Guinea Pig Mammary Gland Research Articles

Introduction of the human growth hormone gene into the guinea pig mammary gland by in vivo transfection promotes sustained expression of human growth hormone in the milk throughout lactation

We tested the feasibility of transfecting mammary tissue in vivo with an expression plasmid encoding the human growth hormone (hGH) gene, under the control of the cytomegalovirus promoter. Guinea pig mammary glands were transfected with plasmid DNA infused through the nipple canal and expression was monitored in control and transfected glands by radioimmunoassay of milk samples for hGH. Sustained expression of hGH throughout lactation was attained with a polyion transfection complex shown to be optimal for the transfection of bovine mammary cells, in vitro. However, contrary to expectations, hGH expression was consistently 5- to 10-fold higher when DEAE–dextran was used alone for transfection. Thus polyion complexes which are optimal for the transfection of cells in vitro may not be optimal in vivo. The highest concentrations of hGH in milk were obtained when glands were transfected within 3 days before parturition. This method may have application for studying the biological role or physical properties of recombinant proteins expressed in low quantities, or for investigating the regulation of gene promoters without the need to construct viral vectors or produce transgenic animals.

Histamine in Mammary Gland: Pregnancy and Lactation

Abstract 1. Histamine concentration, histidine decarboxylase (HDC), histamine N-methyltransferase (HMT) and transglutaminase (TG-ase) activities have been followed in mouse and guinea pig mammary gland during pregnancy and lactation. 2. The most significant changes concerned histidine decarboxylase (HDC) activity and their pattern was similar for the two species. In either species 2nd trimester of pregnancy was associated with stimulation whereas early lactation was associated with reduction of the tissue HDC activity. 3. Hormonal control of HDC is suggested on the basis of variations in the enzyme activity and histamine concentration in mammary gland during estrous cycle. 4. The circumstantial evidence is provided indicating that histamine might be involved in pregnancy associated growth of mammary gland and in excretory activity of lactating gland.

Localization of metallothionein in female reproductive organs of rat and guinea pig.

We demonstrated the localization of metallothionein (MT) in rat uterus and ovaries and in guinea pig mammary glands. During the cyclic changes from one estrous period to the next, strong MT immunostaining was found in the glandular epithelium of the endometrium and weak immunostaining was observed in the simple columnar epithelium. Interestingly, during estrus, the intensity of MT immunostaining decreased in the cytoplasm, whereas during metestrus, diestrus, and proestrus the intensity of strong and similar immunostaining was observed in both the cytoplasm and nucleus. During proestrus and estrus, the number of vaginal epithelial cells containing MT increased on the luminal side of the epithelium and inside the lumen. In rat ovary, strong immunostaining was observed in the cytoplasm and nucleus of granulosa-lutein cells of the corpus luteum and in the cytoplasm of the ovum. In mammary gland of non-pregnant guinea pig, very strong but scattered MT immunostaining was demonstrated in both cytoplasm and nucleus of some epithelial cells of the lactiferous ducts. The mammary tissue of the pregnant guinea pig showed an increase in MT staining in alveolar cells that had proliferated due to pregnancy. The presence of MT in the female reproductive organs, the tissues of which actively grow under the control of female sex hormones, indicates some as yet unknown association of MT with cell proliferation and differentiation.

The activity of phosphoenolpyruvate carboxykinase throughout the lactation cycle of the guinea pig mammary gland.

The enzyme phosphoenolpyruvate carboxykinase (PEPCK) has been measured in the guinea pig mammary gland throughout the pregnancy-lactation cycle. This is of interest since the primary importance of PEPCK is thought to be its role in gluconeogenesis and it is questionable whether or not gluconeogenesis occurs in the mammary gland. The enzyme activity, present in both the cytosol and mitochondria, was shown to follow the lactation profile. During the transition into lactation, cytosolic PEPCK activity increases 11-fold and mitochondrial PEPCK activity 43-fold while tissue weight increases 4-fold. Fructose 1,6-bisphosphatase was found to increase at a rate only slightly greater than that of the tissue weight. The increase in mitochondrial PEPCK activity is thus about 10 times greater than that of general tissue expansion, whereas the cytosolic PEPCK activity increase is only 2-fold greater. The activity of fructose 1,6-bisphosphatase appears to be merely keeping pace with general tissue expansion. The mitochondrial enzyme constitutes 59 +/- 3% of the total gland PEPCK activity in the prepartum state and 86 +/- 2% at midlactation. Therefore, mitochondrial PEPCK is the isoenzyme undergoing the greater increase during the transition into lactation in the guinea pig mammary gland and thus would appear to play the more important role in the conversion of oxalacetate to phosphoenolpyruvate in this tissue.

