Absorption of testosterone-7alpha-3H by reproductive tract tissues of male-embryo rats in vitro]

The dynamics of testosterone-7alpha-3H consumption by the reproductive tract rudiments (tubules, urogenital sinus and genital tubercle) and the muscles has been studied in the in vitro experiments immediately after the isolation of embryos (16-21 days of development), as well as after the preliminary two days cultivation in the diffusion chambers (13-20 days of development). In the first series of experiments, the selective, specific consumption of the labelled androgen was observed at all stages under study, whereas in the second series of experiments the specific and selective consumption in the organs-targets was noted from the 15th day of development (in the genital tubercle and urogenital sinus). No selective consumption in the tubules was noted.     

Absorption of testosterone-7 alpha-3H by tissues of organ-targets in male rat embryos]

The dynamics of consumption of testosterone-7alpha-3H by different tissues of the differentiating reproductive system, as well as by suprarenals, brain, hypothalamus and muscles was studied in the in vivo experiments with the rat male embryos beginning from the 14th and till the 20th days of development. The specific consumption of the labelled androgen related to the metabolic processes was observed at all developmental stages under study. The selective consumption of testosterone-7alpha-3H was noted prior to the beginning and during the morphogenesis of the organs-targets' rudiments. The radioactivity in tissues increased already 1--2 days following the castration of 18 days old embryos. Estrone, testosterone, testosterone-propionate, cyproterone-acetate competitively decreased the consumption of testosterone-7alpha-3H. The problem of reception of tissues of the rat male embryos to the male sex hormone is discussed.     

Effect of testosterone on the rate of total protein synthesis in the reproductive tract of guinea pig embryos in vitro

The influence of testosterone on the rate of total protein synthesis in the reproductive tract tissues and muscles was studied in the guinea pig male embryos from 26 days of development until birth. The rate of protein synthesis was estimated by the intensity of 3H-leucine incorporation per 1 mg protein for 30 min. Testosterone increased the rate of protein synthesis at all studies stages. A higher rate of protein synthesis was revealed at the stages of active morphogenesis and growth of rudiments and organs of genital system and correlated with their ability of 3H-testosterone uptake (Ivanova, 1982).     

Effect of testosterone on the rate of total protein synthesis in the reproductive tract of rabbit embryos in vitro

The total protein synthesis rate in the reproductive tract and muscles of rabbit fetuses was evaluated at the 18th-22nd day of development by the intensity of 3H-leucine incorporation in experiments in vitro. Testosterone increased the total protein synthesis rate in the tissues of the reproductive tract at all the developmental stages under study. The highest rate of protein synthesis was recorded on the 18th day of development.     

Uptake and metabolism of [3H]norepinephrine in the cerebral hemispheres of chick embryos

Abstract— The uptake and metabolism of [³H]norepinephrine were studied in slices of cerebral hemispheres removed from chick embryos at 10, 15 and 20 days of embryonic age, as well as from 90-day-old hens. Brain tissue from all age groups concentrated [³H]norepinephrine to much greater levels at 37°C than at 0°C. There was a marked increase in the rate of accumulation of [³H]norepinephrine in tissues from 10 to 15 days of embryonic age, with no further increase in the rate observed from 15 to 20 days of embryonic age. Tissue slices were incubated for 20 min with [³H]norepinephrine, and the deaminated metabolites of norepinephrine were separated by paper chromatography. In tissues from all age groups, the neutral metabolites were produced in greater amounts than the acid metabolites. A significant increase in the amounts of deaminated metabolites formed was observed in the period from 10 to 15 days of embryonic age and a significant decrease in the amounts formed was observed in the period from 15 to 20 days of embryonic age. The deamination at 20 days was very similar to that observed in the adult. A significant decrease in the level of the deaminated metabolites was noted in all age groups in response to cocaine (an inhibitor of neuronal uptake mechanisms), an observation suggesting that mechanisms for neuronal uptake of NE are functional by 10 days of embryonic development in the chick. However, a significant increase in the level of deaminated metabolites in response to reserpine (an inhibitor of uptake of NE into storage granules) was observed only in slices taken from 20-day embryos and from the 90-day-old hen. The effect in the hen was more prominent than in the 20-day embryo. These results were interpreted to indicate that mechanisms for the uptake of NE develop at an earlier embryonic age than mechanisms for the storage of NE and that mechanisms for storage continue to develop after hatching.     

