INHIBITION OF LACTOGENIC ACTIVITIES OF BOVINE MAMMARY GLAND EXPLANTS BY THE WHEY FRACTION OF BOVINE MILK

The regulation of milk constituents, synthesis and secretion in tissue cultures of the bovine mammary gland was altered by a whey fraction of bovine milk. α-Casein gene expression, casein secretion and fatty acid synthesis were inhibited by the whey fraction in a dose-dependent manner. The whey fraction inhibited the enhancement activity of prolactin on α-casein gene expression and fatty acid synthesis, and also inhibited casein secretion to the medium, in explants cultured in a medium with or without prolactin. No effect on the expression of the β-lactoglobulin gene was found.     

SIGNIFICANCE OF THE CHEMICAL COMPOSITION OF THE SECRETING AND DRY MAMMARY GLAND TO MILK SECRETION

The results herein presented furnish exact critical evidence for one more stage in milk secretion. Cows producing up to 30 pounds of milk at one milking are shown to have the lactose equivalent of all this milk in the udder when milking commences. The average excess of lactose found in the udder after subtracting the amount necessary for the contained milk is equal to 2.1 pounds. This represents the milk retained in the udder when the cow is believed to be dry. These conclusions are further supported by the fact that no sugar is found in the udder in the quiescent state. The study of the total composition of the udder as fat, ash, nitrogen, and lactose, and of the contained milk shows that there is a large excess of fat, ash, and nitrogen in proportion to that necessary for milk formation. The excess of udder lactose over the milk lactose is much less. The lactose would therefore appear to be formed from some element in the blood, probably dextrose, only as needed for the formation of milk. The composition of the dry udder is quite different in certain respects from that of the actively secreting gland. It builds up a fat reserve of a quite different Reichert-Meissl number from that of butter-fat. It has no sugar, its ash content is reduced, and the nitrogen content is like that of the secreting gland.     

TOPOGRAPHY OF DNA SYNTHESIS IN THE MAMMARY GLAND OF THE C3H MOUSE AND ITS CONTROL BY OVARIAN HORMONES: AN AUTORADIOGRAPHIC STUDY

Tritium-labelled thymidine was injected 45 min before sacrifice into virgin female C3H/HeJ mice 7–23 weeks of age, as well as into 10-week-old mice which had been ovariectomized and treated daily with 1 μg of oestradiol-17β and/or 1 mg of progesterone. Autoradiographs were made of squash preparations of the mammary glands, stained by Feulgen's method. The following results were obtained: (1) During normal development of the gland, cells synthesizing DNA are abundant in terminal buds and virtually absent in duct epithelium. Hence ductal growth takes place by the addition of cells produced in the terminal end structures. (2) At 5–6 months, when mammary growth has ceased, a considerable number of cells synthesizing DNA can still be found in alveoli, though not in duct epithelium. Hence the alveolar cells constitute a renewal population in the adult virgin. Because they maintain the potentiality to divide, duct cells are a G₀ population. (3) Ovariectomy results in arrest of DNA synthesis within 3–5 days. Both oestradiol and progesterone restore DNA synthesis in alveoli but only progesterone is able to induce DNA synthesis in duct epithelium, and the differentiation of terminal buds into alveoli. This finding provides an explanation for the resumed proliferation of duct cells in pregnancy. (4) The number of cells engaged in DNA synthesis varies considerably among identical structures within the same gland. This may be due either to synchrony of cell replication and/or to fluctuations of proliferative activity in the gland.     

THE FORMATION OF CHOLINE AND OF ACETYLCHOLTNE BY BRAIN IN VITRO

Abstract— Free choline and acetylcholine (ACh) in mouse or rat brain were assayed biologically. The subcellular distribution of ACh in brain slices that had been incubated in the presence of eserine was compared to that in control brain; during incubation, the ACh outside nerve endings increased four-fold, the ACh released from synaptosomes by osmotic shock doubled but the ACh bound firmly within nerve endings did not increase. The two nerve ending stores of ACh were labelled to similar specific radioactivities when slices were incubated with [³H]choline, but the specific radioactivity of the ACh formed was much lower than that of the added choline. Tissue incubated in the presence of eserine released choline and ACh into the medium and the tissue levels of both substances increased. Brain tissue exposed to Na⁺-free medium lost 84 per cent of its ACh and 66 per cent of its free choline; the amounts of both substances returned towards control values during subsequent incubation in a normal-Na⁺ medium (choline-free). Both the ACh outside nerve endings and the ACh associated with synaptosomes were depleted when tissue was incubated in Na⁺-free medium.     

