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Biomedical subjects

D T Krieger

Publications and source records attributed to D T Krieger.

At least 19 recordsLinked to original sources
PubMed1988

Effects of the antiglucocorticoid RU 486 on adrenal function in dogs.

We studied the antiglucocorticoid effects of RU 486 given orally in doses of 5 (low), 20 (intermediate), and 50 mg/kg (high) daily for 10 days to seven female mongrel dogs. No changes in plasma ACTH or cortisol levels were produced by the 5 mg/kg dose. Plasma ACTH levels increased 3-fold with both intermediate and high dosages. Plasma cortisol levels rose 4-fold (P less than 0.05) within 2 days of commencing the high dose schedule and within 3 days with the intermediate dosage. Plasma aldosterone concentrations increased significantly only with the highest dose schedule. Plasma RU 486 levels rose progressively during the 10 days of RU 486 administration and with the highest dose remained elevated for 7 days after it was stopped. Plasma RU 486 levels measured by RIA and high pressure liquid chromatography were comparable. The monodemethylated metabolite of RU 486 changed in parallel to the parent compound. During the high dose of RU 486, there was a 4% increase in body weight, with a reduction in hematocrit and plasma protein concentration. Plasma electrolyte levels and osmolality did not change. We conclude that in dogs a daily RU 486 dose of 5 mg/kg does not alter adrenal function, whereas higher doses (20 and 50 mg/kg) induce increases in plasma ACTH and cortisol concentrations. Despite the blockade of glucocorticoid receptors by RU 486, the presence of isotonic hypervolemia suggests that there was no functional deficiency of cortisol at the renal tubule or it was overshadowed by augmented mineralocorticoid production and action.

Administration, Oral↗
PubMed1988

Isolation and characterization of a corticotropin-releasing hormone-like peptide from human placenta.

Immunoreactive CRH was detected in extracts of human term placentae [5.2 +/- 0.8 (+/- SE) pmol/g wet wt; n = 9]. Molecular sieve chromatography revealed three size classes of immunoreactive CRH. The major species eluted with the Kav of synthetic rat CRH; the minor species had apparent mol wt (MW) of 18,000 and 8,000. A placental CRH-(1-41)-sized peptide was isolated by fractional acetone precipitation, molecular sieve chromatography, and sequential reverse phase high performance liquid chromatography steps. This peptide had the same chromatographic behavior as did rat CRH in all high performance liquid chromatographic isolation steps as well as the same UV absorbance to immunoreactive CRH ratio after the final purification step. Purified placental CRH stimulated ACTH release from anterior pituitary tissue in a dose-dependent manner and was equipotent with synthetic rat CRH. Partial sequencing indicated that 32 amino acids of this peptide are identical to those of rat and human CRH (sequence deduced from genomic sequence), and comparative peptide mapping with rat CRH provided further evidence that the placental CRH-like peptide is very homologous if not identical to CRH. The high mol wt placental CRH fractions also were partially purified by acetone precipitation, immune affinity chromatography, and gel filtration. Neither of these materials [big form (MW, 18,000) or intermediate form (MWr, 8,000)] stimulated ACTH release from rat pituitary tissue in vitro. Limited trypsin digestion of the highest MW CRH, followed by gel filtration analysis, resulted in conversion to the smaller [8,000 MW-sized and CRH-(1-41)-sized] forms. The detection of a CRH-like peptide in placenta together with our previous demonstration of plasma immunoreactive CRH in pregnant women suggest that the placenta synthesizes and secretes CRH into the maternal circulation.

Amino Acid Sequence↗
PubMed1986

Serotonin-containing elements of the rat pituitary intermediate lobe.

The source and location of serotonin in the intermediate lobe of the rat pituitary were determined by high performance liquid chromatography and immunohistochemistry. Serotonin is present in the intermediate lobe in nerve fibers and terminals, in mast cells and in elements in the blood circulating in vessels on the surface of the lobe.

