scholarly journals Uptake and metabolism of tryptophan by the isolated perfused guinea-pig mammary gland

1976 ◽  
Vol 158 (3) ◽  
pp. 659-662 ◽  
Author(s):  
B Mepham ◽  
A R Peters ◽  
S R Davis
Keyword(s):  
Amino Acids ◽  
Mammary Gland ◽  
Guinea Pig ◽  
Blood Plasma ◽  
Milk Protein ◽  
Absorption Studies ◽  
Uptake And Metabolism ◽  
Tryptophan Uptake

Tryptophan uptake by the isolated perfused lactating guinea-pig mammary gland was 46.5+/-4.6 mug/h per g. Results of absorption studies and the use of [methylene-14C]tryptophan suggest that tryptophan is one of the group of amino acids that are transferred almost quantitatively from blood plasma to milk protein.

scholarly journals Relative distribution of post-nuclear poly(A)-containing RNA abundance groups within the nuclear and post-nuclear polyadenylated and non-polyadenylated RNA populations of the lactating guinea-pig mammary gland

1980 ◽  
Vol 192 (2) ◽  
pp. 489-498 ◽  
Author(s):  
Ian C. Bathurst ◽  
Roger K. Craig ◽  
David G. Herries ◽  
Peter N. Campbell
Keyword(s):  
Mammary Gland ◽  
Guinea Pig ◽  
Milk Protein ◽  
Regulation Of Gene Expression ◽  
Milk Proteins ◽  
Relative Distribution ◽  
Free System ◽  
Low Salt ◽  
Nuclear Rna ◽  
System 2

1. RNA isolated from the post-nuclear supernatant of the lactating guinea-pig mammary gland was fractionated with oligo(dT)–cellulose into three populations; those that bound at ‘low salt’ [long poly(A) tracts, 78–32 nucleotides]; those that bound at ‘high salt’ [shorter poly(A) tracts, 48–21 nucleotides]; and those that did not bind [no poly(A) or short poly(A) tracts, <20 nucleotides]. Nuclear RNA was fractionated into two populations, those that bound in ‘low salt’ and those that did not bind. All the post-nuclear RNA fractions directed the synthesis of milk proteins in a Krebs II ascites cell-free system. 2. 3H-labelled DNA complementary to the post-nuclear poly-(A)-containing RNA population (low-salt fraction) was fractionated into abundant (milk-protein mRNA), moderately abundant and scarce sequences. This complementary DNA was then used to investigate the distribution of the mRNA sequences in the different RNA populations. This showed that all sequences were present in polyadenylated and non-polyadenylated fractions, but that major quantitative differences were apparent. The abundant milk-protein mRNA sequences predominated in the ‘low-salt’ post-nuclear poly(A)-containing RNA fraction, whereas the moderately abundant sequences predominated in the non-polyadenylated post-nuclear RNA fraction. In total cellular RNA, those sequences deemed initially to be moderately abundant within the ‘low-salt’ poly(A)-containing RNA population were present at a concentration very similar to those of the abundant milk-protein mRNA (approx. 6×105 copies of each sequence/cell). Similarly, analysis of the nuclear RNA populations showed that the ‘abundant’ and so-called ‘moderately abundant’ sequences were present in essentially identical concentrations (2×103 copies of each sequence/cell). The majority of these (90–95%) were non-polyadenylated. 3. The results are discussed in terms of the post-transcriptional mechanisms involved in the regulation of gene expression in the lactating guinea-pig mammary gland.

Amino acid transportation, sensing and signal transduction in the mammary gland: key molecular signalling pathways in the regulation of milk synthesis

2020 ◽  
Vol 33 (2) ◽  
pp. 287-297
Author(s):  
Zhihui Wu ◽  
Jinghui Heng ◽  
Min Tian ◽  
Hanqing Song ◽  
Fang Chen ◽  
...  
Keyword(s):  
Amino Acids ◽  
Mammary Gland ◽  
Milk Protein ◽  
Building Blocks ◽  
Branched Chain Amino Acids ◽  
Signalling Pathways ◽  
Recent Advances ◽  
Branched Chain ◽  
Molecular Signalling Pathways

AbstractThe mammary gland, a unique exocrine organ, is responsible for milk synthesis in mammals. Neonatal growth and health are predominantly determined by quality and quantity of milk production. Amino acids are crucial maternal nutrients that are the building blocks for milk protein and are potential energy sources for neonates. Recent advances made regarding the mammary gland further demonstrate that some functional amino acids also regulate milk protein and fat synthesis through distinct intracellular and extracellular pathways. In the present study, we discuss recent advances in the role of amino acids (especially branched-chain amino acids, methionine, arginine and lysine) in the regulation of milk synthesis. The present review also addresses the crucial questions of how amino acids are transported, sensed and transduced in the mammary gland.