Prostaglandin (PG) accumulation in vitro by mouse ovaries, oviducts, and uteri

Uteri, ovaries and oviducts from mice were collected at autopsy. Tissue slices were incubated with [3H]-PGE2 in the presence or absence of a large excess (100 fold) of nonradioactive PGE2 using 0.01M sodium phosphate buffer (pH 7.2). Bound and free PGs were separated by a filtration technique. PGE2 accumulation by the uteri was evaluated as a function of incubation time, wet weight of tissues, and reproductive state. The tissue to medium ratio (T/M) was greater than 1.0 for uteri as the time of incubation increased. This suggests the presence of PGE2 binding sites in mouse uterine tissue. Also, PGE2 accumulation was not observed in oviducts or in ovaries.     

54Mn uptake by the ovaries and reproductive tract of cycling and anestrous ewes.

The uptake of 54Mn by the ovaries and reproductive tract of cycling and anestrous ewes has been investigated following intravenous injection of a single dose of 54MnCl2 and sacrifice of the ewes 6 h later. The uptake of 54Mn was greater in the Graafian follicle and the corpus luteum (CL) of the cycle than in the other components of the ovary. An increased uptake of radioactivity was recorded in the CL of the 11th day of the cycle when compared with that of the 4th day. The uptake of 54Mn was lower in the corpus albicans and follicles. A low uptake of radiomanganese was found also in the various tissues of the reproductive tract. These findings indicate that manganese may play a role in the normal functioning of ovarian activity in the ewe.     

Tissue specificities of Thelohania duorara,Agmasoma penaei, and Pleistophora sp., microsporidian parasites of pink shrimp, Penaeus duorarum

The pathology of pink shrimp, Penaeus duorarum, infected with the microsporidians Thelohania duorara, Agmasoma penaei, and Pleistophora sp. was described. Infections of T. duorara were widespread in most tissues; spores were located throughout the hemocoel, at the periphery of all striated muscle bundles, and in muscle and connective tissue surrounding the digestive tract. A. penaei infections invaded only dorsal abdominal muscles, muscles adjacent to blood vessels, and ovaries. Infected muscles and ovaries were eventually completely destroyed. Masses of A. penaei spores were often engulfed by hemocytes. Pleistophora sp. infected the interior of all striated muscles. Infected muscles were never completely destroyed but were often atrophied.     

Trypanosoma cruzi: morphogenesis in diffusion chambers in the mouse peritoneal cavity and attempted stimulation of host immunity

Sequential morphologic changes and antigen producing capacity of Trypanosoma cruzi in peritoneally implanted diffusion chambers were studied. Diffusion chambers were equipped with two Nuclepore filters (0.20 μm pore size) sandwiched between three Lucite rings. Epimastigotes or trypomastigotes and amastigotes were placed in diffusion chambers and surgically implanted into the peritoneal cavity of mice, or placed in in vitro cell culture, or in various types of culture media and incubated at 26 or 37 C.Epimastigotes maintained in diffusion chambers in mice changed into trypomastigotes as evidenced by the presence of numerous transitional stages and the concomitant decrease in the percentage of the former and increase in the percentage of the latter in chambers removed and examined at 16, 24, 36, 48, 72, and 84 hr after implantation. The maximum of 68% trypomastigotes was noted in chambers examined at 84 hr. Amastigotes subsequently appeared, apparently arising from trypomastigotes and reached the highest percentage (49%) obtained at 132 hr. The total number of parasites in chambers decreased slightly during the first 36 hr (20%). Little change in the total number of parasites was noted during the interval of 36–108 hr. A subsequent decrease in numbers of parasites was noted until by 280 hr after implantation, chambers contained less than 2% of the original number of organisms present in the chambers. No similar transformation of epimastigotes was noted in diffusion chambers maintained in cell culture at 37 C or in a cell culture growth medium or LIT medium at 37 or 26 C.No detectable morphological change was noted when trypomastigotes and amastigotes were implanted in diffusion chambers in the peritoneal cavity of mice. The total number of these parasites decreased notably (82%) after 24 hr.Mice receiving diffusion chambers containing epimastigotes implanted at two different intervals (21 days apart), developed only marginal protective immunity when challenged with virulent T. cruzi three weeks after the second implant of chambers, and no protection was afforded those mice implanted with chambers containing trypomastigotes and amastigotes. Sera collected from mice 6 wk after the second implantation of diffusion chambers containing parasites were observed to have antibody titers to T. cruzi as demonstrated by the fluorescent antibody technique and direct agglutination procedure.     

