max), integrated secretory area (ISA) and percent increase of ISA (delta A) in respect to basal values for both P and LH. LH-RH elicited a secretory response of both hormones in all cases. ISA of LH was significantly greater after LH-RH administration in respect to basal values (p less than 0.001) and delta max accounted to 475 +/- (SE) 36% of the basal concentration. Luteal responsiveness varied from about 115-130% to more marked increments. ISA of P differed from basal to stimulated conditions (p less than 0.05) and delta max was 166 +/- (SE) 14%. The analysis of temporal relationship between P and LH secretion showed that LH promptly rose after LH-RH, while the enhancement of P plasma levels occurred within 31 +/- 19 min after LH rise. Then P levels reached a plateau, values of which were statistically different from those observed before LH-RH administration. In two cases where luteal function was blunted or absent, in spite of marked increments of LH, P secretion did not occur. These data are consistent with the presence of close relationships between hypothalamic, pituitary and luteal functions and strengthen the contention about the usefulness of LH-RH during luteal phase for the lifespan and maintenance of corpus luteum.     

Regression and resurgence of the CL following PGF2alpha treatment 3 days after ovulation in mares

The present study was designed to characterize and compare the physiology and ultrasonographic morphology of the corpus luteum (CL) during regression and resurgence following a single dose of native prostaglandin F2alpha (PGF) given 3 days after ovulation, with a more conventional treatment given 10 days after ovulation. On the day of pre-treatment ovulation (Day 0), horse mares were randomly assigned to receive PGF (Lutalyse; 10 mg/mare, i.m.) on Day 3 (17 mares) or Day 10 (17 mares). Beginning on either Days 3 or 10, follicle and CL data and blood samples were collected daily until post-treatment ovulation. Functional and structural regression of the CL in response to PGF treatment were similar in both the Day 3 and 10 groups, as indicated by an abrupt decrease in circulating concentrations of progesterone, decrease in luteal gland diameter and increase in luteal tissue echogenicity. As a result, the mean +/- S.E.M. interovulatory interval was shorter (P < 0.0001) in the Day 3 group (13.2 +/- 0.9 days) than in the Day 10 group (19.2 +/- 0.7 days). Within the Day 3 group, functional resurgence of the CL was detected in 75% of the mares (12 of 16) beginning 3 days after PGF treatment, as indicated by transient major (6 mares) and minor (6 mares) increases (P < 0.05 and < 0.1, respectively) in progesterone. Correspondingly, mean length of the interovulatory interval was longer (P < 0.03) in mares with major resurgence (15.8 +/- 1.6 days) than in mares with minor (11.2 +/- 1.2 days) and no resurgences (13.5 +/- 0.3 days) in progesterone. Structural resurgence of the CL in the Day 3 group and functional and structural resurgence in the Day 10 group were not detected. In conclusion, PGF treatment 3 days after ovulation resulted in structural and functional regression of the CL and hastened the interval to the next ovulation, despite post-treatment resurgences in progesterone.     

Ultrasonic evaluation of the preovulatory follicle in the mare

Ultrasonically visible characteristics of preovulatory follicles in mares which single ovulated were studied daily for 79 preovulatory periods in 40 mares. The preovulatory follicle became the largest follicle in the ovary from which ovulation later occurred six or more days before ovulation in 65 of 79 (82%) preovulatory periods; the mean was day -7 (range, day -14 to day -4). The increase in mean diameter of the preovulatory follicle was linear (R(2)=99.5%) over day -7 (29.4 +/- 0.8 mm) to day -1 (45.2 +/- 0.5 mm; growth rate, 2.7 mm/day). Follicles which double-ovulated were smaller (P<0.05) on day -1 (36 +/- 1.6 mm; n=12 follicles). Preovulatory follicles exhibited a pronounced change in shape from a spherical to a conical or pear-shaped structure in 84% of the preovulatory periods. Remaining follicles retained a spherical shape. Scores representing thickness of the follicular wall increased (P<0.05) as the interval to ovulation decreased. There was no significant difference among days in mean gray-scale value of the follicular wall or in echogenicity of the follicular fluid. Although diameter and shape of the follicle and thickness of the follicular wall changed during the preovulatory period, no reliable ultrasonically visible predictor of impending ovulation was found.     