Milk Protein Precursors in the Goat During Late Lactation

1993 ◽  
Vol 1993 ◽  
pp. 1-1
Author(s):  
B.J. Bequette ◽  
F.R.C. Backwell ◽  
A.G. Calder ◽  
J.A. Metcalf ◽  
D. Wray-Cahen ◽  
...  
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Mammary Gland ◽  
Milk Protein ◽  
Protein S ◽  
Dairy Goats ◽  
Protein Precursor ◽  
Previous Hypothesis ◽  
Casein Protein ◽  
Isotope Kinetics

Previously, we have reported on work in dairy goats using stable isotope kinetics to examine the precursors for milk protein synthesis (1). Contrary to a previous hypothesis (2), these results suggested that blood free amino acids (AA) are not simply transported into the mammary gland and incorporated directly into milk protein. Although the latter may still occur, a substantial amount of the AA for milk protein synthesis appears to be channelled through constitutive mammary gland protein(s) first. Moreover, the data indicated that a proportion (12-20%) of the casein protein precursor may be derived from extra-mammary sources other than blood free AA, e.g. peptides and/or proteins. It may be possible therefore to alter milk protein synthesis by the provision of different forms of precursor amino acids. Since the previous study was in goats during early lactation (day 61 ± 11), the present study reports on the precursors for milk protein synthesis in goats during late lactation, and allows a comparison between stages of lactation.

Effects of restriction of amino acid supply to the isolated perfused guinea-pig mammary gland

1979 ◽  
Vol 46 (1) ◽  
pp. 69-73 ◽  
Author(s):  
T. Ben Mepham ◽  
Andrew R. Peters ◽  
Stephen Alexandrov
Keyword(s):  
Amino Acids ◽  
Mammary Gland ◽  
Amino Acid ◽  
Guinea Pig ◽  
Essential Amino Acids ◽  
Substrate Solution ◽  
Group 2 ◽  
Group 1

SUMMARYWhen individual essential amino acids were omitted for periods of 40–100 min from the infusate substrate solution in isolated perfused guinea-pig mammary gland experiments, uptake of methionine, tyrosine, phenylalanine, histidine and tryptophan (group 1) was significantly depressed by a mean of 49·8%, whereas the remaining essential amino acids (group 2) showed no significant decrease in uptake. During depletion periods oxidation of [14C\amino acids was increased. The possible significance of the differences in absorption between the 2 groups of amino acids is discussed.

15N enrichment of casein amino acids in the milk from goats given a single intravenous dose of l-[15N]leucine

1999 ◽  
Vol 66 (2) ◽  
pp. 283-288 ◽  
Author(s):  
JOAQUIN RUBERT-ALEMÁN ◽  
GUIDO RYCHEN ◽  
FLORENCE CASSERON ◽  
FRANCOIS LAURENT ◽  
GERARD JEAN MARTIN
Keyword(s):  
Amino Acids ◽  
Mammary Gland ◽  
Milk Protein ◽  
Carbon Sources ◽  
Amino Nitrogen ◽  
Milk Proteins ◽  
15N Enrichment ◽  
N Enrichment ◽  
Branched Chain ◽  
Plasma Leucine

Branched-chain amino acids (AA) are mostly metabolized in the splanchnic area, but some are metabolized within the mammary gland and thus could contribute to the synthesis of non-essential AA (Wohlt et al. 1977). The extraction rate of leucine from plasma by the mammary gland is particularly high (64% in the goat; Roets et al. 1983), in excess of that used for the synthesis of milk proteins (Davis & Mepham, 1976; Wohlt et al. 1977; Roets et al. 1983). Thus, although mammary leucine is mainly used for milk protein, it also provides amino nitrogen and carbon sources for the synthesis of non-essential AA.To our knowledge, no information is available on the transfer and distribution of plasma leucine amino nitrogen to milk protein AA. Using the technique for chromatographic fractionation of AA recently developed by Casseron et al. (1997), we studied the specific 15N enrichment of casein (CN) AA in the goat given a single intravenous injection of [15N]leucine.

scholarly journals Whole-body fluxes and partitioning of amino acids to the mammary gland of cows fed fresh pasture at two levels of intake during early lactation

2003 ◽  
Vol 90 (2) ◽  
pp. 271-281 ◽  
Author(s):  
D. Pacheco ◽  
M.H. Tavendale ◽  
G. W. Reynolds ◽  
T. N. Barry ◽  
J. Lee ◽  
...  
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Mammary Gland ◽  
Dairy Cows ◽  
Milk Protein ◽  
Essential Amino Acids ◽  
Whole Body ◽  
Early Lactation ◽  
Isoleucine Leucine ◽  
Ad Libitum