THE IN VITRO FORMATION OF ACETYL-ASPARTYL-PEPTIDO-SEROTONIN COMPLEXES BY PIG HYPOTHALAMIC TISSUE EXTRACTS

Peptido-serotonin complexes are formed on incubation of pig hypothalamic tissue extracts with acetyl-aspartate, the amino acids glu. ser, gly and ala. an ATP-regenerating system and serotonin. The purification and partial characterisation of two acetyl-aspartyl-peptido-serotonin complexes is described.     

CONNEXIN EXPRESSION AND GAP JUNCTIONS IN THE MAMMARY GLAND

Gap junctional communication plays a vital role in embryogenesis, cell differentiation and the co-ordination of tissue responses. Gap junctions are formed by a family of closely-related proteins called connexins which show tissue-specific patterns of expression. The role of gap junctions in the mammary gland remains unclear. The lumena of mammary gland ducts are lined by luminal cells with an outer layer of basal cells. In rodents, the luminal cells express connexin26 only during pregnancy and lactation and the basal cells, in some reports, express connexin43. In the normal human breast the basal cells express connexin43, although human mammary epithelial cellsin vitrohave been reported to express both connexin26 and connexin43. Analysis of connexin expression at the molecular level is now bringing new insights into the structure and function of gap junctions in a range of normal and pathological cell systems.     

大豆黄酮对大鼠乳腺发育作用的实验研究

本文研究了大豆黄酮对大鼠乳腺发育及其对垂体生长激素（ＧＨ）和催乳素（ＰＲＬ）分泌的影响，结果表明：１）胃饲或皮下注射大豆黄酮能显著提高去卵巢大鼠或正常处女大鼠乳腺重量和乳腺ＤＮＡ、ＲＮＡ含量；２）胃饲或皮下注射大豆黄酮能显著提高正常处女大鼠血清ＧＨ和ＰＲＬ含量，３）大豆黄酮能显著提高大鼠乳腺，垂体和下丘脑胞浆雌二醇受体的数目，并显著提高乳腺孕酮受体的数目，体外受体竞争结合试验结果表明：大豆黄酮与大     

MELANIN FORMATION BY HUMAN BRAIN IN VITRO

Abstract— A quantitative radiometric assay utilising incorporation of ¹⁴C from labelled precursors as a measure of melanin formation by human brain in vitro is described. The assay was validated by comparison with various criteria of melanin formation.
Catecholamines, DOPA and 5HT were precursors for brain melanin formation. Melanin formation was detected in all brain regions studied and was highest in substantia nigra and striatum.
The assay was used to evaluate various hypotheses of brain melanin formation. No evidence for enzymic activity was found and it is concluded that brain melanin formation may be a largely non-enzymic process.     

FETAL DEVELOPMENT: THE EFFECTS OF MATURATION ON IN VITRO PROTEIN SYNTHESIS BY MOUSE BRAIN TISSUE

—The elucidation of the translational regulatory events which function during the critical fetal and neonatal period is an important prerequisite to our understanding of normal, as well as abnormal, brain growth and differentiation. Brain cell suspensions and cell-free homogenates were employed to study the protein synthetic activity during the maturation of fetal- neural tissue. The results clearly demonstrated that while neural tissue from 1-day postnatal mice was 10 times more active in protein synthesis than brain tissue from adult mice, the former was many fold less active in translational events than fetal neural tissue from 13-day post-zygotic mice. Fetal polypeptide synthetic activity was found to decrease from the 13th day to the 19th day post-zygotic. This decrement in the translational activity was not due to amino acid availability or pools, or to differences, quantitatively or qualitatively, in polysome concentrations. The enhanced rate of protein synthetic activity measured with neural tissue from 13-day post-zygotic mice was shown to be due to an increase in rate of protein synthesis and not to an enhanced rate of protein degradation.     

REGULATION OF PROTEIN SYNTHESIS IN THE VENOM GLAND OF VIPERID SNAKES

Morphological changes in the venom gland of V. ammodytes were studied after the removal of the venom from the gland lumina (milking) It was found that the height of the secretory cells was changed during the secretory cycle. The patterns of the rough endoplasmic reticulum and of the Golgi complex were changed as well Milking induced an increased incorporation of [¹⁴C]amino acids into total and venom proteins In V ammodytes, during the first day after milking, 25% of the total counts in protein were precipitable by anti-venom serum, while at 8 days, 80% of the proteins synthesized were venom proteins At this stage, the incorporation was 10- and 20-fold that of unmilked glands for total and venom proteins, respectively. Venom was accumulated (secreted) in the gland lumina of V. ammodytes at a relatively high rate up to 2 wk after milking and leveled off afterwards. Intact glands and gland slices of V ammodytes and V palaestinae, taken from snakes a few days after milking, incorporated [¹⁴C]amino acids into proteins in vitro at a rate higher than that of unmilked glands. The activity of two exportable enzymes (phosphodiesterase and benzoyl arginyl ethyl esterase) was assayed in gland homogenates of V. ammodytes. It was found that 2–3 wk after milking, the intracellular level of these enzymes was up to 2-fold that of unmilked glands.     