Compatibility of chick embryo eye anlagen with the ectoderm of the early amphibian gastrula in vitro

Eye vesicles were isolated from the early chick embryos (stage 9+ after Hamburger and Hamilton, 1951) and combined with the Rana temporaria early gastrula ectoderm (EGE) in vitro. The tissues were jointly incubated in medium 199 diluted twice with deionized water at 22 +/- 1 degree for 7-8 days or the eye vesicles were removed from the EGE ectoderm within 16-18 h. At the joint long-term incubation of these tissues, a toxic effect of the chick embryonic tissues on the EGE cells was noted. In none of the experiments, the inducing effect of the eye vesicle on the EGE was found. Similar data were obtained when the EGE was jointly cultivated with the brain (stage 9-10) and retina (stage 15) of chick embryos. The brain of the chick embryos at stage 15 exerted a weak neuralizing effect on the EGE. In the control experiments, the eye vesicles explanted with the chick embryonic ectoderm remained viable till the end of cultivation but no lentoids formed in the ectoderm. The absence of lens-inducing effect at the joint cultivation of the chick embryonic eye vesicles with the EGE is considered as a result of disturbance of the synthesis or secretion of the corresponding agents rather than a sequence of the species "incompatibility" of the inductor and reacting tissue. Hence, the use of "xenogenic" tissue recombinants is not justified when analyzing the lens-inducing activity of the eye vesicles.     

Characterization of the estrous cycle in Octodon degus

We characterized the reproductive cycle of Octodon degus to determine whether reproductive maturation is spontaneous in juveniles and if ovarian cyclicity and luteal function are spontaneous in adults. Laboratory-reared prepubertal and adult females were monitored for vaginal patency and increased wheel-running. Sexual receptivity was assessed by pairing adult females with a male 1) continuously, 2) at the time of vaginal patency, or 3) following estradiol treatment. Blood samples were assayed for estradiol and progesterone concentrations on Days 1, 4, 8, and 16 relative to vaginal opening. Ovarian tissues were collected 6 and 16 days after behavioral estrus and 6 days after copulation for histology. In juveniles, the onset of cyclic vaginal patency and increased wheel-running activity was spontaneous, occurred in the absence of proximal male cues, and appeared at regular intervals (17.5 ± 1.4 days). In adults, vaginal patency and increased wheel-running occurred cyclically (21.2 ± 0.6 days) in the absence of proximal male cues, and these traits predicted the time of sexual receptivity. Corpora lutea develop spontaneously and are maintained for 12-14 days. The ovaries had well-developed corpora lutea 6 days after mating and 6 days after estrus without mating. Progesterone concentrations were highest in the second half of the cycle when corpora lutea were present and estradiol concentrations peaked on the day of estrus. Thus, female degus appear to exhibit a spontaneous reproductive cycle consistent with other Hystricognathi rodents. Octodon degus is a novel model with which to examine the mechanisms underlying different reproductive cycles.     