The utilisation of essential amino acids (EAA) by the mammary gland of lactating dairy cows fed fresh forages was studied to provide basic information useful in designing strategies to increase the production of milk protein from pasture-fed dairy cows. The relationship between the flux of EAA in the whole body and their uptake by the mammary gland was determined in four cows in early lactation (length of time in milk 44 (SD 14·5) d) producing 21 (SD 4·0) kg milk/d. The cows were maintained in metabolism stalls and fed fresh perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) pasturead libitumor restricted to 75 %ad libitumintake. The whole-body fluxes of amino acids (AA) were measured using an arterio-venous infusion of universally13C-labelled AA. Whole-body fluxes of fourteen AA were estimated. Isotope dilution indicated that mammary utilisation accounted for one-third of the whole-body flux of EAA, with individual AA ranging between 17 and 35 %. Isoleucine, leucine, valine and lysine were the EAA with the greatest partitioning towards the mammary gland (up to 36 % of the whole-body flux), which could reflect a potentially limiting effect on milk protein synthesis. In the case of AA with low partitioning to the mammary gland (for example, histidine), it is suggested that non-mammary tissues may have priority over the mammary gland and therefore the supply of this AA may also limit milk protein synthesis.

scholarly journals Metabolism of l-(U-14C)valine, l-(U-14C)leucine, l-(U-14C)histidine and l-(U-14C) phenylalanine by the isolated perfused lactating guinea-pig mammary gland

1976 ◽  
Vol 156 (3) ◽  
pp. 553-560 ◽  
Author(s):  
S R Davis ◽  
T B Mepham
Keyword(s):  
Amino Acids ◽  
Guinea Pig ◽  
Milk Protein ◽  
Mean Transit Time ◽  
Mammary Tissue ◽  
Plasma Component ◽  
Ruminant Species ◽  
The Mean ◽  
Phenyl Alanine

1. The incorporation of L-[U-14C]leucine, L[U-14C]histidine and L-[U-14C]phenylalanine into casein secreted during perfusion of isolated guinea-pig mammary glands was demonstrated. 2. The extent of incorporation of label into casein residues was consistent with their being derived from free amino acids of the perfusate plasma. 3. The mean transit time of the amino acids from perfusate into secreted casein was approx. 100 min. 4. Whereas radioactive histidine and phenylalanine were incorporated solely into milk protein, radioactivity from [U-14C]valine was also transferred to CO2 and to an unidentified plasma component, and from [U-14C]leucine to plasma glutamic acid. 5. Evidence from experiments with [U-14C]phenylalanine suggests that, as in rats, but in contrast with ruminant species, guinea-pig mammary tissue does not possess phenyl alanine hydroxylase activity. 6. The results are discussed in relation to the possible role of essential amino acid catabolism in the control of milk-protein synthesis.

scholarly journals Effects of a High-Grain Diet With a Buffering Agent on Milk Protein Synthesis in Lactating Goats

2021 ◽  
Vol 8 ◽  
Author(s):  
Meilin He ◽  
Xintian Nie ◽  
Huanhuan Wang ◽  
Shuping Yan ◽  
Yuanshu Zhang
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Mammary Gland ◽  
Amino Acid ◽  
Milk Production ◽  
Milk Protein ◽  
Time Of Flight ◽  
Lactating Goats ◽  
Rumen Acidosis ◽  
Buffering Agent

Chinese dairy industries have developed rapidly, providing consumers with high-quality sources of nutrition. However, many problems have also appeared during the development process, especially the low quality of milk. To improve milk quality, a large amount of concentrated feed is usually added to the diet within a certain period of time, which increases the milk production to a certain extent. However, long-term feeding with high-concentration feed can lead to subacute rumen acidosis. Therefore, the present study aimed to determine the effect of adding a buffer on subacute rumen acidosis, and the improvement of milk production and milk quality. We also aimed to study the mechanism of promoting mammary gland lactation. A total of 12 healthy mid-lactating goats were randomly divided into two groups, they were high-grain diet group (Control) and buffering agent group. To understand the effects of high-grain diets with buffers on amino acids in jugular blood and the effects of amino acids on milk protein synthesis, Milk-Testing™ Milkoscan 4000, commercial kits, and high-performance liquid chromatography (HPLC) measurements were integrated with the milk protein rate, the amino acid concentration in jugular venous blood samples, quantitative real-time PCR, comparative proteomics, and western blotting to study differentially expressed proteins and amino acids in mammary gland tissues of goats fed high-grain diets. Feeding lactating goats with buffering agent increased the percentage of milk protein in milk, significantly increased the amino acid content of jugular blood (p &lt; 0.05), and increase the amino acid transporter levels in the mammary gland. Compared with the high-grain group, 2-dimensional electrophoresis technology, matrix-assisted laser desorption/ionization-time of flight/time of flight proteomics analyzer, and western blot analysis further verified that the expression levels of beta casein (CSN2) and lactoferrin (LF) proteins in the mammary glands of lactating goats were higher when fed a high-grain diets and buffers. The mechanism of increased milk protein synthesis was demonstrated to be related to the activation of mammalian target of rapamycin (mTOR) pathway signals.