TIMING OF DNA SYNTHESIS IN THE MITOTIC CYCLE IN VITRO

A study was made of the timing of DNA synthesis in the mitotic cycle under conditions where the average mitotic cycle of populations of human amnion and kitten lung cells in culture was variable. Three types of experiments were performed: (a) Autoradiographs were made of incorporated tritiated thymidine in cells whose mitotic histories were recorded microcinematographically allowing the measurement of telophase + G1 along with the total length of the mitotic cycle. (b) Measurement of the G2 + prophase part of the mitotic cycle was performed under various conditions by exposing cells to tritiated thymidine and observing the increase in labeled metaphases plus anaphases as a function of time. (c) The effect of a change in pH on parts of the mitotic cycle was tested by continuously photographing a single colony of cells first at pH 7.8 and then at pH 7.1. All of our data point to the same conclusion; namely, that within a population of cells with a given generation time, the length of each of the measurable parts of the mitotic cycle has a particular distribution of values and that, when there is a change in the generation time, under our conditions only the T + G1 distribution changes.     

七星瓢虫的卵黄发生:体外培养的脂肪体中卵黄原蛋白的合成

为了深入研究七星瓢虫的卵黄发生及其激素调节机理,我们建立了脂肪体的体外培养方法,证明了脂肪体在体外培养条件下能够正常进行卵黄原蛋白的合成与分泌。在体外合成的卵黄原蛋白与体内合成的具有相同的电泳迁移率、免疫学特性和部分水解肽谱。放射性氨基酸在体外参人卵黄原蛋白的动力学与在体内的相似。用脂肪体的体外培养方法,结合放射免疫沉淀、SAS-聚丙烯酰胺凝胶电泳、放射自显影等技术,研究了不同发育期脂肪体的合成能力,以及取食人工饲料的雌虫中保幼激素类似物对卵黄原蛋白合成的促进作用。     

INFLUENCE OF EXTERNAL POTASSIUM ON THE SYNTHESIS AND DEPOSITION OF MATRIX COMPONENTS BY CHONDROCYTES IN VITRO

The effect of a high external potassium concentration on the synthesis and deposition of matrix components by chondrocytes in cell culture was determined. There is a twofold increase in the amount of chondroitin 4- and 6-sulfate accumulated by chondrocytes grown in medium containing a high potassium concentration. There is also a comparable increase in the production of other sulfated glycosaminoglycans (GAG) including heparan sulfate and uncharacterized glycoprotein components. The twofold greater accumulation of GAG in the high potassium medium is primarily the result of a decrease in their rate of degradation. In spite of this increased accumulation of GAG, the cells in high potassium fail to elaborate appreciable quantities of visible matrix, although they do retain the typical chondrocytic polygonal morphology. Although most of the products are secreted into the culture medium in the high potassium environment, the cell layer retains the same amount of glycosaminoglycan as the control cultures. The inability of chondrocytes grown in high potassium to elaborate the typical hyaline cartilage matrix is not a consequence of an impairment in collagen synthesis, since there is no difference in the total amount of collagen synthesized by high potassium or control cultures. There is, however, a slight increase in the proportion of collagen that is secreted into the medium by chondrocytes in high potassium. Synthesis of the predominant cartilage matrix molecules is not sufficient in itself to ensure that these molecules will be assembled into a hyaline matrix.     

大鼠颌下腺及其离体培养细胞脱氢表雄酮（DHEA）的免疫组织化学定位

用免疫组织化学ABC法,研究了颌下腺及无血清培养的颌下腺上皮细胞DHEA的定位,结果显示,大鼠颌下腺的浆液性腺泡的上皮细胞及各级导管上皮细胞均呈DHEA免疫反应阳性,无血清培养腺上皮细胞也呈DHEA免疫反应阳性,阳性物质分布于胞质,胞核呈阴性反应,此结果提示：大鼠颌下腺能自身合成DHEA,DHEA对消化功能可能具有重要的调节作用。