Prostaglandin (PG) accumulation by mouse ovaries, oviducts, and uteri

Uteri, ovaries and oviducts from mice were collected at autopsy. Tissue slices were incubated with [³H]-PGE₂ in the presence or absence of a large excess (100 fold) of nonradioactive PGE₂ using 0.01M sodium phosphate buffer (pH 7.2). Bound and free PGs were separated by a filtration technique. PGE₂ accumulation by the uteri was evaluated as a function of incubation time, wet weight of tissues, and reproductive state. The tissue to medium ratio (T/M) was greater than 1.0 for uteri as the time of incubation increased. This suggests the presence of PGE₂ binding sites in mouse uterine tissue. Also, PGE₂ accumulation was not observed in oviducts or in ovaries.     

A comparison of the ribosomal proteins of Drosophila ovary, adult, and embryo

Summary The proteins in the 80S ribosomes of Drosophila melanogaster ovaries and adults have been characterized by two-dimensional polyacrylamide gel electrophoresis. When ribosomal proteins of ovaries and adults were compared with those from embryos, all 3 tissues showed a similar number of proteins. In addition, qualitatively, the electrophoretograms of proteins extracted from the ribosomes of these 3 tissues were found to be indistinguishable. However, apparent quantitative differences in certain acidic proteins were observed between tissues. Using ribosomes from embryos as a standard for comparison, ribosomes from adult flies that were more than 14 days old appeared to have relatively larger amounts of acidic protiens S7 and S9, and relatively smaller amounts of acidic proteins S14 and S25/S27. The transition period occured during the ninth to thirteenth day of adult fly development. Significant differences were not detected between ovarian and embryonic acidic ribosomal proteins. In contrast to the differential ratio of acidic proteins in ovaries, adults, and embryos, a similar distribution of basic proteins was found in these tissues.     

Distribution of radioactivity in subcellular fractions of genital tract tissue and skeletal muscles of guinea pig embryos after administration of 1 alpha,2 alpha-3H(n) testosterone in vitro

The dynamics of radioactivity in subcellular fractions of the reproductive system and muscle has been studied in the guinea pig embryos from the 21st day of development till birth after the preincubation in a medium containing 1 alpha,2 alpha-3H(n) testosterone (T-3H). The total uptake of T-3H per mg tissue is higher in the reproductive system than in the muscle and attains the highest values during the period of differentiation of target organs by the male type (30-33 days). The nucleocytoplasmic ratio of radioactivity is shifted towards its increase in the nucleus with maximum on the 41st-45th days and by the moment of birth. Radioactivity in the microsomal fraction is higher than in the cytosol (soluble proteins). From the 27th day of development on, when the hormone uptake becomes selective, the radioactivity in the cytosol from the reproductive organs is higher than in the muscle tissue. The data obtained suggest the importance of processes in microsomes for differentiation and organogenesis of reproductive system.     

Characterization and regulation of monocarboxylate cotransporters Slc16a7 and Slc16a3 in preimplantation mouse embryos

Concurrent with compaction, preimplantation mouse embryos switch from the high pyruvate consumption that prevailed during cleavage stages to glucose consumption against a constant background of pyruvate uptake. However, zygotes exposed to and subsequently deprived of glucose can form blastocysts by increasing pyruvate uptake. This metabolic switch requires cleavage-stage exposure to glucose and is one aspect of metabolic differentiation that normally occurs in vivo. Monocarboxylates, such as pyruvate and lactate, are transported across membranes via the SLC16 family of H(+)-monocarboxylate cotransporter (MCT) proteins. Thus, the increase in pyruvate uptake in embryos developing without glucose must involve changes in activity and localization of MCT. In mouse embryos, continued expression of Slc16a1 (MCT1) requires glucose supply. Messenger RNA for Slc17a7 (MCT2) and Slc16a3 (MCT4) has been detected in mouse preimplantation embryos; however, protein function, localization, and regulation of expression at the basis of these net pyruvate uptake changes remain unclear. The expression and localization of SLC16A7 and SLC16A3 have therefore been examined to clarify their respective roles in embryos derived from the reproductive tract and cultured under varied conditions. SLC16A3 appears localized to the plasma membrane until the morula stage and also maintains a nuclear distribution throughout preimplantation development. However, continued Slc16a3 mRNA expression is dependent on prior exposure to glucose. SLC16A7 localizes to apical cortical regions with punctate, vesicular expression throughout blastomeres, partially colocalizing in peroxisomes with peroxisomal catalase (CAT). In contrast to SLC16A3 and SLC16A1, SLC16A7 and CAT demonstrate upregulation in the absence of glucose. These striking differences between the two isoforms in expression localization and regulation suggest unique roles for each in monocarboxylate transport and pH regulation during preimplantation development, and implicate peroxisomal SLC16A7 as an important redox regulator in the absence of glucose.     

