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Biomedical subjects

H A Bern

Publications and source records attributed to H A Bern.

At least 109 records · Page 6Linked to original sources

Prolactin receptors in tilapia (Sarotherodon mossambicus) tissues: binding studies using 125I-labeled ovine prolactin.

The binding of 125I-labeled ovine prolactin (oPRL) to membrane preparations of tissue from freshwater-adapted tilapia (Sarotherodon mossambicus) was examined. Liver, ovary, and testis showed a relatively high specific binding (5-10%). A lower specific binding occurred consistently in intestine and gill tissue, and inconsistently in urinary bladder and kidney preparations. Desaturation experiments with MgCl2 indicated that a majority of the PRL receptors were already occupied by endogenous PRL. Scatchard analysis of liver binding gave a dissociation constant of 0.6 X 10(-9) M and a capacity of 207 fmol/mg protein.

Animals↗

Bioassay for salmon prolactin using hypophysectomized Fundulus heteroclitus.

A bioassay for salmon prolactin (PRL) is described. This assay which is based on the sodium-retaining action of PRL in the hypophysectomized killifish, Fundulus heteroclitus, has proved to be rapid, sensitive (250 pg PRL per gram of fish), and specific. The procedure has been used to characterize the biological activity of a highly purified PRL from the pituitaries of the chinook salmon, Oncorhynchus tschawytscha, and a similar PRL isolated (by acid buffer polyacrylamide disc gel electrophoresis) from pituitaries of coho salmon (O. kisutch) (MW ca. 22,000; isoelectric point greater than 9).

Animals↗

Neuropeptide-induced contraction and relaxation of the mouse anococcygeus muscle.

Isometric tension responses to neuropeptides were recorded from anococcygeus muscles isolated from male mice. This smooth muscle tissue is innervated by inhibitory nonadrenergic, noncholinergic nerves that resemble, ultrastructurally, the peptidergic neurons of the gastrointestinal tract; the physiological function of the anococcygeus is not known. Slow sustained contractions were produced by oxytocin (0.2-20 nM), [Arg8]vasopressin (0.4-200 nM), and [Arg]-vasotocin (0.4-100 nM); the mouse anococcygeus is, therefore, one of the few examples of nonvascular smooth muscle from male mammals to respond to low concentrations of oxytocin and related peptides. Substance P (0.5-8 microM) caused distinctive, biphasic increases in muscle tone of some, but not all, preparations. Other neuropeptides producing contractions were neurotensin (2-100 microM) and thyrotropin-releasing hormone (2-100 microM); the responses were of similar time course and displayed selective cross-desensitization, suggesting that these two peptides act through a common distinct mechanism. Tetradecapeptide somatostatin (10-80 microM) and its analog urotensin II (0.1-5 microM), a dodecapeptide from the urophysis of the teleost fish Gillichthys mirabilis, produced similar slowly developing relaxations of carbachol-induced tone. Piscine urotensin II, of which there are no reported effects on nonvascular mammalian systems, was 20-50 times more potent than somatostatin, a well-established mammalian hormone. Of the peptides studied, only vasoactive intestinal polypeptide (0.05-1 microM) caused rapid powerful relaxations in low concentrations; this is consistent with its proposed involvement in nonadrenergic, noncholinergic neurotransmission in the mouse anococcygeus.

Animals↗

Development-associated changes in thyroxine kinetics in juvenile salmon.

Smoltification is a transformation that occurs in some species of salmon, during which solitary fish in fresh water become schooling fish and migrate to the sea. This process is accompanied by large increases in plasma T4. T4 secretion rate and other parameters of T4 metabolism in juvenile coho salmon were estimated by applying kinetic analyses to measurements of the disappearance of injected T4 radiotracer from plasma. Studies were performed at the beginning (March) and end (May) of the increase in T4 concentration in fresh water and seawater. Early and intensive sampling permitted characterization of a very fast initial component of the T4 disappearance curve when analyses included a zero time datum derived from an independent estimate of plasma volume. The plasma volume, equal to 1.77% of body weight, was obtained by measuring the disappearance of radiolabeled albumin from the plasma in two other groups of animals in fresh water and seawater. There were 3- to 7-fold changes in T4 production, distribution, and metabolism between March and May, whereas environment (fresh water vs. seawater) had relatively minor effects on T4 kinetics. In fresh water, the T4 secretion rate was 4.48 ng/h in March and 1.50 ng/h in May. The total T4 pool size was 37.8 ng in March and 12.2 ng in May. Plasma-tissue T4 fluxes were 3- to 7-fold greater in May. Relatively less T4 was distributed in tissue in May (63% vs. 83%), and T4 spent much less time in tissue in May than in March during each pass through the tissue space (11 min vs. 3.1 h). We propose that the difference in secretion rate and a redistribution of T4 between blood and tissues contribute to both the rise and fall in the plasma T4 concentration between March and May. Changes in T4 kinetics during salmonid smoltification resemble those occurring during amphibian metamorphosis and mammalian gestation and neonatal life, and may reflect an increased requirement and an important role for thyroid hormones during periods of rapid development in vertebrates in general.