Control of mammary gland fibroblast growth by insulin, growth hormone and prolactin

Fibroblasts isolated from guinea pig mammary glands were cultured in 96 well culture plates in the presence of various concentrations of insulin, growth hormone and prolactin. Insulin (30 μg/ml increased uptake of tritiated thymidine by 30%. Higher concentrations of insulin did not result in any further increase in thymidine uptake. Growth hormone alone did not alter thymidine uptake in concentrations of 0 to 250 ng/ml. 300 ng/ml gave thymidine uptake of 136% of controls. In the presence of 20 g/ml insulin, growth hormone (250 ng/ml) increased thymidine uptake to approximately double that of controls. Prolactin alone (300 ng/ml decreased thymidine uptake by 19%. Insulin increased thymidine uptake, but the negative effect of prolactin was still evident above 150 ng/ml.

The effect of local and parenteral vaccination on the response of the guinea pig mammary gland to staphylococcal challenge

Preparturient guinea pigs vaccinated locally (orally and itramammarily) or parenterally (intradermally) with killed Staphylococcus aureus (KS), were challenged intramammarily (IMM) with KS or viable S. aureus during the next lactation. The number and types of leukocytes emigrating into the milk were determined before and after IMM chalenge. The milk leukocytosis after challenge with KS was the greatest in the intradermally (ID) vaccinated animals, moderate in the IMM vaccinated animals, and insignificant in the unvaccinated or orally vaccinated animals. The polymorphonuclear (PMN) leukocyte predominated in the milk of the IMM and ID vaccinated animals during the initial 30 h after challenge with KS. Later (30–102 h postchallenge), the mononuclear leukocyte (marcophage and lymphocytes) was the major cell-type. No significant changes in the number or types of leukocytes occurred in the milk of the unvaccinated or orally vaccinated animals after challenge. Intramammary challenge with viable S. aureus caused a large increase in the number of leukocytes in the milk of all animals. The milk leukocytosis occurred more rapidly in locally vaccinated guinea pigs than unvaccinated or ID vaccinated guinea pigs. The PMN leukocyte predominated in the milk of all animals during the period of maximum response. The incidence and severity of staphylococcal mastitis were less in guinea pigs vaccinated locally than ID vaccinated or unvaccinated animals.

Effect of estradiol and relaxin on collagen and non-collagen protein synthesis by mammary fibroblasts

In order to determine the effect of relaxin and estradiol on collagen and noncollagen synthesis by mammary gland fibroblasts, fibroblasts isolated from guinea pig mammary glands were grown on plastic or Cytodex-3 collagen coated microcarriers. On plastic, estradiol (600 pg/ml) increased the incorporation of tritiated glycine into collagen. Non-collagenous protein synthesis was increased at all concentrations of estradiol, but it was greatest with 200 pg/ml estradiol. On Cytodex-3, 400 pg/ml estradiol increased the synthesis of collagen and non-collagenous protein. Relaxin (1 μg/ml) did not affect collagen synthesis but decreased the synthesis of non-collagenous protein.

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

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.

Cyclic AMP changes in guinea pig mammary gland and milk.

Cyclic AMP was measured in guinea pig mammary gland biopsies from midpregnancy through lactation and in daily milk samples throughout lactation. The results indicate that mammary gland cAMP levels rise sharply in late pregnancy to a prepartum peak and then drop abruptly at partus. This is similar to the pattern observed by others in rat and mouse although the guinea pig does not undergo a prepartum progesterone withdrawal. In animals with a 3-wk lactation period, milk cAMP concentration decreases approximately 40% between days 10 and 20, whereas the mammary gland level increases 22% per unit wet tissue weight. Although milk concentrations of lactose and cAMP are weakly correlated, a strong correlation exists between the total collected daily outputs of these two substances. The results suggest that, as lactation proceeds, mammary gland cAMP levels gradually increase, whereas milk cAMP levels decrease. Concurrently, lactose output decreases as lactation proceeds. These observations are consistent with first, a milk secretory route for regulating mammary gland cAMP levels (in addition to synthesis and hydrolysis), and second, an inhibitory action of cAMP on lactose synthesis.

Signal sequences, secondary modification and the turnover of miscompartmentalized secretory proteins in Xenopus oocytes.