Changes of fatty-acid structure of common lipids and contents of peroxidation products in tissues of embryos depending on the level of vitamins A, D3 and E in a diet of geese during the reproductive period

Results concerning the contents of retinol in the liver, residual yoke of 25-day embryos and yoke of eggs depending on the level of vitamins A, D3 and E in the diet of geese by grey Obroshin breeds in reproductive period are presented in the paper. It is established, that vitamin D3 reduces the level of retinol deposition in the tissues of embryos and yoke of eggs of geese, and addition of vitamins A and E to a diet of geese raises the level of retinol both in the liver and residual yoke of embryos, and in yokes of geese eggs. Besides the data about changes of fatty-acid spectrum of common lipids and contents of lipid peroxidations products in tissues of the liver and pectoral muscles of 25-day embryos are presented in the paper depending on the level of vitamins A, D3 and E in geese diet during their reproductive period. Introduction of vitamin A--in quantity of 10000 IU, vitamin D3--in quantity of 3000 IU, in the composition of mixed fodder of geese during the reproductive period and vitamin E in quantity 35 IU on 1 kg to mixed fodder optimizes fatty-acid structure of the common lipids and the level of peroxidations lipids products in the liver and pectoral muscles of embryos.     

Developmental and age-related changes in proteins in the female reproductive tract of Heligmosomoides polygyrus (Nematoda)

Proteins in the female reproductive tract of Heligmosomoides polygyrus at days 8, 16, 35, 90, and 140 postinfection (PI) were examined using polyacrylamide gel electrophoresis. Sixteen-day-old and 140-day-old worms also were examined histochemically. Egg production of these worms was assessed for each age group. In analyzing proteins using electrophoresis, the reproductive tracts were separated into 3 sections: the tip, or anteriormost part of tract, containing oogonia; the middle region, containing developing oocytes; and the posterior region, containing the uterus with fertilized eggs. Three major reproductive tract proteins were identified as having molecular weights of greater than 140 kDa, 115 kDa, and 82 kDa. These were found in all parts of the reproductive tract from worms of all ages except those at 8 days PI (which are too young to produce eggs) and are believed to be yolk proteins. Four low molecular weight proteins (L1-4) are believed to be nucleoproteins. L4 was absent from the posterior section of the reproductive tracts and L3 was limited to the posterior sections and may be associated with sperm stored in the uterus. Of 5 high molecular weight proteins the second heaviest, designated H2, appeared to be relatively more concentrated in the posterior sections of the reproductive tract. An 85-kDa protein was limited to the tip and middle sections of reproductive tracts. Histochemical tests on sectioned H. polygyrus showed strong positive reactions for protein in cytoplasmic granules in developing oocytes and in eggs of younger worms (16 days) but a reduced reaction in older worms (140 days). Strains for collagen showed a slight positive reaction in and between developing oocytes and a strong reaction in the egg shells. Stains for nucleoproteins particularly reacted with sperm stored in the uterus, and slightly reacted with fertilized eggs and the nucleoli of the intestinal and ovarian epithelium. Egg production by H. polygyrus increased to 123 eggs/female/day by 16 days PI but declined from 121 eggs/female/day at 35 days PI to 64 eggs/female/day in worms 140 days old. Electrophoresis indicated no loss in the different types of proteins in the reproductive tract of older worms, but histochemistry and protein content assays suggest that older worms that produce fewer eggs contain a relatively smaller amount of protein in the female reproductive tract.     