Animals↗
PubMed1986

Expression and regulation of proopiomelanocortin-like gene in the ovary and placenta: comparison with the testis.

Proopiomelanocortin (POMC), a precursor protein for ACTH, beta-endorphin, and the MSHs, has been identified in the reproductive tracts of both male and female. With rat pituitary POMC complementary DNA (cDNA) as a hybridization probe, POMC-like messenger RNA (mRNA) was identified in the ovaries of rat, mouse, and monkey. The molecular size of POMC-like mRNA in the ovary was 150-200 bases smaller than in the pituitary and hypothalamus but identical to that in the testis and epididymis. The size heterogeneity of POMC mRNA observed in various tissues is not due to differences in the lengths of the poly(A) tail, as measured by RNase H digestion. S1 nuclease mapping analysis revealed that POMC mRNAs isolated from pituitary, testis, or ovary share the nucleotide sequences coding for ACTH, beta-lipotropin, and the 3'-untranslated region. The regulation of ovarian POMC-like mRNA was also investigated. Treatment of 25-day-old immature female rats with PMSG resulted in profound increases in the ovarian content of total RNA, poly(A) RNA, and POMC-like mRNA. The concentration of ovarian POMC-like mRNA during pregnancy increased increased to 3-4 times that in immature or normally cycling animals. POMC-derived peptides are present in the human placenta and are synthesized de novo in cultured placental cells. In this report we also demonstrate POMC-like mRNA in the placenta of rat, mouse, and human. The size of POMC-like mRNA in the placenta was similar to that observed in the testis, epididymis, and ovary and different from that found in the pituitary or hypothalamus. The concentration of placental POMC-like mRNA did not change throughout pregnancy. In conclusion, we have demonstrated that 1) POMC-like mRNA is present in the ovary and placenta of rodents and primates; 2) the size of POMC-like mRNA in the ovary and placenta, like that in the testis and epididymis, is smaller than that in the pituitary and hypothalamus, probably owing to a shortening of the 5'-ends; and 3) the expression of this gene is regulated by gonadotropins in the ovary but probably not in the placenta.

Animals↗
PubMed1985

Induced expression of the glucocorticoid receptor in the rat intermediate pituitary lobe.

Synthesis and release of pro-opiomelanocortin-derived peptides are under differential regulation in the anterior and intermediate lobes of the pituitary. Glucocorticoids inhibit synthesis of pro-opiomelanocortin-related peptides in the anterior lobe but not in the intermediate lobe. These two lobes are also characterized by differences in neural innervation and blood flow, both of which may represent routes of access for regulatory factors (the intermediate lobe is avascular). Immunoreactive glucocorticoid receptor, which can be demonstrated in many tissues, is absent from the intermediate lobe. Immunocytochemistry was used to demonstrate the presence of immunoreactive glucocorticoid receptor in the intermediate lobe after pituitary stalk transection, neurointermediate lobe grafts to kidney capsule, or monolayer culture of neurointermediate pituitary cells. This appearance of the glucocorticoid receptor is presumably a consequence of removal of intermediate pituitary cells from neural influences that may be responsible for inhibiting their expression under normal conditions in vivo.

Animals↗
PubMed1985

Implantation of normal fetal preoptic area into hypogonadal mutant mice: temporal relationships of the growth of gonadotropin-releasing hormone neurons and the development of the pituitary/testicular axis.