The cytoplasm of the Xenopus oocyte can be altered by the microinjection of proteins and the regulatory responses to such perturbations can then be studied. We have investigated proteolytic systems within the oocyte which may be involved in the maintenance of the integrity of the different subcellular compartments. Thus primary translation products, made in the wheat germ system under the direction of frog liver, chicken oviduct, rat liver rapidly sedimenting endoplasmic reticulum, rat seminal vesicle, guinea pig mammary gland or honey been venom gland RNA, were injected into oocytes. Their stability in the frog cell cytosol was in general low compared to that of their processed counterparts. The latter were usually obtained by collecting the heterologous proteins exported by RNA-injected oocytes. Electrophoretic analysis of oocytes injected with particular primary and processed polypeptides permitted measurement of the stabilities of proteins differing only by the presence or absence of a detachable signal sequence, or by the presence of a specific secondary modification. The effect of the latter on protein stability appears slight. However, the presence of a detachable signal sequence destabilizes those miscompartmentalized secretory proteins which are otherwise stable. Indeed all other results are consistent with this concept for they show that primary translation products are in general much less and are never more stable than their processed counterparts. Thus we provide evidence that errors of compartmentation can be corrected in living cells and that this process is often facilitated by the properties conferred on a protein by a detachable signal sequence.

Mammary growth patterns in guinea pigs during puberty, pregnancy and lactation.

Mammary gland growth patterns were studied in 110 guinea pigs during the growth phase, pregnancy and lactation. Body weight changes were studied and, in addition, mammary indices were wet weight, dry fat-free tissue (DFFT), deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Statistical analyses were mathematical regression models to best fit the actual data. These included linear, quadratic, cubic, and several forms of exponential regression models. Data were separated into growth phase (60 guinea pigs in 10 age groups), pregnancy (20 guinea pigs in 4 groups), and lactation (30 guinea pigs in 6 groups). Data during pregnancy and the first 5 days of lactation were pooled and analyzed also because mammary growth continued beyond pregnancy to Day 5 of lactation. Mammary wet weight increased according to a cubic expression in the growth phase, while mammary DFFT, DNA and RNA were rectilinear through 200 days of age. During pregnancy and the first 5 days of lactation, mammary growth parameters followed the pattern of an exponential equation. Daily rates of increase for mammary DFFT and DNA were twice the rate for mammary wet weight. During lactation, mammary gland indices increased to Day 5 and then decreased gradually from Day 10 to Day 20. The best mathematical models for these change were those which are used to describe lactation curves, but all mammary gland indices decreased later and more gradually than milk production. Comparisons in growth rates of guinea pig mammary glands were made with those published for dairy goats and dairy cows. Rates of mammary DNA changed inversely to lengths of gestation in these 3 species.

Characterisation of a membrane-bound serine-specific casein kinase isolated from lactating guinea-pig mammary gland.

Serine-specific and threonine-specific casein kinase activities have been identified in a Golgi-enriched membrane fraction isolated from the lactating guinea-pig mammary gland. The serine-specific casein kinase has been purified 2000-fold by affinity chromatography on ATP-agarose. The enzyme has an estimated Mr of 100000 as determined by sucrose gradient centrifugation and phosphorylates the serine residues of dephosphorylated guinea-pig caseins A and B in a qualitatively and quantitatively identical manner to caseins A and B secreted by lactating mammary gland explants in organ culture. The enzyme also phosphorylates casein C at serine, but not threonine residues. Studies on the relative location of the enzyme within a Golgi-enriched membrane fraction show that it is an integral component of the membrane, either in the form of a transmembrane protein or exposed on the luminal side of the membrane. Although casein kinase activity is not associated with the endoplasmic reticulum, it remains to be proven whether it is truly a Golgi enzyme, since analysis of subcellular membrane components fractionated by sucrose gradient centrifugation shows that the particulate protein kinase activity of the lactating mammary gland does not cosediment with galactosyl transferase, possibly a reflection of the heterogeneous nature of mammary gland Golgi apparatus. It seems likely that the serine-specific casein kinase activity described is responsible for the phosphorylation of caseins in the lactating guinea-pig mammary gland, and that this occurs after the sequestration of processed but unphosphorylated caseins within the lumen of the endoplasmic reticulum.

Differential expression of alpha-lactalbumin and casein genes during the onset of lactation in the guinea-pig mammary gland.