Early Morphogenesis of Chick Gonad in the Absence of Mesonephros

The development of the gonads in male and female chick embryos with induced unilateral mesonephric agenesis was studied using grafting, histoenzymology, and electron microscopy. As in embryos with a mesonephros, proliferation of the coelomic epithelium and its interaction with mesenchymal cells to form the medullary cords take place in the amesonephric gonads. In a similar manner, gonadal sexual differentiation and the differentiation of steroidogenic tissue, detectable by the presence of Δ5-3β-hydroxysteroid dehydrogenase, do not appear to be affected by the absence of an organized mesonephros. However, the initiation of gonadal development, further growth, and the onset of meiosis observable in developing ovaries are retarded. This delay appears to be reversible, as was demonstrated by experiments in which ovaries from chicks with complete mesonephric agenesis were transplanted into the coelomic cavity of male and female 3 1/2-day-old embryos. Meiosis finally occurred in the oocytes of all ovaries, regardless of the sex of the host. Therefore, the presence of a differentiated mesonephros in chick embryos is not required for the establishment of an undifferentiated gonad and sexual differentiation, or for initiation of meiosis.     

A comparative study of high affinity choline uptake and choline utilization in cholinergic and non-cholinergic tissues

Comparative studies of [3H]choline accumulation were done in the Limulus corpora pedunculata, abdominal ganglia and cardiac ganglion. Dual uptake processes for choline were found in all three tissues. In acute experiments, the corpora pedunculata high affinity choline uptake system showed exclusive sensitivity to ouabain. Prolonged exposure to ouabain revealed that the HAChUS of all three tissues were significantly inhibited. The metabolism of [3H]choline transported via the high affinity process in the three tissues was studied. [3H]Acetylcholine was a major product of the [3H]choline taken up by the corpora pedunculata and the abdominal ganglia. Phosphorylcholine was the major product seen in cardiac ganglion extracts and occurred in significant proportions in abdominal ganglia extracts. [3H]Acetylcholine was not detected in cardiac ganglion extracts. Treatment with either lithium chloride or hemicholinium-3 markedly inhibited high affinity uptake of [3H]choline in all three tissues.     

In vivo uptake and metabolism of [3H]ecdysone in the vitellogenic follicles of Sarcophaga bullata (Diptera) and its localisation by autoradiography

Within 4 h after injection of [³H]ecdysone, almost all tritiated material has disappeared from the haemolymph, indicating that the uptake by the tissues is very fast. After only 15 min, 19% of the label was found in the ecdysterone fraction and 4% in the highly polar products (HPP) fraction. The uptake of [³H]ecdysone by the ovary (mid-vitellogenic) is almost complete within 1 h after injection. The pattern of [³H]ecdysteroids in the ovaries follows a well ordered sequence: firstly, [³H]ecdysone is the major component of the [³ H]ecdysteroids but it disappears within 2 h, next a peak value of [³H]ecdysterone was found at 1 h, whereafter this also disappeared, and from 2 h on, there was a considerable increase in HPP. The HPP consisted of 3 fractions (A, B and C). Glusulase treatment revealed that apparently only fraction B consisted of glucuronide and/or sulphate-conjugates of ecdysteroids. Autoradiographic experiments confirmed that the uptake of [³H]ecdysone was a very rapid process. In ovaries fixed 1 h after injection, the silver grains were abundant in the ooplasm but were also found in the follicle cell cytoplasm and in trophocytes. In follicles examined 16 h after injection, only a few silver grains were observed in the trophocytes and follicle cells. However, the cytoplasm of the oocyte was labelled. The border cells also accumulated label.

The major results indicate that all cell types of the follicle seem to be able to absorb ecdysone from the haemolymph and that there seems to be a rather selective uptake of ecdysone. In the ooplasm, ecdysone is converted to highly polar conjugates.