Effect of inhibition of estrogen synthesis during the luteal phase on function of the corpus luteum in rhesus monkeys

It has been hypothesized that estrogen synthesized by the corpus luteum initiates luteal regression during the nonfertile menstrual cycle in primates. To study the role of endogenous estrogens in functional regression of the monkey corpus luteum, we administered the aromatase inhibitor 1,4,6-androstatriene-3,17-dione (ATD) to rhesus monkeys during the luteal phase of the menstrual cycle. Twice-daily oral administration of ATD suppressed systemic and intraluteal estrogen levels by 80-90%. The midluteal phase rise in estradiol concentrations that occurs in rhesus monkeys was completely abolished by ATD treatment. Despite suppression of estrogen synthesis during the luteal phase, mean menstrual cycle length and length of the luteal phase were not different than in control monkeys treated with vehicle only. Progesterone levels were lower in the ATD-treated group on the second and third day of treatment, but did not differ from control levels during the remainder of the cycle. These data suggest that elevated estrogen synthesis during the luteal phase of the menstrual cycle is not a prerequisite for spontaneous luteolysis in rhesus monkeys.     

B-mode ultrasonographic examination of the accessory sex glands of boars

Thorough examinations of the reproductive system of boars are generally not performed on normal boars to be used for breeding; only boars with problems undergo a form of a breeding soundness examination. In order for veterinarians to identify pathological conditions, the normal architecture of the accessory sex glands needs to be described. The purpose of this study was to use B-mode ultrasonography to describe the accessory sex glands in the boar and to see if transrectal ultrasonography would be a viable option in which to obtain this data. Initially, cross-sectional saline bath examinations of accessory sex glands were performed on crossbred boar reproductive tracts (n = 4) using B-mode ultrasonography equipped with a 5 MHz dual frequency linear array transducer. In situ examinations were also performed on terminal line crossbred boars (n = 16) ranging in age from 10 to 23 months old using the same ultrasound methodology; four boars were under general anesthesia and the remaining 12 were standing in crates. Eight boars were abstinent for 2 days and the other eight had ejaculates collected 2 h prior to examination. The paired bulbourethral glands are best described as a long oval gland with a uniformly echogenic appearance with a large anechoic space in the center of the gland extending most of its length. The walls of the vesicular glands were found to be thin, with the parenchyma having multiple small echolucent areas that appeared to merge and form a central canal. The prostate gland was best identified as a pecan-sized gland with a uniform echogenic appearance. Visualization of the prostate gland was accomplished with more proficiency using the saline bath ultrasonography as compared to in situ examinations. All of the accessory sex glands could be examined using both methodologies of ultrasonographic examination with a 5 MHz frequency linear array transducer. It was determined that each accessory sex gland could be recognized, and differences between ejaculated and nonejaculated boars could be identified. The results of this study demonstrate that transrectal ultrasonography can be used as a diagnostic aid in assessing the accessory sex glands of boars.     

Anatomy and histology of the babirusa (Babyrousa babyrussa) stomach

Stomachs from six adult and one 12-month-old babirusa (Babyrousa babyrussa) were collected from zoological gardens. The babirusa stomach was larger than that of the domestic pig (Sus scrofa), and possessed a large diverticulum ventriculi. Its gastric glands were confined to a small, easily identifiable unit at the end of the corpus ventriculi, and the connections between the different stomach parts were wide and unrestricted, with the exception of the well-defined connection between the diverticulum ventriculi and the fundus ventriculi. Microscopically, the structure of the cardiac, gastric and pyloric glands was similar to that of Sus scrofa. However, the mucusproducing cardiac glands of babirusa occupied a larger area within the stomach (> 70% versus 33%). The pH in the lumen of the cardiac gland area lay between 5.3 and 6.4 and micro-organisms were found here. It was hypothesised that the babirusa is a nonruminant foregut-fermenting frugivore/concentrate selector.     