1. The expression of alpha-lactalbumin and casein genes was examined in guinea-pig mammary tissue taken from animals both pre- and post-partum. 2. Analysis of total RNA by RNA excess hybridization with sequence-specific complementary DNA probes demonstrated that alpha-lactalbumin mRNA was present late in pregnancy, and that maximum concentrations were present at parturition. Casein gene transcripts were absent late in pregnancy (62 days), but by parturition were present at concentrations identical to those found at all time points examined throughout lactation. 3. Studies using mammary explants in organ culture showed that tissue from pregnant animals, or animals at parturition, synthesized and secreted only alpha-lactalbumin. After parturition, at the onset of casein synthesis, differential rates of secretion of alpha-lactalbumin and the caseins were observed. 4. The results are discussed in terms of the multiple intracellular mechanisms involved in the regulation of milk protein gene expression in the guinea-pig mammary gland.

The construction, identification and partial characterization of plasmids containing guinea-pig milk protein complementary DNA sequences.

A complementary DNA (cDNA) plasmid library has been constructed in the plasmid pAT153, using poly(A)-containing RNA isolated from the lactating guinea-pig mammary gland as the starting material. Double stranded cDNA was inserted into the EcoRI site of the plasmid using poly(dA . dT) tails, then transformed into Escherichia coli HB101. From the resulting colonies we have selected and partially characterized plasmids containing cDNA copies of the mRNAs for casein A, casein B, casein C and alpha-lactalbumin. However, the proportion containing casein C cDNA was exceptionally low, and these contained at best 60% of the mRNA sequence.

The identification and localization of actin and actin-like filaments in lactating guinea pig mammary gland alveolar cells.

Cytochalasin B, a microfilament-altering drug, inhibits lactose synthesis in lactating guinea pig mammary gland [Biochim. Biophys. Acta 392:20, 1975] but not primary by inhibiting glucose transport [Eur. J. Cell Biol. 20:150, 1979]. In order to study the possible role of microfilaments in lactose synthesis and secretion, we isolated both the alveolar (milk-secreting) and myoepithelial (contractile) cells from lactating mammary gland. Light microscopy shows that the alveolar cell fraction (viability approximately 71%) is homogenous and that the cells retain strong polarity of secretory structures in the apical region. Two proteins were extracted from the alveolar cell fraction. One (mol wt 42,000) comigrates with skeletal muscle actin on SDS-PAGE gels. The other, a high-molecular-weight (180,000) protein (HMWP) may be analogous to actin-binding protein or clathrin. An extract from the myoepithelial cell fraction also contains a protein that comigrates with actin but no HMWP. Whole tissue extract contains the 42K protein, and a 185K HMWP. Examination of the alveolar cell extract by electron microscopic (EM) negative staining revealed meshworks of multistranded, interconnecting filaments, with attached globular structures (100-200 A) (possibly the HMWP) and single filaments (40-60 A diameter) branching off. To localize these filamentous structures in situ, whole tissue was glycerinated and incubated with rabbit skeletal muscle heavy meromyosin (HMM). Masses of filaments in myoepithelial cells served as convenient standards for HMM decoration. Decorated filaments have cross-arms or projections, unlike the narrow, smooth filaments of control tissue. Decorated filaments in alveolar cells are located beneath the plasma membrane, in close association with secretory vacuoles, and near the Golgi apparatus; filaments near the latter two are often oriented perpendicular to the plasma membrane. Microvesicles are embedded in meshworks under the plasmalemma and near the Golgi apparatus. Intermediate-sized (85-115 A diameter), non-decorated filaments diverge from the meshworks of decorated filaments. Micro-vesicles are associated with intermediate-sized filaments as well. The association of actin-like filaments with secretory vacuoles and microvesicles and their location in areas of the cell concerned with biosynthetic activities suggest a possible function in the intracellular transport of secretory products.

Post-transcriptional regulation of gene expression in guinea pig tissues.

The formation of individual functional mRNA sequences in higher organisms requires many steps in addition to transcription. These include RNA splicing, polyadenylation, base modification, transport from nucleus to cytoplasm and assembly into polyribosomes. Various control mechanisms must also operate. These will function on a quantitative basis to account for the differing frequency of the various classes of cytoplasmic mRNAs, and also on a qualitative basis, because in higher organisms not all the nuclear poly(A)-containing RNA molecules are found in a cytoplasmic poly(A)-containing RNA population from the same tissue. During our studies on the mechanisms controlling the accumulation of the poly(A)-containing RNA sequences which occur with high and moderately high frequency in the cytoplasm of the lactating guinea pig mammary gland, it became apparent that > 75% of the 20,000 or so poly(A)-containing nuclear RNA sequences were not found in the cytoplasmic poly(A)-containing RNA fraction. Here we demonstrate that many of the poly(A)-containing RNA sequences retained in the nucleus of the lactating guinea pig mammary gland are also present in the nucleus and cytoplasm of the liver of the male guinea pig. These observations provide new evidence for a predominant role of post-transcriptional mechanisms in the regulation of structural gene expression in guinea pig tissue.