Central nervous system tissue which included the preoptic area (an area rich in gonadotropin-releasing hormone neurons) was taken from normal 17-day fetal mice and transplanted into the infundibular recess of the third ventricle of the hypothalamus of 90-day male mutant hypogonadal mouse hosts that are unable to synthesize the neurohormone, gonadotropin-releasing hormone. The growth and development of gonadotropin-releasing hormone neurons and fibers in the donor and host tissue as well as recovery of the pituitary-testicular axis were followed from 10 to 120 days post-implantation. Testicular growth was evident in 94% of the hypogonadal animals within 30 days post-implantation, continued for 90 days but showed no further increase during the remainder of the experiment. Increases in seminal vesicle weight, an index of testosterone secretion, were measurable at 30 days and continued through to the end of the experiment. Pituitary concentrations of gonadotropins were doubled at 30 days over that seen in the control mutant mouse and were maintained thereafter at normal or supranormal concentrations. In contrast plasma levels of gonadotropins, although above baseline at 30 days, never reached normal circulating levels. Nevertheless, it appeared that the concentration of luteinizing hormone achieved was sufficient to initiate and maintain testicular growth and testosterone secretion for the entire duration of the experiment. Immunocytochemical analysis of brain tissue was used to determine the presence and numbers of gonadotropin-releasing hormone neurons in the transplant and the distribution of their fibers in the donor and host tissue. The numbers of immunoreactive gonadotropin-releasing hormone neurons present at the time of sacrifice ranged from 3 to 140. Fiber outgrowth from the donor cells into the host was noted as early as 10 days post-implantation and the density of outgrowth continued to increase over the course of the experiment. Positive fibers tended to accumulate over the tuberoinfundibular sulci as they do in normal animals. In those instances where the transplant was placed a long distance from the median eminence, the gonadotropin-releasing hormone axons grew on the internal surface of the third ventricle until they reached these specific exit zones. These studies indicate that in the mutant hypogonadal mouse, central nervous system transplants from normal fetal mice can maintain the function of the pituitary-gonadal axis for periods of up to 120 days post-implantation. Outgrowth of the neurosecretory fibers begins very soon after implantation and the axons tend to follow pathways seen in normal tissue.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗
PubMed1985

Streptozotocin-induced diabetes is associated with reduced immunoreactive beta-endorphin concentrations in neurointermediate pituitary lobe and with disrupted circadian periodicity of plasma corticosterone levels.

Lower concentrations of immunoreactive (IR) beta-endorphin were present in the neurointermediate pituitary lobes of streptozocin-induced diabetic versus control animals at both 2 and 4 weeks after the onset of diabetes. The forms of beta-endorphin-like material present appeared to be similar in both groups when studied with cation-exchange chromatography. Insulin therapy via minipump for 2 weeks did not alter this finding of lowered beta-endorphin concentrations in diabetic animals, despite normalization of blood glucose levels and body weight gain. Lower IR beta-endorphin levels were also found in neurointermediate lobes of weight-restricted rats, but this group had increased plasma IR beta-endorphin concentrations compared to diabetic animals. Concentrations of IR beta-endorphin in microdissected brain regions and in anterior pituitaries of the diabetic animals failed to show consistent changes; in addition, ACTH concentrations in pituitary lobes and plasma did not differ among groups. Circadian rhythmicity of plasma insulin and corticosterone concentrations was absent in the diabetic animals, although food and water intake, while elevated, showed the normal nocturnal pattern of increased ingestion. Furthermore, adrenal hypertrophy was present in the diabetic animals and was accompanied by an elevation of mean plasma corticosterone levels. The present findings indicate that diabetes is associated with a decrease of neurointermediate pituitary lobe synthesis of beta-endorphin, while not affecting the processing of the peptide in this lobe, and confirm previous reports of altered adrenal function in diabetic animals.

Adrenocorticotropic Hormone↗
PubMed1984

Mating and pregnancy can occur in genetically hypogonadal mice with preoptic area brain grafts.

Adult female hypogonadal mice, in whom hypogonadism is secondary to a genetic deficiency in hypothalamic gonadotropin-releasing hormone (GnRH), are infertile. Mating, pregnancy, and delivery of healthy litters were achieved after transplantation of normal fetal preoptic area tissue, a major site of GnRH-containing cell bodies, into the third ventricle of adult female hypogonadal mice. Immunocytochemistry revealed GnRH-containing neurons in the grafts and GnRH-containing processes extending to the lateral median eminence of the host brains.