Noninvasive monitoring of follicle development,ovulation, and corpus luteum formation in the marmoset monkey (Callithrix jacchus) by ultrasonography

This is the first paper to describe ovarian changes associated with follicular growth, ovulation, and corpus luteum (CL) formation as monitored by ultrasonography in a multiovular primate, the marmoset monkey (Callithrix jacchus). Examinations were carried out transabdominally on unsedated females using a 10 MHz probe. Cycles were monitored by plasma progesterone and controlled by administration of prostaglandin F2α (PGF). The reliability of ultrasound was validated by comparing findings with direct observation of the ovaries at laparotomy. In eight females, 25 follicles were counted, of which 92% were depicted correctly by ultrasound. Of 14 CLs in five females, number and position were confirmed at laparotomy for 78%. Ultrasound examinations of ovaries throughout the follicular and luteal phase were performed in eight cycles and related to plasma profiles of luteinizing hormone (LH) and progesterone. One of these cycles was anovulatory. In the remaining seven cycles, 19 follicles were considered ovulatory follicles since they were seen on consecutive days and found again as CLs. Growth of individual follicles was monitored by measurements of follicle diameter from day 7 onward. Disappearance of follicles or changes in echogenicity were noted between days 9 and 11, preventing further measurements. Mean follicle size increased from 2.1 mm (range 1.6 mm–2.7 mm) on day 7 to 3.2 mm (range 2.7 mm–4.0 mm) on the day last seen. With one exception, the day follicles were last seen by ultrasound was consistent with the day of the preovulatory LH surge (day 9–11). The postovulatory rise in progesterone occurred 1–2 days later (day 11–13). These findings suggest that the day of ovulation as observed by ultrasound was characterized by either disappearance of follicles or increased follicular echogenicity. In conclusion, ultrasonography provides a reliable, noninvasive method for examinations of the ovarian cycle in the marmoset monkey. © 1996 Wiley-Liss, Inc.     

Autocrine activation of DNA synthesis in prothoracic gland cells of the silkworm, Bombyx mori

Autocrine activation of DNA synthesis in prothoracic gland cells in last instar larvae of the silkworm, Bombyx mori, was studied using both a long-term in vitro organ culture system and immunocytochemical labeling with 5-bromo-2'-deoxyuridine (BrdU). When prothoracic glands were incubated in a small volume of culture medium (10 microl/gland), the numbers of DNA-synthesizing cells per gland increased significantly, and DNA synthesis was stimulated less by hemolymph, as compared with glands incubated in a large volume (50 microl/gland). Moreover, glands cultured in groups (6 glands per group in a 50-microl drop) also resulted in much higher levels of DNA synthesis than those cultured individually in a 50-microl drop. The mechanism by which alternation of the volume of the incubation medium results in changes in the levels of DNA synthesis was further examined. When prothoracic glands were incubated in medium (50-microl drop per gland) that was preconditioned with glands (in a 10-microl drop individually), a dramatic increase in DNA synthesis activity was also observed, indicating that prothoracic glands may release a factor that stimulates their own DNA synthesis. The growth-promoting factor was further characterized and it was found that the factor is heat stable, and its molecular weight was estimated to be between 1,000 and 3,000 Da. Moreover, the factor also stimulated corpus allatum cell DNA synthesis in vitro. Injection of concentrated putative growth-promoting factor into day 4 last instar-ligated larvae greatly increased cell DNA synthesis of the prothoracic glands, indicating the in vivo function of the present autocrine factor.     