The effect of free amino acid supply on the contribution of peptide-bound phenylalanine and tyrosine to casein synthesis in late lactation goats

1998 ◽  
Vol 1998 ◽  
pp. 3-3
Author(s):  
B.J. Bequette ◽  
F.R.C. Backwell ◽  
C.E. Kyle ◽  
A.G. Calder ◽  
L.A. Crompton ◽  
...  
Keyword(s):  
Amino Acids ◽  
Mammary Gland ◽  
Free Amino ◽  
Milk Protein ◽  
Labelling Technique ◽  
Lactating Goats ◽  
Net Uptake ◽  
Supplemental Protein ◽  
Nu Method ◽  
Casein Synthesis

The mammary gland of lactating ruminants (Guinard & Rulquin 1994) does not appear to extract sufficient quantities of free amino acids (AA) to account for their output as milk protein. Based upon application of a precursor (blood or plasma free AA):product (casein) labelling technique (Backwell et al. 1996) in goats, results suggest that blood peptides or proteins taken up by the gland probably account for this deficit. However, the deficiency appears to be alleviated when supplemental protein or AA are infused (Guinard & Rulquin 1994), suggesting that uptake of peptide bound AA is reduced while that of the free AA is increased. The objective of the current study was to corroborate these findings, thus the precursonproduct labelling technique was applied in lactating goats to determine whether arterial free phenylalanine supply affects the contribution of peptide bound phenylalanine and tyrosine to casein synthesis and compare the results to the net uptake (NU) method.

scholarly journals Identification and subsequent phosphorylation of sequestered partially processed caseins in the lactating guinea-pig mammary gland

1984 ◽  
Vol 222 (2) ◽  
pp. 501-510 ◽  
Author(s):  
A P Boulton ◽  
J C Pascall ◽  
R K Craig
Keyword(s):  
Endoplasmic Reticulum ◽  
Mammary Gland ◽  
Guinea Pig ◽  
Milk Protein ◽  
Secretory Pathway ◽  
Short Pulse ◽  
Early Event ◽  
Post Translational Modification ◽  
Electrophoretic Mobilities ◽  
Membrane Bound

Golgi and endoplasmic-reticulum fractions were prepared from the lactating guinea-pig mammary gland. The endoplasmic-reticulum fraction was highly active in the processing and sequestration of milk-protein primary translation products. Explants from the lactating gland in organ culture were used to identify milk-protein intermediates present in the secretory pathway, and the timing of the events leading to their post-translational modification. With [35S]methionine, the milk proteins labelled after a short pulse (3 min) were represented by the partially processed (but not phosphorylated) caseins and alpha-lactalbumin sequestered within membrane-bound vesicles. After a 30 min labelling period, higher-Mr caseins with electrophoretic mobilities identical with those of the phosphorylated caseins isolated from milk were identified in the incubation medium, and sequestered within membrane-bound vesicles. Pulse-chase experiments established a precursor-product relationship between these forms. Secretion is apparent approx. 30 min after sequestration. Caseins are highly phosphorylated; removal of the phosphate residues with acid phosphatase results in proteins with increased electrophoretic mobility, similar to those of the partially processed early casein intermediates found sequestered in explants after a 3 min pulse with [35S]methionine, and those sequestered within microsomal membranes after mRNA-directed cell-free protein synthesis. A comparison of the proteins labelled during both short (5 min) and long (30 min) pulses with [35S]methionine and [32P]Pi shows that, in contrast with the 35S-labelled caseins, those labelled with [32P]Pi exhibit only electrophoretic mobilities identical with those of the mature caseins isolated from milk and those identified after long labelling periods with [35S]methionine. No phosphorylated early intermediate forms of caseins were identified. We conclude that the synthesis and post-translational modification of guinea-pig caseins occurs in two stages, (i) an early event involving synthesis and sequestration within the endoplasmic reticulum, an event that involves signal-peptide removal, followed (ii) 10-20 min later by phosphorylation at a different point in the secretory pathway, probably in the Golgi complex. Secretion of the phosphorylated caseins occurs 10-20 min later.

Sign in / Sign up

Export Citation Format

Share Document