Animals↗
PubMed1984

Characterization and localization of proopiomelanocortin messenger RNA in the adult rat testis.

Northern blot analysis of total RNA and polyadenylated RNA isolated from adult rat testes showed that a proopiomelanocortin (POMC)-like messenger RNA molecule is present in these extracts. The testicular POMC messenger RNA is comparable in length to amygdala and midbrain POMC messenger RNA and appears to be at least 200 nucleotides shorter than POMC messenger RNA found in the hypothalamus and anterior and intermediate lobes of the pituitary gland. Hybridization in situ showed that POMC messenger RNA is located in Leydig cells, which are the only testicular cells that contain immunostainable POMC-derived peptides. These results suggest that local synthesis of POMC occurs in the testis.

Animals↗
PubMed1984

On the origin of the serotonergic input to the intermediate lobe of the rat pituitary.

Serotonin-containing nerve fibers have been visualized immunocytochemically in the intermediate lobe of the rat pituitary. A 50% depletion of the serotonin level in the intermediate lobe was obtained in our previous experiment in rats with pituitary stalk transection, which may represent the total neuronally derived serotonin there. In the present studies we have attempted to determine the source of these fibers by examining the effect of hypothalamic and midbrain lesions or fiber transections on serotonin levels and serotonin-containing neuronal elements. Complete hypothalamic deafferentation resulted in a significant reduction of immunostained fibers in the intermediate lobe. A 27% fall in the serotonin content (measured by HPLC and electrochemical detection) and a significant disappearance of immunostained fibers were observed after transecting the ascending fibers from the raphe nuclei towards the hypothalamus. The transection combined with the lesioning of the hypothalamic dorsomedial nuclei resulted in a 50% decrease of serotonin level in the intermediate lobe. The present data therefore suggest that serotonin fibers in the intermediate lobe may originate from cells both in the midbrain raphe and hypothalamic dorsomedial nuclei.

Animals↗
PubMed1984

Identification and possible function of pro-opiomelanocortin-derived peptides in the testis.

Using antibodies against peptides derived from different portions of the POMC molecule, immunocytochemical evidence suggests that this precursor and/or the peptides present within it are localized in testicular Leydig cells of at least five species. There is no evidence for the localization of these peptides or their precursor in any other cell type in this organ. Examination of testicular extracts by gel filtration, SDS-PAGE, and RP-HPLC indicate that the testis contains low concentrations of POMC-derived peptides relative to brain. Further analysis indicates that POMC is processed to alpha-MSH and beta-endorphin similar to its processing in intermediate pituitary lobe and brain. The relative mobilities of immunoreactive alpha-MSH and beta-endorphin on RP-HPLC columns indicate that they are in the unacetylated state as in brain and in contrast to the acetylated forms in the intermediate pituitary lobe. The potential for Leydig cells to synthesize POMC and its peptides was suggested by the demonstration of POMC-like mRNA in total testis and Leydig cell cultures. The size of the POMC-like mRNA is approximately 150 base pairs shorter than anterior or intermediate pituitary POMC mRNA. POMC-like mRNA activity has also been localized to Leydig cells in sections of testes using in situ hybridization. Immunostainable beta-endorphin and other POMC-derived peptides are present in testicular Leydig cells during fetal life and following puberty at times when testosterone secretion is maximal. The accumulation of immunostainable POMC-derived peptides in Leydig cells is dramatically increased by LH and hCG. A variety of observations suggests that testicular cells can respond to POMC-derived peptides. ACTH and the MSHs stimulate growth and cAMP accumulation in Sertoli cells. By contrast, studies using antagonists suggested that beta-endorphin and/or another testicular opioid inhibit Sertoli cell proliferation and ABP secretion. These observations are consistent with the postulate that different portions of the POMC molecule may have opposite effects on Sertoli cell function and suggest a mechanism by which Leydig cells could modulate Sertoli cell activity. Intratesticular administration of opiate antagonists inhibits testosterone secretion both in vivo and in vitro. These observations suggest that Leydig cell-derived beta-endorphin may facilitate testosterone secretion either directly or indirectly. The finding of POMC and its derivative peptides in testis, ovary, adrenal, and placenta suggests that all steroid hormone-secreting organs in mammals may utilize this peptidergic system.