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.

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. (3)H-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. 6x10(5) 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 (2x10(3) 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.

Tubulin content and assembly states in guinea pig mammary gland during pregnancy, lactation, and weaning.

AbstractThe microtubule (MT) protein, tubulin, was measured in mammary gland biopsies from 23 guinea pigs averaging four samples per animal from about 20 days antepartum through weaning. [3H]Colchicine binding assays were carried out on supernatants from tissues homogenized sequentially in MT-stabilizing and MT-depolymerizing buffers which yielded estimates, respectively, of free (S-1) and microtubular (S-2) tubulin as well as total tubulin (S-1 + S-2). There is almost a threefold increase in total tubulin which begins about 20 days antepartum and reaches a peak by 11 to 15 days postpartum. There is an initial increase in free tubulin, also beginning about 20 days antepartum; however, about 1 week before birth, free tubulin levels off and microtubular tubulin begins to increase, reaching a peak in midlactation after a sevenfold increase from pregnancy and then declining at weaning. The results show that free tubulin is synthesized in late pregnancy and then converted to the assembled (microtubular) state,...

The Xenopus Oocyte as a Surrogate Secretory System The Specificity of Protein Export

Combining messenger RNA from one kind of secretory cell with the cytoplasm of another such cell can reveal the nature and specificity of protein export mechanisms. We show that messenger RNAs from secretory cells of chickens, rats, mice, frogs, guinea-pigs, locusts and barley plants, when injected into Xenopus oocytes, direct the synthesis and export of proteins. Chicken ovalbumin, Xenopus albumin, mouse thyroid-stimulating hormone, locust vitellin and guinea-pig milk proteins were identified using specific antibodies, whilst chicken lysozyme and ovomucoid, rat albumin, Xenopus vitellogenin and rat seminal vesicle basic proteins were identified provisionally from their molecular weights. Certain endogenous proteins are sequestered and secreted although most oocyte proteins are not exported. Similarly the major polyoma viral protein and the simian virus 40 and polyoma tumour antigens are retained within the oocyte. Radioactive proteins exported by oocytes programmed with chicken oviduct or Xenopus liver RNA are not re-exported in detectable amounts when injected into fresh oocytes, nor is there secretion of chicken oviduct or guinea-pig mammary gland primary translation products prepared using wheat germ extracts. Thus the export of secretory proteins from oocytes cannot be explained by leakage and may require a cotranslational event. The secretory system of the oocyte is neither cell-type nor species-specific yet is highly selective. We suggest that the oocyte can be used as a general surrogate system for the study of gene expression, from transcription through translation to the final subcellular or extracellular destination of the processed protein.

Differential distribution of poly(A)-containing RNA sequences between the nucleus and post-nuclear supernatant of the lactating guinea-pig mammary gland.

RNA complexity analysis of poly(A)-containing RNA isolated from the lactating guinea-pig mammary gland demontrates that the complexity within the nuclear population was three--four-fold greater than that in the equivalent post-nuclear polyribosomal population [see Craig et al., Biochem. J. (1979) 181, 737-756]. Up to 21 000 different sequences distributed in two abundance groups were detected in the nucleus, whereas 5000 sequences distributed in three abundance groups were present in the post-nuclear population. All poly(A)-containing RNA sequences present in the post-nuclear fraction could be detected in the nuclear poly(A)-containing population. Analysis of the relative distribution of the three post-nuclear abundance populations within the nuclear poly(A)-containing RNA population demonstrates that the abundant and moderately abundant post-nuclear sequences were present a similar concentrations within the nucleus, and comprised the abundant nuclear population. The abundant and moderately abundant post-nuclear sequences were present at 62200 and 785 copies of each sequence/cell in the post-nuclear fraction respectively, and 44 and 118 copies of each sequence/cell in the nuclear fraction respectively. The scarce post-nuclear sequences (18.5 copies of each sequence/cell) were also present at low levels in the nuclear fraction (0.1 copy of each sequence/cell). the results are discussed in terms of the post-transcriptional regulation of gene expression in the lactating guinea-pig mammary gland.