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Effects of long-term implantation of vaginal concretions on the cervicovaginal epithelium of mice.

In two experiments, neonatal female BALB/cCrgl or BALB/cfC3HCrgl mice were given subcutaneous injections of 5 micrograms 17 beta-estradiol or sesame oil for the first 3 days of life and were ovariectomized at 60 days of age, at which time vaginal concretions (Experiments I and II) or silica (Experiment II) were implanted intravaginally. Mice were examined at 12 months of age. Three abnormal cervicovaginal epithelial responses were noted: persistent vaginal stratification/cornification (PVS); prominent vaginal squamocolumnar junction (SCJ); epithelial pegs, downgrowths, or lesions (dysplasias). PVS, not present in unimplanted controls, occurs in at least half of the members of the neonatally estrogen-treated groups; implants of concretions or silica did not increase its incidence significantly. Although SCJ was observed in implanted but not in unimplanted controls, its incidence was significantly higher in neonatally estrogen-treated mice than in either control group. The elevated incidence in neonatally estrogen-treated mice was not increased further by implantation of concretions or silica. In neonatally estrogenized mice, the subsequent implantation of a concretion significantly increased the incidence of cervicovaginal abnormalities. Increased PVS and SCJ are teratological consequences of neonatal exposure to a small amount of estrogen; on the other hand, increased dysplasias may, in part, be responses of the estrogenized vaginal epithelium to the concretions.

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Effect of immunosuppression on neonatally diethylstilbestrol-induced genital tract lesion and tumor development in female mice.

Neonatal treatment of female rodents with the synthetic estrogen diethylstilbestrol [(DES) CAS: 56-53-1; alpha, alpha'-diethyl-4,4'-dimethoxystilbene] results in immunodeficiency that persists into adulthood, along with progressive development of genital tract lesions. DES-induced immunosuppression was examined as a possible promoter of genital tract lesion and tumor development in BALB/cCrgl female mice. Secondary suppression of T-cell immunity after neonatal DES treatment failed to increase lesion incidence or to promote genital tract tumor development in 8-month-old mice. Although hyperplastic lesions were present in 70% of the genital tracts from DES-treated mice at 8 months of age, apparently few neoplastic cells are present at this time, since transplantation of genital tracts from these animals into syngeneic hosts did not yield tumors. To reduce the possibility that potential tumors were too immunogenic to survive in the syngeneic hosts, genital tract pieces from 12-month-old DES-treated female mice were transplanted into immunosuppressed hosts; only 1 tumor resulted. The tumor was a mixed adenocarcinoma, squamous cell carcinoma, and sarcoma and was immunogenic. These results suggest that immunosurveillance by T-cells is relatively unimportant in DES-induced genital tract lesion development.

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Growth effect of lithium on mouse mammary epithelial cells in serum-free collagen gel culture.

The effect of lithium on the growth of mammary epithelial cells from adult virgin and midpregnant BALB/c or BALB/cfC3H mice was tested in a serum-free collagen gel culture system. The serum-free medium consisted of a 1:1 mixture of Ham's F12 and Dulbecco's Modified Eagle's medium supplemented with insulin, transferrin, cholera toxin, epidermal growth factor (EGF), and bovine serum albumin fraction V (BSA V). A multifold increase in cell number occurred during 10-12 days of culture in this medium. In dose-response studies in which the concentration of each component of this serum-free medium was varied in turn, the addition of LiCL (10 mM) enhanced growth at most concentrations of each factor. However, LiCl could not enhance growth in the absence of insulin or BSA V, but could replace EGF. The optimal concentration of LiCl was 5-10 mM; higher concentrations (20-80 mM) were toxic. KCl (1-10 mM) when added to the serum-free medium slightly stimulated growth; the addition of NaCl to the medium had little effect on growth. LiCl did not enhance the growth of cells from spontaneous mammary tumors of BALB/cfC3H mice.