Adrenocorticotropic Hormone↗
PubMed1984

The ontogeny of immunoreactive beta-endorphin in fetal, neonatal, and pubertal testes from mouse and hamster.

UNLABELLED: Derivatives of proopiomelanocortin (POMC), physicochemically similar to beta-endorphin and desacetyl alpha MSH, have been identified in adult testes, where these peptides were localized to Leydig cells. In the present study, the presence of immunostainable derivatives of POMC was established in fetal, neonatal, and pubertal testes with the unlabeled antibody peroxidase-antiperoxidase method. Specificity of staining was established by absorption of primary antisera with excess antigen. In the mouse, immunoreactive beta-endorphin was detectable in a few primitive interstitial cells on day 14 of gestation, the day after testicular differentiation. Thereafter, the number of immunostainable cells progressively increased throughout fetal life, so that at birth, they comprised 55% of the total interstitial cells. After birth, the number of immunostaining cells declined, so that they were only 12% of interstitial cells by 5 days of age. After 10 days of age, the number of immunopositive cells progressively increased, and by 40 days, interstitial cells showed intense staining comparable to that in adult mice. At 10 days of age, when the number of immunostainable cells was low, hCG treatment increased both the number and staining intensity of beta-endorphin-positive cells to those seen in adult testes. Antibodies directed against gamma MSH, a peptide within the N-terminal segment of POMC, also produced specific staining of fetal and adult interstitial cells in the mouse. In the hamster, the pattern of staining with anti-beta-endorphin in fetal, neonatal, and pubertal interstitial cells was similar to that observed in mice; the number and staining intensity of immunostainable cells increased during fetal life, declined after birth, and rose again at puberty. IN CONCLUSION: 1) the number and staining intensity of immunostainable interstitial cells have two peaks in mouse and hamster, at birth and after puberty; 2) the number and staining intensity of mouse interstitial cells can be increased by hCG; and 3) the development of immunostainable beta-endorphin activity correlates with the previously reported spontaneous and hCG-induced maturation of morphology and enzyme activities of Leydig cells.

Animals↗
PubMed1984

Preoptic area brain grafts in hypogonadal (hpg) female mice abolish effects of congenital hypothalamic gonadotropin-releasing hormone (GnRH) deficiency.

Normal fetal preoptic area (POA), a site of GnRH production, was implanted into the third ventricle of adult female hypogonadal (hpg) mice. When the grafts were successful, the mice (genetically deficient in hypothalamic GnRH) responded with vaginal opening, cornified vaginal cells, ovarian and uterine development, and increased pituitary FSH content and plasma LH concentrations. Similar results were obtained with fetal POA tissue, whether derived from males or females. Two of four hpg mice with POA grafts mated when caged overnight with males. Hpg females that received cortical tissue from fetuses or from 16-day-old pups, or POA tissue from 16-day-old pups, showed none of these changes, remaining similar to untreated hpg females.

Animals↗
PubMed1984

Demonstration of immunoreactive beta-endorphin- and gamma 3-melanocyte-stimulating hormone-related peptides in the ovaries of neonatal, cyclic, and pregnant mice.