Growth and cellular proliferation of pig corpora lutea throughout the oestrous cycle

Corpora lutea were obtained from gilts on days 2, 4, 8, 12, 15 or 18 after oestrus. Luteal fresh masses and DNA contents increased linearly (P < 0.01) from day 2 to day 12 and day 2 to day 15, respectively. Changes in the ratio of protein:DNA were greatest between days 2 and 4 and days 15 and 18, whereas changes in DNA content were relatively small during the same intervals. Thus, a major component of changes in the size of the corpus luteum during the early and late periods of the luteal phase was cellular hypertrophy. Proliferation of luteal cells in vivo (nuclear incorporation of 5-bromo-2-deoxyuridine, a thymidine analogue) was greatest on day 2 and decreased exponentially (P < 0.01) throughout the oestrous cycle. Results from co-localization of 5-bromo-2-deoxyuridine and factor VIII (von Willebrand factor), a marker of endothelial cells, or 5-bromo-2-deoxyuridine and 3 beta-hydroxysteroid dehydrogenase, a marker of steroidogenic cells, indicated that some of the luteal steroidogenic cells proliferated early in luteal development. However, during early and mid-cycle, most of the luteal cell proliferation occurred in the endothelial cells. Thus, during growth of the pig corpus luteum, which is extremely rapid, most of the proliferating luteal cells are vascular endothelial cells. This observation is consistent with the high vascularity and blood flow of the mature corpus luteum and implies a critical role for angiogenesis in luteal development in the pig, as has been proposed for several other mammalian species.     

Metamorphosis of the corpus allatum and degeneration of the prothoracic glands during the larval-pupal-adult transformation of Drosophila melanogaster: a cytophysiological analysis of the ring gland

The degeneration of the prothoracic glands of Drosophila melanogaster during pupal-adult metamorphosis was analyzed by light microscopy, scanning, and transmission electron microscopy. The ultrastructural observations were correlated with the ability of the ring gland to synthesize ecdysteroids in vitro. The ring gland is prominent during larval life and is identifiable until just before adult eclosion but undergoes dramatic changes in location, shape, size, ultrastructure, and function during pupal-adult development. Prothoracic gland degeneration is characterized by: a gradual decrease in its ability to synthesize ecdysteroids; a decreasing quantity of smooth endoplasmic reticulum (SER) and mitochondria; the absence of intercellular channels; cytoplasmic fragmentation; and the separation of the prothoracic gland from the corpus allatum and corpus cardiacum. An ultrastructural analysis of the corpus allatum during larval-pupal-adult metamorphosis and adult life was also correlated with function, i.e., juvenile hormone biosynthesis, using a radiochemical assay of ring glands and adult corpora allata in vitro. A relatively high concentration of SER, mitochondria, and mitochondrion-scalariform junction complexes are typical features of an active corpus allatum cell. The migration of the corpus allatum from the ring gland to its position as a separate gland in the adult fly was studied in detail. The capacity of the corpus allatum to synthesize juvenile hormone is at its peak in the ring gland of the early wandering third instar larva, whereas the corpus allatum of 2-day-old female adults displayed the greatest synthetic activity during adult life. The physiological significance of the alterations in gland activity is discussed.     

The rabbit submandibular gland: sexual dimorphism, effects of gonadectomy, and variations across the female reproductive cycle

Sex differences and the effect of various endocrine conditions on the histology of the rabbit (Oryctolagus cuniculus L.) submandibular (chin) gland were investigated. The female gland contained significantly more acini/field than the male gland. The diameter of acini was significantly smaller than those of the male gland. Gonadectomy reduced the number of acini/field and increased their diameter in females but provoked the opposite effect in males. Gonadectomy drastically reduced the percent of acini with crystal bodies in both sexes, and the percent of acini with apocrine secretion only in females. Estrous does showed a significantly higher number of acini/field than pregnant (days 20 and 29) and lactating (day 6) does. Acini containing crystal bodies declined from 22% in estrous females to 8% and 3% in pre-parturient (gestation day 31) and lactating (day 6) does, respectively. By contrast, acini showing apocrine secretion increased from 12% in estrous females to 43% in pre-parturient does and declined to 23% on lactation day 6. In all glands glycoproteins were noted in crystal bodies but not in apocrine secretion. Results show a sexual dimorphism in the rabbit chin gland histology and support the participation of gonadal steroids in its physiological regulation.Abbreviations GNX gonadectomized - PAS periodic acid Schiff - PASD periodic acid Schiff-diastase