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Specificity of response of intestinal ion transport systems to a pair of natural peptide hormone analogs: somatostatin and urotensin II.

Specificity of two intestinal ion transport systems toward the natural peptide hormone analogs somatostatin and urotensin II has been demonstrated by electrophysiological and radiotracer studies in vitro. Somatostatin inhibits active C1 secretion across the theophylline-treated rat colon but urotensin II, a dodecapeptide somatostatin analog from the teleost caudal neurosecretory system, is without effect. Conversely, urotensin II stimulates active Na and C1 absorption across the posterior intestine of the 5% seawater-adapted goby, Gillichthys mirabilis, but somatostatin is ineffective. From these and others' studies, it appears that in both systems, increased net absorption results from increased mucosal-to-serosal unidirectional ion fluxes. Based on structure-activity relationships in these and other systems, it is speculated that the difference in amino acid residues at position 4 (somatostatin-Lys, urotensin II-Ala) may contribute to the observed specificity.

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Growth of mammary epithelial cells from neonatally sex hormone-exposed mice in serum-free collagen gel culture.

Neonatal sex hormone treatments are known to cause an increase in mammary tumors in female mice with expressed mammary tumor virus (MTV). The growth of mammary epithelial cells from mice treated neonatally with sex hormones was studied in response to growth-stimulatory factors in a serum-free collagen gel culture system which sustains the growth of normal mammary epithelial cells. Animals were treated with hormones or oil-vehicle for the first 5 days after birth. Cells from control mice at 2 and 3 months of age showed a maximal growth response to insulin at 5-10 micrograms/ml and LiCl at 5-20 mM. Cells responded to epidermal growth factor at all concentrations used (1, 10 and 50 ng/ml). In contrast, mammary epithelial cells from mice treated neonatally with estrogen (estradiol and diethylstilbestrol (DES] showed a reduced growth response to the growth factors tested.

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Growth of normal mouse vaginal epithelial cells in and on collagen gels.

Sustained growth in primary culture of vaginal epithelial cells from ovariectomized adult BALB/cCrgl mice embedded within or seeded on collagen gel matrix was achieved in a serum-free medium composed of Ham's F-12 medium/Dulbecco's modified Eagle's medium, 1:1 (vol/vol), supplemented with insulin, bovine serum albumin fraction V, epidermal growth factor, cholera toxin, and transferrin. Three-dimensional growth of vaginal epithelial cells occurred inside the collagen gel matrix. Cell numbers increased 4- to 8-fold in collagen gel and about 4-fold on collagen gel after 9-10 days in culture. The effect of 17 beta-estradiol (0.00018-180 nM in gel or 0.018-180 nM on gel) and diethylstilbestrol (DES; 0.0186-186 nM in gel) on the growth of vaginal epithelial cells was examined. The addition of estrogen did not enhance the growth of vaginal epithelial cells during this time period either in the complete medium or in a suboptimal medium. Cultures on floating collagen gels in the serum-free medium are composed of 1-3 cell layers with superficial cornification. Estrogen does not appear to be a direct mitogen for vaginal epithelial cells, at least in this system.

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Control of ion transport by Gillichthys mirabilis urinary bladder.

The columnar epithelial cell region of the goby Gillichthys mirabilis urinary bladder is the region responsible for active Na and Cl reabsorption from bladder urine. In 5% seawater-adapted fish, reabsorption occurs via an electrically silent coupled NaCl transport mechanism. Bladder reabsorption is increased in seawater-adapted fish above that observed in 5% seawater-adapted fish; incremental reabsorption results from an electrogenic Na transport in addition to the neutral component. Hypophysectomy of seawater-adapted fish reduces the electrogenic Na transport (measured as short-circuit current, ISC) and increases the transepithelial resistance (R) to values near those of 5% seawater-adapted fish. Cortisol restores the ISC and R to normal seawater-adapted values and will initiate electrogenic Na transport in 5% seawater-adapted fish. Ovine prolactin will also restore the ISC and R of columnar cell regions of hypophysectomized seawater-adapted fish to normal seawater-adapted levels; this effect appears to be due to contamination or to inherent stimulatory activity of the ovine prolactin preparation, since endogenous prolactin is ineffective in the restoration of ISC or R.