Antisera against proopiomelanocortin (POMC)-derived peptides have previously been employed to demonstrate immunostainable materials in the male reproductive tract and in the corpus luteum of rat ovary. The present study was designed to determine how the distribution of such stainable materials varies in mouse ovary as a function of the reproductive status of the animal. Peptide-like activities were localized with the unlabeled antibody peroxidase-antiperoxidase (PAP) technique in ovaries removed from mice during fetal and neonatal development, during different stages of estrous cycle, and during pregnancy, with antisera against beta-endorphin, gamma 3MSH, and an extended N-terminal portion of POMC (16 K). beta-endorphin-like activity was also quantified in ovarian extracts by RIA. Immunostainable beta-endorphin, gamma 3MSH, and 16 K fragment-like activities were present in ovaries of pregnant and normally cycling (but not immature) mice. Intense staining was found predominantly in the corpora lutea. Less intense staining was observed in the interstitium and in the following parts of large follicles: parietal granulosa, corona radiata, and cumulus oophorus. When neonatal mice were injected with hCG, immunostainable beta-endorphin-like material in the ovarian interstitial area increased. Treatment with PMSG increased staining in both secondary follicles and the interstitium. Immunoassayable beta-endorphin-like activity was twice as high (per g wet wt) at pregnancy as during the cycle. We conclude that peptides similar or identical to POMC and/or its components are present in ovarian cells and that the concentration of such material appears to be regulated by gonadotropins.

Aging↗
PubMed1984

The luteinizing hormone-releasing hormone (LHRH) agonist [D-Trp6-Pro9-NEt]LHRH increased rather than lowered LH and alpha-subunit levels in a patient with an LH-secreting pituitary tumor.

Episodic secretion of LH, and the responses of serum LH, alpha-subunit, and testosterone concentrations to the acute administration of LHRH and the chronic administration of the LHRH agonist analog [D-Trp6-Pro9-NEt]LHRH (D-Trp6-Pro9) were evaluated in a 33-yr-old man previously reported to have an LH-secreting pituitary tumor unaccompanied by FSH hypersecretion. Basal serum LH and alpha-subunit concentrations were elevated [57 +/- 0.7 (SEM) mIU/ml (range, 45-71) and 26 ng/ml, respectively]. Frequent sampling revealed six LH secretory spikes over a 24-h period with increments above basal levels varying from 23-40% and interspike intervals ranging from 1.5-5 h. The concentrations of LH or alpha-subunit after iv administration of 150 micrograms LHRH did not increase above these intrinsic LH secretory increments (delta LH: 23%; delta alpha-subunit: 21%). The low basal serum FSH concentrations (3.5 mIU/ml) and elevated basal serum testosterone levels (1480 ng/dl) were unchanged after LHRH. Administration of clomiphene citrate produced no increase in serum LH, FSH, or testosterone concentrations. An attempt was made to decrease LH secretion in this patient using D-Trp6-Pro9. Administration of 200 micrograms daily sc of this LHRH analog for 21 days was associated with increases in serum LH and alpha-subunit concentrations. Mean serum LH and alpha-subunit levels for the 21 days of analog administration were 110 +/- 5.4 (SEM) mIU/ml (range, 70-170) and 64 +/- 3 (SEM) ng/ml (range, 32-84), respectively. During the 9-day period after discontinuance of the LHRH analog, levels of both serum LH and alpha-subunit declined precipitously and mean serum LH and alpha-subunit levels were 58 +/- 7 (SEM) mIU/ml (range, 18-90) and 22 +/- 3 (SEM) ng/ml (range, 12-44), respectively. We conclude that this patient's pituitary tumor has diminished responsiveness to acute LHRH administration and that the effect of chronic D-Trp6-Pro9 is stimulatory rather than inhibitory, as occurs after chronic administration of this analog to normal subjects. The blunted responsiveness to LHRH administration and the lack of response to clomiphene citrate suggest tumor autonomy. The presence of modest paradoxical responsiveness of serum LH and alpha-subunit concentrations during the course of daily D-Trp6-Pro9 administration suggests that central regulatory mechanisms, if present, are abnormal.

Adenoma↗