Amiloride↗

Chloride cells and the hormonal control of teleost fish osmoregulation.

Teleost fish osmoregulation is largely the result of integrated transport activities of the gill, gut and renal system. The basic 'epithelial fabric' in each of these tissues is adapted to provide the appropriate transport mechanisms depending upon whether the fish is in fresh water or sea water. Net NaCl transport by the branchial epithelium reverses direction when euryhaline species migrate between the two media, providing a useful focus in experiments designed to elucidate mechanisms of differentiation and integration of transport function. Isolated opercular membranes and skins from certain seawater-adapted species are good models to study branchial salt extrusion mechanisms. These heterogeneous tissues generate short-circuit currents equal to net chloride secretion. The vibrating probe technique has allowed localization of all current and almost all conductance to the apical crypt of chloride cells. Area-specific surface current and conductance of chloride cells are 18 mA cm-2 and 580 mS cm-2 (1.7 omega cm2), ranking them as one of the most actively transporting and conductive cells known. There is no net sodium transport under short-circuit conditions but the chloride secretion process is sodium-dependent and ouabain and 'loop'-diuretic sensitive. Sodium fluxes through chloride cells are large (PNa = 5.2 X 10(-4) cms-1) nd appear passive and rate-limited by a single barrier. A link may exist between the active transport and leak pathways since sodium fluxes always account for 50% of chloride cell conductance. The sodium pathway is probably the chloride cell-accessory cell tight junction, although this is still unresolved. Chloride secretion can be rapidly modulated by several hormones, including catecholamines, somatostatin, glucagon, vasoactive intestinal polypeptide and urotensins I and II. Regulation by these hormones may be by rapid alterations of cellular cAMP levels. Differentiation of chloride cells and chloride secretion may be controlled by cortisol and prolactin. Cortisol stimulates chloride cell proliferation and differentiation and appears to interact with NaCl to initiate salt secretion. Prolactin appears to cause chloride cell dedifferentiation by reducing both the active-transport and leak pathways proportionately.(ABSTRACT TRUNCATED AT 400 WORDS)

Age Factors↗

Critical period for neonatal estrogen exposure in occurrence of mammary gland abnormalities in adult mice.

There exists a critical period for the development of cervicovaginal lesions in both mice and humans exposed neonatally and antenatally to sex hormones. Mammary glands from year-old female BALB/c mice exposed neonatally to 20 micrograms estradiol for 5 days commencing at 1 day of age showed the most mammary abnormalities, significantly greater than in controls (P less than 0.005). The incidence of abnormalities declined when treatment was begun after Day 1. Treatments begun after Day 3 did not result in this structural pattern. Mice ovariectomized after treatment all had inactive mammary glands with no abnormalities. There is a critical exposure period for the later occurrence of mammary gland abnormalities. However, the aberrant secretory state which accompanies these mammary gland alterations may be a consequence of permanent alteration in ovarian function or its endocrine control.

Aging↗

Chloride secretion and conductance of teleost opercular membrane: effects of prolactin.

Effects of prolactin on transport properties of opercular membranes from seawater-adapted tilapia, Sarotherodon mossambicus, have been examined. These membranes are high conductance (average Gt approximately 4 mS.cm-2) tissues with short-circuit currents (I) equal to net chloride secretion. Despite high Gt, nonlinear current-voltage relationships suggest that opercular membranes cannot be classified as "leaky" tissues. Variability among membranes is reflected in a linear relationship between I and Gt with a slope equal to 26 mV and the zero-current Gt intercept equal to 0.45 mS.cm-2. Prolactin injections decrease I and Gt in a dose-dependent manner. Phosphodiesterase inhibition, without effect on I in untreated fish, often partially reverses these prolactin effects. Gt-I data from prolactin-treated fish yield a slope of 18 mV and a Gt intercept of 0.10 mS.cm-2. The effects of prolactin are discussed in terms of conventional equivalent circuit analysis. Discrepancies between predictions based on this model and the actual data indicate that an alternative interpretation, based on a heterogeneous cell population, is more accurate. Analysis of this circuit suggests that the ratio of paracellular to active transport pathway conductances associated with chloride cells is constant and that differences in Gt and I are due to parallel changes in these conductances. Prolactin may effectively "remove" chloride cells from these membranes as well as inhibit (reversible by elevated cellular cAMP levels) active transport pathway conductance of remaining cells.

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